СПОСОБ ОЧИСТКИ ДЛЯ УДАЛЕНИЯ ВАРИАНТОВ АНТИТЕЛ С СУЛЬФАТИРОВАНИЕМ ТИРОЗИНА; ОЧИЩЕННЫЕ КОМПОЗИЦИИ

Изобретение относится к области биотехнологии. Предложен способ удаления антитела с сульфатированным тирозином из водной смеси, включающей антитело, в котором тирозин является сульфатированным, и антитело, в котором тирозин не является сульфатированным. Указанные антитела представляют собой антитела против LAG3 человека. Сульфатирование тирозина имеет место в CDR-L1 области. Также предложена композиция для терапии, включающая антитела против LAG3 человека. Композиция имеет менее чем 0,5% анти-hLAG3 антител, которые содержат сульфатированный тирозин на CDR-L1. Изобретение обеспечивает способ для очистки композиции антител от загрязняющих ее вариантов антитела, включающих сульфатированный тирозин. 2 н. и 12 з.п. ф-лы, 18 ил., 1 табл., 1 пр.

САЙТ-СПЕЦИФИЧЕСКАЯ КОНЪЮГАЦИЯ АНТИТЕЛО-ЛЕКАРСТВЕННОЕ СРЕДСТВО ПОСРЕДСТВОМ ГЛИКОИНЖЕНЕРИИ

Изобретение относится к области биотехнологии. Описана группа изобретений, включающая конъюгат антитела или его антигенсвязывающего фрагмента и цитотоксина для доставки цитотоксина субъекту (варианты), композицию для терапевтического применения, содержащую вышеуказанный конъюгат, способ лечения нуждающегося в этом пациента, включающий введение эффективного количества композиции, выделенный полинуклеотид, кодирующий антитело для получения вышеуказанного конъюгата, вектор экспрессии, клетку-хозяина для продуцирования вышеуказанного конъюгата, способ получения конъюгата, конъюгат антитело-лекарственное средство для доставки группы лекарственного средства субъекту. В одном из вариантов реализации конъюгат антитела или его антигенсвязывающего фрагмента и цитотоксина содержит по меньшей мере один гликан, содержащий по меньшей мере одну группу формулы -Gal-Sia-C(H)=N-Q-CON-X. Изобретение расширяет арсенал средств для доставки цитотоксинов субъекту. 13 н. и 29 з.п. ф-лы, 36 ил., 16 табл., 15 пр ...

ПРЕПАРАТ АНТИТЕЛА

Изобретение относится к области биотехнологии и иммунологии. Раскрыт стабильный водный препарат, включающий антитело или его фрагмент, которые специфично связываются с α-интерфероном человека. Препарат содержит антитело, гистидин, трегалозу и полисорбат 80. Также описывается способ лечения субъекта с помощью такого препарата и способ стабилизации антитела 13Н5 к α-интерферону человека. Изобретение может быть использовано в медицине. 4 з.п. ф-лы, 18 ил., 2 табл., 2 пр.

ФЕРМЕНТАТИВНАЯ ОДНОРЕАКТОРНАЯ РЕАКЦИЯ ДВОЙНОГО КОНЪЮГИРОВАНИЯ ПОЛИПЕПТИДОВ ЗА ОДНУ СТАДИЮ С ИСПОЛЬЗОВАНИЕМ СОРТАЗЫ

Изобретение относится к способу получения конъюгата Fc области антитела. Способ предусматривает одновременное инкубирование i) первого полипептида, содержащего аминокислотную последовательность LPXTG (SEQ ID NO: 20, где Х может быть любым аминокислотным остатком), ii) второго полипептида, содержащего аминокислотную последовательность LPXTА (SEQ ID NO: 31, где Х может быть любым аминокислотным остатком), iii) третьего полипептида, который имеет два N-конца, при этом данный полипептид имеет олигоглициновую Gm (m равен 2 (SEQ ID NO: 22), или 3 (SEQ ID NO: 23), или 4 (SEQ ID NO: 24), или 5 (SEQ ID NO: 25)) аминокислотную последовательность на его первом N-конце и олигоаланиновую Am (m равен 2 (SEQ ID NO: 26), или 3 (SEQ ID NO: 27), или 4 (SEQ ID NO: 28), или 5 (SEQ ID NO:29)) аминокислотную последовательность на его втором N-конце, iv) четвертого полипептида с сортазной активностью, полученного из сортазы А Staphylococcus aureus и имеющего аминокислотную последовательность SEQ ID NO: 21, и ...

СОСТАВЫ СО СНИЖЕННЫМ ОКИСЛЕНИЕМ

Группа изобретений относится к химико-фармацевтической промышленности и представляет собой жидкий состав, содержащий белок и соединение, предотвращающее окисление белка в жидком составе, выбранное из группы, состоящей из N-ацетил-L-триптофанамида, 5-амино-DL-триптофана, 5-гидроксииндол-3-уксусной кислоты и мелатонина, или их фармацевтически приемлемой соли. Изобретения также включают способ создания указанного белкового состава и способ предотвращения окисления белка в данном составе. Группа изобретений позволяет предотвратить окисление белка в жидкой композиции без образования активных форм кислорода во время длительного хранения и обработки в окружающей среде посредством применения указанных ранее соединений, которые менее светочувствительны и более склонны к окислению, чем L-триптофан. 3 н. и 28 з.п. ф-лы, 10 ил., 7 табл., 3 пр.

АНТИТЕЛА ПРОТИВ TENB2, СКОНСТРУИРОВАННЫЕ С ЦИСТЕИНОМ, И КОНЪЮГАТЫ АНТИТЕЛО - ЛЕКАРСТВЕННОЕ СРЕДСТВО

Изобретение относится к биотехнологии. Предложено антитело, специфичное к TENB2, содержащее легкую и тяжелую цепи. Тяжелая цепь содержит замену на свободный (реакционноспособный) цистеин А121С, что соответствует А114С (нумерация по Кабату) или А118С (нумерация по Eu). Предложены варианты конъюгата для лечения рака предстательной железы, содержащего антитело, ковалентно связанное с ауристатином, в том числе посредством линкера. Описаны: фармацевтическая композиция для лечения рака предстательной железы, использующая в качестве активного начала антитело или его конъюгат; изделие для лечения рака предстательной железы на основе такой композиции. Предложены: способ определения белка TENB2 в образце - на основе антитела, а также анализ для выявления клеток рака предстательной железы у млекопитающего и способ ингибирования клеточной пролиферации - на основе конъюгата антитела и ауристатина. Описан способ получения конъюгата антитела (Аb) и ауристатина (D) с формулой Ab-(L-D), где р равно от 1 ...

КОНЪЮГИРОВАННЫЕ ИММУНОГЛОБУЛИНЫ С C-КОНЦЕВЫМ ЛИЗИНОМ

Группа изобретений относится к биотехнологии, в частности к способам получения конъюгированных иммуноглобулинов с использованием микробной трансглутаминазы. Осуществляют инкубацию иммуноглобулина с микробной трансглутаминазой и терапевтическим или диагностическим агентом, содержащим ацил-донорный субстрат, включающий остаток глутамина, где трансглутаминаза катализирует конъюгацию K447 иммуноглобулина с остатком глутамина ацил-донорного субстрата. Причем иммуноглобулин содержит по меньшей мере один аминокислотный остаток, но менее чем 13 аминокислотных остатков после лизина 447 (K447) на тяжелой цепи согласно системе нумерации ЕС. Когда иммуноглобулин содержит один аминокислотный остаток после K447, указанный один аминокислотный остаток не является пролином, аспарагиновой кислотой, глутаминовой кислотой, лизином или аргинином. Когда иммуноглобулин содержит более чем один аминокислотный остаток после K447, первый аминокислотный остаток не является аспарагиновой кислотой, глутаминовой кислотой ...

КОНЪЮГИРОВАННЫЕ С ЛИЗИНОМ ИММУНОГЛОБУЛИНЫ

НОВОЕ АНТИТЕЛО ПРОТИВ Igβ ЧЕЛОВЕКА

КОНЪЮГАТЫ АНТИТЕЛО-ЛЕКАРСТВЕННОЕ СРЕДСТВО НА ОСНОВЕ ЭРИБУЛИНА И СПОСОБЫ ПРИМЕНЕНИЯ

АНТИ-CDH6-АНТИТЕЛО И КОНЪЮГАТ АНТИ-CDH6-АНТИТЕЛО-ЛЕКАРСТВЕННОЕ СРЕДСТВО

КЭПИРОВАННЫЕ И НЕКЭПИРОВАННЫЕ ЦИСТЕИНЫ АНТИТЕЛА И ИХ ПРИМЕНЕНИЕ В КОНЪЮГАЦИИ АНТИТЕЛО-ЛЕКАРСТВЕННОЕ СРЕДСТВО

ФУНКЦИОНАЛИЗИРОВАННЫЕ ПОЛИПЕПТИДЫ

... 1. Полипептид, содержащий два домена антитела, связанные аминокислотным линкером, где линкер включает два цистеина, способных к образованию внутрицепочечной дисульфидной связи, и где аминокислоты между цистеинами в последовательности линкера образуют петлевой фрагмент, когда два цистеина связаны друг с другом дисульфидной связью. ! 2. Полипептид по п.1, где линкер включает последовательность аминокислот, представленную в SEQ ID No.1. !3. Полипептид по п.1, где линкер включает последовательность аминокислот, представленную в SEQ ID No.2. ! 4. Полипептид по п.1, где петлевой фрагмент связывается с молекулой-мишенью. ! 5. Полипептид по п.4, где молекула-мишень представляет собой РК-модификатор. ! 6. Полипептид по п.5, где РК-модификатор представляет собой сывороточный альбумин. ! 7. Полипептид по п.5, где РК-модификатор представляет собой гиалуроновую кислоту. ! 8. Полипептид по п.1, где линкер включает последовательность аминокислот, представленную в SEQ ID No.3. ! 9. Полипептид по п.1, где ...

КОМПОЗИЦИЯ, СОДЕРЖАЩАЯ АВЕЛУМАБ

АНТИТЕЛА, СПЕЦИФИЧНЫЕ К CD22 ЧЕЛОВЕКА, И ИХ ПРИМЕНЕНИЯ В ТЕРАПИИ И ДИАГНОСТИКЕ

... 1. Молекула антитела, обладающая специфичностью в отношении CD22 человека, содержащая тяжелую цепь, в которой вариабельный домен содержит CDR, имеющую по меньшей мере одну из последовательностей, приведенную в виде HI на фиг.1 (SEQ ID NO:1) для CDR-H1, в виде Н2 на фиг.1 (SEQ ID NO:2) или Н2' (SEQ ID NO:13) или Н2'' (SEQ ID NO:15) или Н2''' (SEQ ID NO:16) для CDR-H2 или в виде Н3 на фиг.1 (SEQ ID NO:3) для CDR-H3. 2. Молекула антитела, обладающая специфичностью в отношении CD22 человека, содержащая легкую цепь, в которой вариабельный домен содержит CDR, имеющую по меньшей мере одну из последовательностей, приведенных в виде L1 на фиг.1 (SEQ ID NO:4) для CDR-L1, в виде L2 на фиг.1 (SEQ ID NO:5) для CDR-L2 или в виде L3 на фиг.1 (SEQ ID NO:6) для CDR-L3. 3. Молекула антитела, обладающая специфичностью в отношении CD22 человека, содержащая тяжелую цепь, в которой вариабельный домен содержит CDR, имеющую по меньшей мере одну из последовательностей, приведенных в SEQ ID NO:1 для CDR-H1, SEQ ...

САЙТ-СПЕЦИФИЧЕСКАЯ КОНЪЮГАЦИЯ АНТИТЕЛО-ЛЕКАРСТВЕННОЕ СРЕДСТВО ПОСРЕДСТВОМ ГЛИКОИНЖЕНЕРИИ

Изобретение относится к области биотехнологии. Описан способ получения связывающего полипептида, конъюгированного с эффекторной группой (варианты). В одном из вариантов реализации способ включает получение модифицированного гликана путем обработки природного или сконструированного гликана связывающего полипептида сиалилтрансферазой и/или галактозилтрансферазой, реакцию концевого остатка сиаловой кислоты и/или остатка галактозы с окисляющим средством и последующую реакцию остатка окисленной сиаловой кислоты и/или остатка окисленной галактозы с эффекторной группой. Изобретение расширяет арсенал способов получения связывающего полипептида, конъюгированного с эффекторной группой. 2 н. и 22 з.п. ф-лы, 36 ил., 16 табл., 15 пр.

Ion channel inhibitors

Biparatopic abeta binding polypeptides

ANTI-CDH6 ANTIBODY DRUG CONJUGATES

LIPOSOME ENCAPSULATED AFFINITY DRUG

Biparatopic abeta binding polypeptides

Compositions and methods for antibodies targeting factor P

Compositions and methods for antibodies targeting factor P

LIPOSOME ENCAPSULATED AFFINITY DRUG

ANTI-CDH6 ANTIBODY DRUG CONJUGATES

Biparatopic abeta binding polypeptides

LIPOSOME ENCAPSULATED AFFINITY DRUG

ANTI-CDH6 ANTIBODY DRUG CONJUGATES

ANTIBODY AGAINST HUMAN CD22 AND THEIR THERAPEUTIC AND DIAGNOSTIC USES

IMMOBILIZED ANTIGEN-BINDING MOLECULES FROM A DOMAIN

Purification process for removal of tyrosine sulfation antibody variants; purified compositions

The present invention relates to purified antibody and antigen-binding fragment compositions that lack sulfated tyrosine on one or more tyrosine residues in the immunoglobulin chains. Purification methods for removing sulfated tyrosine variants from antibody and antigen-binding fragment compositions are also provided.

Glucuronylation as acidic post-translational modification on therapeutic monoclonal antibodies

Compositions and methods for identifying glucuronylated protein drug products are provided.

Compositions and methods for enhancing the killing of target cells by NK cells

The present disclosure provides immunotherapeutic compositions and methods for enhancing an immune response and for treating cancer or inflammatory conditions mediated by autoreactive B cells in a subject. In some aspects, multispecific antigen-binding constructs are provided that recognize at least one tumor antigen or B-lineage cell antigen and NKp30 and/or another activating NK receptor. In some aspects, multispecific antigen-binding constructs are provided that recognize at least two tumor antigens or two antigens expressed by B-lineage cells, NKp30, and another activating NK receptor. The multispecific antigen-binding constructs and methods disclosed herein can be used for the treatment of cancer, even a cancer characterized by a CD16 deficient microenvironment and/or characterized by target cells ( ...

Site-specific antibody-drug conjugation through glycoengineering

The current disclosure provides binding polypeptides (e.g., antibodies), and effector moiety conjugates thereof (e.g., antibody-drug conjugates or ADCs), comprising a site- specifically engineered drug-glycan linkage within native or engineered glycans of the binding polypeptide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided.

Modified Antibody Compositions, Methods of Making and Using Thereof

The present disclosure provides modified antibodies which contain an antibody or antibody fragment (AB) modified with a masking moiety (MM). Such modified antibodies can be further coupled to a cleavable moiety (CM), resulting in activatable antibodies (AAs), wherein the CM is capable of being cleaved, reduced, photolysed, or otherwise modified. AAs can exhibit an activatable conformation such that the AB is more accessible to a target after, for example, removal of the MM by cleavage, reduction, or photolysis of the CM in the presence of an agent capable of cleaving, reducing, or photo lysing the CM. The disclosure further provides methods of making and using such modified antibodies and activatable antibodies.

Compositions for modulation of a TREM or TREML protein and methods of use

The present invention provides compositions and methods for reducing inflammation. In one aspect, the present invention provides a protein inhibitor comprising an ectodomain of at least one TREM or TREML protein. The invention also relates to inhibiting the interaction between a TREM or TREML protein and mitochondria released or derived from necroptotic cells; or between a TREM or TREML protein and cardiolipin.

Process for preparing maytansinoid antibody conjugates

Modified FC Molecules

Disclosed is a process for preparing a pharmacologically active compound, in which at least one internal conjugation sit of an Fc domain sequence is selected that is amenable to conjugation of an additional functional moiety by a defined conjugation chemistry through 5 the side chain of an amino acid residue at the conjugation site. An appropriate amino acid residue for conjugation may be present in a native Fe domain at the conjugation site or may be added by insertion (i.e., between amino acids in the native Fe domain) or by replacement (i.e., removing amino acids and substituting different amino acids). In the latter case, the number of amino acids added need not correspond to the number of amino acids removed 10 from the previously existing Fc domain. This technology may be used to produce useful compositions of matter and pharmaceutical compositions containing them. A DNA encoding the inventive composition of matter, an expression vector containing the DNA, and a host cell containing ...

Aldehyde tags, uses thereof in site-specific protein modification

Improved anti-TNFR1 polypeptides, antibody variable domains & antagonists

The invention relates to anti-TNFR1 polypeptides, antibody single variable domains (dAbs), antagonists and multispecific ligands, as well as methods and uses of these. The anti-TNFR1 polypeptides, antibody single variable domains (dAbs), antagonists and multispecific ligands are useful for treating and/or preventing inflammatory disease, such as arthritis or COPD, as well as for pulmonary administration, oral administration, delivery to the lung and delivery to the GI tract of a patient.

Functionalized polypeptides

Enzymatic conjugation of polypeptides

The present application relates to methods for the functionalization of immunoglobulins, in particular with drugs. Also disclosed herein are linking reagents, functionalized antibodies, pharmaceutical compositions, and method of treating disease and/or conditions.

Covalently linked antigen-antibody conjugates

Herein is reported a conjugate comprising an antigen and an antibody that specifically binds to the antigen wherein a covalent bond is formed between the antigen and an amino acid residue in the CDR2 of the antibody, an antibody comprising a cysteine residue at amino acid position 53 (according to Kabat) in the heavy chain CDR2 and uses thereof.

Covalently linked antigen-antibody conjugates

Herein is reported a conjugate comprising an antigen and an antibody that specifically binds to the antigen wherein a covalent bond is formed between the antigen and an amino acid residue in the CDR2 of the antibody, an antibody comprising a cysteine residue at amino acid position 53 (according to Kabat) in the heavy chain CDR2 and uses thereof.

MODIFIED PROTEINS AND PEPTIDES

The present invention relates to modified proteins and peptides that have reduced ability to bind to pre-existing antibodies. Such modified protein/peptide molecules can comprise C-terminal additions, extensions or tags and/or certain amino acid substitutions. Such modified molecules (e.g. 5 fusions and conjugates) comprise proteins, peptides, antigen binding molecules, antibodies or antibody fragments such as single variable domains e.g. human immunoglobulin (antibody) single variable domains, and also single variable domains derived from non-human sources such as a llama or camel, e.g. a VHH including a nanobody TM (described in e.g. WO 94/04678 and WO 95/04079 inter alia). The invention further relates to uses, formulations, compositions comprising such 0 modified C terminally extended and/or amino acid substituted molecules and also to methods of production and expression of these molecules.

Functionalized polypeptides

The invention provides functionalized polypeptides, especially therapeutic polypeptides (e.g., scFv), comprising a linker sequence that can be rapidly and specifically functionalized by the addition of one or functional moieties (e.g., PEG) or binding specificities (e.g., an amino acid sequence with a particular binding specificity). Such functionalized polypeptides are advantageous in that they have improved pharmacokinetic properties (e.g., improved in vivo half-life, tissue penetration and tissue residency time) over non-functionalized polypeptides. Methods for the rapid and reproducible generation of functionalized polypeptides are also provided.

Anti-CDH6 antibody drug conjugates

The present invention relates to anti-CDH6 antibodies, antibody fragments, antibody drug conjugates, and their uses for the treatment of cancer.

Multivalent meditopes, meditope-binding antibodies and uses thereof

Provided are methods for altering the distribution of a cell surface antigen. Also provided are compositions for use in the methods, including multivalent meditopes, and methods of producing, using, testing, and screening the same, including therapeutic and diagnostic methods and uses.

HUMANIZED ANTI-N2 ANTIBODIES

The present invention encompasses humanized antibodies that specifically bind N2 peptide, methods for the preparation thereof and methods for the use thereof.

Covalently linked antigen-antibody conjugates

Abstract Herein is reported a conjugate comprising an antigen and an antibody that specifically binds to the antigen wherein a covalent bond is formed between the antigen and an amino acid residue in the CDR2 of the antibody, an antibody comprising a cysteine residue at amino acid position 53 (according to Kabat) in the heavy chain CDR2 and uses thereof. 8848704_1 (GHMatters) P98412.AU.1 ...

Antibodies and methods of use

The presently disclosed subject matter provides antibodies that bind KLB and FGFR1, and methods of using the same. In certain embodiments, an antibody of the present disclosure includes a bispecific antibody that binds to an epitope present on FGFR1 and binds to an 5 epitope present on KLB. 17911767_1 (GHMatters) P103043.AU.2 ...

IGG4 FC Fragment Comprising Modified Hinge Region

The present invention relates to a modified IgG4 Fc fragment useful as a drug carrier. When the modified IgG4 Fc fragment of the present invention is combined with an arbitrary drug, the resulting drug conjugate can minimize the effector functions of the IgG4 Fc and the chain exchange with in vivo IgG while maintaining in vivo activity and improving in vivo duration of the drug conjugate.

SINGLE DOMAIN ANTIBODIES AGAINST TNFR1 AND METHODS OF USE THEREFOR

ANTI-CD137 ANTIBODIES

The present application relates to antibody molecules that bind CD137. The antibody molecules find application in the treatment and diagnosis of diseases and disorders, such as cancer and infectious diseases.

ANTIBODIES BINDING TO ILT4

The present application relates to antibodies specifically binding to immunoglobulin-like transcript 4 (ILT4), which is also known as LILRB2, LIR2, MIR10, and CD85d, and corresponding nucleic acids, host cells, compositions, and uses. In some embodiments, the antibodies bind specifically to human ILT4, but do not significantly bind to ILT2, ILT3, or ILT5, or to other members of the LILRA or LILRB families.

C-TERMINAL ANTIBODY VARIANTS

The invention generally relates to anti-sclerostin antibodies having C-terminal modifications, and compositions comprising such antibodies.

GLUCURONYLATION AS A NEW ACIDIC POST-TRANSLATIONAL MODIFICATION ON THERAPEUTIC MONOCLONAL ANTIBODIES

Compositions and methods for identifying glucuronylated protein drug products are provided.

LYSINE CONJUGATED IMMUNOGLOBULINS

Provided herein are conjugated immunoglobulins and methods for generating conjugated immunoglobulins using a microbial transglutaminase.

COMBINATION THERAPY WITH A MEK INHIBITOR, A PD-1 AXIS INHIBITOR, AND A VEGF INHIBITOR

A combination therapy comprising a MEK inhibitor, a PD-1 axis inhibitor, and a VEGF inhibitor is provided for the treatment of colorectal cancer and metastatic colorectal cancer.

METHOD OF CONJUGATION OF CYS-MABS

The present disclosure relates to a method of capping, reducing, and oxidizing cys-mAbs in order to provide homogenous material for subsequent conjugation reactions. The present method demonstrates robust ways to manufacture conjugates of cysteine-engineered antibodies that offer high yield and consistent product quality.

CONJUGATES OF AN ANTI-TNF-ALPHA ANTIBODY

Conjugates of an anti-TNF antibody and one or more nonpeptidic water soluble polymers are provided. Typically, the nonpeptidic water soluble polymer is poly(ethylene glycol) or a derivative thereof. Also provided, among other things, are compositions comprising conjugates, methods of making conjugates, and methods of administering compositions to a patient.

ANTIBODY FRAGMENTS AGAINST THE INSULIN RECEPTOR AND USES THEREOF TO TREAT HYPOGLYCEMIA

The present disclosure relates, in general, to methods of treating or preventing hypoglycemia using a negative modulator antibody fragment that binds to the insulin receptor and modulates the action of insulin at the insulin receptor.

METHODS AND COMPOSITIONS FOR TREATING AUTOIMMUNE AND INFLAMMATORY CONDITIONS

Described herein are compositions and methods for inhibiting an inflammatory or autoimmune response and for inducing immune tolerance in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an antigen presenting cell (APC)-targeted antibody operatively linked to IL-10 or a fragment thereof. The compositions and methods described herein are useful for treating inflammatory and autoimmune disorders.

NOVEL ANTI-HUMAN TSLP RECEPTOR ANTIBODY

... [Problem] To provide an anti-human TSLP receptor antibody for specifically bonding to the human TSLP receptor and blocking the effect of human TSLP receptor-mediated human TSLP. [Solution] The present inventors studied the anti-human TSLP receptor antibody and have provided an anti-human TSLP receptor antibody that includes a heavy-chain variable region composed of an amino acid sequence from amino acid number 1 to 118 of sequence number 1, and a light-chain variable region composed of an amino acid sequence from amino acid number 1 to 108 of sequence number 3. The present inventors perfected the present invention having found that the anti-human TSLP receptor antibody blocks TSLP-induced expression of TARC mRNA and production of MDC protein, and that allergic reaction is inhibited in a simian ascaris antigen sensitization model.

NOVEL ANTI-HUMAN IG.BETA. ANTIBODY

... [Problem] To provide an anti-human Igß antibody in which BCR and Fc?RIIb are crosslinked and which has a more enhanced immunosuppressive function compared to conventional antibodies. [Solution] An anti-human Igß antibody including: a heavy-chain variable region that includes a CDR 1 comprising an amino acid sequence from amino acid numbers 31-35 of SEQ ID NO: 2, a CDR 2 comprising an amino acid sequence from amino acid numbers 50-65 of SEQ ID NO: 2, and a CDR 3 comprising an amino acid sequence from amino acid numbers 98-108 of SEQ ID NO: 2; a light-chain variable region that includes a CDR 1 comprising an amino acid sequence from amino acid numbers 24-38 of SEQ ID NO: 4, a CDR 2 comprising an amino acid sequence from amino acid numbers 54-60 of SEQ ID NO: 4, and a CDR 3 comprising an animo acid sequence from amino acid numbers 93-101 of SEQ ID NO: 4; and a heavy-chain constant region, said region being a human Ig?1 constant region having S239D, H268D, and L328W amino acid variations.

ANTIBODY HAVING INFECTION-INHIBITING ACTIVITY AGAINST HEPATITIS C VIRUS

The present invention provides an anti-hepatitis-C-virus-E2-protein antibody or antigen-binding antibody fragment having infection-inhibiting activity against the hepatitis C virus (HCV). The present invention also provides an anti-hepatitis-C-virus-E2-protein antibody or antigen-binding antibody fragment having a specific variable region, having infection-inhibiting activity against the hepatitis C virus (HCV), and presenting the ability to suppress the appearance of escape variants.

TARGETING OF INNATE IMMUNE RESPONSE TO TUMOR SITE

The invention provides microparticles or nanoparticles for treatment of tumors comprising: (i) a targeting agent to the tumor or the tumor environment; and (ii) at least one inducer that stimulates a desired immune response in the tumor environ-ment, leading to tumor apoptosis, wherein components (i) and (ii) are non-covalently or covalently attached to the surface of said microparticles or nanoparticles. The targeting agent is an agent that recognizes and binds to an antigen, a receptor or other molecules found on the surface of tumor cells or in the tumor environment and are preferably antibodies.

IMPROVED ANTIBODIES OF THE CLASS IGG4

The present invention provides an antibody of the class IgG4 comprising at least one heavy chain which comprises a CH1 domain and a hinge region, wherein in each heavy chain: a. the inter-chain cysteine at position 127, numbered according to the Kabat numbering system, in the CH1 domain is substituted with another amino acid; and b. one or more of the amino acids positioned in the upper hinge region is substituted with cysteine.

MEDITOPES AND MEDITOPE-BINDING ANTIBODIES AND USES THEREOF

Antibodies and meditopes that bind to the antibodies are provided, as well as complexes, compositions and combinations containing the meditopes and antibodies, and methods of producing, using, testing, and screening the same, including therapeutic and diagnostic methods and uses.

PROCESS FOR PREPARING ANTIBODY MAYTANSINOID CONJUGATES

ANTIBODIES TO TRANSGLUTANINASE 2

BIPARATOPNYE POLYPEPTIDES - ANTAGONISTS OF SIGNAL TRANSMISSION OF WNT IN TUMOR CELLS

MODIFIED PROTEINS AND PEPTIDES

PIRROLOBENZODIAZEPINY AND THEIR CONJUGATES, CONNECTED DISULFIDE BOND WITH ANTIBODIES

MODIFIED PROTEINS AND PEPTIDES

BIPARATOPIC POLYPEPTIDES ANTAGONIZING WNT SIGNALING IN TUMOR CELLS

ENCLOSED IN LIPOSOME AFFINE MEDICINAL AGENT

Antibodies to MAdCAM

METHOD FOR PRODUCTION OF CONJUGATES WITH IMPROVED HOMOGENEITY

biparatopnye and - beta - binding polypeptides

METHODS FOR TREATMENT OF GRAFT REJECTION WITH USE OF BLAST-FURNACE ANTIBODIES, DIRECTED AGAINST CD40L

Antibody formulation

The present invention provides high concentration liquid formulations of antibodies or fragments thereof that specifically bind to a human interferon alpha polypeptide.

Method for oxime conjugation to a ketone-modified polypeptide class

Anti- triptase antibodies, and compositions, thereof and uses thereof

리포솜 캡슐화 친화 약제

... 본 개시는 내부 공간을 갖는 리포솜, 상기 내부 공간 내에 배치된 생활성 항엽산제, 상기 리포솜의 외부에 부착된 입체 안정제 및 하나 이상의 엽산 수용체에 대한 특이적 친화도를 갖는 단백질을 포함하는 표적 잔기를 포함하는 리포솜 항엽산 조성물로서, 상기 표적 잔기가 상기 입체 안정제 및 상기 리포솜의 외부 중의 하나 이상에 부착되는 리포솜 항엽산 조성물을 제공한다.

ANTIBODIES ANTICD79B AND INMUNOCONJUGADOS AND METHODS OF USE OF THE SAME

Composiciones de anticuerpos inmunoconjugados que los comprenden para el tratamiento de tumores hematopoyéticos en mamíferos, así como los métodos para usar dichas composiciones de sustancias con el mismo objetivo. Reivindicacion 1: Un anticuerpo anti-CD79b que incluye: (a) al menos una secuencia HVR seleccionada entre el grupo formado por: (i) HVR-L1 que comprende la secuencia A1-A15, donde A1-A15 es KASQSVDYDGDSFLN (identificador de secuencia n°: 131), (ii) HVR-L2 que comprende la secuencia B1-B7, donde B1-B7 es AASNLES (identificador de secuencia n°: 132), (iii) HVR-L3 que comprende la secuencia C1-C9, donde C1-C9 es QQSNEDPLT (identificador de secuencia n°: 133), (iv) HVR-H1 que comprende la secuencia D1-D10, donde D1-D10 es GYTFSSYWIE (identificador de secuencia n°: 134), (v) HVR-H2 que comprende la secuencia E1-E18, donde E1-E18 es GEILPGGGDTNYNEIFKG (identificador de secuencia n°: 135); (vi) HVR-H3 que comprende la secuencia F1-F10, donde F1-F10 es TRRVPVYFDY (identificador de secuencia ...

Anti CDH6 antibodies and anti CDH6 antibody drug conjugates

The present invention addresses the problem of providing: an antibody having an internalization activity of binding to CDH6; an antibody-drug conjugate consisting of the aforesaid antibody and a drug having an antitumor activity; a medicine using the antibody-drug conjugate, said medicine having a therapeutic effect on tumors; a method for treating tumors using the antibody, the antibody-drug conjugate or the medicine; etc. Provided are: an anti-CDH6 antibody having an internalization activity; an antibody-drug conjugate consisting of the aforesaid antibody and a drug having an antitumor activity; and a medicine and a method for treating tumors using the same.

Composition comprising AVELUMAB

The present invention relates to avelumab antibody compositions with an elevated deamidation level, as well as methods for using such compositions to treat a disorder, e.g., a disorder in which the interaction between PD-1 and PD-L1 is detrimental.

ANTI-CD25 FOR TUMOUR SPECIFIC CELL DEPLETION

NOVEL BISPECIFIC ANTIBODY BINDING TO HUMAN TLR2 AND HUMAN TLR4

COMPOSITIONS AND METHODS FOR ANTIBODIES TARGETING FACTOR P

The present invention relates to antibodies or antigen binding fragments thereof that bind to complement Factor P and used thereof as well as combinations of anti-Factor P antibodies with antibodies or antigen binding fragments thereof that bind to complement component 5 (C5).

COMPOSITION COMPRISING AVELUMAB

The present invention relates to avelumab antibody compositions with an elevated deamidation level, as well as methods for using such compositions to treat a disorder, e.g., a disorder in which the interaction between PD-1 and PD-L1 is detrimental. 122-. (canceled)23. A composition comprising avelumab , wherein a fraction of the avelumab molecules is deamidated.24. The composition according to claim 23 , wherein the proportion of deamidated avelumab variants is more than 6% or more than 8%.25. The composition according to claim 24 , wherein the proportion of deamidated avelumab variants is within a range of 6-30% or within a range of 8-25%.26. The composition according to claim 23 , wherein the composition exhibits any one or a combination of an increased PD-L1 binding activity on the cell surface claim 23 , increased ADCC and/or increased therapeutic efficacy as compared to a composition comprising avelumab with an amount of deamidated variants outside said range.27. The composition according to claim 23 , wherein the composition has a LMW species content of less than 10%.28. The composition according to claim 23 , wherein the proportion of avelumab variants that are oxidated at HC Met255 is less than 10%.29. The composition according to claim 23 , wherein the proportion of deamidated avelumab variants is within a range of 8-25% claim 23 ,wherein the composition has a LMW species content within a range of 0.1-10%; andwherein the proportion of avelumab variants that are oxidated at FTC Met255 is within a range of 0.1-10%.30. The composition according to claim 23 , wherein less than 20% of the avelumab molecules are afucosylated.31. The composition according to claim 23 , wherein less than 20% of the avelumab molecules have high mannose N-glycans with five or more mannose residues.32. A method of treating cancer comprising administering a composition according to to a patient having cancer.33. The method according to claim 32 , wherein the cancer is selected from the ...

METHOD FOR THE PRODUCTION OF VARIABLE DOMAINS

The present invention provides methods for the expression and/or production of variable domains with a C-terminal extension that can be used for coupling of the variable domain to one or more further groups, residues or moieties. In the method of the invention a yield of at least 80% of variable domains with a cysteine containing C-terminal extension is obtained. Also variable domains are provided and polypeptides comprising one or more variable domains obtainable by the methods of the present invention, as well as compounds that comprise such variable domains and/or polypeptides coupled to one or more groups, residues or moieties.

ANTAGONISTS, USES & METHODS FOR PARTIALLY INHIBITING TNFR1

The present invention relates to anti-Tumor Necrosis Factor 1 (TNFR1, p55, CD120a, P60, TNF receptor superfamily member 1A, TNFRSF1A) antagonists for partially inhibiting TNFR1 useful for the treatment and/or prevention of TNFR1-mediated diseases or conditions such as arthritis, psoriasis, Crohn's disease, COPD, lung inflammatory conditions and asthma. The invention further relates to methods, uses, formulations, compositions and devices comprising or using such anti-TNFR1 antagonists.

ANTI-CD79B ANTIBODIES AND IMMUNOCONJUGATES AND METHODS OF USE

The present invention is directed to compositions of matter useful for the treatment of hematopoietic tumor in mammals and to methods of using those compositions of matter for the same.

Fc Variants with Increased Affinity for FcyRIIc

The present invention relates to Fc variants having increased affinity for FcgammaRIIc, methods for their generation, Fc polypeptides comprising optimized Fc variants, and methods for using optimized Fc variants.

Anti-CD94 antibodies and methods of use thereof

The present disclosure relates to antibodies that bind to human CD94, as well as methods, uses, polynucleotides, vectors, host cells, and pharmaceutical compositions related thereto. In some embodiments, the antibodies are human or humanized antibodies that bind to human CD94 and do not block binding between CD94 and HLA-E. In some embodiments, the antibodies cross-react with cynomolgus CD94. In some embodiments, the antibodies do not promote internalization of surface-expressed CD94 to the extent of existing antibodies. In some embodiments, the antibodies promote ADCC targeting cells that express human CD94, e.g., on the cell surface.

АНТИ-CD79b АНТИТЕЛА И ИММУНОКОНЪЮГАТЫ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

Настоящее изобретение относится к области биотехнологии. Предложены варианты гуманизированного анти-CD79b антитела, каждый из которых характеризуется наличием легкой и тяжелой цепи и набором 6 CDR с установленной аминокислотной последовательностью и по меньшей мере одним свободным цистеиновым аминокислотным остатком, выбранным из А118С (по Европейской нумерации) в тяжелой цепи и V205C (по нумерации Кэбат) в легкой цепи. Рассмотрены: варианты соединения-конъюгата антитела с лекарственным средством, где антитело связано с лекарственным средством через свободный цистеин; фармацевтический состав для лечения рака на основе антитела; способы детекции CD79b или раковых клеток, а также способ ингибирования клеточной пролиферации, использующие соединение-конъюгат. Описан способ получения соединения-конъюгата. Данное изобретение может найти дальнейшее применение в терапии раковых заболеваний, ассоциированных с CD79b, в том числе при лечении гемопоэтических опухолей у млекопитающих. 8 н. и 62 з.п.

Modified Fc Molecules

Disclosed is a process for preparing a pharmacologically active compound, in which at least one internal conjugation site of an Fc domain sequence is selected that is amenable to conjugation of an additional functional moiety by a defined conjugation chemistry through the side chain of an amino acid residue at the conjugation site. An appropriate amino acid residue for conjugation may be present in a native Fc domain at the conjugation site or may be added by insertion (i.e., between amino acids in the native Fc domain) or by replacement (i.e., removing amino acids and substituting different amino acids). In the latter case, the number of amino acids added need not correspond to the number of amino acids removed from the previously existing Fc domain. This technology may be used to produce useful compositions of matter and pharmaceutical compositions containing them. A DNA encoding the inventive composition of matter, an expression vector containing the DNA, and a host cell containing the expression vector are also disclosed.

Aldehyde-Tagged Immunoglobulin Polypeptides and Methods of Use Thereof

The present disclosure provides aldehyde-tagged immunoglobulin (Ig) polypeptides that can be converted by a formylglycine-generating enzyme to produce a 2-formylglycine (FGly)-modified Ig polypeptide. An FGly-modified Ig polypeptide can be covalently and site-specifically bound to a moiety of interest to provide an Ig conjugate. The disclosure also encompasses methods of production of such aldehyde-tagged Ig polypeptides, FGly-modified Ig polypeptides, and Ig conjugates, as well as methods of use of same.

Process for manufacturing conjugates of improved homogeneity

The invention provides processes for manufacturing cell-binding agent-cytotoxic agent conjugates of improved homogeneity comprising performing the modification reaction at a lower temperature. The inventive processes comprise contacting a cell-binding agent with a bifunctional crosslinking reagent at a temperature of about 15° C. or less to covalently attach a linker to the cell-binding agent and thereby prepare a mixture comprising cell-binding agents having linkers bound thereto.

Method for the production of immunoglobulin single variable domains

The present disclosure relates to an improved method for the manufacture of immunoglobulin single variable domains. More specifically, the present disclosure relates to a method of producing immunoglobulin single variable domains in which the proportion of carbamylated variants is strongly reduced or absent and to improved immunoglobulin single variable domains obtainable by methods of the present disclosure.

Methods and Compositions for Increasing Arylsulfatase A Activity in the CNS

Provided herein are methods and compositions for treating a subject suffering from a deficiency in arylsulfatase A in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an arylsulfatase A.

Immunoconjugates with an Intracellularly-Cleavable Linkage

The present invention relates to therapeutic conjugates with improved ability to target various diseased cells containing a targeting moiety (such as an antibody or antibody fragment), a linker and a therapeutic moiety, and further relates to processes for making and using the conjugates.

OPTIMIZED Fc VARIANTS

The present invention relates to Fc variants having decreased affinity for FcγRIIb, methods for their generation, Fc polypeptides comprising optimized Fc variants, and methods for using optimized Fc variants.

MODIFIED Fc MOLECULES

Disclosed is a process for preparing a pharmacologically active compound, in which at least one internal conjugation site of an Fc domain sequence is selected that is amenable to conjugation of an additional functional moiety by a defined conjugation chemistry through the side chain of an amino acid residue at the conjugation site. An appropriate amino acid residue for conjugation may be present in a native Fc domain at the conjugation site or may be added by insertion (i.e., between amino acids in the native Fc domain) or by replacement (i.e., removing amino acids and substituting different amino acids). In the latter case, the number of amino acids added need not correspond to the number of amino acids removed from the previously existing Fc domain. This technology may be used to produce useful compositions of matter and pharmaceutical compositions containing them. A DNA encoding the inventive composition of matter, an expression vector containing the DNA, and a host cell containing the expression vector are also disclosed. 1. A composition of matter comprising:a monomeric or multimeric Fc domain having a cysteine or non-canonical amino acid substitution at one or more specifically selected conjugation site(s) selected from D46, S48, H49, E50, E53, K55, D61, G62, Q76, Y81, K107, K121, G122, Q123, E126, R136, D137, T140, K141, N142, E169, N170, N171, K173, L179, S181, G183, D194, K195, R197, Q199, Q200, G201, N202, or S223, relative to reference sequence SEQ ID NO:599; and(ii) at least one additional functional moiety, wherein the functional moiety is conjugated to the Fc domain through the side chain of the cysteine residue or non-canonical amino acid residue substituted at said one or more conjugation site(s).2. The composition of matter of claim 1 , wherein the additional functional moiety is a pharmacologically active moiety.3. The composition of matter of claim 2 , wherein the pharmacologically active moiety is a polypeptide claim 2 , a peptide claim 2 , or a ...

RADIOLABELED ANTI-GLYPICAN-3 IMMUNOCONJUGATES FOR IMMUNO-PET IMAGING OF HEPATOCELLULAR CARCINOMA

Provided are immunoconjugate probes useful for detecting hepatocellular carcinoma (HCC) lesions. The probes comprise a glypican-3 (GPC3)-specific monoclonal antibody or fragment thereof conjugated to a radionuclide such as Zr, Cu, and the like. The probes are useful for obtaining PET images with high tumor-to-liver ratios and targeting for diagnostic imaging of HCC lesions or cells in vitro and in vivo. 1. An immunoconjugate probe specific for glypican-3 (GPC3) , the probe comprising an anti-GPC3-specific antibody (mAb) or a target-specific fragment thereof , and a detectable label attached thereto , wherein the detectable label is detectable by positron emission tomography (PET) or SPECT.2. The probe of claim 1 , wherein the detectable label is a radionuclide selected from the group consisting of: Cu claim 1 , Cu claim 1 , Zr claim 1 , I claim 1 , Y claim 1 , Y claim 1 , In claim 1 , I claim 1 , Lu claim 1 , F claim 1 , and Tc.3. The probe of claim 2 , wherein the detectable label is detectable by positron emission tomography (PET) and is zirconium(Zr) or copper(Cu).4. The probe of claim 1 , wherein the detectable label is attached to the anti-GPC3-specific antibody (mAb) claim 1 , or a target-specific fragment thereof claim 1 , by a linker.5. The probe of claim 4 , wherein the linker is DFO.6. A pharmaceutically acceptable composition comprising:an immunoconjugate probe specific for glypican-3 (GPC3), the probe comprising an anti-GPC3-specific antibody (mAb) or a target-specific fragment thereof, and a detectable label attached thereto, wherein the detectable label is detectable by positron emission tomography (PET) or SPECT, and further comprising a pharmaceutically acceptable carrier.7. A method of obtaining an image of a hepatocellular carcinoma in a subject animal or human claim 4 , the method comprising the steps of:(a) delivering to a subject animal or human a pharmaceutically acceptable composition comprising an immunoconjugate probe specific for glypican-3 ...

Activated Formylglycine-Generating Enzymes and Methods of Producing and Using the Same

The present disclosure provides activated formylglycine-generating enzymes (FGE), methods of producing activated FGE, and their use in methods of producing a protein comprising a formylglycine (FGly) residue. The methods of producing activated FGE, as well as methods of use of activated FGE in producing FGly-containing proteins, include both cell-based and cell-free methods. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure are also provided. 186.-. (canceled)87. An activated formylglycine-generating enzyme (FGE) produced by a method comprising treating an FGE with Cu to produce an activated FGE.88. A cell-free composition comprising:an activated formylglycine-generating enzyme (FGE); anda buffer.89. The composition of claim 88 , further comprising a protein comprising an FGE recognition site.90. The composition of claim 88 , wherein the FGE is an N-terminally truncated FGE.91. The composition of claim 90 , wherein the FGE is a N-terminally truncated human FGE.92. A kit comprising:an activated formylglycine-generating enzyme (FGE); andinstructions for using the activated FGE to convert a cysteine residue or a serine residue present in an FGE recognition site of a protein to a formylglycine residue.93. The kit of claim 92 , wherein the FGE is an N-terminally truncated FGE.94. The kit of claim 93 , wherein the FGE is a N-terminally truncated human FGE.95. A kit comprising:a nucleic acid that encodes a formylglycine-generating enzyme (FGE); and{'sup': 2+', '2+, 'Cu or a source of Cu.'}96. The kit of claim 95 , further comprising cells suitable for expressing the FGE encoded by the nucleic acid. This application is a continuation of U.S. application Ser. No. 16/886,187, filed May 4, 2020, now U.S. Pat. No. 11,053,529, which is a divisional of U.S. application Ser. No. 16/414,202, filed May 16, 2019, now U.S. Pat. No. 10,683,527, which is a continuation of U.S. application Ser. No. 15/862,312, filed Jan. 4, 2018, now U.S. Pat. ...

Anti-garp antibody

The present invention relates to an antibody that binds to GARP and is useful as a therapeutic agent for a tumor, and a method for treating a tumor using the aforementioned antibody. It is an object of the present invention to provide an antibody, which inhibits the function of Treg in a tumor and is thereby used as a pharmaceutical product having therapeutic effects, a method for treating a tumor using the aforementioned antibody, and the like. An anti-GARP antibody that binds to GARP and exhibits inhibitory activity to Treg function and exhibits ADCC activity is obtained, and moreover a pharmaceutical composition for use in tumor therapy, comprising the aforementioned antibody, etc. is obtained.

ANTI-FGFR2 ANTIBODY

The present invention provides an antibody which binds to a fibroblast growth factor receptor. 1. An antibody or a functional fragment thereof that binds to a human fibroblast growth factor receptor (FGFR) , wherein the antibody or functional fragment thereof comprisesa heavy chain comprising CDRH1 comprising SEQ ID NO: 58; CDRH2 comprising SEQ ID NO: 59; and a CDRH3 comprising amino acids 3-9 of SEQ ID NO: 60; anda light chain comprising CDRL1 comprising SEQ ID NO: 67; CDRL2 comprising SEQ ID NO: 68; and CDRL3 comprising SEQ ID NO: 69.2. The antibody or functional fragment thereof according to claim 1 , wherein the CDRH3 region comprises amino acids 118-126 of an amino acid sequence selected from the group consisting of SEQ ID NOS: 97 claim 1 , 89 claim 1 , 91 claim 1 , 95 and 83.3. The antibody or functional fragment thereof according to claim 2 , wherein the antibody or functional fragment thereof is humanized.4. The antibody or functional fragment thereof according to claim 2 , wherein the heavy chain variable region comprises amino acids 20-137 of an amino acid sequence selected from the group consisting of SEQ ID NOS: 97 claim 2 , 89 claim 2 , 91 claim 2 , 95 and 83.5. The antibody or functional fragment thereof according to claim 2 , wherein the light chain variable region comprises amino acids 21-130 of SEQ ID NO: 73.6. The antibody or functional fragment thereof according to claim 2 , wherein the heavy chain comprises amino acids 20-467 of an amino acid sequence selected from the group consisting of SEQ ID NOS: 97 claim 2 , 89 claim 2 , 91 claim 2 , 95 and 83.7. The antibody or functional fragment thereof according to claim 2 , wherein the light chain comprises amino acids 21-235 of SEQ ID NO: 73.8. The antibody or functional fragment thereof according to claim 2 , wherein the antibody comprises a heavy chain amino acid sequence encoded by a nucleotide sequence selected from the group consisting of SEQ IDS NO: 96 claim 2 , 88 claim 2 , 90 claim 2 , 94 and ...

C-TERMINAL LYSINE CONJUGATED IMMUNOGLOBULINS

Provided herein are conjugated immunoglobulins and methods for generating conjugated immunoglobulins using a microbial transglutaminase. 138.-. (canceled)40. (canceled)41. The conjugated immunoglobulin of claim 39 , wherein the C-terminal lysine is Lysine 447 (K447) on a heavy chain of the immunoglobulin.42. The conjugated immunoglobulin of claim 39 , wherein the immunoglobulin comprises one amino acid residue after the C-terminal lysine claim 39 , and wherein the one amino acid residue after the C-terminal lysine is glycine claim 39 , alanine claim 39 , valine claim 39 , leucine claim 39 , isoleucine claim 39 , methionine claim 39 , phenylalanine claim 39 , tyrosine claim 39 , tryptophan claim 39 , serine claim 39 , threonine claim 39 , cysteine claim 39 , asparagine claim 39 , glutamine claim 39 , or histidine; or the one amino acid residue after the C-terminal lysine is not proline claim 39 , aspartic acid claim 39 , glutamic acid claim 39 , lysine claim 39 , or arginine.43. (canceled)45. The conjugated immunoglobulin of claim 44 , wherein the functional agent comprising the acyl donor substrate:{'sub': 2', '2', '2', '2', '2', '3, '(i) is according to formula (I), and wherein Z is a CBZ group; wherein L is a polyethylene glycol moiety (PEG) (—O((CH))—), ethyl amine (—NH((CH))—) or propyl amine (—NH((CH))—); and wherein n is 0, 1, 2, 3, 4 or 5; or'}(ii) is according to formula (I), wherein Z is a CBZ group, and wherein L is an amino acid; or(iii) is according to formula (II), wherein Z is a CBZ group; m is 1; n is 1, 2 or 3; and at least one L is Gly.4649.-. (canceled)51. The conjugated immunoglobulin of claim 50 , wherein the acyl donor substrate:{'sub': 2', '2', '2', '2', '2', '3, '(i) is according to formula (III), and wherein Z is a CBZ group; wherein each L is independently a polyethylene glycol moiety (PEG) (—O((CH))—), ethyl amine (—NH((CH))—) or propyl amine (—NH((CH))—); and wherein n is 0, 1, 2, 3, 4 or 5; or'}(ii) is according to formula (III), wherein ...

MODULATION OF CHARGE VARIANTS IN A MONOCLONAL ANTIBODY COMPOSITION

Combinations of different chromatography modalities with particularly refined conditions significantly reduce acid charge variants in a preparation of monoclonal antibodies. The process for reducing acid charge variants utilizes a combination of anion exchange and hydrophobic interaction chromatography, followed by cation exchange chromatography polishing, whereby the levels of acidic or basic charge species of the monoclonal antibodies may be modulated to a desired level. 1. A process for removing acid charge variants from a monoclonal antibody , comprising(a) loading a mammalian cell-expressed monoclonal antibody preparation onto a Protein A support, and eluting the monoclonal antibody from the Protein A support, thereby producing a first eluate comprising the monoclonal antibody;(b) loading the first eluate from step (a) onto an anion exchange and hydrophobic interaction (AEX/HIC) chromatography support, and allowing the first eluate to flow through the support, thereby producing a flow-through pool comprising the monoclonal antibody;(c) loading the flow-through pool comprising the monoclonal antibody onto a cation exchange (CEX) chromatography support having an antibody binding capacity of from about 25 g/L to about 65 g/L, determining when the absorbance units measured at UV A280 decrease from about 7% to about 14% from the peak absorbance units measured at UV A280, and then washing the CEX chromatography support with a wash buffer having a pH of from about 5.8 to about 6.6 and a conductivity target of from about 6.6 mS/cm to about 7.6 mS/cm; and(d) eluting the monoclonal antibody from the CEX chromatography support in step with an elution buffer having a pH of from about 6.0 to about 6.4 and a conductivity target of from about 10 mS/cm to about 14 mS/cm, thereby producing a second eluate comprising the monoclonal antibody and from about 10% to about 20% by weight of acid charge variants of the monoclonal antibody.2. The method of claim 1 , wherein the ...

ANTI-CD25 FOR TUMOUR SPECIFIC CELL DEPLETION

The present disclosure provides antibody sequences found in antibodies that bind to human CD25, in particular an anti CD25-a-634 antibody which do not block the binding of CD25 to IL-2 or IL-2 signalling Antibodies and antigen-binding portions thereof including such sequences can be used in pharmaceutical compositions and methods of treatment, in particular for treating cancer. 1. An antibody or antigen-binding fragment thereof , comprising the aCD25-a-634-HCDR3 amino acid sequence (SEQ ID NO: 4) as variable heavy chain complementarity determining region 3.2. The antibody or antigen-binding fragment thereof of claim 1 , further comprisinga) aCD25-a-634-HCDR1 amino acid sequence (SEQ ID NO: 2) as variable heavy chain complementarity determining region 1; andb) aCD25-a-634-HCDR2 amino acid sequence (SEQ ID NO: 3) as variable heavy chain complementarity determining region 2.3. The antibody or antigen-binding fragment thereof of or claim 1 , further comprising:a) aCD25-a-634-LCDR1 amino acid sequence (SEQ ID NO: 6) as variable light chain complementarity determining region 1;b) aCD25-a-634-LCDR2 amino acid sequence (SEQ ID NO: 7) as variable light chain complementarity determining region 2; andc) aCD25-a-634-LCDR3 amino acid sequence (SEQ ID NO: 8) as variable light chain complementarity determining region 3.4. The antibody or antigen-binding fragment thereof of any preceding claim claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a variable heavy chain comprising aCD25-a-634-HCDR123 amino acid sequence (SEQ ID NO: 5).5. The antibody or antigen-binding fragment thereof of any preceding claim claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a variable light chain comprising aCD25-a-634-LCDR123 amino acid sequence (SEQ ID NO: 9).6. The antibody or antigen-binding fragment thereof of any preceding claim claim 1 , wherein the antibody or antigen-binding fragment thereof is a monoclonal antibody claim 1 , a domain ...

MODIFIED AMINO ACIDS COMPRISING TETRAZINE FUNCTIONAL GROUPS, METHODS OF PREPARATION, AND METHODS OF THEIR USE

Provided herein are modified amino acids comprising a tetrazine groups according to Formula I: polypeptides, antibodies, payloads and conjugates comprising these modified amino acid residues derived from the modified amino acids, and methods of producing the polypeptides, antibodies, payloads and conjugates comprising the modified amino acid residues. The polypeptides, antibodies, payloads and conjugates are useful in methods of treatment and prevention, methods of detection and methods of diagnosis. 2. The compound of claim 1 , wherein V is a single bond claim 1 , —NH— claim 1 , or —CHNH—.12. A polypeptide comprising an amino acid residue corresponding to the compound of any of -.13. A polypeptide comprising at least two amino acid residues corresponding to the compound of any of -.14. The polypeptide of any of - claim 1 , further comprising at least one amino acid residue comprising an azide functional group.16. A conjugate comprising the polypeptide of any of - linked to a payload and optionally comprising a linking moiety between the polypeptide and the payload.17. An antibody comprising an amino acid residue corresponding to the compound of any of -.18. An antibody comprising at least two amino acid residues corresponding to the compound of any of -.19. The antibody of any of - claim 1 , further comprising at least one amino acid residue comprising an azide functional group.20. The antibody of claim 19 , wherein the amino acid residue comprising an azide functional group is selected from the group consisting of a compound according to any of formulas A1-A30 claim 19 , A40 claim 19 , and combinations thereof.21. A conjugate comprising the antibody of - linked to a payload and optionally comprising a linking moiety between the antibody and the payload.22. An orthogonal tRNA aminoacylated with an amino acid residue corresponding to the compound of any of -.23. A method of producing a polypeptide claim 22 , comprising contacting a polypeptide with the orthogonal ...

T cell receptor mimic rl9a

The present invention relates to a methodology of producing antibodies that recognize peptides associated with a tumorigenic or disease state, wherein the peptides are displayed in the context of HLA molecules. These antibodies will mimic the specificity of a T cell receptor (TCR) but will have higher binding affinity such that the molecules may be used as therapeutic, diagnostic and research reagents. The method of producing a T-cell receptor mimic of the present invention includes identifying a peptide of interest, wherein the peptide of interest is capable of being presented by an MHC molecule. Then, an immunogen comprising at least one peptide/MHC complex is formed, wherein the peptide of the peptide/MHC complex is the peptide of interest. An effective amount of the immunogen is then administered to a host for eliciting an immune response, and serum collected from the host is assayed to determine if desired antibodies that recognize a three-dimensional presentation of the peptide in the binding groove of the MHC molecule are being produced. The desired antibodies can differentiate the peptide/MHC complex from the MHC molecule alone, the peptide alone, and a complex of MHC and irrelevant peptide. Finally, the desired antibodies are isolated.

MODIFIED PROTEINS AND PEPTIDES

The present invention relates to modified proteins and peptides that have reduced ability to bind to pre-existing antibodies. Such modified protein/peptide molecules can comprise C-terminal additions, extensions or tags and/or certain amino acid substitutions. Such modified molecules (e.g. fusions and conjugates) comprise proteins, peptides, antigen binding molecules, antibodies or antibody fragments such as single variable domains e.g. human immunoglobulin (antibody) single variable domains, and also single variable domains derived from non-human sources such as a llama or camel, e.g. a VHH including a Nanobody™ (described in e.g. WO 94/04678 and WO 95/04079 inter alia). The invention further relates to uses, formulations, compositions comprising such modified C terminally extended and/or amino acid substituted molecules and also to methods of production and expression of these molecules. 1. A single immunoglobulin variable domain (dAb) , wherein said dAb comprises a sequence selected from a sequence having at least 90% sequence identity to SEQ ID NO 4; a sequence having at least 90% sequence identity to SEQ ID NO 5; and a sequence having at least 90% sequence identity to SEQ ID NO 6; and wherein said dAb comprisesa C-terminal extension which comprises an amino acid extension of from one amino acid to 5 amino acids.2. The single immunoglobulin variable domain (dAb) according to claim 1 , wherein said dAb is derived from a Camelid heavy chain (VHH).3. The single immunoglobulin variable domain (dAb) according to claim 1 , which comprises a C-terminal extension of from one amino acid to 4 amino acids.4. The single immunoglobulin variable domain (dAb) according to claim 1 , wherein said C-terminal extension comprises an alanine residue.5. The single immunoglobulin variable domain (dAb) according to claim 1 , wherein said C terminal extension comprises an amino acid extension selected from: (a) A (b) AS claim 1 , (c) AST (d) ASTK claim 1 , (e) ASTKG (f) AAA or (g) T.624 ...

METHODS AND COMPOSITIONS FOR INCREASING ENZYME ACTIVITY IN THE CNS

Provided herein are methods and compositions for treating a subject suffering from an enzyme deficiency in the central nervous system (CNS). The bifunctional fusion antibodies provided herein comprise an antibody to an endogenous blood brain barrier (BBB) receptor and an enzyme deficient in mucopolysaccharidosis III (MPS-III). The fusion antibodies provided herein comprise N-sulfoglucosamine sulfohydrolase (SGSH), alpha-N-acetylgulcosaminidase (NAGLU), heparin-alpha-glucosaminide N-acetyltransferase (HGSNAT), or N-acetylglucosamine-6-sulfatase (GNS). The methods of treating an enzyme deficiency in the CNS comprise systemic administration of a fusion antibody provided herein. 161.-. (canceled)62. A fusion antibody comprising: (a) a fusion protein comprising the amino acid sequences of an immunoglobulin heavy chain and a SGSH , and (b) an immunoglobulin light chain; wherein the fusion antibody crosses the blood brain barrier (BBB).63. The fusion antibody of claim 62 , wherein the amino acid sequence of the SGSH is covalently linked to the carboxy terminus of the amino acid sequence of the immunoglobulin heavy chain.64. The fusion antibody of claim 62 , wherein the fusion antibody catalyzes hydrolysis of sulfate groups from heparan sulfate.65. The fusion antibody of claim 62 , wherein the fusion antibody is post-translationally modified by a sulfatase modifying factor type 1 (SUMF1).66. The fusion antibody of claim 65 , wherein the post-translational modification comprises a cysteine to formylglycine conversion.67. The fusion antibody of claim 62 , wherein fusion antibody comprises a formylglycine.68. The fusion antibody of claim 62 , wherein the fusion protein further comprises a linker between the amino acid sequence of the SGSH and the carboxy terminus of the amino acid sequence of the immunoglobulin heavy chain.69. The fusion antibody of claim 62 , wherein the SGSH specific activity of the fusion antibody is at least about 1000 units/mg.70. The fusion antibody of ...

METHOD FOR THE PRODUCTION OF VARIABLE DOMAINS

The present invention provides methods for the expression and/or production of variable domains with a C-terminal extension that can be used for coupling of the variable domain to one or more further groups, residues or moieties. In the method of the invention a yield of at least 80% of variable domains with a cysteine containing C-terminal extension is obtained. Also variable domains are provided and polypeptides comprising one or more variable domains obtainable by the methods of the present invention, as well as compounds that comprise such variable domains and/or polypeptides coupled to one or more groups, residues or moieties. 131.-. (canceled)32. Polypeptide comprising one or more single variable domains linked to a C-terminal extension of maximal 10 amino acid residues in which at least one amino acid residue is a cysteine residue , wherein the C-terminal extension is different from SEQ ID NO: 1 and/or wherein the C-terminal extension is different three glycine residues followed by a C-terminal cysteine residue.33. The polypeptide according to claim 32 , wherein the at least one cysteine residue is positioned at the C-terminal end of the C-terminal extension.34. The polypeptide according to claim 32 , wherein the at least one cysteine residue is positioned at a site in the C-terminal extension different from the C-terminal end.35. The polypeptide according to claim 32 , wherein the C-terminal extension claim 32 , in addition to the at least one cysteine residue claim 32 , consists of glycine residues.36. The polypeptide according to claim 32 , wherein the C-terminal extension claim 32 , in addition to the at least one cysteine residue claim 32 , consists of alanine residues.37. The polypeptide according to claim 32 , wherein the C-terminal extension is selected from any of SEQ ID NO's: 2-7 or wherein the C-terminal amino acid of the one or more single variable domains is removed prior to addition of a C-terminal cysteine (e.g. claim 32 , as shown in SEQ ID NO ...

RECOMBINANT POLYPEPTIDE PRODUCTION

Disclosed herein is a method for producing a recombinant polypeptide in a mammalian cell culture in which the mammalian cells have a modified microRNA activity level. In one embodiment, a microRNA activity level is increased. In another embodiment, a microRNA activity level is decreased. In a more particular embodiment, the mammalian cells have a reduced miRNA-let-7a activity level. 1. A method of producing a recombinant polypeptide in a mammalian cell culture , the method comprising:(a) obtaining mammalian cells having reduced miRNA-let-7a activity;(b) culturing the mammalian cells to produce the recombinant polypeptide; and(c) recovering the protein.2. The method of claim 1 , wherein the miRNA-let-7a activity is reduced by a microRNA inhibitor.3. The method of claim 2 , wherein the microRNA inhibitor comprises an antisense oligonucleotide inhibitor of miRNA-let-7a.4. The method of claim 3 , wherein the mammalian cell culture comprises mammalian cells that are transfected with a synthetic antisense oligonucleotide inhibitor of miRNA-let-7a.5. The method of claim 4 , wherein the oligonucleotide inhibitor is chemically modified to improve nuclease resistance claim 4 , to increase resistance to miRNA-directed cleavage by RISC and/or to increase binding affinity.6. The method of claim 1 , wherein the mammalian cell culture comprises mammalian cells that are transfected with an expression vector encoding the antisense oligonucleotide inhibitor of miRNA-let-7a.7. The method of claim 1 , wherein the mammalian cell culture comprises mammalian cells that are stably transfected with an antisense oligonucleotide inhibitor of miRNA-let-7a.8. The method of claim 1 , wherein the mammalian cell culture comprises mammalian cells that are transiently transfected with an antisense oligonucleotide inhibitor of miRNA-let-7a.9. The method of claim 1 , wherein the mammalian cell culture comprises mammalian cells selected from: Chinese hamster ovary (CHO) cells claim 1 , mouse myeloma ( ...

NOVEL ANTI-FIBROBLAST ACTIVATION PROTEIN (FAP) BINDING AGENTS AND USES THEREOF

Provided are novel human-derived antibodies specific for Fibroblast Activation Protein (FAP), preferably capable of selectively inhibiting the enzymatic activity of FAP, and chimeric antigen receptors (CARs) directed against the human FAP antigen as well as methods related thereto. In addition, methods of diagnosing and/or monitoring diseases and treatments thereof which are associated with FAP are provided. Assays and kits related to antibodies specific for FAP are also disclosed. The novel anti-FAP antibodies can be used in pharmaceutical and diagnostic compositions for FAP-targeted immunotherapy and diagnostics. 115-. (canceled)16. A monoclonal human B cell-derived anti-Fibroblast Activation Protein (FAP) antibody , or a biotechnological or synthetic derivative thereof , wherein at least one of the complementarity determining regions (CDRs) and/or variable heavy chain and/or variable light chain of the antibody are encoded by a cDNA derived from an mRNA obtained from a human memory B cell which produced an anti-FAP antibody.17. The antibody of claim 16 , which:(a) is capable of binding to captured or directly coated human FAP with an EC50 of ≦0.1 μM;(b) is capable of binding a FAP epitope in a peptide of 15 amino acids in length, wherein the epitope comprises the amino acid sequence of any one of SEQ ID NOs: 30-37, 60, and 61;(c) is capable of binding a FAP epitope in a peptide of 15 amino acids in length, wherein the epitope consists of the amino acid sequence of any one of SEQ ID NOs: 30-37, 60, and 61;(d) is capable of binding to transmembrane FAP;(e) comprises a human constant region, and/or an Fc region or a region equivalent to an Fc region; or(f) comprises a glyco-engineered Fc region and has an increased proportion of non-fucosylated oligosaccharides in the Fc region as compared to a non-glyco-engineered antibody.18. The antibody of claim 16 , comprising in its variable region or binding domain:(a) at least one CDR of any one of SEQ ID NOs: 62-130;(b) a ...

HOMOGENOUS ANTIBODY DRUG CONJUGATES VIA ENZYMATIC METHODS

The present application in one aspect provide Fc-containing polypeptide conjugates comprising an Fc-containing polypeptide conjugated to a conjugate moiety, wherein the Fc-containing polypeptide comprises an N-glycosylated Fc region comprising an acceptor glutamine residue flanked by an N-glycosylation site and wherein the conjugate moiety is conjugated to the Fc-containing polypeptide via the acceptor glutamine residue. Also provided are methods of making such Fc-containing polypeptide conjugates by using a wildtype or engineered transglutaminases. Further provided are engineered transglutaminases specifically designed for carrying out such reactions. 120-. (canceled)22. The method of claim 21 , wherein the antibody drug conjugate is N-glycosylated in the Fc region.23. The method of claim 21 , wherein the antibody is a human antibody.24. The method of claim 21 , wherein the antibody is a humanized antibody.25. The method of claim 21 , wherein both heavy chains of the antibody are conjugated to the conjugation moiety.26. The method of claim 21 , wherein the antibody is an anti-Her2 antibody.27. The method of claim 26 , wherein the antibody is trastuzumab.28. The method of claim 26 , wherein the anti-Her2 antibody is N-glycosylated in the Fc region.29. The method of claim 26 , wherein the anti-Her2 antibody is N-glycosylated at position 297 claim 26 , wherein the anti-Her2 antibody is conjugated to a conjugation moiety via an endogenous acceptor glutamine residue at position 295.30. The method of claim 21 , wherein the cancer is a Her2+ cancer.31. The method of claim 21 , wherein the cancer is selected from the group consisting of pancreatic cancer claim 21 , ovarian cancer claim 21 , colon cancer claim 21 , breast cancer claim 21 , prostate cancer and lung cancer.32. The method of claim 29 , wherein the cancer is a Her2+ cancer.33. The method of claim 21 , wherein the antibody drug conjugate is in a pharmaceutical composition claim 21 , wherein the pharmaceutical ...

Aldehyde Tags, Uses Thereof in Site-Specific Protein Modification

The invention features compositions and methods for site-specific modification of proteins by incorporation of an aldehyde tag. Enzymatic modification at a sulfatase motif of the aldehyde tag through action of a formylglycine generating enzyme (FGE) generates a formylglycine (FGly) residue. The aldehyde moiety of FGly residue can be exploited as a chemical handle for site-specific attachment of a moiety of interest to a polypeptide.

MEDITOPES AND MEDITOPE-BINDING ANTIBODIES AND USES THEREOF

Antibodies and meditopes that bind to the antibodies are provided, as well as complexes, compositions and combinations containing the meditopes and antibodies, and methods of producing, using, testing, and screening the same, including therapeutic and diagnostic methods and uses. 1. (canceled)2. A method comprising:administering to a subject having cancer, a meditope-enabled antibody or antigen-binding fragment thereof and a meditope that binds to a meditope binding site in the meditope-enabled antibody or antigen-binding fragment thereof, wherein:said meditope-enabled antibody or antigen-binding fragment thereof comprises a heavy chain variable (VH) region; a heavy chain constant region (CH) or portion thereof; a light chain variable (VL) region comprising a threonine, serine, or aspartate at position 40, a residue other than glycine at position 41, a residue other than phenylalanine at position 83, and an aspartate or asparagine at position 85, according to Kabat numbering; and a light chain constant region (CL) or portion thereof;said meditope-enabled antibody or antigen-binding fragment thereof is a meditope-enabled variant of a human or humanized template antibody or antigen-binding fragment thereof, said variant comprising a plurality of framework region (FR) modifications compared to the template antibody or antigen-binding fragment thereof; andsaid meditope-enabled antibody or antigen-binding fragment thereof is capable of binding, via the meditope-binding site, to a cyclic peptide comprising the amino acid sequence of SEQ ID NO: 1.3. The method of claim 2 , wherein the meditope is coupled to a therapeutic or diagnostic agent.4. The method of claim 3 , wherein:the meditope is coupled to the therapeutic agent, which therapeutic agent is selected from the group consisting of: chemotherapeutic agents, therapeutic antibodies, toxins, radioisotopes, enzymes, chelators, boron compounds, photoactive agents, dyes, metals, metal alloys, and nanoparticles; orthe ...

OPTIMIZED Fc VARIANTS

The present invention relates to Fc variants having decreased affinity for FcγRIIb, methods for their generation, Fc polypeptides comprising optimized Fc variants, and methods for using optimized Fc variants.

Human Antibodies that Bind Human TNF-Alpha and Methods of Preparing the Same

Methylglyoxal (MGO)-modified recombinant TNF-alpha antibodies (e.g., Adalimumab) are identified. MGO modification decreases binding between Adalimumab and TNF-alpha. Methods are disclosed for reducing the presence of MGO-modified antibodies in the production of Adalimumab TNF-alpha antibodies. 1. A composition comprising a binding protein capable of binding TNF-alpha , wherein said binding protein comprises at least one methylglyoxal (MGO)-susceptible amino acid , and wherein at least a portion of said binding protein comprises one or more MGO-modified amino acids.2. The composition of claim 1 , wherein the portion of the binding protein that comprises at least one MGO-modified amino acid is less than 12%.3. The composition of claim 1 , wherein the portion of the binding protein that comprises at least one MGO-modified amino acid is less than 6%.4. The composition of claim 1 , wherein the MGO-susceptible amino acid is an arginine.5. The composition of claim 1 , wherein the binding protein is a human antibody or an antigen-binding portion thereof claim 1 , wherein the binding protein dissociates from human TNF-alpha with a Kof 1×10M or less and a Krate constant of 1×10sor less claim 1 , both as determined by surface plasmon resonance claim 1 , and wherein the binding protein neutralizes human TNF-alpha cytotoxicity in a standard in vitro L929 assay with an ICof 1×10M or less.6. A composition comprising a binding protein capable of binding TNF-alpha claim 1 , said binding protein comprising a methylglyoxal (MGO)-susceptible amino acid claim 1 , wherein said composition is prepared by substantially removing molecules of said binding protein that comprise at least one MGO-modified amino acid.7. The composition of claim 6 , wherein more than 70% of said molecules that comprise at least one MGO-modified amino acid is removed.8. The composition of claim 6 , wherein more than 90% of said molecules that comprise at least one MGO-modified amino acid is removed.9. The ...

PROCESS FOR PREPARING PURIFIED DRUG CONJUGATES

The invention provides a process for preparing a cell-binding agent chemically coupled to a drug. The process comprises covalently attaching a linker to a cell-binding agent, a purification step, conjugating a drug to the cell-binding agent and a subsequent purification step. 1. A process for preparing a cell-binding agent-cytotoxic agent conjugate comprising the steps of:(a) contacting a cell-binding agent with a bifunctional crosslinking reagent to covalently attach a linker to the cell-binding agent and thereby prepare a first mixture comprising cell-binding agents having linkers bound thereto,(b) subjecting the first mixture to gel filtration and thereby prepare a purified first mixture of cell-binding agents having linkers bound thereto,(c) conjugating a cytotoxic agent to the cell-binding agents having linkers bound thereto in the purified first mixture by reacting the cell-binding agents having linkers bound thereto with a cytotoxic agent in a solution having a pH of about 4 to about 9 to prepare a second mixture comprising (i) cell-binding agent chemically coupled through the linker to the cytotoxic agent, (ii) free cytotoxic agent, and (iii) reaction by-products, and(d) subjecting the second mixture to ion exchange chromatography or tangential flow filtration to purify the cell-binding agents chemically coupled through the linkers to the cytotoxic agent from the other components of the second mixture and thereby prepare a purified second mixture of cell-binding agents chemically coupled through the linkers to the cytotoxic agent.2. The process of claim 1 , wherein the gel filtration is a cross-linked dextran gel column.3. The process of claim 2 , wherein the cross-linked dextran gel column is a cross-linked dextran gel G25F resin column chromatography is.4. The process of claim 1 , wherein the solution in step (c) has a pH of from about 4 to about 6.5. The process of claim 1 , wherein the solution in step (c) has a pH of from about 6.5 to about 9.6. The ...

CAPPED AND UNCAPPED ANTIBODY CYSTEINES, AND THEIR USE IN ANTIBODY-DRUG CONJUGATION

An antibody production process in mammalian cells in which engineered unpaired cysteine residues are post-translationally modified and capped with particular chemical entities, which capped antibodies are well suited to further site-specific conjugation steps to form antibody-drug conjugates (ADCs) or protein drug conjugates; ADCs produced using these capped antibodies including in particular ADCs formed by the selective reduction of the capped antibodies' cysteine residues, and ADCs formed using chemical handles such as aldehyde/azide/alkyne biorthogonal groups, which permit additional drug conjugation chemistry; and uncapped antibodies produced by cells in low cysteine, cysteine and glutathione media, and ADCs produced via direct conjugation to these uncapped antibodies. 1. A method of bonding a predetermined capping moiety onto one or more unpaired cysteine residues on an antibody , said method comprising the step of: growing an antibody-expressing cell line in a culture medium containing said predetermined capping moiety , or a precursor of said predetermined capping moiety , wherein said cell line expresses said antibody , and wherein said predetermined capping moiety is attached by a covalent bond to at least one of said unpaired cysteine residues on said expressed antibody.2. The method of claim 1 , wherein said predetermined capping moiety is selected from the group consisting of 5-thio-2-nitrobenzoic acid (TNB) claim 1 , 2-mercaptopyridine claim 1 , dithiodipyridine (DTDP) claim 1 , 4-thiobenzoic acid claim 1 , 2-thiobenzoic acid claim 1 , 4-thiobenzenesulfonic acid claim 1 , 2-thiobenzenesulfonic acid claim 1 , methyl sulfonate (Ms) claim 1 , p-toluenesulfonate (Ts) and trifluoromethanesulfonate (Tf).3. The method of claim 1 , wherein said predetermined capping moiety is selected from a reactive group consisting of maleimido trioxa-4-formyl benzamide (MTFB) like molecules with an aldehyde handle or maleimido azido-lysine like molecules with an azide handle ...

COMPOSITIONS AND METHODS FOR ANTIBODIES TARGETING BMP6

The present invention relates to antibodies and antigen-binding fragments thereof to human BMP6 and compositions and methods of use thereof. 182.-. (canceled)83. A method of treating functional iron-deficiency anemia , the method comprising administering subcutaneously to a subject in need thereof a pharmaceutical composition comprising an antibody or antigen binding-fragment thereof that binds human BMP6 , wherein the antibody or antigen-binding fragment thereof comprises:(a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 69, 70 and 71, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 79, 80 and 81, respectively; or(b) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 72, 73 and 74, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 82, 83 and 84, respectively.84. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain (VH) sequence of SEQ ID NO: 75.85. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) sequence of SEQ ID NO: 85.86. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof comprises a VH sequence of SEQ ID NO: 75 and a VL of SEQ ID NO: 85.87. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain sequence of SEQ ID NO: 77.88. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof comprises a light chain sequence of SEQ ID NO: 87.89. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain sequence of SEQ ID NO: 77 and a light chain sequence of SEQ ID NO: 87.90. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof binds human BMP6 with a KD of ≤1 nM.91. The method of claim 83 , wherein the antibody or antigen-binding fragment thereof has at least about 100-fold greater affinity for ...

METHOD FOR TREATING SEIZURES

Methods for treating seizures, more particularly treating seizures associated with epilepsy. 1. A method for treating seizures in a subject in need thereof , comprising administering to the subject a therapeutically effective amount of an agent that inhibits the CD40-CD40L interaction and pathway in the brain.2. The method of claim 1 , wherein the subject has increased susceptibility for epilepsy.3. The method of claim 2 , wherein the increased susceptibility for epilepsy is due to a condition selected from the group consisting of traumatic brain injury claim 2 , brain tumors claim 2 , stroke claim 2 , neuroinflammation claim 2 , dementia claim 2 , drug intoxication claim 2 , chemical intoxications claim 2 , neurodevelopmental disorders claim 2 , and metabolic disorders of the nervous system.4. The method of claim 2 , wherein the increased susceptibility for epilepsy is due to a systemic disease selected from the group consisting of diabetes claim 2 , hypertension claim 2 , chronic inflammatory disorders claim 2 , and immunological disorders.5. The method of claim 1 , wherein the subject manifests epilepsy.6. The method of claim 5 , wherein the epilepsy is Temporal Lobe Epilepsy (TLE).7. The method of claim 5 , wherein the epilepsy is status epilepticus.8. The method of claim 1 , wherein the subject does not manifest epilepsy.9. The method of claim 1 , wherein the agent is an antagonist of CD40 or CD40L.10. The method of claim 9 , wherein the antagonist is a CD40L antagonist.11. The method of claim 10 , wherein the CD40L antagonist is an anti-CD40L antibody.12. The method of claim 11 , wherein the anti-CD40L antibody comprises a fully humanized anti-CD40L antibody.13. The method of claim 11 , wherein the anti-CD40L antibody comprises a fragment of a fully functional anti-CD40L antibody.14. The method of claim 11 , wherein the anti-CD40L antibody is PEGylated.15. The method of claim 11 , wherein the anti-CD40L antibody is administered in a pharmaceutically acceptable ...

PROTEIN SCREENING METHODS

The invention provides methods and compositions useful for identifying polypeptides with desired characteristics in vitro. 1. An X-display complex comprising:(a) a first nucleic acid molecule comprising a polypeptide-encoding sequence;(b) a polypeptide encoded by the first nucleic acid molecule; and(c) a second nucleic acid molecule comprising a nucleic sequence complementary to a portion of the first nucleic acid molecule,wherein the first nucleic acid molecule is bound to the second nucleic molecule through complementary nucleic acid base pairing, and wherein the second nucleic acid molecule is non-covalently bound to the polypeptide.2. The complex of claim 1 , further comprising:(a) a high affinity ligand covalently bound to the second nucleic acid molecule; and(b) a ligand acceptor bound to a peptide acceptor,wherein the high affinity ligand is bound to the ligand acceptor and the peptide acceptor is covalently bound to the polypeptide.3. The complex of claim 2 , wherein one or more of the following conditions are met:(a) the complex further comprises a second high affinity ligand, wherein the second high affinity ligand is covalently bound to the peptide acceptor, and wherein the second high affinity ligand is bound to the ligand acceptor; and(b) the ligand acceptor is covalently bound to the peptide acceptor.4. The complex of claim 3 , wherein one or more of the following conditions are met:(a) the peptide acceptor is bound to the second high affinity ligand through a linker;(b) the linker comprises polyethylene glycol; and(c) the linker comprises a polysialic acid linker.57-. (canceled)8. The complex of claim 1 , wherein one or more of the following conditions are met: (i) a ligand acceptor covalently bound to the second nucleic acid molecule; and,', '(ii) a high affinity ligand bound to a peptide acceptor,', 'wherein the ligand acceptor is bound to the high affinity ligand and the peptide acceptor is covalently bound to the C-terminus of the polypeptide;, '( ...

NOVEL ANTI-HUMAN TSLP RECEPTOR ANTIBODY

[Problem] To provide an anti-human TSLP receptor antibody that specifically binds to human TSLP receptor and inhibits an action of human TSLP through human TSLP receptor. 121-. (canceled)22. An anti-human TSLP receptor antibody or an antigen-binding fragment thereof , comprising a heavy chain variable region comprising CDR1 consisting of the amino acid sequence of amino acid numbers 31 to 35 of SEQ ID NO: 1 , CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 66 of SEQ ID NO: 1 , and CDR3 consisting of the amino acid sequence of amino acid numbers 99 to 107 of SEQ ID NO: 1 , and a light chain variable region comprising CDR1 consisting of the amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 3 , CDR2 consisting of the amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 3 , and CDR3 consisting of the amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO: 3.23. The anti-human TSLP receptor antibody or the antigen-binding fragment thereof according to claim 22 , selected from (1) or (2) below:(1) an anti-human TSLP receptor antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region consisting of the amino acid sequence of amino acid numbers 1 to 118 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence of amino acid numbers 1 to 108 of SEQ ID NO: 3.(2) an anti-human TSLP receptor antibody or an antigen-binding fragment thereof which is an antibody or an antigen-binding fragment thereof derived from posttranslational modification of the anti-human TSLP receptor antibody or the antigen-binding fragment thereof of (1).24. The anti-human TSLP receptor antibody or the antigen-binding fragment thereof according to claim 23 , comprising a heavy chain variable region consisting of the amino acid sequence of amino acid numbers 1 to 118 of SEQ ID NO: 1 claim 23 , and a light chain variable region consisting of the amino acid sequence of amino acid numbers 1 ...

Anti-ctla4 antibodies and methods of use thereof

The invention provides anti-CTLA4 binding proteins (e.g., antibodies, bispecific antibodies, and chimeric receptors) and their use in treating and preventing cancer, as well as compositions and kits comprising the anti-CTLA4 binding proteins.

Antibodies and methods of use

The presently disclosed subject matter provides antibodies that bind KLB and FGFR1, and methods of using the same. In certain embodiments, an antibody of the present disclosure includes a bispecific antibody that binds to an epitope present on FGFR1 and binds to an epitope present on KLB.

HUMAN ANTIBODIES THAT BIND HUMAN TNF-ALPHA AND METHODS OF PREPARING THE SAME

Methylglyoxal (MGO)-modified recombinant TNF-alpha antibodies (e.g., Adalimumab) are identified. MGO modification decreases binding between Adalimumab and TNF-alpha. Methods are disclosed for reducing the presence of MGO-modified antibodies in the production of Adalimumab TNF-alpha antibodies. 1. A composition comprising a binding protein capable of binding TNF-alpha , wherein said binding protein comprises at least one methylglyoxal (MGO)-susceptible amino acid , and wherein at least a portion of said binding protein comprises one or more MGO-modified amino acids.2. The composition of claim 1 , wherein the portion of the binding protein that comprises at least one MGO-modified amino acid is less than 12%.3. The composition of claim 1 , wherein the portion of the binding protein that comprises at least one MGO-modified amino acid is less than 6%.4. The composition of claim 1 , wherein the MGO-susceptible amino acid is an arginine.5. The composition of claim 1 , wherein the binding protein is a human antibody or an antigen-binding portion thereof claim 1 , wherein the binding protein dissociates from human TNF-alpha with a Kof 1×10M or less and a Krate constant of 1×10sor less claim 1 , both as determined by surface plasmon resonance claim 1 , and wherein the binding protein neutralizes human TNF-alpha cytotoxicity in a standard in vitro L929 assay with an ICof 1×10M or less.6. A composition comprising a binding protein capable of binding TNF-alpha claim 1 , said binding protein comprising a methylglyoxal (MGO)-susceptible amino acid claim 1 , wherein said composition is prepared by substantially removing molecules of said binding protein that comprise at least one MGO-modified amino acid.7. The composition of claim 6 , wherein more than 70% of said molecules that comprise at least one MGO-modified amino acid is removed.8. The composition of claim 6 , wherein more than 90% of said molecules that comprise at least one MGO-modified amino acid is removed.9. The ...

METHODS OF TREATING TRANSPLANT REJECTION USING A DOMAIN ANTIBODY DIRECTED AGAINST CD40

Methods of treating renal transplant rejection using anti-CD40L domain antibodies are provided. The anti-CD40L dAbs are less likely to cause platelet aggregation and thus cause thromboembolism. Appropriate anti-CD40L dAbs doses and administration regimens are also provided. Combination treatments for transplant rejection, particularly renal transplant rejection, using anti-CD40L dAbs, a CTLA4 mutant molecule (e.g., belatacept) and/or anti-CD28 optionally with conventional immunosuppressive renal transplant therapy are provided. 1: A method of treating renal transplant rejection comprising administering a therapeutically effective amount of BMS2h-572-633-CT-L2 (SEQ ID NO: 1) to a patient in need thereof.2: The method of claim 1 , wherein the transplant rejection is an acute transplant rejection.3: The method of claim 1 , wherein the transplant rejection is a chronic transplant rejection.4: The method of claim 1 , comprising administering a BMS2h-572-633-CT-L2 (SEQ ID NO: 1) dose from about 2 to about 30 mg/kg patient weight.5: The method of claim 1 , comprising administering a BMS2h-572-633-CT-L2 (SEQ ID NO: 1) dose from about 20 to about 30 mg/kg patient weight.6: The method of claim 1 , comprising administering a BMS2h-572-633-CT-L2 (SEQ ID NO: 1) at dose of about 20 mg/kg patient weight.7: The method of claim 1 , wherein BMS2h-572-633-CT-L2 (SEQ ID NO: 1) is administered with an immunosuppressive/immunomodulatory and/or anti-inflammatory agent.8: The method of claim 7 , wherein said immunosuppressive/immunomodulatory and/or anti-inflammatory agent is a CTLA4 mutant molecule.9: The method of claim 8 , wherein the CTLA4 mutant molecule is L104EA29Y-Ig (Belatacept).10: The method of claim 9 , wherein L104EA29Y-Ig (Belatacept) is administered at a dose from about 10 mg/kg to about 20 mg/kg patient weight.11: The method of claim 10 , wherein L104EA29Y-Ig (Belatacept) is administered at a dose of about 20 mg/kg patient weight.12: The method of claim 1 , wherein BMS2h- ...

METHOD FOR THE SEROLOGICAL DIAGNOSIS OF RHEUMATOID ARTHRITIS

This disclosure relates to the field of diagnostic methods. A new marker antibody for rheumatoid arthritis, as well as a new method for detecting rheumatoid arthritis, is disclosed herein. Also provided herein are means and methods for predicting whether a subject will develop rheumatoid arthritis. This disclosure is based on the discovery that patients with rheumatoid arthritis have antibodies in their circulation that react specifically with citrullinated antibodies. In one aspect, the disclosure, therefore, relates to an antibody comprising a citrulline residue. In another aspect, the disclosure provides a method for the detection of antibodies specific for rheumatoid arthritis in a sample from a subject, wherein the sample is contacted with a citrullinated antibody and wherein it is determined whether the sample comprises antibodies specifically reactive with the citrullinated antibody. 1. An antibody comprising a citrulline residue.2. The antibody of claim 1 , wherein the antibody is directed against a component of the outer cell surface of a red blood cell.3. The antibody of claim 2 , wherein the antibody is reactive with glycophorin.4. The antibody of claim 3 , wherein the glycophorin is glycophorin A.5. A method for the detection of antibodies specific for rheumatoid arthritis in a sample from a subject claim 3 , the method comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'contacting the sample with the antibody of ; and'}detecting antibodies specifically reactive with the antibody.6. The method according to claim 5 , wherein the citrullinated antibody is immobilized on a solid support.7. The method according to claim 6 , wherein the solid support is a nitrocellulose membrane.8. The method according to claim 6 , wherein the solid support is a polystyrene support.9. The method according to claim 5 , wherein the detection occurs as part of a hemagglutination assay.10. The method according to claim 9 , the method comprising:{'claim-ref': {'@idref': ...

Mixtures of antibodies

Described herein are antibodies and mixtures of antibodies optionally produced by a host cell line, nucleic acids encoding the antibodies and mixtures of antibodies, host cells containing such nucleic acids, and methods of treatment using the antibodies, mixtures of antibodies, or nucleic acids encoding the antibodies or mixtures of antibodies. Also described are methods of producing mixtures of antibodies in host cells.

Bispecific antibody exhibiting increased alternative fviii-cofactor-function activity

The present invention provides light chain amino acid substitutions that improve the FVIII cofactor function-substituting activity of ACE910 (Emicizumab), novel light chains showing FVIII cofactor function-substituting activity, and heavy chain amino acid substitutions that improve the FVIII cofactor function-substituting activity of novel light chain-containing bispecific antibodies.

IGG4 FC FRAGMENT COMPRISING MODIFIED HINGE REGION

A modified IgG4 Fc fragment useful as a drug carrier is disclosed. When coupled to a drug, the resulting drug-IgG4 Fc conjugate can minimize the effector functions of the IgG4 Fc and the chain exchange with in vivo IgG while maintaining in vivo activity and improving in vivo duration of the drug conjugate. 121-. (canceled)22. A drug conjugate comprising a modified aglycosylated IgG4 Fc fragment and a drug ,wherein the drug is conjugated to the modified aglycosylated IgG4 Fc fragment via a linker, andwherein the modified aglycosylated IgG4 Fc fragment consists of the amino acid sequence of SEQ ID NO: 2.23. The drug conjugate according to claim 22 , wherein the drug is selected from the group consisting of human growth hormone claim 22 , growth hormone-releasing hormone claim 22 , growth hormone-releasing peptide claim 22 , interferon claim 22 , interferon receptor claim 22 , a colony-stimulating factor claim 22 , interleukin claim 22 , interleukin receptor claim 22 , enzyme claim 22 , interleukin-binding protein claim 22 , cytokine-binding protein claim 22 , macrophage-activating factor claim 22 , macrophage peptide claim 22 , B cell factor claim 22 , T cell factor claim 22 , necrosis glycoprotein claim 22 , immunotoxin claim 22 , lymphotoxin claim 22 , tumor necrosis factor claim 22 , tumor suppressor claim 22 , transforming growth factor claim 22 , α-1 antitrypsin claim 22 , albumin claim 22 , α-lactalbumin claim 22 , apolipoprotein-E claim 22 , erythropoietin claim 22 , high-glycosylated erythropoietin claim 22 , angiopoietin claim 22 , hemoglobin claim 22 , thrombin claim 22 , thrombin receptor-activating peptide claim 22 , thrombomodulin claim 22 , blood coagulation factor VII claim 22 , blood coagulation factor VIIα claim 22 , blood coagulation factor VIII claim 22 , coagulation factor IX claim 22 , blood coagulation factor XIII claim 22 , plasminogen activator claim 22 , fibrin-binding peptide claim 22 , urokinase claim 22 , streptokinase claim 22 , hirudin ...

METHOD FOR INCREASING PYRO-GLUTAMIC ACID FORMATION OF A PROTEIN

A method for conversion of an N-terminal glutamine and/or glutamic acid residue of a protein to pyro-glutamic acid within a purification process. 1. A method for purification of a protein containing an N-terminal glutamine and/or an N-terminal glutamic acid , the method comprising a step of incubating said protein under conditions to promote cyclization of the N-terminal glutamine and/or an N-terminal glutamic acid of said protein to N-terminal pyro-glutamic acid.2. The method according to wherein the protein is an antibody.3. The method according to claim 1 , wherein the protein is an anti-CD20 antibody.4. The method of claim 3 , wherein the anti-CD20 antibody comprises a light chain sequence identical to SEQ ID NO:1 and a heavy chain sequence identical to SEQ ID NO:2.5. The method of claim 1 , wherein the conversion takes place at a temperature of 20-45° C.6. The method of wherein the conversion is carried out in the cell media claim 1 , or in a buffer.7. The method of wherein the concentration of the buffer is selected from the range of from 20 mM to 150 mM.8. The method of wherein the pH is in the range of from 3.5-9.0.9. The method of wherein the conversion from N-terminal glutamine and/or glutamic acid to pyro-glutamic acid reaches at least 50%.10. The method of wherein the conversion step comprises non-enzymatic conversion of the N-terminal glutamine and/or the glutamic acid to pyro-glutamic acid.11. The method of wherein the protein has from 1 to 4 N-terminal glutamine residues and/or from 1 to 4 N-terminal glutamic acid residues before conversion.12. The method of further comprising the steps of a protein A chromatography with a suitable resin and an ion exchange chromatography with a suitable resin.13. The method of wherein the method comprises the steps:(a) protein A capture with a suitable resin,(b) a viral inactivation at a pH in the range of from 3.0-4.0,(c) an anion exchange chromatography,(d) the conversion of the N-terminal glutamine and/or glutamic ...

Methods and compositions for increasing arylsulfatase a activity in the cns

Provided herein are methods and compositions for treating a subject suffering from a deficiency in arylsulfatase A in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an arylsulfatase A.

GLYCOENGINEERED ANTIBODY DRUG CONJUGATES

The current disclosure provides binding polypeptides (e.g., antibodies), and targeting moiety conjugates thereof, comprising a site-specifically engineered glycan linkage within native or engineered glycans of the binding polypeptide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided. 1. A method of making an effector moiety conjugated binding polypeptide comprising the steps of:(a) reacting a cytidine monophosphate-sialic acid (CMP-sialic acid) derivative with a glycan of a binding polypeptide to form a sialic acid derivative-conjugated binding polypeptide;(b) reacting the sialic acid derivative-conjugated binding polypeptide with an effector moiety to form the effector moiety conjugated binding polypeptide, wherein an imine bond is formed, and wherein neither the binding polypeptide nor the sialic acid derivative-conjugated binding polypeptide are treated with an oxidizing agent.2. The method of claim 1 , wherein the sialic acid derivative-conjugated binding polypeptide comprises a terminal keto or aldehyde moiety.3. The method of claim 1 , wherein the effector moiety comprises a terminal aminooxy moiety or is bound to a moiety comprising an aminooxy derivative.4. The method of claim 3 , wherein the effector moiety is selected from those in .5. The method of claim 1 , wherein step (b) results in the formation of an oxime bond.6. The method of claim 1 , wherein the effector moiety comprises a terminal hydrazine.7. The method of claim 1 , wherein step (b) results in the formation of a hydrazone linkage.929-. (canceled)31. The method of claim 30 , wherein Ris selected from the group consisting of CH claim 30 , CHCH(C═O)CH claim 30 , CHOH claim 30 , OH or H.34. The method of claim 1 , wherein the binding polypeptide is an antibody or ...

METHOD FOR THE PRODUCTION OF IMMUNOGLOBULIN SINGLE VARIABLE DOMAINS

The present invention relates to an improved method for the manufacture of immunoglobulin single variable domains. More specifically, the present invention relates to a method of producing immunoglobulin single variable domains in which the proportion of carbamylated variants is strongly reduced or absent and to improved immunoglobulin single variable domains obtainable by methods of the present invention. 1Pichia. A method for producing an immunoglobulin single variable domain in a host comprising:{'i': Pichia', 'Pichia, 'i) cultivating a host under conditions that are such that the host will multiply;'}{'i': Pichia', 'Pichia, 'ii) maintaining the host under conditions that are such that the host produces and secretes an immunoglobulin single variable domain; and'}iii) isolating and/or purifying the secreted immunoglobulin single variable domain;whereina) conditions are applied that reduce carbamylation of one or more amino acid residues in the immunoglobulin single variable domain, orb) conditions are applied that remove immunoglobulin single variable domains comprising one or more carbamylated amino acid residues, orc) a combination of (a) and (b).2. The method according to claim 1 , whereina) carbamylation of the one or more amino groups, optionally an N-terminal amino group and/or one or more amino groups in a side-chain of a lysine and/or an arginine residue, in the immunoglobulin single variable domain is reduced, orb) immunoglobulin single variable domains comprising one or more carbamylated amino groups, optionally an N-terminal carbamylamino group and/or one or more carbamylamino group in a side-chain of a lysine and/or an arginine residue, are removed, orc) a combination of (a) and (b).3Pichia. The method according to claim 1 , wherein the conditions in a) comprise a reduced pH as compared to the standard pH conditions for the host.45.-. (canceled)6Pichia. The method according to claim 1 , wherein the conditions of a) comprise a reduced culture time claim ...

CONJUGATED COMPOUNDS COMPRISING CYSTEINE-ENGINEERED ANTIBODIES

This disclosure provides conjugate compounds comprising antibodies and fragments thereof engineered with one or more reactive cysteine residues and more specifically to conjugate compounds with therapeutic or diagnostic applications. The conjugate compounds comprise cysteine-engineered antibodies or fragments thereof conjugated, for example, with chemotherapeutic drugs, toxins, and detection labels such as radionuclides or fluorophores. The disclosure also provides methods of using the disclosed conjugate compounds for in vitro, in situ, ex vivo, and in vivo diagnosis or treatment of mammalian cells, or associated pathological conditions. 1. A method for making a conjugate compound comprising:(i) mutagenizing at least a nucleic acid sequence encoding an antibody or Fc fusion protein by inserting a codon encoding for a cysteine (C) between the codons encoding the amino acids at positions 239 and 240, wherein the amino acid position numbering is according to the EU index as set forth in Kabat;(ii) expressing the cysteine-engineered antibody or Fc fusion protein;(iii) isolating the cysteine-engineered antibody or Fc fusion protein; and(iv) reacting at least one engineered cysteine group of the cysteine-engineered antibody or Fc fusion protein with a heterologous moiety.2. A method for making a conjugate compound comprising:(i) operably linking a nucleic acid sequence encoding a variable heavy chain region or a heterologous protein to a nucleic acid sequence encoding an Fc region protein, wherein the nucleic acid sequence encoding the Fc region protein comprises:at least a codon encoding a cysteine inserted between the codons encoding the amino acid at positions 239 and 240, wherein the amino acid position numbering is according to the EU index as set forth in Kabat;(ii) expressing the cysteine-engineered antibody or Fc fusion protein;(iii) isolating the cysteine-engineered antibody or Fc fusion protein; and(iv) reacting at least one engineered cysteine group of the ...

Methods and Compositions for Analyzing Proteins

Methods, compositions and kits are disclosed for determining one or more target polypeptides in a sample where the target polypeptides have undergone a post-translational modification. A mixture comprising the sample and a first reagent comprising a cleavage-inducing moiety and a first binding agent for a binding site on a target polypeptide is subjected to conditions under which binding of respective binding moieties occurs. The binding site is the result of post-translational modification activity involving the target polypeptide. The method may be employed to determine the target polypeptide itself. In another embodiment the presence and/or amount of the target polypeptide is related to the presence and/or amount and/or activity of an agent such as an enzyme involved in the post-translational modification of the target polypeptide. The interaction between the first binding agent and the binding site brings the cleavage-inducing moiety into close proximity to a cleavable moiety, which is associated with the polypeptide and is susceptible to cleavage only when in proximity to the cleavage-inducing moiety. In this way, an electrophoretic tag for each of the polypeptides may be released. Released electrophoretic tags are separated and the presence and/or amount of the target polypeptides are determined based on the corresponding electrophoretic tags. 144-. (canceled)46. The compound of claim 45 , wherein T comprises an antibody or a fragment thereof.47. The compound of claim 45 , wherein T comprises an antibody specific for a target protein or polypeptide.48. The compound of claim 45 , wherein T comprises a haptenized antibody.49. The compound of claim 45 , wherein T comprises a biotinylated protein.50. The compound of claim 45 , wherein T comprises an antibody derivatized with a functional polymer.51. The compound of claim 45 , wherein L comprises an olefin claim 45 , a thiazole claim 45 , an oxazole claim 45 , an imidazole claim 45 , or a selenoether.52. The ...

COMBINATION THERAPY OF AN AFUCOSYLATED CD20 ANTIBODY WITH A mTOR INHIBITOR

The present invention is directed to the combination therapy of an afucosylated anti-CD20 antibody with a mTOR inhibitor for the treatment of cancer, especially to the combination therapy of CD20 expressing cancers with an afucosylated humanized B-Ly1 antibody and a mTOR inhibitor such as Temsirolimus or Everolimus. 1. Use of an afucosylated anti-CD20 antibody with an amount of fucose of 60% or less , for the manufacture of a medicament for the treatment of cancer in combination with a mTOR inhibitor.2. Use according to characterized in that the amount of fucose of is between 40% and 60%.3. Use according to characterized in that the amount of fucose of is 50% or less.4. Use according to any one of to characterized in that said cancer is a CD20 expressing cancer.5. Use according to characterized in that said CD20 expressing cancer is a B-Cell Non-Hodgkin's lymphoma (NHL).6. Use according to any one of to claim 4 , characterized in that said afucosylated anti-CD20 antibody is a type II anti-CD20 antibody.7. Use according to or claim 4 , characterized in that said antibody is a humanized B-Ly1 antibody.8. Use according to claim 7 , characterized in that said mTOR inhibitor is rapamycin claim 7 , or a rapamycin analog or derivative.9. Use according to claim 7 , characterized in that said mTOR inhibitor is is Temsirolimus or Everolimus.10. Use according to any one of to claim 7 , characterized in that one or more additional other cytotoxic claim 7 , chemotherapeutic or anti-cancer agents claim 7 , or compounds or ionizing radiation that enhance the effects of such agents are administered.11. A composition comprising an afucosylated anti-CD20 antibody with an amount of fucose of 60% or less and a mTOR inhibitor for the treatment of cancer.12. The composition according to claim 11 , characterized in that said anti-afucosylated CD20 antibody is a humanized B-Ly1 antibody.13. A method of treatment of a patient suffering from cancer by administering an afucosylated anti-CD20 ...

Aldehyde Tags, Uses Thereof in Site-Specific Protein Modification

The invention features compositions and methods for site-specific modification of proteins by incorporation of an aldehyde tag. Enzymatic modification at a sulfatase motif of the aldehyde tag through action of a formylglycine generating enzyme (FGE) generates a formylglycine (FGly) residue. The aldehyde moiety of FGly residue can be exploited as a chemical handle for site-specific attachment of a moiety of interest to a polypeptide.

PYRROLOBENZODIAZEPINES AND ANTIBODY DISULFIDE CONJUGATES THEREOF

A compound of formula I: 4. The compound according to claim 1 , wherein Rand Rare both H.5. The compound according to claim 1 , wherein Rand Rare both methyl.6. The compound according to claim 1 , wherein one of Rand Ris H and the other is methyl.7. The compound according to claim 1 , wherein Y is a single bond.10. The compound according to claim 1 , wherein Rand Rare H.11. The compound according to claim 1 , wherein Rand Rare H.12. The compound according to claim 1 , wherein Rare Rare both OR claim 1 , where Ris optionally substituted Calkyl.13. The compound of claim 12 , wherein Ris Me.14. The compound according to claim 1 , wherein X is O.15. The compound according to claim 1 , wherein Ris H.16. The compound according to claim 1 , wherein there is a double bond between C2 and C3 in each monomer unit.17. The compound according to claim 16 , wherein Rand Rare independently selected from H and R.18. The compound according to claim 17 , wherein Rand Rare independently R.19. The compound according to claim 18 , wherein Rand Rare independently optionally substituted Caryl.20. The compound according to claim 1 , wherein Rand Rare independently selected from ═O claim 1 , ═CH claim 1 , ═CH—R claim 1 , and ═C(R).21. The compound according to claim 20 , wherein Rand Rare ═CH.22. The compound according to claim 1 , wherein R″ is a Calkylene group or a Calkylene group.24. The method of wherein the cell binding agent is an antibody or an active fragment thereof.25. The method of claim 24 , wherein the antibody or antibody fragment is an antibody or antibody fragment for a tumour-associated antigen.26. The method of wherein the antibody or antibody fragment is an antibody which binds to one or more tumor-associated antigens or cell-surface receptors selected from (1)-(53):(1) BMPR1B (bone morphogenetic protein receptor-type IB);(2) E16 (LAT1, SLC7A5);(3) STEAP1 (six transmembrane epithelial antigen of prostate);(4) 0772P (CA125, MUC16);(5) MPF (MPF, MSLN, SMR, megakaryocyte ...

Enzymatic conjugation of polypeptides

The present application relates to methods for the functionalization of immunoglobulins, in particular with drugs. Also disclosed herein are linking reagents, functionalized antibodies, pharmaceutical compositions, and method of treating disease and/or conditions.

C-TERMINAL LYSINE CONJUGATED IMMUNOGLOBULINS

Provided herein are conjugated immunoglobulins and methods for generating conjugated immunoglobulins using a microbial transglutaminase. 138.-. (canceled)39. A conjugated immunoglobulin comprising an immunoglobulin and an acyl donor substrate , whereina1) the immunoglobulin comprises at least one amino acid residue but less than 13 amino acid residues after Lysine 447 (K447) on a heavy chain constant region of the immunoglobulin, wherein the amino acid residues are numbered according to the EU numbering system, and wherein, when the immunoglobulin comprises one amino acid residue after K447, the one amino acid residue after the K447 is not proline, aspartic acid, glutamic acid, lysine, or arginine; and when the immunoglobulin comprises more than one amino acid residue after K447, the first amino acid residue after the K447 is not aspartic acid, glutamic acid or proline and the last amino acid residue after the K447 is selected from the group consisting of phenylalanine, leucine, isoleucine, methionine, valine, serine, proline, threonine, alanine, tyrosine, histidine, glutamine, asparagine, aspartic acid, glutamic acid, cysteine, tryptophan, and glycine, andb1) the acyl donor substrate comprises a glutamine residue and a therapeutic or a diagnostic agent,wherein the K447 of the immunoglobulin is conjugated to the glutamine residue of the acyl donor substrate; or wherein:a2) the immunoglobulin comprises at least one amino acid residue but less than 13 amino acid residues after Lysine 447 (K447) on a heavy chain of the constant region of the immunoglobulin, wherein the amino acid residues are numbered according to the EU numbering system, and wherein, when the immunoglobulin comprises one amino acid residue after K447, the one amino acid residue after the K447 is not proline, aspartic acid, glutamic acid, lysine, or arginine; and when the immunoglobulin comprises more than one amino acid residue after K447, the first amino acid residue after the K447 is not aspartic ...

GLYCOENGINEERED ANTIBODY DRUG CONJUGATES

The current disclosure provides binding polypeptides (e.g., antibodies), and targeting moiety conjugates thereof, comprising a site-specifically engineered glycan linkage within native or engineered glycans of the binding polypeptide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided. 116-. (canceled)17. A method of making an effector moiety conjugated binding polypeptide comprising the steps of:(a) reacting a CMP-sialic acid derivative with a glycan of a binding polypeptide to form a sialic acid derivative-conjugated binding polypeptide;(b) reacting the sialic acid derivative-conjugated binding polypeptide with an effector moiety to form the effector moiety conjugated binding polypeptide, wherein a thioether bond is formed.18. The method of claim 17 , wherein neither the binding polypeptide nor the sialic acid derivative-conjugated binding polypeptide are treated with an oxidizing agent.19. The method of claim 17 , wherein the sialic acid derivative comprises a terminal thiol moiety.20. The method of claim 17 , wherein the effector moiety comprises a maleimide moiety.22. The method of claim 17 , wherein the effector moiety comprises one or more proteins claim 17 , nucleic acids claim 17 , lipids claim 17 , carbohydrates claim 17 , or combinations thereof.23. The method of claim 17 , wherein the effector moiety comprises a glycan.24. The method of claim 17 , wherein the effector moiety comprises one or more glycoproteins claim 17 , glycopeptides claim 17 , or glycolipids.25. The method of claim 17 , wherein the binding protein has one or more native or engineered glycosylation sites.26. The method of claim 17 , further comprising achieving or modifying the glycosylation of the binding protein using one or more glycosyltransferases ...

Manufacturing Methods To Control C-Terminal Lysine, Galactose And Sialic Acid Content In Recombinant Proteins

Provided herein is a method for producing an antibody, such as an anti-TNFα antibody (e.g., infliximab) having a C-terminal lysine content of about 20% to about 70%, and a sialic acid content of about 1% to about 20%, comprising culturing a zinc-responsive host cell transfected with DNA encoding the antibody in a culture medium comprising at least 0.5 μM zinc; and controlling the concentration of zinc in the culture medium, thereby producing the antibody.

SITE-SPECIFIC ANTIBODY-DRUG CONJUGATION THROUGH GLYCOENGINEERING

The current disclosure provides binding polypeptides (e.g., antibodies), and effector moiety conjugates thereof (e.g., antibody-drug conjugates or ADCs), comprising a site-specifically engineered drug-glycan linkage within native or engineered glycans of the binding polypeptide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided. 1. A binding polypeptide comprising at least one modified glycan comprising at least one moiety of Formula (IV):{'br': None, '-Gal-Sia-C(H)═N-Q-CON—X\u2003\u2003 Formula (IV),'} A) Q is NH or O;', 'B) CON is a connector moiety; and', 'C) X is an effector moiety;', 'D) Gal is a component derived from galactose;', 'E) Sia is a component derived from sialic acid; and', 'wherein Sia is present or absent., 'wherein222-. (canceled)24. An isolated polynucleotide encoding the binding polypeptide of .2526-. (canceled)27. A method of making the binding polypeptide of claim 1 , the method comprising reacting an effector moiety of Formula (I):{'br': None, 'sub': '2', 'NH-Q-CON—X\u2003\u2003 Formula (I),'} A) Q is NH or O;', 'B) CON is a connector moiety; and', 'C) X is an effector moiety,, 'whereinwith an altered binding polypeptide comprising an oxidized glycan.2833-. (canceled)34. The method of claim 23 , wherein the binding polypeptide is an antibody.35. The method of claim 23 , wherein the binding polypeptide is an immunoadhesin.36. The method of claim 23 , wherein the binding polypeptide comprises a human Fc domain.37. The method of claim 23 , wherein the modified glycan is N-linked to the binding polypeptide via an asparagine residue at amino acid position 297 of the Fc domain claim 23 , according to EU numbering.38. The method of claim 23 , wherein the modified glycan is N-linked to the binding polypeptide via an ...

Hyperglycosylated binding polypeptides

Provided are binding polypeptides (e.g., antibodies), and effector moiety conjugates thereof, comprising a CH1 domain (e.g., a human IgG1 CH1 domain), wherein the CH1 domain has an engineered N-linked glycosylation site at amino acid position 114, according to Kabat numbering. Also provided are nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided.

MOLECULAR IMPRINTING OF WEST NILE ANTIBODIES WITH PHYSIOLOGICAL PH MATCHING

Methods and devices for molecular imprinting include a molecular imprinting synthesis and matching a physiological pH of a template utilized in the molecular imprinting synthesis to achieve molecular imprinting. Molecular imprinting can be achieved by matching the physiological pH of the template used in a molecular imprinting synthesis. Furthermore, electrostatic charges can be complementary matched to the template, by obtaining crystallographic data of a protein template. Particularly, positively and negatively charged amino acids can be counted and matched by an oppositely charged monomer. For hydrophobic amino acids, isoleucin, leucin, and valine amino acids are counted. Since not all hydrophobic amino acids are exposed, the hydrophobic amino acid and hydrophobic monomer ratio can be determined experimentally by varying ratios from 1:1 to 1:10. 1. A method of molecular imprinting , comprising:configuring a molecular imprinting synthesis; andmatching a physiological pH of a template utilized in said molecular imprinting synthesis to achieve said molecular imprinting.2. The method of wherein said matching further comprises complementary matching electrostatic charges to said template by obtaining crystallographic data of a protein template.3. The method of wherein said matching further comprises counting and matching positively and negatively charged amino acids by an oppositely charged monomer.4. The method of further comprises if said amino acids comprise hydorphobic amino acids claim 3 , counting isoleucin claim 3 , leucin claim 3 , and valine amino acids.5. The method of determining a hydroph amino acid and hydrophobic monomer ratio experimentally by varying ratios from a 1:1 ratio to a 1:10 ratio because not all of said hydrophobic amino acids are exposed.6. The method of wherein said molecular imprinting synthesis comprises a physiological pH matched synthesis wherein a molecular imprinting pH at synthesis is matched to a physiological pH of said template.7. ...

PREVENTION OF PROTEIN DISULFIDE BOND REDUCTION

Disclosed are methods for testing the presence and/or activity of glutathione system components and thioredoxin system components during the manufacturing process of disulfide bond-containing proteins. Also disclosed are methods for mitigating reduction of disulfide bonds during the manufacturing process, and for lowering the reduction potential of disulfide bond-containing proteins. Provided are compositions, kits, and methods for mitigating reduction and diminishing reduction potential of disulfide bond-containing proteins during protein manufacturing processes. 1. A method for increasing the yield of an intact disulfide bond-containing protein of interest in a cell culture or solution , comprising manufacturing the disulfide bond-containing protein of interest in the presence of a glutathione reductase inhibitor , a thioredoxin reductase inhibitor , or both a glutathione reductase inhibitor and a thioredoxin reductase inhibitor , whereby the amount of intact disulfide bond-containing protein is higher in the cell culture or solution manufactured in the presence of a glutathione reductase inhibitor , a thioredoxin reductase inhibitor , or a glutathione reductase inhibitor and a thioredoxin reductase inhibitor as compared to the amount of intact disulfide bond-containing protein in a cell culture or solution not manufactured in the presence of a glutathione reductase inhibitor , a thioredoxin reductase inhibitor , or a glutathione reductase inhibitor and a thioredoxin reductase inhibitor.2. The method of where increasing the yield of an intact disulfide bond-containing protein of interest in a cell culture or solution is controlled via the online measurement of the cell culture redox potential.3. The method of where increase of the yield of an intact disulfide bond-containing protein of interest in a cell culture or solution is monitored online via measurement of the cell culture redox potential.4. The method of claim 1 , further comprising: detecting the presence ...

Anti-pacap antibodies

The present invention is directed to antagonistic antibodies and antigen binding fragments thereof having binding specificity for PACAP. These antibodies inhibit, block or neutralize at least one biological effect associated with PACAP, e.g., vasodilation. In exemplary embodiments these antibodies and antigen binding fragments thereof may comprise specific VH, VL, and CDR polypeptides described herein. In some embodiments these antibodies and antigen binding fragments thereof bind to and/or compete for binding to specific epitope(s) on human PACAP. The invention is further directed to using these antagonistic anti-PACAP antibodies, and binding fragments thereof, for the diagnosis, assessment, and treatment of diseases and disorders associated with PACAP and conditions where antagonism of PACAP-related activities, such as vasodilation, mast cell degranulation, and/or neuronal activation, are therapeutically beneficial, e.g., headache and migraine indications.

Antibodies Binding to ILT4

The present application relates to antibodies specifically binding to immunoglobulin-like transcript 4 (ILT4), which is also known as LILRB2, LIR2, MIR10, and CD85d, and corresponding nucleic acids, host cells, compositions, and uses. In some embodiments, the antibodies bind specifically to human ILT4, but do not significantly bind to ILT2, ILT3, or ILT5, or to other members of the LILRA or LILRB families. 1. (canceled)2. (canceled)3. (canceled)4. (canceled)5. The isolated antibody of that specifically binds to human ILT4 (hILT4 or ILT4) , which comprises(a) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 9G4 (SEQ ID Nos: 125-127) and a VL comprising the amino acid sequence of the VL CDR1, CDR2 and CDR3 of 9G4 (SEQ ID Nos: 128-130);(b) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 9C8 (SEQ ID Nos: 131-133) and a VL comprising the amino acid sequence of the VL CDR1, CDR2 and CDR3 of 9C8 (SEQ ID Nos: 134-136);(c) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 2H2 (SEQ ID Nos: 137-139) and a VL comprising the VL CDR1, CDR2 and CDR3 of 2H2 (SEQ ID Nos: 140-142);(d) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 2E5 (SEQ ID Nos: 143-145) and a VL comprising the amino acid sequence of the VL CDR1, CDR2 and CDR3 of 2E5 (SEQ ID Nos: 146-148);(e) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 24E5 (SEQ ID Nos: 149-151) and a VL comprising the amino acid sequence of the VL CDR1, CDR2 and CDR3 of 24E5 (SEQ ID Nos: 152-154);(f) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 21D9 (SEQ ID Nos: 155-157) and a VL comprising the amino acid sequence of the VL CDR1, CDR2 and CDR3 of 21D9 (SEQ ID Nos: 158-160);(g) a VH comprising the amino acid sequence of the VH CDR1, CDR2 and CDR3 of 21D9.b (SEQ ID Nos: 155-157) and a VL comprising the amino acid sequence of the VL CDR1, CDR2 and CDR3 of 21D9.b (SEQ ID Nos: 158-160);(h) a VH ...

Anti-cd79b antibodies and immunoconjugates and methods of use

The present invention is directed to compositions of matter useful for the treatment of hematopoietic tumor in mammals and to methods of using those compositions of matter for the same.

MODIFIED AMINO ACIDS COMPRISING TETRAZINE FUNCTIONAL GROUPS, METHODS OF PREPARATION, AND METHODS OF THEIR USE

Provided herein are modified amino acids comprising a tetrazine groups according to Formula I: polypeptides, antibodies, payloads and conjugates comprising these modified amino acid residues derived from the modified amino acids, and methods of producing the polypeptides, antibodies, payloads and conjugates comprising the modified amino acid residues. The polypeptides, antibodies, payloads and conjugates are useful in methods of treatment and prevention, methods of detection and methods of diagnosis. 111-. (canceled)13. The polypeptide of comprising at least two amino acid residues of the compound of Formula I.14. The polypeptide of any of claim 12 , further comprising at least one amino acid residue comprising an azide functional group.16. A conjugate comprising the polypeptide of linked to a payload claim 12 , where the link is formed by a reaction of the tetrazole of the at least one amino acid residues claim 12 , with a payload comprising a strained alkene functional group claim 12 , and optionally comprising a linking moiety between the polypeptide and the payload.18. The antibody of comprising at least two amino acid residues of the compound of Formula I.19. The antibody of claim 17 , further comprising at least one amino acid residue comprising an azide functional group.21. A conjugate comprising the antibody of linked to a payload claim 17 , where the link is formed by a reaction of the tetrazole of the at least one amino acid residues claim 17 , with a payload comprising a strained alkene functional group claim 17 , and optionally comprising a linking moiety between the antibody and the payload.23. A method of producing a polypeptide claim 22 , comprising contacting a first polypeptide with the orthogonal tRNA of under conditions suitable for incorporating the amino acid residue into the first polypeptide.24. The method of claim 23 , wherein the orthogonal tRNA base pairs with a codon that is not normally associated with an amino acid.25. The method of ...

B7-H4 ANTIBODIES AND METHODS OF USE THEREOF

The present disclosure provides antibodies and antigen-binding fragments thereof that specifically bind to human B7-H4 (and optionally cynomolgus monkey, mouse, and/or rat B7-H4) and compositions comprising such antibodies or antigen-binding fragments thereof. In a specific aspect, the antibodies or antigen-binding fragments thereof that specifically bind to human B7-H4 increase T cell proliferation, increase interferon-gamma production, and/or deplete B7-H4 expressing cells via ADCC activity. The present disclosure also provides methods for treating disorders, such as cancer, by administering an antibody or antigen-binding fragment thereof that specifically binds to human B7-H4. 1. An isolated antibody or antigen-binding fragment thereof that specifically binds to human B7-H4 , comprising heavy chain variable region (VH) complementarity determining region (CDR) 1 , VH CDR2 , VH CDR3 and light chain variable region (VL) CDR1 , CDR2 , and CDR3 sequences selected from the group consisting of:(a) SEQ ID NOs:458-463, respectively;(b) SEQ ID NOs:5-10, respectively;(c) SEQ ID NOs:15-20, respectively;(d) SEQ ID NOs:25-30, respectively;(e) SEQ ID NOs:35-40, respectively;(f) SEQ ID NOs:45-50, respectively;(g) SEQ ID NOs:55-60, respectively;(h) SEQ ID NOs:65-70, respectively;(i) SEQ ID NOs:75-80, respectively;(j) SEQ ID NOs:85-90, respectively;(k) SEQ ID NOs:95-100, respectively;(l) SEQ ID NOs:105-110, respectively;(m) SEQ ID NOs:115-120, respectively;(n) SEQ ID NOs:125-130, respectively;(o) SEQ ID NOs:135-140, respectively;(p) SEQ ID NOs:145-150, respectively;(q) SEQ ID NOs:155-160, respectively;(r) SEQ ID NOs:165-170, respectively;(s) SEQ ID NOs:175-180, respectively;(t) SEQ ID NOs:185-190, respectively; and(u) SEQ ID NOs:195-200, respectively.2. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:11 claim 1 , 21 claim 1 , 31 claim 1 , 41 claim 1 ...

COMPOSITIONS AND METHODS RELATED TO ANTI-EGFR ANTIBODY DRUG CONJUGATES

In some aspects, the invention relates to an antibody-drug conjugate, comprising an anti-epidermal growth factor receptor (“EGFR”) antibody; a linker; and an active agent. The antibody-drug conjugate may comprise a self-immolative group. The linker may comprise an O-substituted oxime, e.g., wherein the oxygen atom of the oxime is substituted with a group that covalently links the oxime to the drug; and the carbon atom of the oxime is substituted with a group that covalently links the oxime to the antibody. 1. An antibody-drug conjugate , comprising an anti-EGFR antibody; a linker; and an active agent , wherein:the linker comprises an O-substituted oxime; wherein:a) the oxygen atom of the oxime is substituted with a group that covalently links the oxime to the active agent; andthe carbon atom of the oxime is substituted with a group that covalently links the oxime to the antibody; orb) the carbon atom of the oxime is substituted with a group that covalently links the oxime to the active agent; andthe oxygen atom of the oxime is substituted with a group that covalently links the oxime to the antibody.4. The antibody-drug conjugate of claim 2 , wherein the sugar or sugar acid is a monosaccharide.6. (canceled)7. The antibody-drug conjugate of claim 2 , wherein W is —C(O)— claim 2 , —C(O)NR′— claim 2 , —C(O)O— claim 2 , —S(O)NR′— claim 2 , —P(O)R″NR′— claim 2 , —S(O)NR′— claim 2 , or —PONR′— claim 2 , in each case where the C(O) claim 2 , S claim 2 , or P is directly bound to the phenyl ring claim 2 , and R′ and R″ are each independently hydrogen claim 2 , (C-C)alkyl claim 2 , (C-C)cycloalkyl claim 2 , (C-C)alkoxy claim 2 , (C-C)alkylthio claim 2 , mono- or di-(C-C)alkylamino claim 2 , (C-C)heteroaryl claim 2 , or (C-C)aryl.8. The antibody-drug conjugate of claim 2 , wherein W represents —C(O)NR′— claim 2 , where C(O) is bonded to the phenyl ring and NR′ is bonded to L.911-. (canceled)12. The antibody-drug conjugate of claim 1 , wherein the linker comprises an alkylene ...

Dock-and-Lock (DNL) Complexes for Therapeutic and Diagnostic Use

Disclosed herein are methods and compositions dock and lock (DNL) complexes comprising an AD moiety selected from an AKAP protein and a DDD moiety selected from a protein kinase A regulatory subunit. Also disclosed are fusion proteins comprising an AD moiety or DDD moiety attached to an effector moiety. The DDD moieties form dimers that bind to the AD moiety to form the DNL complexes. The effector moieties may be selected from a wide range of known effector moieties that produce one or more physiological effects, including but not limited to cell death. The DNL complexes may further comprise one or more diagnostic and/or therapeutic agents. The DNL complexes are of use for treating and/or diagnosing a variety of diseases or conditions. 2. The complex of claim 1 , wherein the amino acid sequence of the AD moiety is selected from the group consisting of SEQ ID NO:3 claim 1 , SEQ ID NO:4 claim 1 , SEQ ID NO:7 claim 1 , SEQ ID NO:32 claim 1 , SEQ ID NO:33 claim 1 , SEQ ID NO:34 claim 1 , SEQ ID NO:35 claim 1 , SEQ ID NO:36 claim 1 , SEQ ID NO:37 claim 1 , SEQ ID NO:38 claim 1 , SEQ ID NO:39 claim 1 , SEQ ID NO:40 claim 1 , SEQ ID NO:41 claim 1 , SEQ ID NO:42 claim 1 , SEQ ID NO:43 claim 1 , SEQ ID NO:44 claim 1 , SEQ ID NO:45 claim 1 , SEQ ID NO:46 claim 1 , SEQ ID NO:47 claim 1 , SEQ ID NO:48 claim 1 , SEQ ID NO:49 claim 1 , SEQ ID NO:50 claim 1 , SEQ ID NO:51 claim 1 , SEQ ID NO:52 claim 1 , SEQ ID NO:53 claim 1 , SEQ ID NO:54 claim 1 , SEQ ID NO:55 claim 1 , SEQ ID NO:56 claim 1 , SEQ ID NO:57 claim 1 , SEQ ID NO:58 claim 1 , SEQ ID NO:59 claim 1 , SEQ ID NO:60 claim 1 , SEQ ID NO:61 claim 1 , SEQ ID NO:62 claim 1 , SEQ ID NO:63 claim 1 , SEQ ID NO:64 claim 1 , SEQ ID NO:65 claim 1 , SEQ ID NO:66 claim 1 , SEQ ID NO:67 claim 1 , SEQ ID NO:68 claim 1 , SEQ ID NO:69 claim 1 , SEQ ID NO:70 claim 1 , SEQ ID NO:71 claim 1 , SEQ ID NO:72 claim 1 , SEQ ID NO:73 claim 1 , SEQ ID NO:74 claim 1 , SEQ ID NO:75 claim 1 , SEQ ID NO:76 claim 1 , SEQ ID NO:77 claim 1 , SEQ ID NO:78 ...

NOVEL HUMAN CONTROL ANTIBODIES AND USES THEREFOR

The present invention provides novel, rationally designed human control antibodies for use in various in vivo and in vitro applications. The antibodies of the present invention have well characterized variable domains that have been designed to minimize or eliminate antigen binding without altering gross antibody structure. Using the antibodies of the present invention in various assays allows researchers to distinguish effects that result from specific antigen-antibody interactions from other, non-specific antibody effects. 2. The isolated antibody or antibody fragment of claim 1 , wherein at least two of the heavy chain complementary determining regions are mutated.3. The isolated antibody or antibody fragment of claim 1 , wherein the CDRH1 region contains a mutation relative to the Avastin reference antibody.4. The isolated antibody or antibody fragment of claim 3 , wherein the CDRH1 is SEQ ID NO: 2.5. The isolated antibody or antibody fragment of claim 1 , wherein the CDRH2 region contains a mutation relative to the Avastin reference antibody.6. The isolated antibody or antibody fragment of claim 5 , wherein the CDRH2 is SEQ ID NO: 4 or SEQ ID NO: 5.7. The isolated antibody or antibody fragment of claim 1 , wherein the CDRH3 region contains a mutation relative to the Avastin reference antibody.8. The isolated antibody or antibody fragment of claim 7 , wherein the CDRH3 is SEQ ID NO: 7.9. The isolated antibody or antibody fragment of claim 2 , wherein the CDRH1 and CDRH3 regions are mutated relative to the Avastin reference antibody.10. The isolated antibody or antibody fragment of claim 9 , wherein the CDRH1 is SEQ ID NO: 2 and the CDRH3 is SEQ ID NO: 7.11. The isolated antibody or antibody fragment of claim 1 , wherein the isolated antibody or antibody fragment exhibits at least 50 fold reduced specific binding affinity to the target antigen as compared to the Avastin reference antibody.12. The isolated antibody or antibody fragment of claim 1 , wherein the ...

PROTEIN SCREENING METHODS

The invention provides methods and compositions useful for identifying polypeptides with desired characteristics in vitro. 128-. (canceled)29. A method of producing a library of nucleic acid-polypeptide (X-display) complexes , comprising the steps of:(a) providing a library of mRNA sequences comprising a sequence element complementary to a first nucleic acid linker;(b) providing a first nucleic acid linker operably linked to a first high affinity ligand such that the first nucleic acid linker binds to the mRNA through complementary nucleic acid base pairing;(c) providing a second high affinity ligand operably linked to a peptide acceptor;(d) providing a ligand acceptor with at least two binding sites or providing at least such that the ligand acceptor binds to the first high affinity ligand and the second high affinity ligand; and(e) allowing translation of the mRNA to occur such that the peptide acceptor binds to the translated protein, thereby forming a nucleic acid-polypeptide complex linking the mRNA to the protein.30. The method of claim 29 , further comprising allowing reverse transcription of the mRNA using the first nucleic acid linker in the nucleic acid-polypeptide complex as a primer such that a DNA/RNA hybrid is formed.31. The method of claim 30 , further comprising degrading the mRNA and synthesizing a complementary DNA strand thereby forming a DNA/DNA hybrid in the nucleic acid-polypeptide complex.32. The method of wherein the mRNA further comprises a TMV enhancer.33. The method of wherein the mRNA further comprises a Cμ sequence.34. The method of wherein the mRNA further comprises a FLAG tag.35. The method of wherein the mRNA further comprises a streptavidin (SA) display sequence.36. The method of wherein the mRNA further comprises an A20 tail.37. A library of nucleic acid-polypeptide complexes produced by the method of .3856-. (canceled) This application is a continuation of U.S. Utility application Ser. No. 13/009,500, filed Jan. 19, 2011 which is a ...

TAG FOR LABELING BIOMOLECULES

Provided is a tag for labeling a biomolecule, comprising a polymer backbone, one or more pendant moieties, and an end group capable of binding to the biomolecule, wherein each of the pendant moieties is attached to the polymer backbone and capable of chelating with an element. Also provided is a conjugate, comprising a biomolecule covalently coupled with the tag. 1125.-. (canceled)126. A tag for labeling a biomolecule comprising a polymer backbone terminating into an end group capable of stoichiometrically binding to the biomolecule , and one or more pendant moieties each attached to the polymer backbone and capable of chelating with an element.127. The tag of claim 126 , wherein the polymer backbone is selected from the group consisting of polypeptide claim 126 , polypeptoid claim 126 , poly β-peptide claim 126 , poly γ-peptide claim 126 , poly δ-peptide claim 126 , and a derivative thereof.128. The tag of claim 126 , wherein the polymer backbone is a homopolymer or a copolymer.129. The tag of claim 126 , wherein the polymer backbone is formed by polymerization of a monomer selected from the group consisting of NCA claim 126 , NTAα-amino acid claim 126 , β-amino acid claim 126 , γ-amino acid claim 126 , δ-amino acid claim 126 , N-substituted amino acid claim 126 , and a derivative thereof.130. The tag of claim 126 , wherein the pendant moiety is selected from the group consisting of EDTA claim 126 , DTPA claim 126 , DCTA claim 126 , DOTA claim 126 , TETA claim 126 , NOTA and a derivative thereof.131. A tag for labeling a biomolecule comprising a polymer backbone terminating into an end group capable of binding to the biomolecule claim 126 , and one or more pendant moieties each attached to the polymer backbone and capable of chelating with an element claim 126 , wherein the tag is bio-compatible.132. The tag of claim 131 , wherein the polymer backbone is selected from the group consisting of polypeptide claim 131 , polypeptoid claim 131 , poly β-peptide claim 131 , ...

PROCESS FOR MANUFACTURING CONJUGATES OF IMPROVED HOMOGENEITY

The invention provides processes for manufacturing cell-binding agent-cytotoxic agent conjugates of improved homogeneity comprising performing the modification reaction at a lower temperature. The inventive processes comprise contacting a cell-binding agent with a bifunctional crosslinking reagent at a temperature of about 15° C. or less to covalently attach a linker to the cell-binding agent and thereby prepare a mixture comprising cell-binding agents having linkers bound thereto. 1. A process for preparing a cell-binding agent having a linker bound thereto , which process comprises contacting a cell-binding agent with a bifunctional crosslinking reagent at a temperature of about 15° C. or less to covalently attach a linker to the cell-binding agent and thereby prepare a mixture comprising cell-binding agents having linkers bound thereto.2. The process of claim 1 , wherein the contacting occurs in a solution having a pH of about 7.5 to about 9.3. The process of claim 2 , wherein the pH is about 7.8.4. The process of claim 2 , wherein the solution comprises a buffering agent selected from a citrate buffer claim 2 , an acetate buffer claim 2 , a succinate buffer claim 2 , and a phosphate buffer.5. The process of claim 2 , wherein the solution comprises a buffering agent selected from the group consisting of HEPPSO (N-(2-Hydroxyethyl)piperazine-N′-(2-hydroxypropanesulfonic acid)) claim 2 , POPSO (Piperazine-1 claim 2 ,4-bis-(2-hydroxy-propane-sulfonic acid) dehydrate) claim 2 , HEPES (4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid) claim 2 , HEPPS (EPPS) (4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid) claim 2 , TES (N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid) claim 2 , and a combination thereof.6. The process of claim 1 , wherein the contacting occurs at a temperature of about −10° C. to about 15° C.7. The process of claim 6 , wherein the temperature is about 10° C.8. The process of claim 1 , wherein the cell-binding agent is selected from the ...

Anti-Transglutaminase 2 Antibodies

The invention provides antibodies and antigen-binding fragments thereof that selectively bind to an epitope within the core region of transglutaminase type 2 (TG2). Novel epitopes within the TG2 core are provided. The invention provides human TG2 inhibitory antibodies and uses thereof, particularly in medicine, for example in the treatment and/or diagnosis of conditions including Celiac disease, scarring, fibrosis-related diseases, neurodegenerative/neurological diseases and cancer.

ANTIBODIES AND METHODS OF USE

The presently disclosed subject matter provides antibodies that bind KLB and FGFR1, and methods of using the same. In certain embodiments, an antibody of the present disclosure includes a bispecific antibody that binds to an epitope present on FGFR1 and binds to an epitope present on KLB. 1. A nucleic acid encoding a bispecific antibody , or an antigen-binding portion thereof , that binds to beta-Klotho (KLB) and Fibroblast Growth Factor Receptor 1c (FGFR1c) , wherein the bispecific antibody , or an antigen-binding portion thereof , binds to a KLB epitope within a fragment of KLB consisting of the amino acid sequence set forth in SEQ ID NO: 142.2. The nucleic acid of claim 1 , wherein the bispecific antibody claim 1 , or an antigen-binding portion thereof claim 1 , comprises (a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 132 and (b) a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 134.3. The nucleic acid of claim 1 , wherein the bispecific antibody claim 1 , or an antigen-binding portion thereof claim 1 , comprises:(a) a heavy chain variable region CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 136;(b) a heavy chain variable region CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 137;(c) a heavy chain variable region CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 138;(d) a light chain variable region CDR1 domain comprising the amino acid sequence set forth in SEQ ID NO: 139;(e) a light chain variable region CDR2 domain comprising the amino acid sequence set forth in SEQ ID NO: 140; and(f) a light chain variable region CDR3 domain comprising the amino acid sequence set forth in SEQ ID NO: 141.4. The nucleic acid of claim 2 , wherein the bispecific antibody claim 2 , or an antigen-binding portion thereof claim 2 , comprises a heavy chain and a light chain claim 2 , wherein the heavy chain comprises amino acids ...

Methods and Compositions for Treating Autoimmune and Inflammatory Conditions

Described herein are compositions and methods for inhibiting an inflammatory or autoimmune response and for inducing immune tolerance in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an antigen presenting cell (APC)-targeted antibody operatively linked to IL-10 or a fragment thereof. The compositions and methods described herein are useful for treating inflammatory and autoimmune disorders. 1. A method of inducing immune tolerance in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an APC-targeted antibody operatively linked to IL-10 or a fragment thereof.2. A method for preventing or treating graft versus host disease in a subject in need thereof comprising administering to the subject a therapeutically effective amount of DC-ASGPR linked to IL-10 or a fragment thereof.3. The method of claim 2 , wherein the anti-DC-ASGPR antibody is a humanized antibody having three heavy chain CDRs and three light chain CDRs from the variable regions of an anti-DC-ASGPR heavy chain and light chain variable region pairs selected from SEQ ID NO:3 and 8; SEQ ID NO:58 and 60; SEQ ID NO:62 and 64; or SEQ ID NO:66 and 68; or is a humanized antibody having three heavy chain CDRs and three light chain CDRs from the heavy and light chains of an anti-DC-ASGPR heavy chain and light chain pair selected from SEQ ID NO:69 and 70 and SEQ ID NO:71 and 72.4. A method for inhibiting an inflammatory or autoimmune response in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an antigen presenting cell (APC)-targeted antibody operatively linked to IL-10 or a fragment thereof.5. A method of suppressing a T cell response in a subject in a subject having or at risk of developing an inflammatory response by administering to the subject a therapeutically effective amount of an APC-targeted antibody operatively linked to IL-10 or a fragment ...

SINGLE CHAIN PROTEINS WITH C-TERMINAL MODIFICATIONS

The invention pertains to an isolated Vsingle domain antibody comprising a C-terminal modification, where the C-terminal modification comprises a deletion of at least one amino acid residue that eliminates the interaction of a pre-existing antibody with the single domain antibody without interfering with the binding of the single domain antibody with its target. 1. A Vsingle domain antibody comprising a C-terminal deletion of at least two amino acid residues such that the deletion of at least two amino acid residue in the single domain antibody eliminates the interaction of a pre-existing antibody with the single domain antibody without interfering with the binding of the single domain antibody with its target.2. (canceled)3. (canceled)4. The single domain antibody of claim 1 , wherein the single domain antibody is a human V.5. The single domain antibody of claim 4 , wherein the human Vis selected from the group consisting of SEQ ID NO: 8 claim 4 , SEQ ID NO: 9 claim 4 , SEQ ID NO: 10 claim 4 , SEQ ID NO: 11 claim 4 , SEQ ID NO: 12 claim 4 , and SEQ ID NO: 13.6. (canceled)7. (canceled)8. The single domain antibody of claim 1 , wherein the C-terminal deletion comprises at least three amino acid residues from the C-terminus of the single domain antibody.9. The single domain antibody of claim 1 , wherein the C-terminal deletion comprises at least four claim 1 , at least five claim 1 , at least six claim 1 , at least seven claim 1 , at least eight claim 1 , at least nine amino acid residues from the C-terminus of the single domain antibody.10. The single domain antibody of claim 1 , wherein the C-terminal modification is the deletion of an amino acid sequence selected from the group consisting of Arg-Thr claim 1 , Arg-Thr-Val claim 1 , Gly-Gln claim 1 , and Gly-Gln-Pro.11. (canceled)12. The single domain antibody of claim 1 , wherein the Vcomprises SEQ ID NO: 8 claim 1 , and wherein the C-terminal modification comprises a deletion of two to three amino acids.13. The ...

ANTIBODY HAVING INFECTION-INHIBITING ACTIVITY AGAINST HEPATITIS C VIRUS

An anti-hepatitis C virus E2 protein antibody or antigen-binding antibody fragment thereof has infection inhibiting activity against hepatitis C virus (HCV). An anti-hepatitis C virus E2 protein antibody or antigen-binding antibody fragment thereof includes a certain variable region, which have infection inhibiting activity against hepatitis C virus (HCV) and exhibits an escape mutant emergence suppressive property. 112-. (canceled)13. An anti-hepatitis C virus E2 protein antibody or antigen-binding antibody fragment thereof , which has infection inhibiting activity against hepatitis C virus (HCV).14. The antibody or antigen-binding antibody fragment according to claim 13 , which exhibits an escape mutant emergence suppressive property.15. The antibody or antigen-binding antibody fragment according to claim 13 , wherein said antibody or antigen-binding antibody fragment comprises a heavy chain variable region of the following (a) or (b):(a) a heavy chain variable region comprising a CDR1 consisting of the amino acid sequence of SEQ ID NO: 5, a CDR2 consisting of the amino acid sequence of SEQ ID NO: 6, and a CDR3 consisting of the amino acid sequence of SEQ ID NO: 7; or(b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2.16. The antibody or antigen-binding antibody fragment according to claim 13 , wherein said antibody or antigen-binding antibody fragment comprises a light chain variable region of any one of the following (c) to (f):(c) a light chain variable region comprising a CDR1 consisting of the amino acid sequence of SEQ ID NO: 8, a CDR2 consisting of the amino acid sequence of SEQ ID NO: 9 and a CDR3 consisting of the amino acid sequence of SEQ ID NO: 10;(d) a light chain variable region comprising a CDR1 consisting of the amino acid sequence of SEQ ID NO: 23, a CDR2 consisting of the amino acid sequence of SEQ ID NO: 24 and a CDR3 consisting of the amino acid sequence of SEQ ID NO: 25;(e) a light chain variable region ...

HUMANIZED ANTIBODIES TO TNF-LIKE LIGAND 1A (TL1A) AND USES THEREOF

Described herein are humanized anti-TL1A antibodies and pharmaceutical compositions for the treatment of inflammatory bowel disease (IBD), such as Crohn's Disease (CD) and ulcerative colitis (UC). 1. An antibody or antigen binding fragment thereof that binds to TL1A , comprising a heavy chain variable framework region comprising a human IGHV1-46*02 framework or a modified human IGHV1-46*02 framework , and alight chain variable framework region comprising a human IGKV3-20 framework or a modified human IGKV3-20 framework; wherein the heavy chain variable framework region and the light chain variable framework region collectively comprise no or fewer than nine amino acid modification(s) from the human IGHV1-46*02 framework and the human IGKV3-20 framework.2. The antibody or antigen binding fragment of claim 1 , wherein the amino acid modification(s) comprise: (a) a modification at amino acid position 45 in the heavy chain variable region; (b) a modification at amino acid position 47 in the heavy chain variable region; (c) a modification at amino acid position 55 in the heavy chain variable region; (d) a modification at amino acid position 78 in the heavy chain variable region; (e) a modification at amino acid position 80 in the heavy chain variable region; (f) a modification at amino acid position 82 in the heavy chain variable region; (g) a modification at amino acid position 89 in the heavy chain variable region; or (h) a modification at amino acid position 91 in the heavy chain variable region claim 1 , per Kabat numbering; (i) or a combination of two or more modifications selected from (a) to (h).3. The antibody or antigen binding fragment of claim 1 , wherein the amino acid modification(s) comprise (a) R45K claim 1 , (b) A47R claim 1 , (c) M55I claim 1 , (d) V78A claim 1 , (e) M80I claim 1 , (f) R82T claim 1 , (g) V89A claim 1 , or (h) M91L in the heavy chain variable region claim 1 , per Kabat numbering; (i) or a combination of two or more modifications selected ...

ENGINEERED SITE-SPECIFIC ANTIBODIES AND METHODS OF USE

Provided are novel antibody drug conjugates (ADCs), and methods of using such ADCs to treat proliferative disorders. 1. An engineered antibody comprising one or more unpaired cysteine residue(s) wherein the unpaired cysteine residue(s) are exclusive of cysteines that form native interchain disulfide bonds.2. An engineered antibody according to wherein the engineered antibody comprises an IgG1 antibody.3. An engineered antibody according to or wherein the engineered antibody comprises a CL domain and the CL domain comprises a kappa CL domain.4. An engineered antibody according to or wherein the engineered antibody comprises a CL domain and the CL domain comprises a lambda CL domain.5. An engineered antibody according to any one of to wherein the engineered antibody comprises a monoclonal antibody.6. An engineered antibody according to any one of to wherein the engineered antibody comprises a humanized antibody or a CDR grafted antibody.7. An engineered antibody according to any one of to wherein the engineered antibody comprises two unpaired cysteine residues.8. An engineered antibody according to comprising two light chains claim 7 , wherein each light chain comprises one of the two unpaired cysteine residues.9. An engineered antibody according to wherein each of the two unpaired cysteine residues is present in the CL domains of the light chains.10. An engineered antibody of wherein the each of the two unpaired cysteine residues is present in the C-terminal region of the CL domains of the light chains.11. An engineered antibody of wherein each of the two unpaired cysteine residues is positioned in the exposed loop structures of the CL domain.12. An engineered antibody of wherein each of two unpaired cysteine residues is at a position selected from any one of residues 121-128 claim 8 , residues 182-191 or residues 201-213 of the CL domain wherein the residues are numbered according to Kabat.13. An engineered antibody of wherein the antibody light chain comprises a CL ...

Analysis of Disulfide Bonds

The present disclosure relates to methods of evaluating, identifying, and/or producing (e.g., manufacturing) pharmaceutical products (e.g., protein therapeutics). In some instances, methods herein allow highly resolved evaluation of the disulfide bond profiles of protein therapeutics. 1. (canceled)2. A method of manufacturing a pharmaceutical product , said method comprising:obtaining a sample of a batch of a test biologic;determining a disulfide bond profile for said sample;acquiring an assessment made by comparing said determined disulfide bond profile with a disulfide bond profile of a target protein (e.g., a specification including a disulfide bond profile of a target protein), wherein said target protein is a biologic approved under a primary approval pathway;processing said batch of said test biologic into a pharmaceutical product if said assessment reveals said determined disulfide bond profile conforms with said disulfide bond profile of said target protein;thereby manufacturing a pharmaceutical product.3. The method of claim 2 , wherein said determining step comprises digesting said sample with one or more protease enzymes in a digestion buffer.4. The method of claim 3 , wherein said digestion buffer comprises trypsin claim 3 , flavastacin claim 3 , LysC claim 3 , and/or GluC.5. The method of claim 2 , wherein said determining step comprises digesting said sample with no more than one protease enzyme in a digestion buffer.6. The method of claim 5 , wherein said digestion buffer comprises trypsin.7. The method of claim 2 , wherein said determining step comprises digesting said sample with at least two protease enzymes in a digestion buffer.8. The method of claim 7 , wherein said digestion buffer comprises no more than two protease enzymes.9. The method of claim 8 , wherein said digestion buffer comprises trypsin and GluC.10. The method of claim 9 , wherein said digestion buffer further comprises one or more glycosidase enzymes.11. The method of claim 10 , ...

Aldehyde Tags, Uses Thereof in Site-Specific Protein Modification

The invention features compositions and methods for site-specific modification of proteins by incorporation of an aldehyde tag. Enzymatic modification at a sulfatase motif of the aldehyde tag through action of a formylglycine generating enzyme (FGE) generates a formylglycine (FGly) residue. The aldehyde moiety of FGly residue can be exploited as a chemical handle for site-specific attachment of a moiety of interest to a polypeptide. 124.-. (canceled)26. The method of claim 25 , wherein the heterologous sulfatase motifis positioned at a C-terminus of the modified non-naturally occurring, recombinant polypeptide;is present in a terminal loop of the modified non-naturally occurring, recombinant polypeptide;is, when the modified non-naturally occurring, recombinant polypeptide is a transmembrane protein, present at an internal site within an extracellular loop or an intracellular loop;is present at an internal site or at the N-terminus of the modified non-naturally occurring, recombinant polypeptide, and is solvent-accessible when the polypeptide is folded; and/oris present at a site of post-translational modification.27. The method of claim 25 , wherein the heterologous sulfatase motif is positioned in the modified non-naturally occurring claim 25 , recombinant polypeptide at a site of post-translational modification of that is native or non-native to the parent of the modified non- naturally occurring claim 25 , recombinant polypeptide.28. The method of claim 25 , wherein the modified non-naturally occurring claim 25 , recombinant polypeptide is selected from the group consisting of an Fc fragment claim 25 , an antibody claim 25 , an antigen-binding fragment of an antibody claim 25 , a blood factor claim 25 , a fibroblast growth factor claim 25 , a protein vaccine claim 25 , and an enzyme.29. The method of claim 25 , wherein the modified non-naturally occurring claim 25 , recombinant polypeptide comprises an Fc fragment.30. The method of claim 25 , wherein the ...

T CELL RECEPTOR-LIKE ANTIBODY AGENTS SPECIFIC FOR EBV LATENT MEMBRANE PROTEIN 2A PEPTIDE PRESENTED BY HUMAN HLA

Described herein are antibodies, fragments thereof and multi-specific binding agents that bind an Epstein-Barr virus (EBV) latent membrane protein 2 (LMP2) peptide presented by HLA class I molecules, in particular, HLA-A02. Also provided herein are methods of using the same or compositions thereof for the detection, prevention and/or therapeutic treatment of diseases characterized by expression of an EBV-LMP2 peptide presented by HLA-A02, in particular, Burkit's lymphoma, Hodgkin's lymphoma and nasopharyngeal carcinoma. 1. A human antibody agent that binds an EBV-LMP2/HLA peptide complex , wherein the human antibody agent comprises:(a) a heavy chain variable region having a sequence at least 95% identical to a heavy chain variable region sequence as set forth in any one of SEQ ID NOs: 5, 9, 13, 17, 21, 25, and 29, and(b) a light chain variable region having a sequence at least 95% identical to a light chain variable region sequence as set forth in any one of SEQ ID NOs: 3, 7, 11, 15, 19, 23, and 27.2. The human antibody agent of claim 1 , wherein the human antibody agent has a Kof about 2.0 to about 170 nM.3. The human antibody agent of claim 1 , wherein the EBV-LMP2/HLA peptide complex comprises an EBV-LMP2 peptide that has an amino acid sequence that is or comprises CLGGLLTMV (SEQ ID NO: 1).4. The human antibody agent of claim 3 , wherein the human antibody agent interacts directly with amino acid residues at one or more positions selected from the group consisting of position 1 claim 3 , position 5 claim 3 , position 8 claim 3 , and combinations thereof claim 3 , in the EBV-LMP2 peptide.513.-. (canceled)14. The human antibody agent of claim 1 , wherein the human antibody agent is a human monoclonal antibody or a fragment thereof.15. The human antibody agent of claim 14 , wherein the human monoclonal antibody is an IgG1.16. The human antibody agent of claim 14 , wherein the human monoclonal antibody fragment is an scFv.17. An isolated nucleic acid molecule ...

MUTATED ANTI-TNFa ANTIBODIES AND METHODS OF THEIR USE

The present invention is directed to modified antibodies, including anti-TNFα antibodies, in which C-terminal amino acids of heavy chain sequences are modified from a native sequence of proline-glycine-lysine (“PGK”) to one that includes a proline positioned between the glycine and lysine, resulting in a C-terminal sequence of proline-glycine-proline-lycine (“PGPK”). The invention further provides methods of producing and using such antibodies.

IGG4 FC FRAGMENT COMPRISING MODIFIED HINGE REGION

The present invention relates to a modified IgG4 Fc fragment useful as a drug carrier. When the modified IgG4 Fc fragment of the present invention is combined with an arbitrary drug, the resulting drug conjugate can minimize the effector functions of the IgG4 Fc and the chain exchange with in vivo IgG while maintaining in vivo activity and improving in vivo duration of the drug conjugate. 2. The modified IgG4 Fc fragment of claim 1 , wherein in vivo chain exchange and monomer formation do not occur.3. The modified IgG4 Fc fragment of claim 1 , wherein the hinge region was mutated by the deletion of 1 to 8 continuous or discontinuous amino acid(s) comprising a Cys residue at the 8position.4. The modified IgG4 Fc fragment of claim 1 , wherein the hinge region was mutated by the deletion of 1 to 8 continuous or discontinuous amino acid(s) comprising a Cys residue at the 11position.5. The modified IgG4 Fc fragment of claim 1 , wherein the hinge region was mutated by the deletion of 1 to 5 continuous or discontinuous amino acid(s) comprising a Cys residue at the 8position.6. The modified IgG4 Fc fragment of claim 1 , wherein the hinge region was mutated by the deletion of 1 to 5 continuous or discontinuous amino acid(s) comprising a Cys residue at the 11position.7. The modified IgG4 Fc fragment of claim 1 , wherein the hinge region was mutated by the deletion of 1 to 3 continuous or discontinuous amino acid(s) comprising a Cys residue at the 8position.8. The modified IgG4 Fc fragment of claim 1 , wherein the hinge region was modified by the deletion of 1 to 3 continuous or discontinuous amino acid(s) comprising a Cys residue at the 11position.9. The modified IgG4 Fc fragment of claim 1 , which is aglycosylated.10. The modified IgG4 Fc fragment of claim 1 , which is modified by a nonpeptidyl polymer.11. A nucleic acid encoding the IgG4 Fc fragment of .12. A vector comprising the nucleic acid of .13. A microorganism introduced with the vector of .14. A method of preparing ...

Immunoconjugates with an Intracellularly-Cleavable Linkage

The present invention relates to therapeutic conjugates with improved ability to target various diseased cells containing a targeting moiety (such as an antibody or antibody fragment), a linker and a therapeutic moiety, and further relates to processes for making and using the conjugates.

ANTIBODY LIBRARY AND ANTIBODY SCREENING METHOD USING SAME

The present invention relates to a novel antibody library and an antibody-screening method using same. Having a human sequence-derived specific VH or VL scaffold, the antibody library according to the present invention exhibits high thermodynamic stability and enjoys the advantages of allowing high soluble expression as well as reversible folding. In addition, the antibody according to the present invention includes a variety of rationally controlled CDRs so as to exhibit high specificity and high affinity to all antigens and thus can be advantageously used for selecting an adequate candidate antibody against a target antigen. 1. A set of antibodies or fragments thereof ,wherein each antibody or fragment thereof comprises a pair of a heavy-chain variable region and a light-chain variable region,wherein the heavy-chain variable region comprises:a framework region included in a heavy-chain variable region selected from the group consisting of VH3-15 (SEQ ID NO: 1), VH3-23 (SEQ ID NO: 6) and VH1-69 (SEQ ID NO: 11); anda combination of a heavy-chain complementarity-determining region 1 (CDRH1), a heavy-chain complementarity-determining region 2 (CDRH2), and a heavy-chain complementarity-determining region 3 (CDRH3), which are different for each heavy-chain variable region, andthe light-chain variable region comprises:a framework region included in a light-chain variable region selected from the group consisting of Vκ1-39 (SEQ ID NO: 16), Vκ3-20 (SEQ ID NO: 21), Vκ3-20-2 (SEQ ID NO: 26) and Vλ1-51 (SEQ ID NO: 31); anda combination of a light-chain complementarity-determining region 1 (CDRL1), a light-chain complementarity-determining region 2 (CDRL2), and a light-chain complementarity-determining region 3 (CDRL3), which are different for each light-chain variable region.2. The set of antibodies or fragments thereof according to claim 1 , wherein the set of antibodies or fragments thereof comprises:a framework region included in a pair of heavy- and light-chain variable ...

Purification Process for Removal of Tyrosine Sulfation Antibody Variants; Purified Compositions

The present invention relates to purified antibody and antigen-binding fragment compositions that lack sulfated tyrosine on one or more tyrosine residues in the immunoglobulin chains. Purification methods for removing sulfated tyrosine variants from antibody and antigen-binding fragment compositions are also provided. 2. The composition of wherein the antibody or antigen-binding fragment further lacks detectable levels of sulfated tyrosine in one or more members selected from the group consisting of FR-L1 claim 1 , FR-L2 claim 1 , CDR-L2 claim 1 , FR-L3 claim 1 , CDR-L3 claim 1 , FR-L4 claim 1 , FR-H1 claim 1 , CDR-H1 claim 1 , FR-H2 claim 1 , CDR-H2 claim 1 , FR-H3 claim 1 , CDR-H3 claim 1 , FR-H4 and a constant domain.3. The composition of wherein the antibody is Ab6 comprising a heavy chain amino acid sequence of SEQ ID NO: 52 and a light chain amino acid sequence of SEQ ID NO: 51.4. The composition of wherein the antibody or antigen-binding fragment comprises CHO N-linked glycans.5. The composition of wherein the antibody is Ab6 and the composition comprises one or more glycosylated antibody Ab6 species having molecular weights of about 148590 Da claim 3 , 148752 Da or 148914 Da.6. The composition of wherein the antibody is Ab6 with the N-terminal heavy chain glutamine converted to pyroglutamate and/or C-terminal heavy chain lysine removed.7. A method for removing tyrosine sulfated antibodies or antigen-binding fragments thereof from an aqueous mixture comprising tyrosine sulfated and non-tyrosine sulfated antibodies or antigen-binding fragments thereof comprising adjusting the pH of the mixture to about 6.5 to about 7.0 claim 3 , contacting the aqueous mixture with an anion exchange resin claim 3 , and retaining a non-resin bound aqueous fraction of the mixture from the resin.8. The method of which further comprises washing the resin with an aqueous composition under isocratic conditions and removing and retaining the wash composition from the resin.9. ( ...

GLUCURONYLATION AS A NEW ACIDIC POST-TRANSLATIONAL MODIFICATION ON THERAPEUTIC MONOCLONAL ANTIBODIES

Compositions and methods for identifying glucuronylated protein drug products are provided. 16-. (canceled)7. A protein drug product comprising a therapeutic protein , wherein less than 10% of the therapeutic protein's amino acids are glucuronylated.8. The protein drug product of claim 7 , wherein less than 5% of the therapeutic protein's amino acids are glucuronylated.9. The protein drug product of claim 7 , wherein the amino acids are lysine amino acids.10. The protein drug product of claim 7 , wherein the therapeutic protein is an antibody or fusion protein.11. The protein drug product of claim 10 , wherein the antibody is a monoclonal antibody or an antigen binding fragment thereof.12. A protein drug product comprising a therapeutic protein claim 10 , wherein less than 3% of lysine amino acids of the therapeutic protein are glucuronylated.13. A method for increasing purity of a protein drug product comprising:analyzing the protein drug product to identify glucuronylation on a primary amine of one or more amino acids of the protein drug product; andremoving glucuronylated proteins from the protein drug product to produce a purified protein drug product.14. The method of claim 13 , wherein the glucuronylated proteins are removed by high performance liquid chromatography techniques.15. The method of claim 14 , wherein the chromatography techniques are selected from the group consisting of size exclusion chromatography claim 14 , ion exchange chromatography claim 14 , affinity chromatography claim 14 , and combinations thereof.16. The method of claim 15 , wherein the chromatography technique is coupled with mass spectroscopy analysis.17. The method of claims 13 , wherein the amino acids are lysine amino acids.18. The method of claim 13 , wherein the therapeutic protein is an antibody or fusion protein.19. The method of claim 18 , wherein the antibody is a monoclonal antibody or an antigen binding fragment thereof.20. A method for identifying post-translationally ...

Deglycosylation Reagents and Methods

Compositions and methods are provided for efficiently preparing a completely deglycosylated antibody where efficiency is measured in relative amounts of reagents in soluble or lyophilized form, and time and temperature of the reaction. Compositions and methods are also provided for separating substantially all N-linked glycans from a glycosylated antibody and for preserving functionality of the antibody. The methods are compatible with glycan labeling and protease digestion without the need for prior purification steps. 1. A composition , comprising:(i) a bile salt or a dialyzable non-cleavable carboxylated anionic surfactant(ii) peptide-N-glycosidase F (PNGase F);(iii) a completely deglycosylated antibody as determined by mass spectrometry; and(iv) glycan cleavage products.2. The composition according to claim 1 , wherein the composition comprises a bile.3. The composition according to claim 1 , wherein the completely deglycosylated antibody has antigen binding activity.4. The composition according to claim 3 , wherein the deglycosylated antibody is selected from deglycosylated human IgG1 claim 3 , human IgG2 claim 3 , human IgG3 claim 3 , human IgG4 claim 3 , human IgM claim 3 , human IgA1 claim 3 , human IgA2 claim 3 , human IgE claim 3 , murine IgG1 claim 3 , murine IgG2a and murine IgA.5. The composition according to claim 3 , wherein the completely deglycosylated antibody is suitable for use in an immunoassay.6. The composition according to claim 1 , wherein the glycan cleavage products and/or the antibody are labeled with a fluorescent label claim 1 , a radioisotope claim 1 , methyl acetyl claim 1 , an antibody or a combination thereof.7. The composition according to claim 1 , further comprising a protease.8. The composition according to claim 7 , wherein the protease is trypsin.9. The composition according to claim 7 , wherein the protease is selected from trypsin claim 7 , GluC claim 7 , AspN claim 7 , proteinase K claim 7 , Factor Xa claim 7 , Enterokinase ...

MODIFIED HUMAN VARIABLE DOMAINS

The present invention provides polypeptides comprising modified human, humanized, or chimeric immunoglobulin heavy chain variable domains. Corresponding antibodies, variants, fragments, nucleic acids, vectors, phages, libraries, methods and kits are also provided. 1. A polypeptide comprising a human , humanized , or chimeric immunoglobulin heavy chain variable domain , wherein the variable domain comprises:(a) an N-terminal amino acid selected from the group consisting of: arginine, asparagine, aspartate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, and tyrosine; or(b) an N-terminal sequence selected from the group consisting of: alanine-proline, alanine-aspartate, alanine-glutamate, alanine-threonine, alanine-valine, alanine-serine, and alanine-leucine.2. The polypeptide of claim 1 , wherein the human claim 1 , humanized claim 1 , or chimeric immunoglobulin heavy chain variable domain comprises the N-terminal sequence alanine-proline.3. The polypeptide of claim 1 , wherein the polypeptide comprises a signal peptide upstream of the N-terminal amino acid of the human claim 1 , humanized claim 1 , or chimeric immunoglobulin heavy chain variable domain.4. An antibody claim 1 , antibody variant or antibody fragment comprising the polypeptide of .5. A nucleic acid encoding the polypeptide of or an antibody claim 1 , antibody variant or antibody fragment comprising the polypeptide of claim 1 , optionally wherein the nucleic acid is within a vector or a phage.6. A library comprising at least about 10distinct nucleic acids claim 5 , vectors claim 5 , or phages according to .8. The method of claim 7 , wherein each amplified nucleic acid encodes a human claim 7 , humanized claim 7 , or chimeric immunoglobulin heavy chain variable domain comprising an N-terminal alanine claim 7 , optionally wherein each amplified nucleic acid encodes a human claim 7 , humanized claim 7 , or chimeric immunoglobulin heavy ...

Engineered Antibodies for the Detection of Phosphorylated Tyrosine

Presented herein are novel phosphotyrosine binding compositions, including antibodies and antibody fragments. The inventors of the present disclosure have resolved the crystal structures of two widely utilized pan-specific pY antibodies, PY20 and 4G10. These two known antibodies, although developed independently from animal immunizations, have surprisingly similar modes of recognition of the phosphate group, and revealed a generic binding structure among pan-specific pY antibodies. Based on this newly discovered convergent structure, engineered CDR-L3 loops were developed that impart greatly improved affinity to pY binding antibodies and other pY binding compositions. The inventions disclosed herein include antibodies and antibody fragments bearing these novel CDR-L3 sequences and methods of using such pY binding compositions for the detection of phosphotyrosine-bearing proteins. 1. A phosphotyrosine binding composition , comprisingan antibody, or phosphotyrosine-binding fragment thereof, wherein the antibody or antibody fragments comprises:a body scaffold comprising heavy and light chain sequences;a CDR L1 sequence, a CDR L2 sequence, a CDR H1 sequence, a CDR H2 sequence and a CDR H3 sequence, wherein one or more of the CDR L1 sequence, CDR L2 sequence, CDR H1 sequence, CDR H2 sequence and CDR H3 sequences imparts phosphotyrosine binding affinity; anda CDR L3 sequence comprising a sequence selected from the group consisting of: SEQ ID NO: 1-SEQ ID NO: 4 and SEQ ID NO: 6-SEQ ID NO: 23.2. The phosphotyrosine binding composition of claim 1 , wherein{'sub': '2', 'the antibody or fragment thereof comprises a whole antibody, a Fab, a Fab′, a F(ab), or a scFv.'}3. The phosphotyrosine binding composition of claim 1 , whereinthe antibody or fragment thereof comprises one or more functional moieties.4. The phosphotyrosine binding composition of claim 3 , whereinthe one or more functional moieties comprises an affinity tag, a fluorescent label, or an epitope for binding by ...

METHOD AND MEANS FOR DETECTING THE LEVEL OF TOTAL VEGF-A

The present invention relates to a method for measuring the level of VEGF-A in the presence of a VEGF-A antagonist, kits comprising means for detecting VEGF-A in the presence of a VEGF-A antagonist, compositions of matter comprising a first and a second antibody suitable for detecting the level of VEGF-A in the presence of a VEGF-A antagonist, as well as methods of detecting a complex comprising human VEGF-A and a non-human or chimeric protein. 1. A method for measuring the level of VEGF-A , in the presence of a VEGF-A antagonist , the method comprising: wherein said first and said second antibody, are capable of binding to VEGF-A in the presence of the VEGF-A antagonist, and wherein the binding of said first and of said second antibody does not interfere with each other,', 'wherein one of said antibodies is detectably labeled, thereby forming a detectably labeled complex comprising the first antibody, VEGF-A, and the second antibody, and, 'incubating a sample with a first and a second antibody,'}detecting the complex formed, thereby measuring the level of VEGF-A in the presence of a VEGF-A antagonist.2. The method of claim 1 , wherein the VEGF-A antagonist is a VEGF-A-binding polypeptide.3. The method of claim 1 , wherein the VEGF-A antagonist is bevacizumab.4. The method of claim 1 , wherein the first and/or the second antibody binds to an epitope which are covered by or bound by a VEGF receptor.5. The method of claim 1 , wherein the first or the second antibody comprises CDRs consisting of amino acids 46-51 of SEQ ID NO: 2 claim 1 , amino acids 69-71 of SEQ ID NO: 2 claim 1 , amino acids 108-116 of SEQ ID NO: 2 claim 1 , amino acids 44-52 of SEQ ID NO: 3 claim 1 , amino acids 70-76 of SEQ ID NO: 3 claim 1 , amino acids 115-125 of SEQ ID NO: 3 claim 1 , orcomprises CDRs consisting of amino acids 47-52 of SEQ ID NO: 4, amino acids 70-72 of SEQ ID NO: 4, amino acids 109-117 of SEQ ID NO: 4, amino acids 45-52 of SEQ ID NO: 5, amino acids 70-77 of SEQ ID NO: 5, and ...

LYSINE CONJUGATED IMMUNOGLOBULINS

Provided herein are conjugated immunoglobulins and methods for generating conjugated immunoglobulins using a microbial transglutaminase. 2. The conjugated immunoglobulin of claim 1 , wherein the heavy chain further comprises an amino acid residue which has been added to its C-terminus at position 448 claim 1 , and wherein said amino acid residue is not proline or an acidic amino acid residue.3. The conjugated immunoglobulin of claim 2 , wherein the at least one amino acid residue which has been added to the C-terminus at position 448 is leucine.4. The conjugated immunoglobulin of claim 1 , wherein the immunoglobulin claim 1 , or antigen-binding portion thereof claim 1 , further comprises a second engineered lysine residue claim 1 , wherein the second engineered lysine residue is a second lysine residue insertion or a second natural amino acid residue which has been mutated to a lysine residue claim 1 , and wherein the second engineered lysine residue is conjugated to the glutamine residue of the acyl donor substrate.5. The conjugated immunoglobulin of claim 4 ,wherein the natural amino acid residue which has been mutated to the engineered lysine residue is Threonine 135 (T135K) on a heavy chain of the immunoglobulin, or antigen-binding portion thereof, and the second natural amino acid residue which has been mutated to the second engineered lysine residue is Leusine 201 (L201K) on a kappa light chain of the immunoglobulin, or antigen-binding portion thereof; orwherein the natural amino acid residue which has been mutated to the engineered lysine residue is Threonine 135 (T135K) on a heavy chain of the immunoglobulin, or antigen-binding portion thereof, and the second natural amino acid residue which has been mutated to the second engineered lysine residue is Serine 202 (S202K) on a kappa light chain of the immunoglobulin, or antigen-binding portion thereof;wherein the amino acid residues are numbered according to the EU numbering system.6. The conjugated ...

METHODS FOR OXIME CONJUGATION TO KETONE-MODIFIED POLYPEPTIDES

The invention provides improved methods to prepare protein conjugates from a ketone-modified protein. In one embodiment, the protein is prepared by linking two free cysteines by reaction with a 1,3-dihaloacetone or similar ketone-containing reactant, linking the sulfur atoms of the two cysteines together. The ketone inserted between the sulfur atoms is then used to form an oxime, thus conjugating the protein to a payload. In another embodiment, two cysteine residues are tied together by reaction with a 1,3-dihaloacetone or similar reactant, and the new ketone is used to form an oxime with a suitable payload molecule, thus conjugating the protein to a payload. The method provides improved reaction conditions for the oxime formation, whereby higher yields and improved product homogeneity are achieved. 1. A method for converting a polypeptide comprising a reducible disulfide bond into a ketone-modified polypeptide that further comprises a group of the formula [PP]—S—CH—C(═O)—CH—S—[PP] , where each S is a sulfur from the disulfide bond , and [PP] indicates where the ends of the linking group are attached to the polypeptide ,wherein the method comprises the step of: forming a mixture of a polypeptide containing a reducible disulfide bond, an aqueous buffer, and 1,3-dihaloacetone, then adding a reducing agent capable of reducing the disulfide bond.2. The method of claim 1 , wherein the 1 claim 1 ,3-dihaloacetone is 1 claim 1 ,3-dichloroacetone.3. The method of claim 1 , wherein the reducing agent is a water-soluble phosphine or phosphine salt.4. The method of claim 1 , wherein the polypeptide is an antibody or antibody fragment.5. A method for converting a ketone-modified polypeptide into an oxime-modified polypeptide comprising the step of: contacting the ketone modified polypeptide with a group of the formula R—O—NHin the presence of an amine promoter and at a polypeptide concentration of at least about 1 mg/mL.6. The method of claim 1 , wherein the ketone-modified ...

ANTIBODIES WITH DECREASED DEAMIDATION PROFILES

The present invention relates to antibodies with decreased deamidation profiles, and methods for producing antibodies with decreased deamidation profiles. 1. A method of producing an antibody with a decreased deamidation profile , wherein said antibody would otherwise be predisposed to an elevated deamidation profile.23-. (canceled)4. The method of claim 1 , wherein said antibody comprises an asparagine residue preceding and adjacent to a glycine claim 1 , serine claim 1 , threonine or an aspartic acid residue claim 1 , as read N-terminus to C-terminus.5. The method of claim 4 , wherein said residues are located in at least one of the VHCDR1 claim 4 , VHCDR2 claim 4 , VHCDR3 claim 4 , VLCDR1 claim 4 , VLCDR2 claim 4 , or VLCDR3 regions of said antibody.6. The method of claim 5 , wherein said residues are located in the VHCDR2 of said antibody.7. The method of any of claim 5 , wherein said antibody deamidation profile is decreased by about 60% claim 5 , about 50% claim 5 , about 40% claim 5 , about 30% claim 5 , about 20% claim 5 , or about 10% as compared to a control deamidation profile.8. The method of any of claim 7 , wherein said method comprises production of an antibody from cells grown at a temperature in the range of between about 30° C. to about 37° C.9. The method of any of claim 8 , wherein said temperature is about 34° C.10. The method of any of claim 9 , wherein said method comprises production of an antibody from cells grown in media at a pH from the range of between about 6.0 to about 7.2 pH units.11. The method of any of claim 10 , wherein said pH is about 6.9 pH units.12. The method of any of claim 11 , wherein said method comprises production of an antibody from cells grown in a biphasic culture.13. The method of claim 12 , wherein said biphasic culture comprises at least one temperature shift.14. The method of claim 13 , wherein said temperature shift comprises a shift from about 34° C. to about 32° C.15. (canceled)16. The method of any of claim ...

Fab FRAGMENT SPECIFICALLY BINDING TO EGFR

The present invention relates to a fragment antigen-binding (Fab) fragment specifically binding to epidermal growth factor receptor (EGFR), an expression construct for preparing the Fab fragment, a method for preparing the Fab fragment, and a pharmaceutical composition containing the Fab fragment. The Fab fragment to EGFR of the present invention is smaller than the antibody, and thus can favorably permeate into tissues or tumors and can be prepared in bacteria, resulting in low production costs. Furthermore, the Fab fragment to EGFR of the present invention has an increased in vivo half-life through pegylation. 1. A fragment antigen-binding (Fab) fragment specifically binding to epidermal growth factor receptor (EGFR) , the Fab fragment comprising:{'sub': 'H', '(a) a heavy chain variable region (V) comprising an amino acid sequence of SEQ ID NO: 4;'}{'sub': 'H1', '(b) a heavy chain variable region 1 (C) comprising an amino acid sequence of SEQ ID NO: 5;'}{'sub': 'L', '(a) a light chain variable region (V) comprising an amino acid sequence of SEQ ID NO: 6; and'}(d) a light chain constant region (CO comprising an amino acid sequence of SEQ ID NO: 7.2. The Fab fragment of claim 1 , wherein the Cfurther comprises Cys-Asp-Lys at the C-terminal thereof.3. The Fab fragment of claim 1 , wherein the Cfurther comprises Glu-Cys at the C-terminal thereof.4. The Fab fragment of claim 2 , wherein the Chas Thr-His-Thr-Cys-Ala-Ala further linked to Cys-Asp-Lys at the C-terminal thereof.5. The Fab fragment of claim 1 , wherein the Fab fragment is pegylated.6. The Fab fragment of claim 5 , wherein the Cof the Fab fragment is pegylated.7. The Fab fragment of claim 6 , wherein in the Thr-His-Thr-Cys-Ala-Ala at the C-terminal of the Cof the Fab fragment claim 6 , the Cys residue is pegylated.8. The Fab fragment of claim 5 , wherein the polyethylene glycol (PEG) used in the pegylation has a molecular weight of 5-50 kDa.9. The Fab fragment of claim 8 , wherein the PEG has a molecular ...

BIOLOGICAL PRODUCTS

There is disclosed antibody molecules containing at least one CDR derived from a mouse monoclonal antibody having specificity for human CD22. There is also disclosed a CDR grafted antibody wherein at least one of the CDRs is a modified CDR. Further disclosed are DNA sequences encoding the chains of the antibody molecules, vectors, transformed host cells and uses of the antibody molecules in the treatment of diseases mediated by cells expressing CD22. 118-. (canceled)19. An antibody molecule having specificity for human CD22 comprising a heavy chain and a light chain wherein the variable domains comprise the amino acid sequences of SEQ ID NO:1 for CDR-H1; SEQ ID NO:3 for CDR-H3; SEQ ID NO:4 for CDR-L1; SEQ ID NO:5 for CDR-L2; and SEQ ID NO:6 for CDR-L3.20. The antibody molecule of claim 19 , which is a CDR-grafted antibody molecule.21. The antibody molecule of claim 19 , wherein the variable domain comprises human acceptor framework regions and non-human donor CDRs.22. A composition comprising the antibody molecule of any one of -.23. The composition according to claim 22 , comprising a pharmaceutically acceptable excipient claim 22 , diluent claim 22 , or carrier.24. The composition according to claim 22 , additionally comprising anti-T cell claim 22 , anti-IFNγ claim 22 , anti-LPS antibodies claim 22 , or a non-antibody ingredient. This application is a Continuation of application Ser. No. 15/069,078, filed on Mar. 14, 2016, which is a Continuation of application Ser. No. 11/519,585, filed on Sep. 11, 2006, which is a Continuation of application Ser. No. 10/428,408, filed May 2, 2003, now U.S. Pat. No. 7,355,011 B2 which claims priority under 35 U.S.C. §119(a)-(d) to United Kingdom Application No. GB 0210121.0, filed May 2, 2002, all applications being incorporated by reference herein in their entireties.The present invention relates to an antibody molecule having specificity for antigenic determinants of the B lymphocyte antigen, CD22. The present invention also ...

SUBTYPES OF HUMANIZED ANTIBODY AGAINST INTERLEUKIN-6 RECEPTOR

An antibody subtype (1) which is a subtype of the humanized PM-1 antibody against interleukin-6 receptor (IL-6R) and in which one C-terminal of the heavy chain is Pro-NH(447), and an antibody subtype (2) which is a subtype of the humanized PM-1 antibody against interleukin-6 receptor (IL-6R) and in which both C-terminals of the heavy chain are Pro-NH(447), and a pharmaceutical composition comprising them. 16-. (canceled)7: A method of producing an antibody subtype 1 comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody , wherein said antibody subtype 1 comprises a heavy chain that has amino acids 1-448 of the amino acid sequence set forth in SEQ ID NO: 1 , a heavy chain that has amino acids 1-447 of the amino acid sequence set forth in SEQ ID NO: 1 and a C-terminal that is Pro-NH2 , and a light chain that has the amino acid sequence set forth in SEQ ID NO: 2.8: A method of producing an antibody subtype comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody , wherein said antibody subtype comprises a heavy chain that has amino acids 1-448 of the amino acid sequence set forth in SEQ ID NO: 1 in which an N-terminal glutamine (Gln) is pyroglutamylated , a heavy chain that has amino acids 1-447 of the amino acid sequence set forth in SEQ ID NO: 1 in which an N-terminal glutamine (Gln) is pyroglutamylated and a C-terminal that is Pro-NH2 , and a light chain that has the amino acid sequence set forth in SEQ ID NO: 2.9: A method of producing an antibody subtype 2 comprising culturing a host cell comprising at least one nucleic acid encoding for a heavy chain and at least one nucleic acid encoding for a light chain of an anti-IL-6R antibody , wherein said antibody subtype 2 comprises two heavy chains that have amino acids 1-447 ...

HYPERGLYCOSYLATED BINDING POLYPEPTIDES

Provided are binding polypeptides (e.g., antibodies), and effector moiety conjugates thereof, comprising a CH1 domain (e.g., a human IgG1 CH1 domain), wherein the CH1 domain has an engineered N-linked glycosylation site at amino acid position 114, according to Kabat numbering. Also provided are nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided. 1. An isolated binding polypeptide comprising a CH1 domain , wherein the CH1 domain comprises an asparagine residue at amino acid position 114 , according to Kabat numbering.2. The binding polypeptide of claim 1 , wherein the CH1 domain:further comprises any amino acid residue except proline at position 115, according to Kabat numbering, and a serine or threonine residue at position 116, according to Kabat numbering; oris an IgG1 CH1 domain, or variant thereof, that is optionally a human IgG1 CH1 domain, or variant thereof.3. (canceled)4. (canceled)5. The binding polypeptide of wherein:the side chain of the asparagine residue is linked to a glycan though a β-glycosylamide linkage;the glycan is a biantennary glycan;the glycan is a naturally occurring mammalian glycoform;the glycan comprises a reactive aldehyde group;the glycan comprises an oxidized saccharide residue comprising a reactive aldehyde group; orthe glycan is linked to an effector moiety.69-. (canceled)10. The binding polypeptide of claim 5 , wherein the oxidized saccharide residue is a terminal sialic acid or galactose.11. (canceled)12. The binding polypeptide of claim 5 , wherein the effector moiety:is a cytotoxin that is optionally selected from the group consisting of the cytotoxins listed in Table 1;is a detection agent;is linked through an oxime or hydrazone linkage to saccharide residue of the glycan, wherein the saccharide is optionally a terminal sialic acid ...

SODIUM CHANNEL INHIBITORS

Binding members for sodium channel Nav1.7 and their use in medicine including for treatment of pain or epilepsy. Binding members comprise a fusion protein containing a Nav1.7-binding peptide, e.g., venom toxin peptide or knottin (“donor diversity scaffold domain”) inserted within an antibody variable domain (“recipient diversity scaffold domain”), and a partner domain (e.g., antibody variable domain), optionally wherein the partner domain enhances specificity of binding to Nav1.7 over other sodium channels. 1. A binding member that binds and inhibits human sodium channel Nav1.7 , the binding member comprising a fusion protein and a partner domain , whereinthe fusion protein comprises a donor diversity scaffold domain inserted into a recipient diversity scaffold domain, wherein the donor diversity scaffold domain comprises a donor scaffold and a donor interaction sequence, and wherein the recipient diversity scaffold domain comprises a recipient scaffold and a recipient interaction sequence, whereinthe donor diversity scaffold domain is a cysteine-rich peptide, such as an ion-channel modulating peptide, venom toxin peptide or knottin, which binds to the voltage sensing domain of Nav1.7, whereinthe recipient diversity scaffold domain is an antibody variable domain, whereinthe partner domain is an antibody variable domain.2. A binding member according to claim 1 , wherein the donor diversity scaffold domain is a peptide having at least 95% sequence identity to (i) ProTx-III claim 1 , (ii) ProTx-II claim 1 , (iii) Huwentoxin-IV claim 1 , (iv) Ssm6a claim 1 , or (v) GpTx-1.3. A binding member according to claim 1 , wherein the donor diversity scaffold domain replaces all or part of a complementarity determining region (CDR) of the recipient diversity scaffold antibody variable domain.4. A binding member according to claim 1 , wherein the recipient diversity scaffold domain is an antibody VL domain.5. A binding member according to claim 4 , wherein the donor diversity ...

CYSTEINE ENGINEERED ANTI-TENB2 ANTIBODIES AND ANTIBODY DRUG CONJUGATES

Cysteine engineered anti-TENB2 antibodies are engineered by replacing one or more amino acids of a parent anti-TENB2 antibody with non cross-linked, reactive cysteine amino acids. Methods of design, preparation, screening, and selection of the cysteine engineered anti-TENB2 antibodies are provided. Cysteine engineered anti-TENB2 antibodies (Ab) are conjugated with one or more drug moieties (D) through a linker (L) to form cysteine engineered anti-TENB2 antibody-drug conjugates having Formula I: 1. A cysteine engineered anti-TENB2 antibody comprising one or more free cysteine amino acids and a sequence selected from SEQ ID NOS:8-23.2. The cysteine engineered anti-TENB2 antibody of wherein the cysteine engineered anti-TENB2 antibody binds to a TENB2 polypeptide.3. The cysteine engineered anti-TENB2 antibody of prepared by a process comprising replacing one or more amino acid residues of a parent anti-TENB2 antibody by cysteine.47-. (canceled)10. The cysteine engineered anti-TENB2 antibody of wherein the parent anti-TENB2 antibody is selected from a monoclonal antibody claim 1 , a bispecific antibody claim 1 , a chimeric antibody claim 1 , a human antibody claim 1 , and a humanized antibody.11. The cysteine engineered anti-TENB2 antibody of which is an antibody fragment.1215-. (canceled)16. A method of determining the presence of a TENB2 protein in a sample suspected of containing said protein claim 1 , said method comprising exposing said sample to a cysteine engineered anti-TENB2 antibody of and determining binding of said antibody to said TENB2 protein in said sample claim 1 , wherein binding of the antibody to said protein is indicative of the presence of said protein in said sample.17. The method of wherein said sample comprises a cell suspected of expressing said TENB2 protein.18. The method of wherein said cell is a prostate claim 16 , ovarian claim 16 , breast claim 16 , lung claim 16 , or pancreatic cancer cell.19. The method of wherein the antibody is ...

DE-N-Acetyl Sialic Acid Antigens, Antibodies Thereto, and Methods of Use in Cancer Therapy

The present invention generally provides compositions methods and composition relating to the diagnosis and/or treatment of cancers having a cell surface de-N-acetylated sialic acid antigen, e.g., an at least partially de-N-acetylated ganglioside and/or a de-N-acetylated sialic acid-modified cell surface protein. 144.-. (canceled)45. An antibody that specifically binds a de-N-acetylated sialic acid (deNAc SA) epitope on an extracellularly accessible surface of a cancerous cell , the deNAc SA epitope minimally defined by a dimer containing at least one de-N-acetylated sialic acid residue having a free amine adjacent an N-acylated sialic acid residue or a sialic acid derivative residue , wherein:the antibody binds to the deNAc SA epitope on the surface of CHP-134 neuroblastoma cells, and [ a VH CDR1 comprising amino acid residues 26 to 35 of SEQ ID NO:7,', 'a VH CDR2 comprising amino acid residues 50 to 66 of SEQ ID NO:7,', 'and', 'a VH CDR3 comprising amino acid residues 101 to 108 of SEQ ID NO:7;'}], 'a heavy chain polypeptide comprising, a VL CDR1 comprising amino acid residues 24 to 39 of SEQ ID NO:3,', 'a VL CDR2 comprising amino acid residues 55 to 61 of SEQ ID NO:3, and', 'a VL CDR3 comprising amino acid residues 94 to 100 of SEQ ID NO:3., 'a light chain polypeptide comprising], 'the antibody competes for binding to the deNAc SA epitope with an antibody comprising46. The antibody of claim 45 , wherein the antibody is selected from the group consisting of: an IgG claim 45 , a F(ab′)2 claim 45 , a F(ab) claim 45 , an Fv claim 45 , or an scFv.47. The antibody of claim 45 , wherein the antibody is a humanized antibody or a fully human antibody.48. The antibody of claim 45 , wherein the antibody comprises: a VH CDR1 comprising amino acid residues 26 to 35 of SEQ ID NO:7,', 'a VH CDR2 comprising amino acid residues 50 to 66 of SEQ ID NO:7, and', 'a VH CDR3 comprising amino acid residues 101 to 108 of SEQ ID NO:7; and, 'a heavy chain polypeptide comprising a VL CDR1 ...