ПОЛУЧЕНИЕ АНТИТЕЛ ИЗ ОДНИХ ТЯЖЕЛЫХ ЦЕПЕЙ В ТРАНСГЕННЫХ ЖИВОТНЫХ

Настоящее изобретение относится к биотехнологии. Описан способ получения VH антител, включающий стадии обеспечения более одного гетерологического VH локуса тяжелых цепей на разных хромосомах в данном млекопитающем. Причем каждый VH локус тяжелых цепей содержит один или более сегментов V-гена, один или более сегментов D-гена, один или более сегментов J-гена и сегмент гена, кодирующий постоянную область тяжелых цепей, которая при экспрессии не включает участок Сн1. В результате антигенной стимуляции только один локус продуктивно рекомбинируется в В-клетке, что приводит к получению VH антитела из одних тяжелых цепей и экспрессии указанного VH антитела из указанного продуктивно рекомбинировавшегося локуса. Изобретение позволяет расшить репертуар ответа антител. 4 н. и 28 з.п. ф-лы, 12 ил., 1 табл.

Способ получения димерной формы мутантного иммуноглобулина IgA2m1-изотипа в клетках млекопитающих

Изобретения относятся к области биотехнологии. Предложена рекомбинантная плазмидная ДНК p3.4-P4A1IgA2m1mut2-J12His-Intht, кодирующая нейтрализующее моноклональное антитело P4A1 к вирусу SARS-CoV-2 мутантного изотипа IgA2m1 и J-цепь человека. Плазмида содержит интронированный ген тяжелой цепи антитела, находящийся под контролем промотора hEF1-HTLV и сигнала полиаденилирования бычьего гормона роста BGH polyA, ген легкой цепи антитела и ген J-цепи человека, находящиеся под контролем промотора CMV и внутреннего сайта связывания рибосом (IRES) вируса энцефаломиокардита (EMCV), посттранскрипционный регуляторный элемент WPRE вируса гепатита сурка (WHV) и сигнал полиаденилирования тимидинкиназы вируса герпеса. Промоторы hEF1-HTLV и CMV размещены в последовательности, обеспечивающей однонаправленную транскрипцию. Также предложен способ получения димера нейтрализующего моноклонального антитела P4A1 против вируса SARS-CoV-2 мутантного изотипа IgA2m1 с J-цепью, включающий культивирование клеток СНО ...

Генетическая конструкция рБТШ70 для экспрессии основного человеческого стресс белка, используемого в терапевтических целях, в молоке и линия трансгенных мышей, содержащих данную конструкцию

МОНОКЛОНАЛЬНОЕ АНТИТЕЛО ПРОТИВ MELK И ЕГО ИСПОЛЬЗОВАНИЕ

СПОСОБЫ ПРЕОДОЛЕНИЯ ИММУНОЛОГИЧЕСКОЙ ТОЛЕРАНТНОСТИ С ИСПОЛЬЗОВАНИЕМ МНОЖЕСТВА НАПРАВЛЯЮЩИХ РНК

СПОСОБЫ И КОМПОЗИЦИИ ДЛЯ ПОЛУЧЕНИЯ СУРФАКТАНТНОГО БЕЛКА D (SP-D)

ХИМЕРНЫЕ РЕЦЕПТОРЫ АНТИГЕНА, НАЦЕЛЕННЫЕ НА PSCA

МОДЕЛЬ НЕСОВМЕСТИМОЙ ПО HLA ГУМАНИЗИРОВАННОЙ МЫШИ NSG С ПОЛУЧЕННЫМ ОТ ПАЦИЕНТА КСЕНОТРАНСПЛАНТАТОМ

ОТЛИЧНЫЕ ОТ ЧЕЛОВЕКА ЖИВОТНЫЕ С ГУМАНИЗИРОВАННЫМ ГЕНОМ СИГНАЛЬНОГО РЕГУЛЯТОРНОГО БЕЛКА

АНТИТЕЛО ПРОТИВ РЕЦЕПТОРА IGF-I

TRANSCHROMOSOMALE TRANSGEN-NAGETIERE ZUR HERSTELLUNG VON HUMANEN ANTIKÖRPERN

Improved recombinant protein production system

Animal models and therapeutic molecules

The present invention relates inter alia to a rodent or rodent cell having a genome comprising: i) one or more companion animal IGH V region genes, one or more companion animal D region genes and one or more companion animal J region genes; and (ii) optionally one or more companion animal IGL kappa V region genes and one or more companion animal IGL kappa J region genes; and/or one or more companion animal IGL lambda V region genes and one or more companion animal IGL lambda J region genes, wherein the rodent or rodent cell is capable of expressing the companion animal variable region gene(s) to form an antibody chain and wherein the companion animal species is not a rodent. The companion animal genes are preferably canine. Also described are methods for producing such cells, methods for producing antibody chains using the cells, and methods of treating diseases of companion animals by administration of the antibodies.

Viral methods of T cell therapy

Methods of producing a population of genetically modified cells using viral or non-viral vectors. Disclosed are also modified viruses for producing a population of genetically modified cells and/or for the treatment of cancer.

Transgenic mice

The invention relates to nucleic acid constructs for expression in mice for the production of heavy chain only antibodies and VH domains, transgenic mice, related methods and uses.

Viral methods of T cell therapy

Disclosed are methods of producing modified T cells or natural killer (NK) cells, comprising: a) obtaining a population of T cells or NK cells; b) introducing a CRISPR system into the immune cells, comprising a nuclease and a guide RNA (gRNA) which binds within 10 bases of a protospacer adjacent motif near the cytokine inducible SH2-containing protein (CISH) gene, where the CRISPR system introduces a double strand break in the CISH gene to suppress its expression; and c) introducing a transgene encoding a T cell receptor (TCR) or chimeric antigen receptor (CAR) into the cells using either an AAV vector or a CRISPR system; and d) culturing the cells for at least 7 days. Also disclosed is the use of the cells as agents for treating cancers.

TRANSCHROMOSOMALE TRANSGEN RODENTS FOR THE PRODUCTION OF HUMÄNEN ANTIBODIES

Anti-VEGF-A and anti-ANG2 antibodies and uses thereof

The present invention relates to bispecific antibodies having activity against a vascular endothelial growth factor (VEGF) and an angiopoietin (ANG), and methods of making and using such bispecific antibodies.

Reconstituted RSV antigen

Disclosed is an antigen comprising reconstituted respiratory syncytial virus (RSV) F protein, where the reconstituted RSV F protein comprises an antigenic region connected at either end to an HRN region and to an HRC region, and the antigenic region comprises one or more antigenic sites of groups consisting of site Ø, site II, and site IV. Also disclosed are a nucleic acid molecule for encoding the antigen and a vaccine composition comprising the antigen and used for initiating an immune response against RSV.

Antibodies recognizing tau

The invention provides antibodies that specifically bind tau. The antibodies inhibit or delay tau-associated pathologies and associated symptomatic deterioration.

Transgenic animal for producing diversified antibodies that have the same light chain i

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.

Antibody and method for producing same

Disclosed is an antibody in which the 80th amino acid residue in a variable region based on the Kabat method and the 171th amino acid residue in a constant region based on the Kabat method are substituted with cysteine in an antibody in which the 80th amino acid residue in the variable region and the 171th amino acid residue in the constant region are not cysteine.

PRODUCTION CELL LINE ENHANCERS

The present invention relates to discovery of the ectopic expression of EDEM2 in a production cell to improve the yield of a useful multi-subunit protein. Thus, the present invention provides for production cell lines, such as the canonical mammalian biopharmaceutical production cell - the CHO cell, containing recombinant polynucleotides encoding EDEM2. Also disclosed is a production cell containing both an EDEM2-encoding polynucleotide as well an XBP1-encoding polynucleotide. Improved titers of antibodies produced by these cell lines are disclosed, as well as the improved cell densities attained by these cells in culture.

Compositions and methods for inhibiting endogenous immunoglobulin genes and producing transgenic human idiotype antibodies

MPHOSPH1 peptides and vaccines including the same

As discussed in greater detail herein, isolated epitope peptides derived from MPHOSPH1 bind to an HLA antigen and induce cytotoxic T lymphocytes (CTL) and thus are suitable for use in the context of cancer immunotherapy, more particularly cancer vaccines. The inventive peptides encompass both the above-mentioned MPHOSPH1-derived amino acid sequences and modified versions thereof, in which one, two, or several amino acids are substituted, deleted, inserted or added, provided such modified versions retain the requisite CTL inducibility of the original sequences. Further provided are polynucleotides encoding any of the aforementioned peptides as well as pharmaceutical agents or compositions that include any of the aforementioned peptides or polynucleotides. The peptides, polynucleotides, and pharmaceutical agents or compositions of this invention find particular utility in either or both of the treatment and prevention of cancers and tumors, including, for example, bladder cancer, breast cancer ...

Humanized antibodies specific to the protofibrillar form of the beta-amyloid peptide

The present application relates to humanized antibodies specific to the protofibrillar form of the beta-amyloid peptide, and to the use of said antibodies in the field of Alzheimer's disease.

Fertile transgenic animals useful for producing antibodies bearing human variable regions

The present invention relates inter alia to fertile non-human vertebrates such as mice and rats useful for producing antibodies bearing human variable regions, in which endogenous antibody chain 5 expression has been inactivated. 17779909_1 (GHMatters) P96880.AU.4 ...

NON-HUMAN ANIMALS HAVING A HUMANIZED SIGNAL-REGULATORY PROTEIN GENE

Genetically modified non-human animals and methods and compositions for making and using the same are provided, wherein the genetic modification comprises a humanization of an endogenous signal-regulatory protein gene, in particular a humanization of a SIRPa gene. Genetically modified mice are described, including mice that express a human or humanized SIRPa protein from an endogenous SIRPa locus.

CHIMERIC ANTIGEN RECEPTORS TARGETED TO PSCA

Chimeric transmembrane immunoreceptors (CAR) targeted to PSCA are described.

FACTOR X VARIANTS

La présente invention concerne une protéine qui est un variant du facteur X comprenant une séquence mutée de SEQ ID NO :1, ladite protéine comprenant à son extrémité N- terminale, le peptide signal de séquence SEQ ID NO :7, et un propeptide différent du propeptide naturel du facteur X.

METHODS OF TREATING MUSCULAR DYSTROPHY

The invention provides for AAV vectors expressing the ANO5 gene and antioxidant therapy as methods of inducing muscle regeneration and a method of treating muscular dystrophy.

ANTIBODY FOR BINDING TO INTERLEUKIN 4 RECEPTOR

Disclosed is an antibody capable of binding to the interleukin 4 (IL-4) receptor (IL-4). Also disclosed are a nucleic acid sequence encoding the antibody, a vector including the nucleic acid sequence, and a host cell transformed or transfected with the vector. Provided are a method for producing the antibody, a medical use of the antibody, and a kit including the antibody.

OX40-BINDING POLYPEPTIDES AND USES THEREOF

Provided herein are VHH-containing polypeptides that bind OX40. In some embodiments, VHH-containing polypeptides that bind and agonize OX40 are provided. Uses of the VHH-containing polypeptides are also provided.

HIGH-THROUGHPUT MOUSE MODEL FOR OPTIMIZING ANTIBODY AFFINITIES

Described herein are compositions (e.g. cells and transgenic animals) and methods relating to engineered Ig loci that permit expression of particular antibodies or antibody segments while still permitting recombination and/or maturation process for antibody optimization.

TRANSGENIC BIRDS THAT PRODUCE CHIMERIC HUMAN IMMUNOGLOBULINS

The invention relates to transgenic birds capable of producing chimeric immunoglobulins, with a combination of human and avian sequence, in their B cells. In some embodiments, the birds are chickens. When challenged with an antigen, the transgenic avians produce antigen-specific functional antibodies. The invention also relates to light chain immunoglobulin transgenes for making such transgenic avians, as well as methods and vectors for disrupting endogenous immunoglobulin loci in birds.

Anti-c5a-antibody and their application

DIRECTED DESTRUCTION OF T-CELL RECEPTOR AND/OR RECEPTOR OF HLA

NEW T CELL RECEPTORS AND IMMUNOTHERAPY WITH THEIR USING

ENKhANSERY PRODUCING CELL LINES

HUMANIZED ANTIBODY SPECIFIC TO THE PROTOFIBRILLAR FORM OF THE BETA-AMYLOID PEPTIDE

enkhansery producing cell lines

METHODS FOR TREATMENT OF MUSCULAR DYSTROPHY

DIRECT SAMPLING THE CELLS, EXPRESSING HIGH LEVELS OF GETEROMERNYKh PROTEINS, USING GLUTAMINSINTETAZNYKh VECTORS VNUTRIGENNOI THE COMPLEMENTATION

ANTIBODY HUMANIZE SPECIFIC THE SHAPE PROTOFIBRILLAIRE OF PEPTIDE BETA-AMYLOIDE.

Anti-c5a antibodies and uses thereof

Non-human animals expressing exogenous terminal deoxynucleotidyltransferase

ANTI-CD25 FOR TUMOUR SPECIFIC CELL DEPLETION

METHODS FOR BREAKING IMMUNOLOGICAL TOLERANCE USING MULTIPLE GUIDE RNAS

TRANSGENIC TRANSCHROMOSOMAL RODENTS FOR MAKING HUMAN ANTIBODIES

The present invention provides novel transgenic nonhuman mammals capable of producing human sequence antibodies, as well as methods of producing and using these antibodies.

АНТИТЕЛА К ТАУ И ИХ ПРИМЕНЕНИЯ

Группа изобретений относится к биотехнологии. Предложены моноклональное антитело или его антигенсвязывающий фрагмент, которые специфически связывают тау-белок человека, его получение и применения. Антитело содержит HCDR1 в соответствии с SEQ ID NO: 732, HCDR2 в соответствии с SEQ ID NO: 734, HCDR3 в соответствии с SEQ ID NO: 736 и LCDR1 в соответствии с SEQ ID NO: 846, LCDR2 в соответствии с SEQ ID NO: 848, LCDR3 в соответствии с SEQ ID NO: 850, как определено в соответствии со способом по Kabat. Или тяжелая цепь содержит HCDR1 в соответствии с SEQ ID NO: 1002, HCDR2 в соответствии с SEQ ID NO: 1004 и HCDR3 в соответствии с SEQ ID NO: 1006 и легкая цепь содержит LCDR1 в соответствии с SEQ ID NO: 1116, LCDR2 в соответствии с SEQ ID NO: 1118 и LCDR3 в соответствии с SEQ ID NO: 1120. Антитела или его антигенсвязывающий фрагмент применимы для снижения уровней саркозил-нерастворимого тау-белка у субъекта и в лечении таупатии. 12 н. и 28 з.п. ф-лы, 33 ил., 15 табл., 19 пр.

ОТЛИЧНЫЕ ОТ ЧЕЛОВЕКА ЖИВОТНЫЕ С ГУМАНИЗИРОВАННЫМ ГЕНОМ СИГНАЛЬНОГО РЕГУЛЯТОРНОГО БЕЛКА

Изобретение относится к области биохимии, в частности к мыши для экспрессии гуманизированного белка SIRPα, содержащей замещение экзонов 2, 3 и 4 гена SIRPα мыши в эндогенном локусе SIRPα мыши на экзоны 2, 3 и 4 гена SIRPα человека с образованием гуманизированного гена SIRPα, а также к клетке и ткани вышеуказанной мыши. Также раскрыт способ получения вышеуказанной мыши. Изобретение также относится к способу трансплантации человеческих гематопоэтических клеток мыши, способу фагоцитоза меченого субстрата, способу изменения фагоцитоза, а также к способу оценки терапевтической эффективности потенциального лекарственного средства с использованием вышеуказанной мыши или ее клетки. Изобретение эффективно в использовании для анализов приживления трансплантата, активации фагоцитоза и сигнальной трансдукции. 8 н. и 9 з.п. ф-лы, 8 ил., 5 табл., 4 пр.

Генетическая конструкция рБТШ70 для экспрессии основного человеческого стресс белка в молоке трансгенных животных

Изобретение относится к биотехнологии, конкретно к рекомбинантному получению белка теплового шока человека 70 (БТШ70), и может быть использовано для получения БТШ70 в молоке трансгенных животных. На основе экспрессионного вектора рВС1 получают экспрессионную плазмиду рБТШ70 размером 16912 п. н., с картой, приведенной на рис. 1, которая содержит ген БТШ70 человека и кодирует сигнальную последовательность лактоферрина. Изобретение позволяет получить высокий титр человеческого БТШ70, обладающего шаперонными свойствами, характерными для нативного человеческого БТШ70, в молоке трансгенных животных. 6 ил.

СПОСОБЫ ПРЕОДОЛЕНИЯ ИММУНОЛОГИЧЕСКОЙ ТОЛЕРАНТНОСТИ С ИСПОЛЬЗОВАНИЕМ МНОЖЕСТВА НАПРАВЛЯЮЩИХ РНК

Изобретение относится к области биохимии, в частности к способу получения антигенсвязывающих белков против представляющего интерес чужеродного антигена, предусматривающему получение генетически модифицированной крысы или мыши, у которой локус-мишень содержит биаллельную делецию всего или части гена, кодирующего аутоантиген. При этом получение вышеуказанной мыши или крысы включает введение в эмбрион мыши или крысы белка Cas9 или нуклеиновой кислоты, кодирующей белок Cas9, а также первой и второй направляющей нуклеотидной последовательности. Изобретение позволяет эффективно сократить сроки получения генетически модифицированной мыши или крысы с пониженной толерантностью к представляющему интерес чужеродному антигену, а также обеспечивает получение большего количества антител против представляющего интерес чужеродного антигена. 2 н. и 34 з.п. ф-лы, 32 ил., 24 табл., 8 пр.

ЖИВОТНЫЕ, НЕ ЯВЛЯЮЩИЕСЯ ЧЕЛОВЕКОМ, ЭКСПРЕССИРУЮЩИЕ ЭКЗОГЕННУЮ ТЕРМИНАЛЬНУЮ ДЕЗОКСИНУКЛЕОТИДИЛТРАНСФЕРАЗУ

Изобретение относится к области биотехнологии, в частности к генетически модифицированной мыши, содержащей последовательность нуклеиновой кислоты, кодирующую терминальную дезоксинуклеотидилтрансферазу (TdT) человека, функционально связанную с элементом контроля транскрипции, и вариабельную область иммуноглобулина, а также к ее ЭС-клетке. Также раскрыто применение вышеуказанной ЭС-клетки мыши для получения генетически модифицированной мыши. Изобретение может быть эффективно использовано для индуцирования экспрессии антитела, содержащего вариабельный домен человека. 4 н. и 31 з.п. ф-лы, 16 ил., 5 табл., 10 пр.

ХИМЕРНЫЕ РЕЦЕПТОРЫ АНТИГЕНА, НАЦЕЛЕННЫЕ НА PSCA

Изобретение относится к области биотехнологии. Описана группа изобретений, включающая молекулу нуклеиновой кислоты, кодирующую химерный рецептор антигена, нацеленный на PSCA (антиген стволовой клетки простаты), популяцию человеческих Т-клеток для лечения рака, экспрессирующего PSCA, композицию, содержащую популяцию аутологичных или аллогенных человеческих Т-клеток, для применения в способе лечения рака, экспрессирующего PSCA у пациента и химерный рецептор антигена, нацеленный на PSCA. В одном из вариантов реализации молекула нуклеиновой кислоты экспрессирует полипептид, содержащий аминокислотную последовательность, выбранную из SEQ ID NO: 26, 28, 30, 32, 34 и 36. Изобретение расширяет арсенал средств, кодирующих химерный рецептор антигена, нацеленный на PSCA. 4 н. и 10 з.п. ф-лы, 23 ил., 4 табл., 14 пр.

ПЕПТИДЫ MPHOSPH1 И ВАКЦИНЫ, ВКЛЮЧАЮЩИЕ ИХ

Изобретение относится к области биотехнологии, конкретно к противоопухолевым вакцинам на основе эпитопных пептидов MPHOSPH1, и может быть использовано в медицине. Получают пептид состоящий из аминокислотной последовательности SEQ ID NO: 120. Пептид может содержать замены С- и/или N-концевой аминокислоты указанной последовательности на лейцин или метионин. Полученный эпитопный пептид обладает способностью индуцировать цитотоксические Т-лимфоциты (CTL) в присутствии антигенпредставляющей клетки (АРС), несущей HLA-A*0201 или HLA-А*0206. Изобретение позволяет индуцировать иммунный ответ против злокачественной опухоли, экспрессирующей MPHOSPH1 у индивидуума, HLA антиген которого представляет собой HLA-A*0201 или HLA-A*0206. 14 н.п. ф-лы, 6 ил., 3 табл., 1 пр.

Способ получения димерной формы иммуноглобулина IgA1-изотипа в клетках млекопитающих

Изобретения относятся к области биотехнологии. Предложена рекомбинантная плазмидная ДНК p3.4-P4A1IgA1-J12His-ht, кодирующая нейтрализующее моноклональное антитело P4A1 к вирусу SARS-CoV-2 изотипа IgA1 и J-цепь человека. Плазмидная ДНК содержит сплайсированный ген тяжелой цепи антитела, находящийся под контролем промотора hEF1-HTLV и сигнала полиаденилирования бычьего гормона роста BGH poly, ген легкой цепи антитела и ген J-цепи человека, находящиеся под контролем промотора CMV и внутреннего сайта связывания рибосом (IRES) вируса энцефаломиокардита (EMCV), посттранскрипционный регуляторный элемент WPRE вируса гепатита сурка (WHV) и сигнал полиаденилирования тимидинкиназы вируса герпеса. Промоторы hEF1-HTLV и CMV размещены в последовательности, обеспечивающей однонаправленную транскрипцию. Также описан способ получения димера нейтрализующего моноклонального антитела P4A1 против вируса SARS-CoV-2 (2019-nCoV) изотипа IgA1 с J-цепью, включающий культивирование клеток СНО, трансфецированных плазмидной ...

Генетическая конструкция для экспрессии функционально-активного человеческого стресс-белка (БТШ70) с мутированными сайтами гликозилирования для наработки в эукариотических экспрессионных системах

Изобретение относится к биотехнологии, конкретно к рекомбинантному получению белка теплового шока человека 70 (БТШ70), и может быть использовано для получения БТШ70 в молоке трансгенных животных. Получен человеческий БТШ70 с SEQ ID NO:1, характеризующийся мутациями в сайтах гликозилирования, которые не влияют на шаперонные свойства белка и позволяют предотвратить не характерную для внутриклеточных форм БТШ70 посттрансляционную модификацию, возникающую обычно в процессе получения рекомбинантного БТШ70 в молоке трансгенных животных. Изобретение позволяет получить БТШ70 человека, обладающий шаперонными свойствами нативного человеческого БТШ70, в молоке трансгенных животных. 2 н.п. ф-лы, 8 ил., 1 табл., 2 пр.

ПЕПТИДЫ MPHOSPH1 И ВАКЦИНЫ, ВКЛЮЧАЮЩИЕ ИХ

... 1. Выделенный пептид согласно следующим ниже (a) или (b):(a) пептид, содержащий аминокислотную последовательность, выбранную из группы, состоящей из SEQ ID NO: 5, 14, 64, 73, 77, 79, 97, 103 и 120;(b) пептид, содержащий аминокислотную последовательность, где 1, 2 или несколько аминокислот заменены, удалены, вставлены и/или добавлены в аминокислотной последовательности, выбранной из группы, состоящей из SEQ ID NO: 5, 14, 64, 73, 77, 79, 97, 103 и 120,и где указанный пептид обладает способностью индуцировать цитотоксические T-лимфоциты (CTL).2. Выделенный пептид по п. 1, где пептид имеет одну или обе из следующих характеристик:(a) вторая аминокислота с N-конца аминокислотной последовательности, выбранной из группы, состоящей из SEQ ID NO: 5, 14, 64, 73, 77, 79, 97, 103 и 120, выбрана из группы, состоящей из лейцина и метионина; и(b) C-концевая аминокислота аминокислотной последовательности, выбранной из группы, состоящей из SEQ ID NO: 5, 14, 64, 73, 77, 79, 97, 103 и 120, выбрана из группы ...

TNF-α- СВЯЗЫВАЮЩИЕ БЕЛКИ

... 1. Гуманизированный связывающий белок, содержащий антигенсвязывающий домен, способный связывать фактор-альфа некроза опухолей человека (TNF-α), причем антигенсвязывающий домен содержит, по меньшей мере, одну CDR, содержащую аминокислотную последовательность остатков 31-35 SEQ ID NO:22; остатков 50-65 SEQ ID NO:22; остатков 98-106 SEQ ID NO:22; остатков 24-34 SEQ ID NO:23; остатков 50-56 SEQ ID NO:23 или остатков 89-97 SEQ ID NO:23, где связывающий белок содержит акцепторный каркас человека.2. Связывающий белок по п.1, где связывающий белок содержит, по меньшей мере, 3 CDR.3. Связывающий белок по п.2, где, по меньшей мере, 3 CDR содержат CDR-набор вариабельного домена, содержащий (a) остатки 31-35 SEQ ID NO:22; остатки 50-65 SEQ ID NO:22 и остатки 98-106 SEQ ID NO:22; и (b) остатки 24-34 SEQ ID NO:23; остатки 50-56 SEQ ID NO:23 и остатки 89-97 SEQ ID NO:23.4. Связывающий белок по п.3, где антигенсвязывающий домен содержит аминокислотную последовательность, содержащую остатки 31-35 SEQ ID ...

СПОСОБЫ ПРЕОДОЛЕНИЯ ИММУНОЛОГИЧЕСКОЙ ТОЛЕРАНТНОСТИ С ИСПОЛЬЗОВАНИЕМ МНОЖЕСТВА НАПРАВЛЯЮЩИХ РНК

ЖИВОТНЫЕ, НЕ ЯВЛЯЮЩИЕСЯ ЧЕЛОВЕКОМ, ЭКСПРЕССИРУЮЩИЕ ЭКЗОГЕННУЮ ТЕРМИНАЛЬНУЮ ДЕЗОКСИНУКЛЕОТИДИЛТРАНСФЕРАЗУ

СПОСОБЫ ЛЕЧЕНИЯ МЫШЕЧНОЙ ДИСТРОФИИ

МЫШИ, У КОТОРЫХ ВЫРАБАТЫВАЮТСЯ СВЯЗЫВАЮЩИЕ БЕЛКИ, СОДЕРЖАЩИЕ VL-ДОМЕНЫ

Transgenic animals

Methods for modulating production profiles of recombinant proteins

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also 5 disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant.

Methods of treating muscular dystrophy

The invention provides for AAV vectors expressing the ANO5 gene and antioxidant therapy as methods of inducing muscle regeneration and a method of treating muscular dystrophy.

Cell line for recombinant protein and/or viral vector production

Cells and cell lines are disclosed that are able to produce therapeutic proteins, antibodies, vectors, and viral vectors such as lentiviral vectors and adeno-associated viral (AAV) vectors. The cells and/or cell lines can have mutations or deletions in either one or both of the endogenous dihydrofolate reductase (DHFR-/-) or glutamine synthetase (GS-/-) genes such that DHFR and/or GS expression or function is substantially reduced or eliminated.

An engineered two-part cellular device for discovery and characterisation of T-cell receptor interaction with cognate antigen

The present invention relates to a two-part device, wherein a first part is an engineered antigen-presenting cell system (eAPCS), and a second part is an engineered TCR-presenting cell system (eTPCS).

Non-human animals expressing exogenous terminal deoxynucleotidyltransferase

Provided herein are methods and compositions related to non-human animals that express exogenous Terminal Deoxynucleotidyltransferase (TdT).

Antibody and method for producing same

Disclosed is an antibody in which the 80th amino acid residue in a variable region based on the Kabat method and the 171th amino acid residue in a constant region based on the Kabat method are substituted with cysteine in an antibody in which the 80th amino acid residue in the variable region and the 171th amino acid residue in the constant region are not cysteine.

Viral methods of making genetically modified cells

The present disclosure provides methods of producing a population of genetically modified cells using viral or non-viral vectors. Also disclosed are modified viruses for producing a population of genetically modified cells and/or for the treatment of cancer.

Compositions and methods for enhancing gene expression

The present disclosure generally relates to nucleic acid molecules for use in regulating gene expression. Disclosed herein include nucleic acid molecules containing one or more structural elements of the viral capsid enhancer operably linked to a coding sequence of a gene of interest. In some embodiments, the viral capsid enhancer comprises a Downstream Loop (DLP) from a viral capsid protein, or a variant of the DLP.

Recombinant protein production system

The present disclosure relates generally to an improved gene expression system in the field of recombinant gene expression. The invention relates to a system showing an improved yield and quality of protein production and methods for increasing production of a protein produced by cultured cells, particularly cultured eukaryotic cells.

Promoter of Hspa5 gene

In order to provide a means for enhancing, production in a host cell such as a mammal-derived cultured cell, of a foreign protein used for protein pharmaceuticals, the present invention provides: a transformed cell having a novel Hspa5 gene promoter; and a method for realizing high-secretion production of a foreign protein by using said transformed cell.

AAV capsid designs

The disclosure in some aspects relates to recombinant adeno-associated viruses having distinct tissue targeting capabilities. In some aspects, the disclosure relates to gene transfer methods using the recombinant adeno-associated viruses. In some aspects, the disclosure relates to isolated AAV capsid proteins and isolated nucleic acids encoding the same.

Non-HLA matched humanized NSG mouse model with patient-derived xenograft

The invention described herein provides non-HLA matched humanized mouse model ( ...

PRODUCTION CELL LINE ENHANCERS

The present invention relates to discovery of the ectopic expression of EDEM2 in a production cell to improve the yield of a useful multi-subunit protein. Thus, the present invention provides for production cell lines, such as the canonical mammalian biopharmaceutical production cell - the CHO cell, containing recombinant polynucleotides encoding EDEM2. Also disclosed is a production cell containing both an EDEM2-encoding polynucleotide as well an XBP1-encoding polynucleotide. Improved titers of antibodies produced by these cell lines are disclosed, as well as the improved cell densities attained by these cells in culture.

MPHOSPH1 peptides and vaccines including the same

As discussed in greater detail herein, isolated epitope peptides derived from MPHOSPH1 bind to an HLA antigen and induce cytotoxic T lymphocytes (CTL) and thus are suitable for use in the context of cancer immunotherapy, more particularly cancer vaccines. The inventive peptides encompass both the above-mentioned MPHOSPH1-derived amino acid sequences and modified versions thereof, in which one, two, or several amino acids are substituted, deleted, inserted or added, provided such modified versions retain the requisite CTL inducibility of the original sequences. Further provided are polynucleotides encoding any of the aforementioned peptides as well as pharmaceutical agents or compositions that include any of the aforementioned peptides or polynucleotides. The peptides, polynucleotides, and pharmaceutical agents or compositions of this invention find particular utility in either or both of the treatment and prevention of cancers and tumors, including, for example, bladder cancer, breast cancer ...

Fertile transgenic animals useful for producing antibodies bearing human variable regions

The present invention relates ...

NON-HUMAN ANIMALS EXPRESSING EXOGENOUS TERMINAL DEOXYNUCLEOTIDYLTRANSFERASE

Provided herein are methods and compositions related to non-human animals that express exogenous Terminal Deoxynucleotidyltransferase (TdT).

ANTI-LAG-3 ANTIBODY

Provided is an anti-LAG-3 antibody that can be frequently administered to a non-rat animal. The anti-LAG-3 antibody comprises: (a) an L chain comprising an L-chain variable region that includes CDR1 having a QSLLDSDGNTY (SEQ ID NO. 16) amino acid sequence, CDR2 having an SVS amino acid sequence, and CDR3 having an MQATHVPFT (SEQ ID NO. 17) amino acid sequence, and an L-chain constant region of a non-rat mammalian antibody; and (b) an H chain comprising an H-chain variable region that includes CDR1 having a GFDFDTYP (SEQ ID NO. 18) amino acid sequence, CDR2 having an ITIKTHNYAT (SEQ ID NO. 19) amino acid sequence, and CDR3 having an NREDFDY (SEQ ID NO. 20) amino acid sequence, and an H-chain constant region of a non-rat mammalian antibody. Also provided is a pharmaceutical composition containing the anti-LAG-3 antibody as an active ingredient. Also provided is a method of producing the anti-LAG-3 antibody.

COMPOSITIONS AND METHODS FOR ENHANCING GENE EXPRESSION

The present disclosure generally relates to nucleic acid molecules for use in regulating gene expression. Disclosed herein include nucleic acid molecules containing one or more structural elements of the viral capsid enhancer operably linked to a coding sequence of a gene of interest. In some embodiments, the viral capsid enhancer comprises a Downstream Loop (DLP) from a viral capsid protein, or a variant of the DLP.

SYSTEM AND METHOD FOR CHARACTERIZING SIZE AND CHARGE VARIANT DRUG PRODUCT IMPURITIES

Systems and methods for characterizing size and charge variant protein drug product impurities are provided.

ANTI-ROBO2 ANTIBODIES, COMPOSITIONS, METHODS AND USES THEREOF

The invention provides antibodies, and antigen-binding fragments thereof, that specifically bind to ROBO2, as well as uses, and methods thereof.

METHODS FOR MODULATING PRODUCTION PROFILES OF RECOMBINANT PROTEINS

The present invention relates to methods and compositions for modulating glycosylation of recombinant proteins expressed by mammalian host cells during the cell culture process. Also 5 disclosed are methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising a disaccharide or a trisaccharide, while keeping the osmolality constant.

TRANSGENIC MICE

The invention relates to nucleic acid constructs for expression in mice for the production of heavy chain only antibodies and VH domains, transgenic mice, related methods and uses.

NON-HUMAN ANIMALS THAT MAKE SINGLE DOMAIN BINDING PROTEINS

Genetically modified non-human animals and methods and compositions for making and using them are provided, wherein the genetic modification comprises (a) a deletion in an immunoglobulin constant region CH1 gene (optionally a deletion in a hinge region) of a heavy chain constant region gene sequence, and (b) replacement of one or all endogenous VH, DH and JH gene segments with at least one unrearranged light chain variable (VL) gene segment and at least one unrearranged light chain joining (JL) gene segment capable of recombining to form a rearranged light chain variable region (VL/JL) nucleotide sequence operably linked to the heavy chain constant region gene sequence comprising a deletion in the CH1 gene and/or insertion of a genetically engineered single rearranged light chain, wherein the mouse is capable of expressing a functional IgM, single domain antigen binding proteins, e.g., VL single domain binding proteins, and a genetically engineered rearranged light chain.

GENETICALLY MODIFIED CELLS, TISSUES, AND ORGANS FOR TREATING DISEASE

Genetically modified cells, tissues, and organs for treating or preventing diseases are disclosed. Also disclosed are methods of making the genetically modified cells and non-human animals. The genetic modification may include a nucleic acid that is transcribed as a human leukocyte antigen G (HLA-G) mRNA comprising a deletion in the 3 ' untranslated region, or a nucleic acid comprising a CD47 gene that is codon-optimized for expression in pig cells.

SEMA5B PEPTIDES AND VACCINES INCLUDING THE SAME

As discussed in detail herein, isolated epitope peptides derived from SEMA5B bind to an HLA antigen and induce cytotoxic T lymphocytes (CTL) and thus are suitable for use in the context of cancer immunotherapy, more particularly cancer vaccines. The inventive peptides encompass both the above mentioned amino acid sequences and modified versions thereof, in which one, two, or several amino acids are substituted, deleted, inserted or added, provided such modified versions retain the requisite HLA binding and/or CTL inducibility of the original sequences. Further provided are polynucleotides encoding any of the aforementioned peptides as well as pharmaceutical agents or compositions that include any of the aforementioned peptides or polynucleotides. The peptides, polynucleotides, pharmaceutical agents or compositions of this invention find particular utility in the treatment and/or prevention of cancers and tumors, including, for example, esophageal cancer, NSCLC, RCC and SCLC.

METHODS OF TREATING MUSCULAR DYSTROPHY

The antibody against VEGF synthesis and use thereof

Targeted disruption T cells and/or HLA receptors

OPTIMIZED RPE65 PROMOTER AND CODING SEQUENCES

Transgenic transchromosomal rodents for making human antibodies

Immunocompromised Ungulates

Porcine animals, tissue and organs as well as cells and cell lines derived from such animals are provided that lack functional endogenous immunoglobulin loci and are deficient in immunoglobulin expression and B-cells. These animals are useful as model systems for research and for development of new pharmaceutical and biological agents. In addition, methods are provided to prepare such animals.

Mice That Make VL Binding Proteins

Genetically modified mice and methods for making an using them are provided, wherein the mice comprise a replacement of all or substantially all immunoglobulin heavy chain V gene segments, D gene segments, and J gene segments with at least one light chain V gene segment and at least one light chain J gene segment. Mice that make binding proteins that comprise a light chain variable domain operably linked to a heavy chain constant region are provided. Binding proteins that contain an immunoglobulin light chain variable domain, including a somatically hypermutated light chain variable domain, fused with a heavy chain constant region, are provided. Modified cells, embryos, and mice that encode sequences for making the binding proteins are provided.

Production of Transgenic Avians Using Improved Retroviral Vectors

A transgenic avian containing in its genome an exogenous nucleotide sequence which includes a promoter component and a vector with reduced promoter interference wherein the exogenous nucleotide sequence is integrated into the genome and the avian.

Mgmt-based method for obtaining high yeilds of recombinant protein expression

The present invention relates to a novel enhancer of protein production in host cells. It discloses a vector for expressing recombinant proteins in these cells, comprising a nucleotide sequence encoding a) a secretion peptidic signal, b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT, EC 2.1.1.63), a mutant or a catalytic domain thereof, and c) a recombinant protein. Said MGMT enzyme is preferably the so-called SNAP protein.

Novel t cell receptors and immune therapy using the same

The present invention pertains to antigen recognizing constructs against tumor associated antigens (TAA), in particular the TAA Serine protease inhibitor Kazal-type 2 (SPINK2). The invention in particular provides novel T cell receptor (TCR) based molecules which are selective and specific for the tumor expressed antigen of the invention. The TCR of the invention, and SPINK2 binding fragments derived therefrom, are of use for the diagnosis, treatment and prevention of SPINK2 expressing cancerous diseases. Further provided are nucleic acids encoding the antigen recognizing constructs of the invention, vectors comprising these nucleic acids, recombinant cells expressing the antigen recognizing constructs and pharmaceutical compositions comprising the compounds of the invention.

MSBI SEQUENCES AS AN EARLY MARKER FOR THE FUTURE DEVELOPMENT OF CANCER AND DISEASES OF THE CNS AND AS A TARGET FOR THE TREATMENT AND PREVENTION OF THESE DISEASES

Described are MSBI (Multiple Sclerosis Brain Isolate) nucleotide sequences as well as probes and primers comprising part of said nucleotide sequences and antibodies against polypeptides encoded by said nucleotide sequences. Said compounds are useful as early markers for the future development of cancer and diseases of the CNS (Multiple sclerosis MS, Prion-linked diseases, amyotrophic lateral sclerosis, transmissible spongiforme encephalitis, Parkinson's disease, Alzheimer disease) and should represent targets for treatment and prevention. 1. An expression vector comprising a Multiple Sclerosis Brain isolate (MSBI) polynucleic acid comprising:{'figref': {'@idref': 'DRAWINGS', 'FIG. 1(A)'}, 'a nucleotide sequence depicted in or (F),'}wherein the vector is adenoviral, vaccinia virus, avipox virus, herpes virus, or a retrovirus vector.2. The expression vector of wherein the retrovirus vector is Moloney murine leukemia virus (MoMuLV) claim 1 , Harvey murine sarcoma virus (HaMuSV) claim 1 , murine mammary tumor virus (MuMTV) claim 1 , Rous sarcoma virus (RSV) claim 1 , gibbon ape leukemia virus (GaLV).3. A host cell transformed with an expression vector according to .4. A host cell transformed with an expression vector according to .5. The host cell of claim 3 , wherein the cell is a Chinese hamster cell claim 3 , a monkey cell claim 3 , a baby hamster kidney cell claim 3 , a pig kidney cell claim 3 , a rabbit kidney cell claim 3 , a human osteosarcoma cell claim 3 , a HeLa cell a human hepatoma cell claim 3 , or an insect cell.6. The host cell of claim 4 , wherein the cell is a Chinese hamster cell claim 4 , a monkey cell claim 4 , a baby hamster kidney cell claim 4 , a pig kidney cell claim 4 , a rabbit kidney cell claim 4 , a human osteosarcoma cell claim 4 , a HeLa cell a human hepatoma cell claim 4 , or an insect cell.7Spodoptera frugiperda. The host cell of claim 5 , wherein the monkey cell is a COS or Vero cell claim 5 , the pig kidney cell is a PK15 cell claim 5 , ...

Synthesis and structure of high potency rna therapeutics

This invention provides expressible polynucleotides, which can express a target protein or polypeptide. Synthetic mRNA constructs for producing a protein or polypeptide can contain one or more 5′ UTRs, where a 5′ UTR may be expressed by a gene of a plant. In some embodiments, a 5′ UTR may be expressed by a gene of a member of Arabidopsis genus. The synthetic mRNA constructs can be used as pharmaceutical agents for expressing a target protein or polypeptide in vivo.

Synthesis of High Molecular Weight Proteins Using Inteins

This disclosure is directed to split intein protein production systems using transgenic target organisms such as . A vector set for transforming a target organism includes: a first vector having a first donor sequence that encodes (i) a first non-native protein and (ii) at least one split intein domain; a second vector having a second donor sequence that encodes (i) a second non-native protein and (ii) at least one split intein domain. The respective split intein domains encoded by the first and second vectors are configured to associate with one another and ligate the first and second non-native proteins to thereby form a fused protein. 1Bombyx mori. A method of producing transgenic , the method comprising:providing a first vector having a first donor sequence that encodes a first non-native protein and at least one split intein domain;providing a second vector having a second donor sequence that encodes a second non-native protein and at least one split intein domain;{'i': 'Bombyx mori', 'incorporating the first vector into one or more cells; and'}{'i': 'Bombyx mori', 'incorporating the second vector into one or more cells,'}wherein the split intein domains encoded by the first and second vectors are configured to associate with one another and ligate the first and second non-native proteins to thereby form a fused protein.2Bombyx mori.. The method of claim 1 , further comprising providing a gene editing assembly that includes a nuclease configured to target one or more locations within a silk protein gene of the3. The method of claim 2 , wherein the gene editing assembly targets the FibH gene.4. The method of claim 1 , wherein the first donor sequence claim 1 , the second donor sequence claim 1 , or both encode for a spider silk protein.5. The method of claim 4 , wherein the spider silk protein comprises an AS28 protein claim 4 , a MaSp1 protein claim 4 , a MaSp4 protein claim 4 , or combination thereof.6. The method of claim 4 , wherein the spider silk protein ...

NUCLEIC ACID MOLECULES AND APPLICATIONS THEREOF IN HUMAN ANTIBODY

Nucleic acid molecules include immunoglobulin genes or parts of immunoglobulin genes. The nucleic acid molecules includes the IgM gene (IgHCμ) and IgM switch region (Sμ). The sequences of the Sμ and the IgHCμ are both derived from a transgenic host animal. In this invention, human antibodies are directly generated and no humanization process is required, and the human antibody druggability is increased. The transgenic human antibody mouse has normal early Bcell development, maturation and the Bcell number in comparison with that of wild type animal, thereby facilitating the differentiation of the Bcells. The specificity and diversity of the produced antibody are improved; and the efficiency for screening the therapeutic antibody is improved. 1. Nucleic acid molecules comprising immunoglobulin genes or parts of the immunoglobulin genes , wherein , the nucleic acid molecules comprises an IgM gene (IgHCμ) and an IgM switch region (Sμ) , and sequences of the Sμ and the IgHCμ are both derived from a transgenic host animal.2. The nucleic acid molecules according to claim 1 , wherein the IgHCμ comprises a CH1 exon claim 1 , a CH2 exon claim 1 , a CH3 exon claim 1 , a CH4 exon claim 1 , and intron sequences between the CH1 exon and the CH2 exon claim 1 , the CH2 exon and the CH3 exon claim 1 , and between the CH3 exon and the CH4 exon claim 1 , a TM1 claim 1 , a TM2 claim 1 , and PolyA signal sequences.3. The nucleic acid molecules according to claim 1 , wherein a nucleotide sequence of the Sμ is listed as positions ranging from (2550) to (4451) in SEQ ID NO:1.4. The nucleic acid molecules according to claim 1 , further comprising an IgH heavy chain 5′-enhancer of the transgenic host animal.5. The nucleic acid molecules according to claim 4 , wherein a nucleotide sequence of the IgH heavy chain 5′-enhancer is listed as positions ranging from (433) to (1444) in SEQ ID NO:1.6. (canceled)7. The nucleic acid molecules according to claim 1 , further comprising IgG genes (Igγ).8. ...

ANTI-CD25 ANTIBODY AGENTS

The present disclosure provides antibody sequences found in antibodies that bind to human CD25. In particular, the present disclosure provides sequences of anti-human CD25 antibodies, 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 composition and methods of treatment, in particular for treating cancer. 146.-. (canceled)47. A composition comprising an antibody or antigen-binding fragment thereof , comprising:a) aCD25-a-646-HCDR3 amino acid sequence (SEQ ID NO: 4) as variable heavy chain complementarity determining region 3;b) a variable heavy chain complementarity determining region 1 (HCDR1) selected from the group consisting of FTFASYGMH (SEQ ID NO: 10) and FTFPSYGMH (SEQ ID NO: 11);c) a a variable heavy chain complementarity determining region 2 (HCDR2) selected from the group consisting of VIWYDASTKYYADSVKG (SEQ ID NO: 12), VIWYDAINKYYADSVKG (SEQ ID NO: 13), VIWYDAVNKYYADSVKG (SEQ ID NO: 14), and VIWYDALNKYYADSVKG (SEQ ID NO: 15);d) aCD25-a-646-LCDR1 amino acid sequence (SEQ ID NO: 6) as variable light chain complementarity determining region 1;e) aCD25-a-646-LCDR2 amino acid sequence (SEQ ID NO: 7) as variable light chain complementarity determining region 2; andf) aCD25-a-646-LCDR3 amino acid sequence (SEQ ID NO: 8) as variable light chain complementarity determining region 3; anda pharmaceutically acceptable carrier or excipient.48. The composition of claim 47 , wherein the antibody or antigen-binding fragment thereof comprises a variable heavy chain comprising a sequence having at least 95% sequence identity to a variable heavy chain region sequence selected from aCD25-a-646-m1-HCDR123 amino acid sequence (SEQ ID NO: 16) claim 47 , aCD25-a-646-m2-HCDR123 amino acid sequence (SEQ ID NO: 17) claim 47 , aCD25-a-646-m3-HCDR123 amino acid sequence (SEQ ID NO: 18) claim 47 , aCD25-a-646-m4-HCDR123 amino acid sequence (SEQ ID NO: 19) claim 47 , ...

GENETICALLY-MODIFIED POULTRY EGG

Provided are a poultry knock-in egg and knock-out egg. The present invention pertains to a knock-out poultry egg in which at least one oviduct-specific gene has been knocked out, said gene being selected from the group consisting of ovalbumin, ovomucoid, ovomucin, ovotransferrin, ovoinhibitor, and lysozyme, and at least one egg allergen protein has been reduced or eliminated, said protein being selected from the group consisting of ovalbumin, ovomucoid, ovomucin, ovotransferrin, ovoinhibitor, and lysozyme. 1. A knock-out poultry egg , wherein at least one oviduct-specific gene is knocked out , the gene being selected from a group consisting of ovalbumin , ovomucoid , ovomucin , ovotransferrin , ovoinhibitor , and lysozyme , and at least one egg allergen protein is reduced or eliminated , the protein being selected from a group consisting of ovalbumin , ovomucoid , ovomucin , ovotransferrin , ovoinhibitor , and lysozyme.2. The knock-out poultry egg according to claim 1 , wherein a base sequence encoding the knocked-out oviduct-specific gene includes deletion claim 1 , substitution claim 1 , or insertion of a base or bases in a region near a 5′ side or 3′ side of a PAM sequence.3. The knock-out poultry egg according to claim 1 , wherein the oviduct-specific gene is homozygously knocked-out and a genotype of the oviduct-specific gene is null (−/−).4. The knock-out poultry egg according to claim 1 , wherein the oviduct-specific gene is ovalbumin.5. The knock-out poultry egg according to claim 4 , wherein a base sequence encoding ovalbumin includes deletion claim 4 , substitution claim 4 , or insertion of a base or bases in a region corresponding to a base sequence represented by SEQ ID NO: 1 (OVATg1) and a vicinity thereof.6. The knock-out poultry egg according to claim 1 , wherein the oviduct-specific gene is an ovomucoid gene.7. The knock-out poultry egg according to claim 6 , wherein a base sequence encoding ovomucoid includes deletion claim 6 , substitution claim 6 ...

VIRAL VECTOR SUITABLE FOR GENE THERAPY ENCODING A VARIANT OF A BPIFB4 PROTEIN

The present invention relates inter alia to a polynucleotide or viral vector encoding a variant of BPIFB4 (Bactericidal/Permeability Increasing protein family B, member 4) protein and to their use for the treatment of pathologies involving impairment of nitric oxide signalling. 1. A method for the treatment of an endothelial dysfunction due to release of NO from endothelial cells below physiological levels or due to a decrease in the activity of eNOS or a method for the treatment of a clinical condition wherein it is beneficial to obtain an increase in the activation of eNOS or production of NO , the method comprising administering to a subject in need of said treatment a therapeutic amount of a polynucleotide or of a viral vector containing a polynucleotide encoding a BPIFB4 protein variant having an aminoacid sequence with at least 95% homology to the aminoacid sequence of SEQ ID NO: 1 and characterised in that said sequence comprises a Valine at the position corresponding to position 229 of SEQ ID NO: 1 , a Threonine at the position corresponding to position 281 SEQ ID NO: 1 , a Phenylalanine at a position corresponding to position 488 of SEQ ID NO: 1 and a Threonine at a position corresponding to position 494 of SEQ ID NO: 1 , said protein variant having activity in increasing the activity of eNOS and/or the production of NO , said vector operatively linked to expression control sequences , and wherein the activity of eNOS and/or the production of NO is increased and results in the amelioration or treatment of a pathology or clinical condition selected from arterial hypertension , renal failure , erectile dysfunction , retinal degeneration , uveoretinitis , vascular retinopathy , glaucoma and pulmonary hypertension.2. The method according to wherein the viral vector is selected from MV serotypes 1-9 vectors.3. The method according to wherein the vector is an adenoviral vector.4. The method according to comprising administering to a subject a viral vector ...

NUCLEIC ACID MOLECULES AND APPLICATIONS THEREOF IN PREPARING HUMAN SINGLE-DOMAIN ANTIBODY

Nucleic acid molecules and single-domain antibody applications are provided. The nucleic acid molecules include a transgenic host animal immunoglobulin genes or parts of immunoglobulin genes. Main characters are: it comprises the transgenic host animal IgH 5′-enhancer, IgM switch region (Sμ), and IgM sequences, specifically, all the IgM sequences are originated from the transgenic host animal, and the CH1 sequences of the IgM is deleted (IgM-dCH1). The regulatory control sequences of IgG are also derived from the transgenic host animal including Sγ, TM1, TM2, PolyA sequences, etc., and IgG Cγ sequences (Hinge, CH2 and CH3) are human sequence (Igγ-dCH1). The present invention ensures normal B-cell development in transgenic animal, and the transgenic animal expresses human single-domain antibodies, reducing the subsequent antibody humanization process and improving the druggability of the antibodies. 1. Nucleic acid molecules comprising immunoglobulin genes or parts of the immunoglobulin genes , wherein , the nucleic acid molecules comprises an IgG gene (Igγ) , an IgG switch region (Sγ) and an IgM gene (IgHCμ) and an IgM switch region (Sμ) , wherein , the IgM gene and the IgG gene do not have a CH1 function.2. The nucleic acid molecules according to claim 1 , wherein the Sμ claim 1 , the Sγ and the IgHCμ are derived from a transgenic host animal.3. The nucleic acid molecules according to claim 1 , wherein the IgHCμ comprises an IgM CH2 exon claim 1 , an IgM CH3 exon claim 1 , an IgM CH4 exon claim 1 , intron sequences between the IgM CH2 exon and the IgM CH3 exon claim 1 , and between the IgM CH3 exon and the IgM CH4 exon claim 1 , a TM1 claim 1 , a TM2 and PolyA signal sequences of the transgenic host animal.4. (canceled)5. The nucleic acid molecules according to claim 3 , wherein the Igγ comprises a Hinge exon claim 3 , a CH2 exon claim 3 , a CH3 exon and intron sequences between the Hinge exon and the CH2 exon claim 3 , and between the CH2 exon and the CH3 exon a ...

Methods and compositions for preparing surfactant protein d (sp-d)

Some embodiments of the methods and compositions provided herein relate to the preparation surfactant protein-D (SP-D). Some embodiments include the expression of human SP-D in certain cell lines, and the purification of human SP-D from such cell lines. Some embodiments include the preparation of certain oligomeric forms of human SP-D.

APPLICATION OF UTX GENE IN PREPARATION OF DRUGS FOR PREVENTING OR TREATING LIPID DISEASES

An application of a UTX gene in preparation of drugs for preventing or treating lipid diseases. The invention further discloses a method for knocking out a UTX gene from a mouse liver. The invention further discloses a UTX overexpression adenovirus as well as a preparation method and an application thereof. The invention further discloses a method for upregulating UTX expression in a mouse liver. The invention further discloses therapeutic action of UTX overexpression on HFD induced hyperlipidemia and NAFLD. The invention provides an available laboratory basis for preparing lipid-lowering drugs, so that the UTX can be used for preparing drugs affecting the lipid, and a new research method is provided for researching the occurrence and development of dyslipidemia. 1. A method for knocking out a UTX gene from a mouse liver which specifically knocks out a UTX gene from a liver tissue by constructing a genetically engineered mouse , comprising the following steps of:{'sup': f/f', 'f/y, '1) mating a female UTXmouse with a male albcre mouse to obtain a male UTX:albcre mouse through gene identification;'}{'sup': f/y', 'f/f', 'f/f', 'f/f, '2) mating the male UTX:albcre mouse with the female UTXmouse to obtain a female UTX:albcre mouse or a male UTX:albcre mouse; and'}{'sup': 'f/f', '3) genetically identifying the female UTX:albcre mouse to obtain a liver-specific UTX knockout mouse.'}2. A preparation method of a UTX overexpression adenovirus , comprising the following steps of:1) obtaining a cDNA: extracting an RNA from a mouse liver tissue and reversely transcribing a cDNA;2) obtaining a UTX gene: amplifying a UTX gene by PCR using a cDNA of the liver tissue as a template;3) constructing a UTX adenovirus overexpression vector: cloning the UTX gene to a pShuttle vector, linearizing a restriction enzyme PmeI, co-transfecting the UTX gene with a pAdEasy-1 vector into a BJ5183 strain, and screening a recombinant positive plasmid to obtain the UTX adenovirus overexpression ...

Anti-tau antibodies and uses thereof

Provided herein are antibodies that specifically bind Tau and methods of using the same.

NON-HUMAN ANIMALS HAVING AN ENGINEERED IMMUNOGLOBULIN LAMBDA LIGHT CHAIN LOCUS

Non-human animals (and/or non-human cells) and methods of using and making the same are provided, which non-human animals (and/or non-human cells) have a genome comprising human antibody-encoding sequences (i.e., immunoglobulin genes). Non-human animals described herein express antibodies that contain human Igλ light chains, in whole or in part. In particular, non-human animals provided herein are, in some embodiments, characterized by expression of antibodies that contain human Igλ light chains, in whole or in part, that are encoded by human Igλ light chain-encoding sequences inserted into an endogenous Igλ light chain locus of said non-human animals. Methods for producing antibodies from non-human animals are also provided. 129-. (canceled)31. The method of claim 30 , wherein the engineered endogenous immunoglobulin λ light chain locus comprises two mouse Eλs.32. The method of claim 31 , wherein the two mouse Eλs are a mouse Eλ and a mouse Eλ3-1.33. The method of claim 30 , wherein the engineered endogenous immunoglobulin λ light chain locus comprises three human Eλs.34. The method of claim 30 , wherein the germline genome of the genetically modified mouse further comprises:{'sub': H', 'H', 'H', 'H', 'H', 'H, '(i) an engineered endogenous immunoglobulin heavy chain locus comprising insertion of one or more human Vgene segments, one or more human Dgene segments, and one or more human Jgene segments, which human V, Dand Jgene segments are operably linked to a mouse immunoglobulin heavy chain constant region; or'}{'sub': H', 'H', 'H', 'H', 'H', 'H, '(ii) an engineered endogenous immunoglobulin heavy chain locus comprising insertion of one or more human Vgene segments, one or more human Dgene segments, and one or more human Jgene segments, which human V, Dand Jgene segments are operably linked to a mouse immunoglobulin heavy chain constant region, and an engineered endogenous immunoglobulin κ light chain locus comprising insertion of one or more human Vκ gene segments ...

Non-human gene-edited mammal, protein crude extract separated from connective tissue of non-human gene-edited mammal, method for protein crude extract and uses of protein crude extract

A non-human gene-edited mammal, a protein crude extract isolated from a connective tissue of the non-human gene-edited mammal, a method for preparing the protein crude extract, and uses of the protein crude extract are provided. The method includes: microinjecting a deoxyribonucleic acid sequence construct (DNA construct) containing SEQ ID NO: 4 or SEQ ID NO: 5 into a rat embryo, transplanting the rat embryo into a female rat of the same species to develop into a mature rat, and isolating a protein crude extract from a connective tissue of the mature rat. The protein crude extract includes human type I collagen and a non-native chimeric protein peptide chain.

APPLICATION OF UTX GENE IN PREPARATION OF DRUGS FOR PREVENTING OR TREATING LIPID DISEASES

An application of a UTX gene in preparation of drugs for preventing or treating lipid diseases. The invention further discloses a method for knocking out a UTX gene from a mouse liver. The invention further discloses a UTX overexpression adenovirus as well as a preparation method and an application thereof. The invention further discloses a method for upregulating UTX expression in a mouse liver. The invention further discloses therapeutic action of UTX overexpression on HFD induced hyperlipidemia and NAFLD. The invention provides an available laboratory basis for preparing lipid-lowering drugs, so that the UTX can be used for preparing drugs affecting the lipid, and a new research method is provided for researching the occurrence and development of dyslipidemia. 1. A method for treating or preventing a lipid disease comprising a step of administering a subject in need with an adenovirus expressing UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X) protein , wherein the lipid disease is hyperlipidemia; the adenovirus is an UTX overexpression adenovirus.2. The method according to claim 1 , wherein the UTX overexpression adenovirus is prepared by the following steps of:1) extracting RNAs from a mouse liver tissue and reverse transcribing the RNAs into cDNAs;2) amplifying an UTX gene by PCR using the cDNAs as a template;3) cloning the UTX gene into a pShuttle vector to yield a pShuttle-UTX vector, linearizing the pShuttle-UTX vector with a restriction enzyme PmeI to yield a linearized pShuttle-UTX vector, co-transfecting the linearized pShuttle-UTX vector with a pAdEasy-1 vector into a BJ5183 strain, and screening a recombinant positive plasmid to obtain an UTX adenovirus overexpression vector; and4) transfecting the UTX adenovirus overexpression vector into cultured AD-293 cells for continuously culturing for 7 days to 10 days, discarding cell culture supernatant, collecting and suspending cells in an EP tube, repeatedly freezing/unfreezing in a ...

OPTIMIZED ENGINEERED MEGANUCLEASES HAVING SPECIFICITY FOR A RECOGNITION SEQUENCE IN THE HEPATITIS B VIRUS GENOME

The present invention encompasses engineered nucleases which recognize and cleave a recognition sequence within a Hepatitis B virus (HBV) genome. The engineered meganucleases can exhibit at least one optimized characteristic, such as enhanced specificity and/or efficiency of indel formation, when compared to the first-generation meganuclease HBV 11-12x.26. Further, the invention encompasses pharmaceutical compositions comprising engineered meganuclease proteins, nucleic acids encoding engineered meganucleases, and the use of such compositions for treating HBV infections or hepatocellular carcinoma. 125.-. (canceled)26. A polynucleotide comprising a nucleic acid sequence encoding an engineered meganuclease that recognizes and cleaves a recognition sequence consisting of SEQ ID NO: 10 within a Hepatitis B virus genome , wherein said engineered meganuclease comprises a first subunit and a second subunit , wherein said first subunit binds to a first recognition half-site of said recognition sequence and comprises a first hypervariable (HVR1) region , wherein said second subunit binds to a second recognition half-site of said recognition sequence and comprises a second hypervariable (HVR2) region , wherein said engineered meganuclease comprises the amino acid sequence of SEQ ID NO: 12 or 13.27. The polynucleotide of claim 26 , wherein said polynucleotide is an mRNA.28. The polynucleotide of claim 27 , wherein said mRNA is a polycistronic mRNA encoding one or more of said engineered meganuclease.29. The polynucleotide of claim 28 , wherein said polycistronic mRNA encodes:(a) at least one of said engineered meganuclease, which recognizes and cleaves a recognition sequence consisting of SEQ ID NO: 10; and(b) a second engineered meganuclease, which recognizes and cleaves a second recognition sequence, which is present in a Hepatitis B virus genome but differs from SEQ ID NO: 10.30. The polynucleotide of claim 29 , wherein said second recognition sequence consists of SEQ ID ...

Method for producing antibody

Disclosed is an antibody in which the 80th amino acid residue in a variable region based on the Kabat method and the 171th amino acid residue in a constant region based on the Kabat method are substituted with cysteine in an antibody in which the 80th amino acid residue in the variable region and the 171th amino acid residue in the constant region are not cysteine.

Inducible disease models methods of making them and use in tissue complementation

Disclosed herein, are inducible immunodeficient animals and methods to make them by adding an IL2Rg/RAG2 rescue cassette (RG-reg) or an IL2Rg/RAG2/FAH rescue cassette (FRG-reg) to a line of IL2Rg/RAG2 knockout (RG-KO) or IL2Rg/RAG2/FAH knockout (FRG-KO) swine. The rescue cassette enables line breeding of immunocompetent (regRG-KO) or (regFRG-KO) swine for rapid propagation. The rescue cassette can be excised, specifically in germ cells of regRG-KO or regFRG-KO swine, such that offspring of animals do not possess the rescue cassette and are immunodeficient. The immunodeficient swine also provide host embryos having genetic ablations to provide a niche for organ complementation by human stem cells.

Non-human animals having a humanized signal-regulatory protein gene

Genetically modified non-human animals and methods and compositions for making and using the same are provided, wherein the genetic modification comprises a humanization of an endogenous signal-regulatory protein gene, in particular a humanization of a SIRPα gene. Genetically modified mice are described, including mice that express a human or humanized SIRPα protein from an endogenous SIRPα locus.

GENETICALLY MODIFIED NON-HUMAN ANIMALS FOR GENERATING THERAPEUTIC ANTIBODIES AGAINST PEPTIDE-MHC COMPLEXES, METHODS OF MAKING AND USES THEREOF

A non-human animal is genetically modified with sequence encoding a human or humanized MHC molecule or associated molecule, e.g., β2 microglobulin, and expression of the sequence by the non-human animal induces tolerance to the corresponding human HLA from which the human or human MHC molecule is derived. The tolerance exhibited by these non-human animals allows these animals to generate specific antibody responses to the corresponding human HLA when such HLA is presenting a peptide that is antigenic to the non-human animal. Such an antibody response, which specifically targets a pMHC complex of interest without binding to the MHC molecule may be useful in immunotherapeutic modalities that target a component of the immunological synapse which provides the specificity of that interaction. 1. A genetically modified non-human animal whose genome comprises(a) a nucleotide sequence encoding a human or humanized MHC molecule or at least a peptide binding portion thereof, and(b) an (un)rearranged human or humanized immunoglobulin heavy locus and/or an (un)rearranged human or humanized immunoglobulin light chain locus, optionally wherein at least one of the (un)rearranged human or humanized immunoglobulin heavy locus and/or an (un)rearranged human or humanized immunoglobulin light chain locus is unrearranged,wherein the genetically modified non-human animal expresses the human or humanized MHC molecule or at least a peptide binding portion thereof,wherein the genetically modified non-human animal expresses immunoglobulins comprising a human or humanized heavy chain variable domain and/or a human or humanized light chain variable domain, andwherein the non-human animal is tolerized to the human or humanized MHC molecule or at least a peptide binding portion thereof such that it generates a specific B-cell response when immunized with an antigenic peptide-MHC (pMHC) complex that comprises (i) a peptide that is heterologous to the non-human animal complexed with (ii) human HLA ...

AGENTS BINDING SPECIFICALLY TO HUMAN CADHERIN-17, HUMAN CADHERIN-5, HUMAN CADHERIN-6 AND HUMAN CADHERIN-20 RGD MOTIF

The disclosure relates to agents binding specifically to human cadherin 17 (CDH17), and/or to human cadherin 5 (CDH5), and/or to human cadherin 6 (CDH6), and/or to human cadherin 20 (CDH2O). The disclosure also relates to the use of these agents in therapy, methods for diagnosis and/or prognosis and/or stratification of a cancer in a subject, and pharmaceutical compositions comprising said agents. The disclosure also relates to cancer markers and markers of metastasis. 1. An agent binding specifically to an epitope comprising residues 603 to 605 of human cadherin 17 (CDH17) , and/or to an epitope comprising residues 236 to 238 or residues 299 to 301 of human cadherin 5 (CDH5) , and/or to an epitope comprising residues 83 to 85 of human cadherin 6 (CDH6) and/or to an epitope comprising residues 89 to 91 of human cadherin 20 (CDH20) , wherein said agent is an immunoglobulin agent or a non-immunoglobulin agent selected from the group consisting of a peptide aptamer , a nucleic acid aptamer , a DARPin , an affibody , and an anticalin.2. The agent according to claim 1 , wherein the epitope comprises the sequence shown in SEQ ID NO: 1 (VSLRGDTRG).3. The agent according to claim 2 , wherein said immunoglobulin agent is an antibody or an antigen-binding fragment of said antibody.4. The agent according to claim 3 , wherein said antigen -binding fragment is selected from the group consisting of Fv claim 3 , Fab claim 3 , F(ab′) claim 3 , and Fab′.5. The agent according to claim 3 , wherein said antibody or said antigen-binding fragment comprises within the heavy chain:a CDR comprising the amino acid sequence shown in SEQ ID NO: 2 [CDR-H1], a CDR comprising the amino acid sequence shown in SEQ ID NO: 3 [CDR-H2], and a CDR comprising the amino acid sequence shown in SEQ ID NO: 4 [CDR-H3], or a functionally equivalent variant of said CDRs,ora CDR comprising the amino acid sequence shown in SEQ ID NO: 5 [CDR-H1], a CDR comprising the amino acid sequence shown in SEQ ID NO: 6 [CDR ...

TCR LIBRARIES

The present invention relates to a library of particles, the library displaying a plurality of different T cell receptors (TCRs), wherein the plurality of TCRs consists essentially of TCRs comprising an alpha chain comprising an alpha chain variable domain and a beta chain comprising a beta chain variable domain and the library comprises more than one TRAY gene product and/or more than one TRBV gene product, wherein the beta chain variable domain does not comprise one or more of a TRBV5-1, 5-3, 5-4, 5-5, 5-6, 5-7 or 5-8 gene product and wherein the plurality of TCRs do not consist essentially of TCRs comprising a TRAV12-2 gene product from a natural repertoire and a TRBV6 gene product from a natural repertoire and TCRs comprising a TRAV21 gene product from a natural repertoire and a TRBV6 gene product from a natural repertoire. 1. A library of particles displaying a plurality of T cell receptors (TCRs) , wherein the plurality of TCRs:i) consists essentially of TCRs comprising an alpha chain comprising as alpha chain variable domain and beta chain comprising a beta chain variable domain that does not comprise one or more of a TRBV5-1, 5-3, 5-4, 5-5, 5-6, 5-7 or 5-8 gene product;ii) comprises more than one TRAV gene product and/or more than one TRBV gene product; andiii) does not consist essentially of TCRs comprising a TRAV12-2 gene product from a natural repertoire and a TRBV6 gene product from a natural repertoire and TCRs comprising a TRAV21 gene product from a natural repertoire and a TRBV6 gene product from a natural repertoire.2. The library according to claim 1 , wherein the beta chain variable domain does not comprise one or either of a TRBV10-2 gene product and a TRBV10-3 gene product.3. The library according to or claim 1 , wherein the beta chain variable domain does not comprise a TRBV2 gene product and/or a TRBV9 gene product and/or a TRBV14 gene product.4. The library according to any one of to claim 1 , wherein the beta chain variable domain does not ...

ADAM6 KNOCKIN MICE

A transgenic mouse that is engineered by one or more genetic modifications that delete endogenous Adam6a and Adam6b genes in a male mouse, which mouse comprises in its genome only one exogenous Adam6 transgene, and expresses an ADAM6 protein comprising at least 90% sequence identity to SEQ ID NO:1 or SEQ ID NO:2, which Adam6 transgene is functional in a male mouse. 1. A transgenic mouse that is engineered by a deletion of endogenous Adam6a and Adam6b genes in a male mouse , which mouse comprises in its genome only one exogenous Adam6 transgene , and expresses an ADAM6 protein comprising at least 90% sequence identity to SEQ ID NO:1 or SEQ ID NO:2 , which Adam6 transgene is functional in a male mouse.2. The mouse of claim 1 , wherein said Adam6 transgene is Adam6a or Adam6b.3. The mouse of claim 1 , wherein said endogenous Adam6a and Adam6b genes are knocked out by said deletion.4. The mouse of claim 1 , which comprises in its genome immunoglobulin gene segments that upon VDJ rearrangement encode immunoglobulin sequences of one or more antibody variable domains.5. The mouse of claim 1 , which comprises an immunoglobulin heavy chain variable region (IHVR) locus comprising human immunoglobulin gene segment (hIGS) coding sequences and murine expression control sequences in operable linkage to control expression of the hIGS coding sequences.6. The mouse of claim 5 , wherein the exogenous Adam6 transgene is embedded in said IHVR locus.7. The mouse of claim 5 , wherein said hIGS coding sequences and said murine expression control sequences are comprised in a heterologous expression cassette further comprising said Adam6 transgene claim 5 , wherein said heterologous expression cassette is integrated within the mouse genome and replaces endogenous immunoglobulin gene segment coding sequences claim 5 , or replaces an immunoglobulin gene segment of the endogenous IHVR locus.8. Use of the mouse of in a method of generating an antibody.9. Use of the mouse of in a method of ...

OX40-BINDING POLYPEPTIDES AND USES THEREOF

Provided herein are VHH-containing polypeptides that bind OX40. In some embodiments, VHH-containing polypeptides that bind and agonize OX40 are provided. Uses of the VHH-containing polypeptides are also provided. 1. A polypeptide comprising at least one VHH domain that binds OX40 , wherein the VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10 , a CDR2 comprising the amino acid sequence of SEQ ID NO: 11 , and a CDR3 comprising the amino acid sequence of SEQ ID NO: 12.2. The polypeptide of claim 1 , wherein the VHH domain is humanized.3. (canceled)4. The polypeptide of claim 1 , wherein the VHH domain comprises a FR2 comprising the amino acid sequence of SEQ ID NO: 22 and a FR3 comprising the amino acid sequence of SEQ ID NO: 23.5. The polypeptide of claim 1 , wherein the VHH domain comprises the amino acid sequence of SEQ ID NO: 9.6. The polypeptide of claim 1 , comprising two VHH domains.7. The polypeptide of claim 1 , comprising three VHH domains.8. The polypeptide of claim 1 , comprising four VHH domains.9. The polypeptide of claim 1 , wherein the polypeptide comprises at least one binding domain that binds a second antigen other than OX40.10. The polypeptide of claim 9 , wherein the second antigen is selected from PD-1 claim 9 , PD-L1 claim 9 , and 41BB.11. The polypeptide of claim 10 , wherein the at least one binding domain that binds a second antigen is an antagonist or an agonist.12. The polypeptide of claim 1 , wherein each VHH domain binds OX40.13. The polypeptide of claim 12 , wherein each VHH domain comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 10 claim 12 , a CDR2 comprising the amino acid sequence of SEQ ID NO: 11 claim 12 , and a CDR3 comprising the amino acid sequence of SEQ ID NO: 12.14. (canceled)15. The polypeptide of claim 13 , wherein each VHH domain comprises a FR2 comprising the amino acid sequence of SEQ ID NO: 22 and a FR3 comprising the amino acid sequence of SEQ ID NO: 23.16. The polypeptide ...

ANIMAL MODELS AND THERAPEUTIC MOLECULES

The present invention relates inter alia to a rodent or rodent cell having a genome comprising: i) one or more companion animal IGH V region genes, one or more companion animal D region genes and one or more companion animal J region genes; and (ii) optionally one or more companion animal IGL kappa V region genes and one or more companion animal IGL kappa J region genes; and/or one or more companion animal IGL lambda V region genes and one or more companion animal IGL lambda J region genes, wherein the rodent or rodent cell is capable of expressing the companion animal variable region gene(s) to form an antibody chain and wherein the companion animal species is not a rodent. 1. A rodent or rodent cell having a genome comprising;i) one or more companion animal IGH V region genes, one or more companion animal D region genes and one or more companion animal J region genes; and(ii) optionally one or more companion animal IGL kappa V region genes and one or more companion animal IGL kappa J region genes; and/or one or more companion animal IGL lambda V region genes and one or more companion animal IGL lambda J region genes,wherein the rodent or rodent cell is capable of expressing the companion animal variable region gene(s) to form an antibody chainand wherein the companion animal species is not a rodent.2. A rodent or rodent cell having a genome comprisingi) one or more companion animal IGL kappa V region genes and one or more companion animal IGL kappa J region genes; and/or one or more companion animal IGL lambda V region genes and one or more companion animal IGL lambda J region genes, andii) optionally one or more companion animal IGH V region genes, one or more companion animal or host D region genes and one or more companion animal or host J region geneswherein the rodent or rodent cell is capable of expressing the companion animal variable region gene(s) to form an antibody chainand wherein the companion animal species is not a rodent.3. The rodent or rodent ...

METHODS, COMPOSITIONS AND CELLS FOR PREPARING SURFACTANT PROTEIN D (SP-D)

Some embodiments of the methods and compositions provided herein relate to the preparation surfactant protein-D (SP-D). Some embodiments include the expression of human SP-D in certain cell lines, and the purification of human SP-D from such cell lines. Some embodiments include the preparation of certain oligomeric forms of human SP-D. 1. A method for producing a human surfactant protein D (SP-D) polypeptide composition comprising:(a) introducing a polynucleotide encoding the SP-D polypeptide into a human mammalian cell;(b) culturing the cell under conditions in which the SP-D polypeptide is expressed; and(c) isolating the expressed SP-D polypeptide from the cell.2. The method of claim 1 , wherein the cell is derived from a human myeloid leukemia cell.3. The method of claim 2 , wherein the cell is selected from the group consisting of NM-H9D8 claim 2 , NM-H9D8-E6Q12 claim 2 , and NM-F9.4. The method of claim 3 , wherein the cell is a NM-H9D8 cell.5. The method of claim 1 , wherein the polynucleotide encodes a wild type SP-D polypeptide leader sequence or a wild type T-cell receptor (TCR) polypeptide leader sequence.6. (canceled)7. (canceled)8. The method of claim 5 , wherein the polynucleotide encodes a polypeptide having an amino acid sequence comprising SEQ ID NO:05 or SEQ ID NO:10.9. The method of claim 5 , wherein the polynucleotide encodes a polypeptide having an amino acid sequence comprising SEQ ID NO:04 or SEQ ID NO:09.10. (canceled)11. (canceled)12. (canceled)13. (canceled)14. (canceled)15. The method of claim 1 , wherein the SP-D polypeptide comprises a residue at a polymorphic position claim 1 , wherein the residue is selected from the group consisting of Met11/31 claim 1 , Thr160/180 claim 1 , Ser 270/290 claim 1 , and Ala 286/306.16. (canceled)17. (canceled)18. The method of claim 1 , further comprising isolating a population of the expressed SP-D polypeptides claim 1 , each expressed SP-D polypeptide comprising a complex-type carbohydrate attached at ...

METHODS AND COMPOSITIONS FOR PREPARING SURFACTANT PROTEIN D (SP-D)

Some embodiments of the methods and compositions provided herein relate to the preparation surfactant protein-D (SP-D). Some embodiments include the expression of human SP-D in certain cell lines, and the purification of human SP-D from such cell lines. Some embodiments include the preparation of certain oligomeric forms of human SP-D. 1. A composition comprising a human surfactant protein D (SP-D) polypeptide prepared by a method comprising:(a) introducing a polynucleotide encoding the SP-D polypeptide into a human mammalian cell;(b) culturing the cell under conditions in which the SP-D polypeptide is expressed; and(c) isolating the expressed SP-D polypeptide from the cell.2. The composition of claim 1 , wherein the cell is derived from a human myeloid leukemia cell.3. The composition of claim 2 , wherein the cell is selected from the group consisting of NM-H9D8 claim 2 , NM-H9D8-E6Q12 claim 2 , and NM-F9.4. The composition of claim 3 , wherein the cell is a NM-H9D8 cell.5. The composition of claim 3 , wherein the cell is a NM-H9D8(8B11) cell.6. The composition of claim 1 , wherein the polynucleotide encodes a wild type SP-D polypeptide leader sequence or a wild type T-cell receptor (TCR) polypeptide leader sequence.7. (canceled)8. (canceled)9. The composition of claim 6 , wherein the polynucleotide encodes a polypeptide having an amino acid sequence comprising SEQ ID NO:05 or SEQ ID NO:10.10. The composition of claim 6 , wherein the polynucleotide encodes a polypeptide having an amino acid sequence comprising SEQ ID NO:04 or SEQ ID NO:09.11. (canceled)12. (canceled)13. (canceled)14. (canceled) 15. (canceled) 16. (canceled) 17. (canceled) 18. (canceled) 19. (canceled)20. The composition of comprising a population of the expressed SP-D polypeptides claim 1 , each expressed SP-D polypeptide comprising a complex-type carbohydrate attached at an N-glycosylation site claim 1 , wherein the population has a glycosylation pattern comprising the following characteristics:(i ...

Transgenic mouse expressing common human light chain

The present disclosure relates to a transgenic non-human mammal (e.g., mouse) that expresses a common light chain, wherein the transgenic non-human mammal comprises a hybrid light chain locus from which a light chain having a human Vκ joined to a mouse Jκ is expressed. Also provided are methods of making a transgenic non-human mammal capable of expressing a common light chain.

METHODS OF DETECTING ALLOANTIBODIES TO HLA CLASS II ANTIGENS

Described herein are materials and methods of incorporating CLIP peptide into the peptide binding groove of HLA Class II antigens and using such HLA Class II antigens for the detection of alloantibodies. 1. A method of preparing an HLA Class II antigen in which a Class II-associated invariant chain peptide (CLIP) is present in the peptide binding groove comprising:transfecting a host cell with a gene encoding an invariant chain andexpressing the HLA Class II antigen.2. The method of wherein the gene encoding the invariant chain is CD74.3. A method of preparing an HLA Class II antigen in which a CLIP peptide is present in the peptide binding groove above a selected threshold for determination of antibodies specific for HLA Class II antigens in which a CLIP is present in the peptide binding groove comprising:transfecting a host cell which expresses an invariant chain with a gene expressing an HLA Class II antigen.47.-. (canceled)8. A method of screening for antibodies that specifically bind an HLA Class II antigen in which a CLIP is present in the peptide binding groove comprising the steps of:preparing a panel comprising a solid-phase substrate having an HLA Class II antigen in which a CLIP is present in the peptide binding groove immobilized or attached hereto;obtaining a serum sample from a human subject,contacting the panel with the serum sample, anddetecting binding of antibodies in said serum sample to the HLA Class II antigen in which a CLIP is present in the peptide binding groove.9. The method of wherein the HLA Class II antigen in which a CLIP is present in the peptide binding groove is prepared by the method of transfecting a host cell with a gene encoding an invariant chain and expressing the HLA Class II antigen.10. The method of wherein the HLA Class II antigen in which a CLIP is present in the peptide binding groove is prepared by the method of transfecting a host cell which expresses an invariant chain with a gene expressing an HLA Class II antigen.11. ...

Animal models and therapeutic molecules

The invention discloses methods for the generation of chimaeric human—non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods.

Transgenic Non-Human Vertebrate for the Expression of Class-Switched, Fully Human, Antibodies

The present invention relates to humanisation of antibodies in vivo. The invention provides non-human vertebrates, cells, populations and methods useful for humanising chimaeric antibodies in vivo. Using the present invention it is possible straightforwardly and rapidly to obtain antigen-specific antibodies that are fully human (ie, comprising human variable and constant regions) and have undergone recombination, junctional diversification, affinity maturation and isotype switching in vivo in a non-human vertebrate system. Furthermore, such antibodies are humanised (eg, totally human)—and selected—totally in vivo, and as such the present invention harnesses in vivo filtering for expressibility, affinity and biophysical characteristics in the context of the desired human variable and constant region pairings. This is avoids problems of down-grading antibody characteristics when humanising the constant region of chimaeric antibodies in vitro. 2. The vertebrate or cell of claim 1 , wherein said vertebrate is a mouse or rat claim 1 , or wherein said vertebrate cell is a mouse cell or rat cell.3. The vertebrate or cell of claim 2 , comprising an antibody heavy chain locus that comprises (in the 5′ to 3′ direction) an unrearranged human variable region claim 2 , a first switch claim 2 , an endogenous mu constant region claim 2 , a second switch and a human constant region of said non-mu isotype; wherein said heavy chain locus is capable of undergoing switching from IgM to said non-mu isotype; and/or wherein the first switch is a rodent Smu or the endogenous Smu of the IgH locus.4. The vertebrate or cell according to claim 3 , wherein said genome comprises endogenous RAG-1 and RAG-2 genes.5. The vertebrate or cell according to claim 4 , wherein said human variable region comprises a plurality of:(a) unrearranged human VH and/or D and/or JH gene segments; and/or(b) a substantially complete human functional repertoire of human VH and/or D and/or JH gene segments.6. The ...

CELL LINE FOR RECOMBINANT PROTEIN AND/OR VIRAL VECTOR PRODUCTION

Cells and cell lines are disclosed that are able to produce therapeutic proteins, antibodies, vectors, and viral vectors such as lentiviral vectors and adeno-associated viral (AAV) vectors. The cells and/or cell lines can have mutations or deletions in either one or both of the endogenous di-hydrofolate reductase (DHFR−/−) or glutamine synthetase (GS−/−) genes such that DHFR and/or GS expression or function is substantially reduced or eliminated. 1. A human embryonic kidney (HEK) cell which does not express functional endogenous di-hydrofolate reductase (DHFR) and/or glutamine synthetase (GS).2. A human embryonic kidney (HEK) cell line which does not express functional endogenous di-hydrofolate reductase (DHFR) and/or glutamine synthetase (GS).3. The HEK cell or cell line of or , stably or transiently transfected with a first heterologous nucleic acid sequence , and optionally stably or transiently transfected with a second heterologous nucleic acid sequence.4. The HEK cell or cell line of or , stably or transiently transfected with a first heterologous nucleic acid sequence and a first selectable marker , and optionally stably or transiently transfected with a second heterologous nucleic acid sequence and a second selectable marker.5. The HEK cell or cell line of or , wherein the first heterologous nucleic acid sequence encodes a therapeutic protein or polynucleotide sequence and optional second heterologous nucleic acid sequence encodes a therapeutic protein or polynucleotide sequence.6. The HEK cell or cell line of claim 4 , wherein the therapeutic protein or polynucleotide sequence encoded by the first heterologous nucleic acid sequence and the therapeutic protein or polynucleotide sequence encoded by the optional second heterologous nucleic acid sequence are the same or are different.7. The HEK cell or cell line of claim 4 , wherein the first or second selectable marker does not provide resistance to an antibiotic.8. The HEK cell or cell line of claim 4 , ...

ANIMAL MODELS AND THERAPEUTIC MOLECULES - Track 1

The invention discloses methods for the generation of chimaeric human-non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods. 1. A method of obtaining an antigen specific antibody or antigen binding fragment thereof , comprising a human immunoglobulin heavy (IgH) chain , wherein said human IgH chain comprises a human IgH chain variable region and a human IgH chain constant region , the method comprising:expressing the antibody or antigen binding fragment thereof, from a cell, wherein the cell comprises a nucleic acid encoding said human IgH chain variable region and said human IgH chain constant region of said antibody,wherein said human IgH chain variable region is of a transgenic mouse contacted with said antigen;wherein the germline of said mouse has a genome with a homozygous immunoglobulin heavy chain (IgH) locus,wherein said homozygous IgH locus comprises unrearranged human immunoglobulin heavy chain (IgH) variable region (VH) gene segments comprising one or more human IgH V gene segments, one or more human D gene segments, one or more human JH gene segments and human intronic DNA contiguous with mouse intronic DNA located at an endogenous IgH locus upstream of an enhancer and a constant (C) region comprising an endogenous CH gene segment,wherein said C region comprises mouse Cμ and said human variable region gene segments are operably linked to said C region to permit rearrangement and expression of chimeric IgM heavy chain polypeptide comprising a human variable region and a mouse μ constant region,wherein in said transgenic mouse expression of IgH heavy chains comprising endogenous mouse variable regions is reduced or prevented;wherein said mouse comprises IgH-VDJCμ transcripts encoding said chimeric ...

SELF-LIMITING CAS9 CIRCUITRY FOR ENHANCED SAFETY (SLICES) PLASMID AND LENTIVIRAL SYSTEM THEREOF

The present invention describes a Self-Limiting Cas9 circuitry for Enhanced Safety (SLiCES) which consists of an expression unit for the Cas9 (SpCas9), a first Cas9 self-targeting sgRNA and a second sgRNA targeting a chosen genomic locus. The self limiting circuit, by controlling Cas9 levels, results in increased genome editing specificity. For its in vivo utilization, SLiCES was integrated into a lentiviral delivery system (lentiSLiCES) via circuit inhibition to achieve viral particle production. Following its delivery into target cells, the lentiSLiCES circuit is switched on to edit the intended genomic locus while simultaneously stepping up its own neutralization through SpCas9 inactivation. By preserving target cells from residual nuclease activity, the present hit and go system increases safety margins for genome editing. 2. The plasmid according to claim 1 , wherein the anti-Cas9 sgRNA is encoded by a sequence of 17-23 nucleotide preferably.3. The plasmid according to claim 2 , wherein anti-Cas9 sgRNA encoding sequence is a sequence having at least a 60% homology with a sequence selected in the group consisting of SEQ ID N. 1-6.4. The plasmid according to claim 1 , whereinthe expression cassette for a Cas9 molecule and/or the nucleotide sequence that encodes for an anti-Cas9 sgRNA comprises at least an intron; andthe expression cassette for the Cas9 molecule and/or the sequence encoding for anti-cas9 sgRNA is preceded by a sequence including an inducible promoter.5. The plasmid according to claim 1 , wherein the expression cassette for the Cas9 molecule and the sequence encoding for anti-cas9 sgRNA are both preceded by a sequence including an inducible promoter.6. A genetically-modified micro-organism comprising the plasmid according to .7. A cell transfected with the plasmid according to .8. A viral or artificial delivery system comprising the plasmid according to .9. (canceled)10. A method of treating a monogenic disorder claim 1 , high cholesterol claim 1 , ...

CIRCULAR RNA FOR TRANSLATION IN EUKARYOTIC CELLS

Disclosed are methods and constructs for engineering circular RNA. Disclosed is a vector for making circular RNA, said vector comprising the following elements operably connected to each other and arranged in the following sequence: 1. A vector for making circular RNA , said vector comprising the following elements operably connected to each other and arranged in the following sequence:a) a 5′ homology arm,b) a 3′ Group I intron fragment containing a 3′ splice site dinucleotide,c) a 5′ spacer sequence,d) an internal ribosome entry site (IRES),e) a protein coding region or noncoding region,f) a 3′ spacer sequence,g) a 5′ Group I intron fragment containing a 5′ splice site dinucleotide, andh) a 3′ homology arm,said vector allowing production of a circular RNA that is translatable or biologically active inside eukaryotic cells.2. (canceled)3Cyanobacterium Anabaena. The vector of claim 1 , wherein the 3′ Group I intron fragment and the 5′ Group I intron fragment are from a sp. pre-tRNA-Leu gene.4. The vector of claim 1 , wherein the 3′ Group I intron fragment and the 5′ Group I intron fragment are from a T4 phage Td gene.5S. cerevisiaeS. cerevisiae. The vector of claim 1 , wherein the IRES is selected from an IRES sequence of Taura syndrome virus claim 1 , Triatoma virus claim 1 , Theiler's encephalomyelitis virus claim 1 , simian Virus 40 claim 1 , Solenopsis invicta virus 1 claim 1 , Rhopalosiphum padi virus claim 1 , Reticuloendotheliosis virus claim 1 , fuman poliovirus 1 claim 1 , Plautia stali intestine virus claim 1 , Kashmir bee virus claim 1 , Human rhinovirus 2 claim 1 , Homalodisca coagulata virus-1 claim 1 , Human Immunodeficiency Virus type 1 claim 1 , Homalodisca coagulata virus-1 claim 1 , Himetobi P virus claim 1 , Hepatitis C virus claim 1 , Hepatitis A virus claim 1 , Hepatitis GB virus claim 1 , foot and mouth disease virus claim 1 , Human enterovirus 71 claim 1 , Equine rhinitis virus claim 1 , Ectropis obliqua picorna-like virus claim 1 , ...

TRANSGENE EXPRESSION IN AVIANS

A transgenic avian containing in its genome an exogenous nucleotide sequence which includes a promoter component and a vector with reduced promoter interference wherein the exogenous nucleotide sequence is integrated into the genome and the avian. 125-. (canceled)26. A method of using a transgenic chicken to produce an exogenous protein comprising:providing a transgenic chicken,wherein the transgenic chicken contains in its genome an exogenous nucleotide sequence comprising (i) an ovalbumin promoter operably linked to an exogenous coding region encoding an exogenous protein and (ii) a self-inactivating avian leukosis viral (ALV) vector comprising a 5′ long terminal repeat that comprises a nucleic acid at least 90% identical to nucleotides 174-346 SEQ ID NO:29,wherein the transgenic chicken makes the exogenous protein in a hard shell egg laid by the chicken.27. A method of using a transgenic chicken to produce an exogenous protein comprising:providing a transgenic chicken,wherein the transgenic chicken contains in its genome an exogenous nucleotide sequence comprising (i) an oviduct specific promoter operably linked to an exogenous coding region encoding an exogenous protein and (ii) a self-inactivating avian leukosis viral (ALV) vector comprising a 5′ long terminal repeat that comprises a nucleic acid at least 90% identical to nucleotides 174-346 SEQ ID NO:29,wherein the transgenic chicken makes the exogenous protein in a hard shell egg laid by the chicken,wherein the oviduct specific promoter comprises a nucleic acid at least 90% identical to a nucleic acid molecule that comprises from 5′ to 3′ the ovalbumin Dnase hypersensitive site (DHS) III, 5′ UTR-3′ portion of Exon 1, and 3′ UTR;whereinthe 5′ UTR-3′ portion from Exon 1 is the nucleotide sequence 8295 through 8311 of SEQ ID NO: 22;the 3′ UTR is the nucleotide sequence 13576 through 14209 of SEQ ID NO: 22; andthe DHS III is the nucleotide sequence 3253 through 3559 of SEQ ID NO: 22.28. The method of wherein the ...

METHODS OF PRODUCING NOR-OPIOID AND NAL-OPIOID BENZYLISOQUINOLINE ALKALOIDS

A method of demethylizing an opioid to a nor-opioid is provided. The method comprises contacting an opioid with at least one enzyme. Contacting the opioid with the at least one enzyme converts the opioid to a nor-opioid. A method of converting a nor-opioid to a nal-opioid is provided. The method comprises contacting a nor-opioid with at least one enzyme. Contacting the nor-opioid with the at least one enzyme converts the nor-opioid to a nal-opioid. 1. A method of demethylating a first opioid to a second opioid , comprising:contacting the first opioid with at least one enzyme, wherein contacting the first opioid with the at least one enzyme converts the first opioid to the second opioid through loss of an O-linked methyl group, andwherein the first opioid is not selected from the group consisting of codeine and thebaine.2. A method of demethylating an opioid to a nor-opioid , comprising:contacting a first opioid with at least one enzyme, wherein contacting the first opioid with the at least one enzyme converts the first opioid to a second opioid through loss of an O-linked methyl group; andcontacting the second opioid with at least one enzyme, wherein contacting the opioid with the at least one enzyme converts the second opioid to a nor-opioid through loss of an N-linked methyl group.3. A method of demethylating an opioid to a nor-opioid , comprising:contacting the opioid with at least one enzyme, wherein contacting the opioid with the at least one enzyme converts the opioid to the nor-opioid through removal of a N-linked methyl group from the opioid,wherein the opioid is not thebaine when the opioid contacts the at least one enzyme in vitro.4. (canceled)5. (canceled)6. The method of claim 1 , wherein the second opioid is produced by culturing an engineered cell comprising a coding sequence for encoding the at least one enzyme.7. The method of claim 2 , wherein the nor-opioid is produced by culturing an engineered cell comprising a coding sequence for encoding the ...

Cellobiohydrolase Variants and Polynucleotides Encoding Same

The present invention relates to cellobiohydrolase variants, polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of producing and using the variants. 1. A cellobiohydrolase variant , comprising a substitution at one or more positions corresponding to positions 201 , 243 , 286 , and 343 of the polypeptide of SEQ ID NO: 1 , wherein the variant has cellobiohydrolase activity and wherein the variant has at least 60% , e.g. , at least 65% , at least 70% , at least 75% , at least 80% , at least 85% , at least 90% , at least 95% , at least 96% , at least 97% , at least 98% , or at least 99% , but less than 100% , sequence identity to a parent cellobiohydrolase.2. The cellobiohydrolase variant of claim 1 , wherein the parent cellobiohydrolase has at least 60% claim 1 , e.g. claim 1 , at least 65% claim 1 , at least 70% claim 1 , at least 75% claim 1 , at least 80% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% claim 1 , at least 99% claim 1 , or 100% sequence identity to SEQ ID NO: 1 claim 1 , 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , 6 claim 1 , 7 claim 1 , 8 claim 1 , or 9.3. The cellobiohydrolase variant of claim 1 , wherein the parent cellobiohydrolase is a fragment of SEQ ID NO: 1 claim 1 , 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , 6 claim 1 , 7 claim 1 , 8 claim 1 , or 9 claim 1 , wherein the fragment has cellobiohydrolase activity.4. The cellobiohydrolase variant of claim 1 , which has at least 60% claim 1 , e.g. claim 1 , at least 65% claim 1 , at least 70% claim 1 , at least 75% claim 1 , at least 80% claim 1 , at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , at least 96% claim 1 , at least 97% claim 1 , at least 98% claim 1 , or at least 99% claim 1 , but less than 100% claim 1 , sequence identity to the polypeptide of SEQ ID NO: 1 claim 1 , 2 claim 1 , 3 claim 1 , ...

MGMT-BASED METHOD FOR OBTAINING HIGH YEILDS OF RECOMBINANT PROTEIN EXPRESSION

The present invention relates to a novel enhancer of protein production in host cells. It discloses a vector for expressing recombinant proteins in these cells, comprising a nucleotide sequence encoding a) a secretion peptidic signal, b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT, EC 2.1.1.63), a mutant or a catalytic domain thereof, and c) a recombinant protein. Said MGMT enzyme is preferably the so-called SNAP protein. 1. A vector for expressing recombinant proteins in host cells , comprising a nucleotide sequence encoding in a single open reading frame , from 5′ to 3′:a) a peptidic secretion signal which is functional in said host cells,b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT, EC 2.1.1.63), a catalytic domain, or a mutant thereof, andc) a recombinant protein.2. The expression vector according to claim 1 , wherein said MGMT enzyme is the protein of SEQ ID NO:4 or an homologous thereof.3. The expression vector according to claim 1 , wherein said MGMT mutant is the SNAP protein of SEQ ID NO:2 or an homologous thereof.4. The expression vector according to any one of to claim 1 , wherein said open reading frame is operatively associated with an inducible promoter which is functional in the same host cell as the peptidic signal is.5Drosophila. The expression vector according to any one of to claim 1 , wherein said secretion peptidic signal and said inducible promoter are functional in non-vertebrate cells claim 1 , preferably in insect cells claim 1 , more preferably in S2 cells6. The expression vector according to any one of to claim 1 , wherein said secretion peptidic signal and said inducible promoter are functional in vertebrate cells claim 1 , preferably in mammal cells claim 1 , more preferably in CHO cells claim 1 , YB2/O cells claim 1 , COS cells claim 1 , HEK cells claim 1 , NIH3T3 cells claim 1 , HeLa cells or derivatives thereof.7. The expression vector according to anyone of to claim 1 , wherein said recombinant protein is selected ...

Mouse artificial chromosome vector and use thereof

A mouse artificial chromosome vector is stable in rodent cells, tissues, and/or individuals, specifically a mouse artificial chromosome vector derived from a mouse chromosome selected from mouse chromosome 10 and mouse chromosome 16. A cell or a non-human animal may include the vector. The vector may be used for producing proteins and human antibodies.

TRANSGENIC CLONED PIGLET EXPRESSING HUMAN PROINSULIN AND METHOD OF PRODUCING THE SAME

A transgenic cloned piglet expressing human proinsulin and a method of preparing the same, and more particularly, to a recombinant vector for human proinsulin expression, a genetically modified cell line into which the recombinant vector is introduced, a transgenic cloned piglet expressing human proinsulin, and a method of producing the same. 1. A recombinant vector for expressing human proinsulin , the recombinant vector comprising a human proinsulin gene represented by a nucleotide sequence of SEQ ID NO: 1 , an enhancer , and a promoter.2. The recombinant vector of claim 1 , wherein the enhancer is a mouse PDX-1 gene enhancer.3. The recombinant vector of claim 1 , wherein the promoter is a rat insulin II gene promoter.4. The recombinant vector of claim 1 , wherein the recombinant vector is represented by a nucleotide sequence of SEQ ID NO: 3.5. The recombinant vector of claim 1 , wherein the recombinant vector is represented by the following vector map.6. A genetically modified cell line prepared by introducing a recombinant vector for expressing human proinsulin according to and a porcine proinsulin gene knockout recombinant vector into a somatic cell.7. The genetically modified cell line of claim 6 , wherein the porcine proinsulin gene knockout recombinant vector includes a sequence encoding sgRNA represented by a nucleotide sequence of SEQ ID NO: 5 or 6; and a Cas9 gene.8. The genetically modified cell line of claim 6 , wherein the porcine proinsulin gene knockout recombinant vector is represented by a nucleotide sequence of SEQ ID NO: 7.9. The genetically modified cell line of claim 6 , wherein the somatic cell is a fibroblast.10. The genetically modified cell line of claim 6 , wherein the transformed cell line is set forth in Accession No. KCLRF-BP-00408.11. A method of producing a transgenic cloned piglet expressing human proinsulin claim 6 , the method comprising:{'claim-ref': {'@idref': 'CLM-00006', 'claim 6'}, 'nuclear-transferring the genetically ...

Animal Models and Therapeutic Molecules

The invention discloses methods for the generation of chimaeric human—non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods.

Animal models and therapeutic molecules

The invention discloses methods for the generation of chimaeric human-non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods.

Animal Models and Therapeutic Molecules

The invention discloses methods for the generation of chimaeric human-non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods. 1. A method of obtaining an antigen specific antibody or antigen binding fragment thereof , said antigen-specific antibody comprising a human immunoglobulin heavy (IgH) chain comprising a human IgH chain variable region and a human IgH chain constant region , said antigen-binding fragment thereof comprising said human IgH chain variable region , the method comprising:expressing said antibody from a cell comprising nucleic acid encoding said human IgH chain variable region and said human IgH chain constant region of said antibody, or expressing said fragment from a cell comprising nucleic acid encoding said human IgH chain variable region,wherein said IgH chain variable region is of a transgenic mouse contacted with said antigen,wherein the germline of said mouse comprises a homozygous immunoglobulin heavy chain (IgH) locus,wherein said homozygous IgH locus comprises unrearranged human immunoglobulin heavy chain (IgH) variable region (VH) DNA comprising one or more human IgH V gene segments, one or more human D gene segments, one or more human JH gene segments and human intronic DNA contiguous with mouse intronic DNA and located at an endogenous IgH locus upstream of an enhancer and a constant (C) region comprising an endogenous CH gene segment,wherein said C region comprises mouse Cu and said human variable region gene segments are operably linked to said C region to permit rearrangement and expression of IgM heavy chain polypeptide comprising a human variable region and a mouse u constant region,wherein in said transgenic mouse expression of IgH heavy chains comprising endogenous mouse ...

METHODS FOR PRODUCING ANTIGEN-BINDING PROTEINS AGAINST FOREIGN ANTIGENS

Methods and compositions are provided for generating antigen-binding proteins against a foreign antigen of interest. 1(a) making a non-human animal with reduced tolerance of a foreign antigen of interest;(b) immunizing the non-human animal produced in step (a) with the foreign antigen of interest; and(c) maintaining the non-human animal under conditions sufficient to initiate an immune response to the foreign antigen of interest, wherein the non-human animal produces antigen-binding proteins against the foreign antigen of interest.. A method of generating antigen-binding proteins against a foreign antigen of interest, comprising: This application is a continuing application of U.S. application Ser. No. 15/600,466, filed May 19, 2017, which claims the benefit of U.S. Application No. 62/339,472, filed May 20, 2016, and U.S. Application No. 62/368,604, filed Jul. 29, 2016, each of which is herein incorporated by reference in its entirety for all purposes.The Sequence Listing written in file 571554SEQLIST.txt is 38.3 kilobytes, was created on Dec. 16, 2021, and is hereby incorporated by reference.Immunization of non-human animals (e.g., rodents, such as mice or rats) with a “non-self” protein is a commonly used method to obtain specific antigen-binding proteins such as monoclonal antibodies. This approach, however, is dependent on a divergence in sequence between native proteins in the non-human animal and the protein being immunized to enable the non-human animal's immune system to recognize the immunogen as non-self (i.e., foreign). The generation of antibodies against antigens having a high degree of homology with self-antigens can be a difficult task due to immunological tolerance. Because functionally important regions of proteins tend to be conserved across species, immunological tolerance to self-antigens often poses a challenge to the generation of antibodies to these key epitopes.Although progress has been made in targeting various genomic loci, there still ...

Transgenic rabbit with common light chain

Herein is reported a transgenic vector comprising a humanized light chain locus, wherein said humanized light chain locus comprises (a) a V gene segment derived from human light chain V segment IGKV1-39-01, (b) 3′ proximal to said light chain gene segment a promoter, and (c) 5′ proximal to said light chain gene segment at least a fragment of the human IGKJ4 J-element. 1. A method of generating a bispecific antibody comprising: use of a common antibody light chain comprising a variable domain that has the amino acid sequence of SEQ ID NO: 01 or is a variant thereof.2. The method according to wherein the method comprises combining two common antibody light chains with a first antibody heavy chain and a second antibody heavy chain claim 1 , wherein the first antibody heavy chain together with a common antibody light chain forms a first antigen binding site and the second antibody heavy chain together with a common antibody light chain forms a second antigen binding site.3. The method according to claim 1 , wherein the common light chain comprises 1 to 11 amino acid mutations within the amino acid sequence of SEQ ID NO: 01.4. The method according to claim 1 , wherein the common light chain comprises 1 to 13 amino acid mutations within the amino acid sequence of SEQ ID NO: 01 claim 1 , whereof at most 11 mutations are in the HVRs.5. A bispecific full-length antibody comprising two heavy chains and two common light chains each comprising a variable domain that has the amino acid sequence of SEQ ID NO: 01 or is a variant thereof.6. A transgenic vector comprising a humanized light chain locus claim 1 , wherein said humanized light chain locus comprises(a) as V gene segment the human light chain V segment IGKV1-39-01,(b) 3′ proximal to said light chain gene segment a promoter, and(c) 5′ proximal to said light chain gene segment the human IGKJ4 J-element or a functional fragment thereof.7. A transgenic rabbit claim 6 , comprising the humanized immunoglobulin locus present in ...

VARIANT OF A BPIFB4 PROTEIN

The present invention relates to a variant of BPIFB4 protein (Bactericidal/Permeability Increasing protein family B, member 4) and to a polynucleotide or a vector encoding said variant of BPIFB4 protein and to their use for the treatment of pathologies involving impairment of nitric oxide signalling. 117.-. (canceled)18. An isolated polynucleotide encoding a BPIFB4 protein variant consisting of the aminoacid sequence of SEQ ID NO: 1 , said protein variant having activity in increasing the activity of eNOS and/or the production of NO.19. The isolated polynucleotide of wherein the encoded BPIFB4 protein variant is linked to a sequence useful for targeting the BPIFB4 protein variant to a specific organ or tissue.20. A vector containing an isolated polynucleotide according to claim 18 , operatively linked to expression control sequences.21. The vector according to wherein the vector is a viral vector.22. The vector according to wherein the vector is a viral vector selected from AAV serotypes 1-9 vectors.23. The vector according to wherein the vector is an adenoviral vector.24. A pharmaceutical composition comprising an isolated polynucleotide according to in admixture with pharmaceutically acceptable carriers and/or excipients.25. A pharmaceutical composition according to which is suitable for endovenous claim 24 , subcutaneous claim 24 , intraocular or retroocular administration.26. A pharmaceutical composition comprising a vector according to in admixture with pharmaceutically acceptable carriers and/or excipients.27. A pharmaceutical composition according to which is suitable for endovenous claim 26 , subcutaneous claim 26 , intraocular or retroocular administration.28. A pharmaceutical composition according to which is a polymeric-based nano-system or a polycomplex nanosystem.29. A method for the treatment of a pathology or clinical condition claim 18 , the method comprising administering to a subject in need of said treatment a therapeutic amount of an isolated ...

Animal Models and Therapeutic Molecules

The invention discloses methods for the generation of chimaeric human-non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanised antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods. 1. A method of obtaining an antigen specific antibody or antigen binding fragment thereof , said antigen-specific antibody comprising a human immunoglobulin heavy (IgH) chain comprising a human IgH chain variable region and a human IgH chain constant region , said antigen-binding fragment thereof comprising said human IgH chain variable region , the method comprising:(A) providing a cell comprising nucleic acid encoding said human IgH heavy chain of said antibody, or a cell comprising nucleic acid encoding said human IgH chain variable region,wherein said IgH chain variable region is of a transgenic mouse contacted with said antigen,wherein the germline of said mouse comprises a homozygous immunoglobulin heavy chain (IgH) locus,wherein said homozygous IgH locus comprises unrearranged human immunoglobulin heavy chain (IgH) variable region (VH) DNA comprising one or more human IgH V gene segments, one or more human D gene segments, one or more human JH gene segments and human intronic DNA contiguous with mouse intronic DNA and located at an endogenous IgH locus upstream of an enhancer and a constant (C) region comprising an endogenous CH gene segment,wherein said C region comprises mouse Cμ and said human variable region gene segments are operably linked to said C region to permit rearrangement and expression of IgM heavy chain polypeptide comprising a human variable region and a mouse μ constant region,wherein in said transgenic mouse expression of IgH heavy chains comprising endogenous mouse variable regions is reduced or prevented,wherein said mouse comprises IgH-VDJCμ transcripts ...

Recombinant Adenovirus-Based Interferon Biotherapeutics in Swine

Disclosed herein is a recombinant adenovirus genome, said adenovirus genome comprising a heterologous nucleic acid inserted into a cloning site of said genome, said heterologous nucleic acid comprising: (a) a first nucleic acid sequence comprising an adenovirus tripartite sequence (e.g., SEQ ID NO:1) operably linked to a second nucleic acid sequence encoding an interferon (e.g., SEQ ID NO:2); (b) a third nucleic acid sequence comprising a bovine growth hormone polyA termination sequence operably linked to said second nucleic acid sequence (e.g., SEQ ID NO:3); (c) a fourth nucleic acid sequence comprising a porcine elongation factor 1-alpha (EF1α) promoter (e.g., SEQ ID NO:4); (d) a fifth nucleic acid sequence operably linked to said fourth nucleic acid sequence, said fifth nucleic acid sequence encoding a suppressor of cytokine signaling 1 (SOCS1) protein (e.g., SEQ ID NO:5). Furthermore, there is disclosed a method of producing interferon in an animal (e.g., swine). 1. A recombinant adenovirus genome , said adenovirus genome comprising a heterologous nucleic acid inserted into a cloning site of said genome , said heterologous nucleic acid comprising:a. a first nucleic acid sequence comprising an adenovirus tripartite sequence operably linked to a second nucleic acid sequence encoding an interferon;b. a third nucleic acid sequence comprising a bovine growth hormone polyA termination sequence operably linked to said second nucleic acid sequence;c. a fourth nucleic acid sequence comprising a porcine elongation factor 1-alpha (EF1α) promoter;d. a fifth nucleic acid sequence operably linked to said fourth nucleic acid sequence, said fifth nucleic acid sequence encoding a suppressor of cytokine signaling 1 (SOCS1) protein.2. The adenovirus genome of claim 1 , wherein the second nucleic acid sequence is codon-optimized for a bacterial host cell.3E. coli.. The adenovirus genome of claim 2 , wherein the bacterial host is4. The adenovirus genome of claim 1 , wherein the ...

COMPOSITIONS AND METHODS FOR PRODUCING EXOSOME LOADED THERAPEUTICS FOR TREATING CARDIOVASCULAR DISEASE

A composition for delivering cargo to cytoplasm of a cell, wherein the cargo manipulates angiogenesis. In one embodiment the composition comprises: an exosome; and cargo, located within the exosome, comprising at least one plasmid. In another embodiment the composition comprises: an exosome; and cargo, located within the exosome, comprising a DNA plasmid bioengineered specifically to self-produce monoclonal neutralizing antibodies. 1. A composition for delivering cargo to cytoplasm of a cell , wherein the cargo manipulates angiogenesis , the composition comprising:an exosome; andcargo, located within the exosome, comprising at least one plasmid.2. The composition of claim 1 , wherein the exosome is isolated from autologous cells of a patient.3. The composition of claim 1 , wherein the exosome is isolated from a cell line claim 1 , a primary cell culture claim 1 , or a combination thereof.4. The composition of claim 1 , wherein the exosome is isolated from a stem cell.5. The composition of claim 1 , wherein the at least one plasmid is a RNA plasmid claim 1 , a DNA plasmid claim 1 , or any combination thereof.6. The composition of claim 1 , wherein the at least one plasmid is a retrovirus or an adeno-associated virus (AAV).7. The composition of claim 1 , wherein the at least one plasmid promotes expression of the VEGF gene.8. The composition of claim 1 , wherein the at least one plasmid is a proprotein convertase subtilisin/kexin type 9 inhibitor.9. The composition of claim 1 , wherein the at least one plasmid is a miR-214 inhibitor.10. The composition of claim 1 , wherein the cargo further comprises a CRISPR-Cas9 system claim 1 , a Zinc finger claim 1 , a single base editor claim 1 , or a combination thereof.11. The composition of claim 1 , wherein the at least one plasmid is a DNA plasmid bioengineered specifically to self-produce monoclonal neutralizing antibodies.12. The composition of claim 1 , wherein the exosome further comprises at least one targeting agent.13 ...

METHODS AND COMPOSITIONS RELATED TO RECOMBINANT NEIL2

Certain embodiments are directed to NEIL2 compositions and method of using NEIL2 composition to treat subjects in need of such treatment. 1. A NEIL2 polypeptide comprising a NEIL2 polypeptide 80% identical to SEQ ID NO:1 or a segment thereof coupled to a lung targeting moiety.2. The NEIL2 polypeptide of claim 1 , wherein the lung targeting moiety is a peptide.3. The NEIL2 polypeptide of claim 2 , wherein the peptide is a carrier peptide.4. The NEIL2 polypeptide of claim 3 , wherein the carrier peptide consists of the amino acid sequence of SEQ ID NO:9.5. The NEIL2 polypeptide of claim 3 , wherein the NEIL2 polypeptide is 95% identical to SEQ ID NO:1.6. The NEIL2 polypeptide of claim 1 , wherein the NEIL2 polypeptide comprises 300 consecutive amino acids of SEQ ID NO:1.7. The NEIL2 polypeptide of claim 1 , wherein the NEIL2 polypeptide consists essentially of the amino acid sequence of SEQ ID NO:1.8. A method of treating an inflammatory condition comprising administering a NEIL2 polypeptide to a subject in need thereof.9. The method of claim 8 , wherein the NEIL2 polypeptide is provided by an expression vector encoding a NEIL2 polypeptide.10. The method of claim 8 , wherein the inflammatory condition is allergic rhinitis claim 8 , allergic asthma claim 8 , or inflammation due to a bacterial infection.11. A method of treating a bacterial infection comprising administering a NEIL2 polypeptide to a subject in need thereof.12. The method of claim 11 , wherein the NEIL2 polypeptide is provided by an expression vector encoding a NEIL2 polypeptide.13Klebsiella. The method of claim 11 , wherein the bacterial infection is a infection.14Acinetobacter. The method of claim 11 , wherein the bacterial infection is an infection. The present application claims priority to U.S. Provisional Application No. 62/738,605 filed Sep. 28, 2019, which is incorporated by reference herein in its entirety.This invention was made with government support under NS073976 awarded by the National ...

Viral expression construct comprising a fibroblast growth factor 21 (fgf21) coding sequence

The invention relates to a viral expression construct and related viral vector and nucleic acid molecule and composition and to their use wherein said construct and vector are suitable for expression in a mammal and comprise a nucleotide sequence encoding a Fibroblast growth factor 21 (FGF21) to be expressed in liver, adipose tissue and/or skeletal muscle.

ENGINEERED POLYPEPTIDES FOR ANTIGEN DELIVERY

Recombinant high affinity Invasin polypeptide are provided herein. Further provided are methods of delivering therapeutics such as vaccines by conjugation to the engineered recombinant Invasin polypeptides. 1. A recombinant polypeptide comprising an Invasin D5 domain coding sequence at least 80% identical to amino acid residues 887-986 of SEQ ID NO: 1 , wherein the Invasin D5 domain coding sequence comprises an amino acid substitution at the position corresponding to Gly 909 and/or an amino acid substitution at the position corresponding to Ser 910.2. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid substitution at the position corresponding to Gly 909 and an amino acid substitution at the position corresponding to Ser 910.3. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid substitution for a residue having a positively charged side chain at the position corresponding to Gly 909.4. The polypeptide of claim 3 , wherein the polypeptide comprises an Arg substitution at the position corresponding to Gly 909.5. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid substitution for an aliphatic residue at the position corresponding to Ser 910.6. The polypeptide of claim 5 , wherein the polypeptide comprises a Gly substitution at the position corresponding to Ser 910.7. The polypeptide of claim 2 , wherein the polypeptide comprises an Arg substitution at the position corresponding to Gly 909 and a Gly substitution at the position corresponding to Ser 910.8. The polypeptide of claim 7 , wherein the polypeptide comprises a RGDmotif.9. The polypeptide of claim 8 , wherein the RGDmotif increases integrin binding affinity by at least 10 fold as compared to wild-type Invasin.10. The polypeptide of claim 8 , wherein the RGDmotif increases integrin binding affinity by at least 50 fold as compared to wild-type Invasin.11. The polypeptide of claim 8 , wherein the RGDmotif increases integrin binding ...

Non-human animals having an immunoglobulin heavy chain variable region that includes an engineered diversity cluster and uses thereof

Non-human animals and methods and compositions for making and using them are provided, wherein said non-human animals have a genome comprising an engineered or recombinant diversity cluster within an immunoglobulin heavy chain variable region, which engineered or recombinant diversity cluster comprises an insertion of one or more coding sequences of a non-immunoglobulin polypeptide of interest. Non-human animals described herein express antibodies characterized by complementary determining regions (CDRs), in particular, CDR3s having diversity that directs binding to particular antigens. Methods for producing antibodies from non-human animals are also provided, which antibodies contain human variable regions and mouse constant regions.

MICE THAT MAKE VL BINDING PROTEINS

Genetically modified mice and methods for making an using them are provided, wherein the mice comprise a replacement of all or substantially all immunoglobulin heavy chain V gene segments, D gene segments, and J gene segments with at least one light chain V gene segment and at least one light chain J gene segment. Mice that make binding proteins that comprise a light chain variable domain operably linked to a heavy chain constant region are provided. Binding proteins that contain an immunoglobulin light chain variable domain, including a somatically hypermutated light chain variable domain, fused with a heavy chain constant region, are provided. Modified cells, embryos, and mice that encode sequences for making the binding proteins are provided. 1. A mouse whose germline genome comprises an endogenous immunoglobulin (Ig) heavy chain locus modified to comprise a human genomic germline kappa (κ) sequence comprising(i) unrearranged functional human Ig light chain variable κ (hVκ) gene segments and(ii) all five unrearranged functional human Ig light chain joining κ (hJκ1-hJκ5) gene segments,wherein the human genomic germline κ sequence(A) replaces at the endogenous Ig heavy chain locus an endogenous genomic sequence comprising endogenous immunoglobulin heavy chain V gene segments, all endogenous immunoglobulin heavy chain D gene segments, and all endogenous immunoglobulin heavy chain J gene segments, and{'sub': 'H', '(B) rearranges in a B cell during B cell development to form a rearranged Ig hVκ/hJκ gene sequence operably linked to the endogenous Ig heavy chain constant region (C) nucleic acid sequence at the endogenous Ig heavy chain locus, and'}{'sup': '+', 'sub': 'H', 'wherein the mouse comprises a CD19 B cell comprising the rearranged Ig hVκ/hJκ gene sequence operably linked to the endogenous Ig Cnucleic acid sequence.'}214.-. (canceled)15. A mouse that expresses from its germline an immunoglobulin comprising a first polypeptide comprising a first human light chain ...

Anti-tau antibodies and uses thereof

Provided herein are antibodies that specifically bind Tau and methods of using the same.

ANTI-TAU ANTIBODIES AND USES THEREOF

Provided herein are antibodies that specifically bind Tau and methods of using the same. 1. A monoclonal antibody , or an antigen-binding fragment thereof , that specifically binds a human Tau , the antibody comprising a heavy chain complementarity determining region 1 (HCDR1) , a heavy chain complementarity determining region 2 (HCDR2) , and a heavy chain complementarity determining region 3 (HCDR3) as set forth in SEQ ID NO: 402 and a light chain complementarity determining region 1 (LCDR1) , a light chain complementarity determining region 2 (LCDR2) , and a light chain complementarity determining region 3 (LCDR3) as set forth in SEQ ID NO: 572.2. The monoclonal antibody or antigen-binding fragment according to claim 1 , wherein:the HCDR1 comprises SEQ ID NO: 732, the HCDR2 comprises SEQ ID NO: 734, the HCDR3 comprises SEQ ID NO: 736, the LCDR1 comprises SEQ ID NO: 846, the LCDR2 comprises SEQ ID NO: 848, and the LCDR3 comprises SEQ ID NO: 850 as defined according to the method of Kabat; orthe HCDR1 comprises SEQ ID NO: 1002, the HCDR2 comprises SEQ ID NO: 1004, the HCDR3 comprises SEQ ID NO: 1006, the LCDR1 comprises SEQ ID NO: 1116, the LCDR2 comprises SEQ ID NO: 1118, and the LCDR3 comprises SEQ ID NO: 1120 as defined by IMGT.3. The monoclonal antibody or antigen-binding fragment according to claim 1 , wherein the residue at position 49 of the light chain according to the method of Kabat is not cysteine.4. The monoclonal antibody or antigen-binding fragment according to claim 3 , wherein the residue at position 49 of the light chain according to the method of Kabat is serine.5. The monoclonal antibody or antigen-binding fragment according to wherein the residue at position 57 of the heavy chain according to the method of Kabat is not cysteine.6. The monoclonal antibody or antigen-binding fragment according to claim 5 , wherein the residue at position 57 of the heavy chain according to the method of Kabat is serine.7. The monoclonal antibody or antigen-binding ...

INNOVATIVE PREPARATION PROCESS FOR BMP-2 STEM CELLS

The present invention discloses an innovative preparation process for recombinant human fat cell peptide (BMP-2 stem cells for short), comprising the following steps: gene synthesis of BMP-2 stem cells, specific expression system construction of mature peptide, expression induction, separation, purification and amplification. The method disclosed by the present invention is simple and flexible; has simple product separation and purification process, high purity, high activity and low production cost; has the functions of preventing clinical virus, resisting cell aging, and promoting wound healing and repair in epidermis, dermis and mucosa; and has broad application prospects in the fields such as medical care, cosmetology and health care. 1. An innovative preparation process for BMP-2 stem cells , characterized in comprising the following steps in sequence:(1) constructing an expression vector while rapidly synthesizing a gene by multiplex PCR;(2) introducing an expression plasmid into a host bacteria to construct an expression engineering bacteria;(3) fermenting the engineering bacteria and inducing expression at 30° C.; and(4) performing enzymatic cleavage to expression products to release BMP-2 stem cells, and purifying the expression products by a nickel column to obtain a BMP-2 stem cell fine product.2E. coli.. The innovative preparation process of claim 1 , characterized in that in step (1) claim 1 , a series of primers are designed claim 1 , the full length of a target gene is obtained by multiple rounds of PCR amplification claim 1 , a vector with homology arms (containing the target gene) is obtained at the same time claim 1 , self-ligation is achieved by PCR to construct a circular DNA sequence containing a gap claim 1 , and complete BMP-2 stem cells of the expression vector are obtained by intracellular repair through transformed3. The innovative preparation process of claim 1 , characterized in that the gene of BMP-2 stem cells in the expression vector ...

METHODS, COMPOSITIONS AND SYSTEMS FOR PRODUCTION OF RECOMBINANT SPIDER SILK POLYPEPTIDES

Disclosed are methods, compositions, and systems for transforming silkworms to produce spider silk and analogs of spider silk. In certain embodiments, the method may include inserting a DNA sequence coding for at least a portion of a spider silk fibroin polypeptide, or an analog of a spider silk fibroin polypeptide, positioned between at least a portion of the 5′ and 3′ ends of a silkworm fibroin gene to generate a fusion gene construct having a sequence that encodes for a polypeptide comprising both spider silk fibroin and silkworm silk fibroin sequences. In certain embodiments, the fused gene is able to replace a native gene present in the silkworm such that the transformed silkworm expresses a polypeptide comprising a spider silk fibroin polypeptide, or an analog thereof, and expresses significantly less of the native silkworm silk. 122.-. (canceled)23. A method of making a transgenic silkworm that produces a silk comprising a spider silk polypeptide , or an analog of a spider silk polypeptide:{'i': Bombyx mori, Bombyx mandarina, Antheraea peryni', 'Antheraea yamamai;, 'a) providing a silkworm, wherein the silkworm is , or'}b) transforming the silkworm of step (a) by inserting a nucleic acid sequence encoding a spider silk fibroin polypeptide, or an analog of a spider silk fibroin polypeptide, into the genome of the silkworm, wherein the nucleic acid comprises at least 2800 nucleotides encoding alternating beta-sheet domains and alpha-helix domains, wherein the amino acid sequence of the beta-sheet domains are each at least 60% identical to at least one of the beta-sheet domains as set forth in SEQ ID NO: 2, and wherein expression of the native silkworm fibroin gene is reduced; andc) selecting for a transgenic silkworm from step (b) that produces a silk that is at least 50% stronger than native silkworm silk.24Bombyx moriBombyx mandarina.. The method of claim 23 , wherein the silkworm is or25Antheraea peryniAntheraea yamamai.. The method of claim 23 , wherein the ...

Humanized antibodies specific to the protofibrillar form of the beta-amyloid peptide

Humanized antibodies specific to the protofibrillar form of the beta-amyloid peptide are provided. Methods of using antibodies specific to the protofibrillar form of the beta-amyloid peptide in the field of Alzheimer's disease are also provided.

HUMANIZED ANTI-RAGE ANTIBODY

This invention provides a humanized, anti-human receptor for advance glycation end-products (RAGE) monoclonal antibody or a fragment of such antibody which binds human RAGE as well as compositions containing, and uses of, such antibody and fragments. 1. A humanized , anti-human receptor for advance glycation end-products (RAGE) monoclonal antibody comprising:(a) consecutive amino acids constituting a heavy chain of such antibody, the sequence of which is set forth in SEQ ID NO: 7; and(b) consecutive amino acids constituting a light chain of such antibody, the sequence of which is set forth in SEQ ID NO: 8,or a fragment of such antibody which binds human RAGE.2. The fragment of the humanized claim 1 , anti-human RAGE antibody of wherein the fragment is a F(ab′)2 fragment.3. A polypeptide comprising consecutive amino acids and capable of functioning as the heavy chain of an antibody claim 1 , wherein the sequence of such consecutive amino acids is set forth in SEQ ID NO: 7.4. A polypeptide comprising consecutive amino acids and capable of functioning as the light chain of an antibody claim 1 , wherein the sequence of such consecutive amino acids is set forth in SEQ ID NO: 8.5. A vector which encodes the polypeptide of .6. The vector of claim 5 , comprising consecutive nucleotides claim 5 , the sequence of which is set for in SEQ ID NO: 9.7. A vector which encodes the polypeptide of .8. The vector of claim 7 , comprising consecutive nucleotides claim 7 , the sequence of which is set for in SEQ ID NO: 10.9. A eucaryotic cell which expresses the monoclonal antibody of .10. The eucaryotic cell of comprising a vector comprising consecutive nucleotides claim 9 , the sequence of which is set forth in SEQ ID NO: 9 claim 9 , and comprising a vector comprising consecutive nucleotides claim 9 , the sequence of which is set forth in SEQ ID NO: 10.11. The eucaryotic cell of claim 9 , wherein the eucaryotic cell is a mammalian cell.12. The mammalian cell of claim 11 , wherein the ...

An engineered two-part cellular device for discovery and characterisation of t-cell receptor interaction with cognate antigen

The present invention relates to a two-part device, wherein a first part is an engineered antigen-presenting cell system (eAPCS), and a second part is an engineered TCR-presenting cell system (eTPCS).

ANTI-PD-1 ANTIBODIES AND USES THEREOF

Provided are anti-PD-1 antibodies or fragments thereof. In various example, the antibodies or fragments thereof includes a heavy chain variable region comprising heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and a light chain variable region comprising light chain complementarity determining regions LCDR1, LCDR2, and LCDR3. Methods of using the antibodies or fragments thereof for treating and diagnosing diseases such as cancer, infection or immune disorders are also provided. 124-. (canceled)25. A method of treating a disease or condition in an individual in need thereof , comprising administering to the individual a composition comprising an effective amount of antibody or fragment thereof that specifically binds to PD-1 , wherein the antibody or fragment thereof comprises a heavy chain variable region comprising heavy chain complementarity determining regions HCDR1 , HCDR2 , and HCDR3 , and a light chain variable region comprising light chain complementarity determining regions LCDR1 , LCDR2 , and LCDR3 , wherein:(a) the HCDR1 comprises an amino acid sequence selected from the group consisting of GFTFSSYT (SEQ ID NO: 1), GYTFTSYT (SEQ ID NO: 7), and GFAFSSYD (SEQ ID NO: 13);(b) the HCDR2 comprises an amino acid sequence selected from the group consisting of ISHGGGDT (SEQ ID NO: 2), INPTTGYT (SEQ ID NO: 8), and ITIGGGTT (SEQ ID NO: 14);(c) the HCDR3 comprises an amino acid sequence selected from the group consisting of ARHSGYERGYYYVMDY (SEQ ID NO: 3), ARDDAYYSGY (SEQ ID NO: 9), and ARHRYDYFAMDN (SEQ ID NO: 15);(d) the LCDR1 comprises an amino acid sequence selected from the group consisting of ESVDYYGFSF (SEQ ID NO: 4), ENIYSNL (SEQ ID NO: 10), and ENVDNYGINF (SEQ ID NO: 16);(e) the LCDR2 comprises an amino acid sequence selected from the group consisting of AAS (SEQ ID NO: 5), AAK (SEQ ID NO: 11), and VSS (SEQ ID NO: 17); and(f) the LCDR3 comprises an amino acid sequence selected from the group consisting of QQSKEVPW (SEQ ID NO: 6), ...

Anti-igf-i receptor antibody

The present invention provides an anti-IGF-I receptor antibody that binds specifically to an IGF-I receptor of a vertebrate and has the proliferation-inducing activity of a vertebrate-derived cell, or a fragment thereof, or derivatives of these.

TRANSGENIC NON-HUMAN VERTEBRATE FOR THE IN VIVO PRODUCTION OF DUAL SPECIFICITY IMMUNOGLOBULINS OR HYPERMUTATED HEAVY CHAIN ONLY IMMUNOGLOBULINS

The invention relates, in one aspect, generally to novel concept of guided selection of antibody variable domains, combination and expression entirely in vivo. An application is to produce multivalent polypeptides. The present invention relates to multivalent (eg, multispecific) antibodies, antibody chains and polypeptides, as well as heavy chain-only antibodies (H2 antibodies) that are devoid of light chains. The invention further relates to the selection, maturation and production of these in vivo in non-human vertebrates and non-human vertebrate cells. To this end the invention also relates to such non-human vertebrates and cells. The invention also relates to the provision of means to produce and select heavy chain-only antibodies and heavy chains comprising variable domains that have undergone affinity maturation. 1. A mouse wherein an Ig heavy or light chain locus of the genome of the mouse comprises (in 5′ to 3′ orientation) (i) an unrearranged human variable region for encoding a first variable domain , (ii) a sequence encoding an optional linker and (iii) a sequence encoding an epitope binding moiety.2. The mouse of claim 1 , wherein the variable region of (i) is capable of undergoing rearrangement claim 1 , whereby the mouse is capable of expressing a polypeptide comprising a binding site specific for a target epitope claim 1 , the binding site having the structure VI-L-E claim 1 , where VI is an antibody variable domain claim 1 , E is an epitope binding moiety and L is an optional peptide linker.3. The mouse of claim 1 , wherein sequence (i) is capable of undergoing rearrangement so that the cell produces antibody chains each having an epitope binding site that comprises first and second variable domains derived from sequences (i) and (iii) claim 1 , wherein there is no further antibody domain between the first and second variable domains.4. A polypeptide comprising a binding site specific for a target epitope claim 1 , the binding site having the ...

NON-HUMAN ANIMALS HAVING A HUMANIZED SIGNAL-REGULATORY PROTEIN GENE

Genetically modified non-human animals and methods and compositions for making and using the same are provided, wherein the genetic modification comprises a humanization of an endogenous signal-regulatory protein gene, in particular a humanization of a SIRPα gene. Genetically modified mice are described, including mice that express a human or humanized SIRPα protein from an endogenous SIRPα locus. 153.-. (canceled)54. A rodent whose genome comprises a humanized SIRPα gene at an endogenous rodent SIRPα locus , wherein the humanized SIRPα gene(i) comprises exon 1 of a rodent SIRPα gene, exons 2, 3 and 4 of a human SIRPα gene, and exons 5, 6, 7 and 8 of the rodent SIRPα gene,(ii) is operably linked to a rodent SIRPα promoter at the endogenous rodent SIRPα locus, and(iii) expresses in the rodent a humanized SIRPα protein comprising an extracellular portion of the human SIRPα protein encoded by the human SIRPα gene and an intracellular portion of the rodent SIRPα protein encoded by the rodent SIRPα gene; andwherein the rodent is a rat.55. The rodent of claim 54 , wherein the extracellular portion of the human SIRPα protein comprises amino acid residues 28-362 of the human SIRPα protein.56. The rodent of claim 54 , wherein the human SIRPα protein comprises the amino acid sequence as set forth in SEQ ID NO: 4.57. The rodent of claim 54 , and wherein the rodent is homozygous for the humanized SIRPα gene.58. The rodent of claim 54 , wherein the rodent does not express an endogenous rodent SIRPα protein.59. A rodent embryonic stem (ES) cell claim 54 , comprising a humanized SIRPα gene at an endogenous rodent SIRPα locus claim 54 , wherein the humanized SIRPα gene(i) comprises exon 1 of a rodent SIRPα gene, exons 2, 3 and 4 of a human SIRPα gene, and exons 5, 6, 7 and 8 of the rodent SIRPα gene,(ii) is operably linked to a rodent SIRPα promoter at the endogenous rodent SIRPα locus, and(iii) encodes a humanized SIRPα protein comprising an extracellular portion of the human SIRP ...

DIRECT SELECTION OF CELLS EXPRESSING HIGH LEVELS OF HETEROMERIC PROTEINS USING GLUTAMINE SYNTHETASE INTRAGENIC COMPLEMENTATION VECTORS

This invention relates to the general field of recombinant expression of polypeptides in animal cell culture. More specifically, the invention concerns improved selection of cells transfected with recombinantly engineered vectors designed to express polypeptides, in particular heteromultimeric polypeptides. 1. A vector comprising:a) a first nucleic acid encoding a first polypeptide, andb) a second nucleic acid encoding a second polypeptide, wherein the second polypeptide is an initial mutant subunit of a selectable marker,wherein the transcription of the first nucleic acid is operably linked to transcription of the second nucleic acid,further comprising:c) a third nucleic acid encoding a third polypeptide wherein the third polypeptide is capable of associating with the first polypeptide to form a heteromeric complex, andd) a fourth nucleic acid which encodes fourth polypeptide, wherein the fourth polypeptide is a complementary mutant subunit of the selectable marker,wherein the transcription of the third nucleic acid is operably linked to transcription of the fourth nucleic acid,wherein the initial mutant subunit and the complementary mutant subunit of the selectable marker interact to provide a selectable activity, and further wherein the vector is capable of being transfected into mammalian cells and improving selection of transfected cells.2. The vector of wherein the heteromeric complex is an immunoglobulin.3. The vector of wherein the first nucleic acid encodes an immunoglobulin heavy chain claim 2 , and the third nucleic acid encodes an immunoglobulin light chain.4. The vector of any one of - claim 2 , wherein the selectable marker is metabolic enzyme is selected from the group consisting of glutamine synthetase claim 2 , arginosuccinate lyase claim 2 , adenylosuccinate synthetase claim 2 , and glutamate dehydrogenase.5. The vector of any one of - claim 2 , wherein an internal ribosomal entry site (IRES) occurs at a site selected from the group consisting of:a ...

BISPECIFIC ANTIBODIES AGAINST CEACAM5 AND CD47

The invention provides bispecific antibodies binding to human carcinoembryonic antigen CEACAM5 and human CD47, polynucleotides encoding such bispecific antibodies and vectors and host cells comprising such polynucleotides. The invention further provides methods for selecting and producing such antibodies and methods of using such antibodies in the treatment of diseases in monotherapy as well in combination. 1. A bispecific antibody comprising a first binding part specifically binding to human CEACAM5 and a second binding part specifically binding to human CD47.2. The bispecific antibody of claim 1 , wherein said antibody comprises a Fc region that has been glycoengineered to have a reduced number of fucose residues as compared to the same bispecific antibody that has not been glycoengineered.3. The bispecific antibody of or claim 1 , wherein said first binding part specifically binds to human CEACAM5 and CEACAM 6.4. The bispecific antibody of claim 3 , wherein the first binding part specifically binds in a balanced manner to human CEACAM5 and human CEACAM6.5. The bispecific antibody of claim 4 , wherein the EC50 values of binding to human CEACAM5 and human CEACAM6 differ by less than a factor of 3 (balanced binding).6. The bispecific antibody according to any one of the preceding claims claim 4 , wherein claim 4 ,a) the first binding part comprises a heavy chain variable region comprising as CDRs a CDRH1 of SEQ ID NO:25, a CDRH2 of SEQ ID NO:26 and a CDRH3 of SEQ ID NO:27 and a light chain variable region comprising as CDRs a CDRL1 of SEQ ID NO: 112, a CDRL2 of SEQ ID NO: 113, and a CDRL3 of SEQ ID NO: 114, andb) the second binding part comprises a heavy chain variable region comprising as CDRs a CDRH1 of SEQ ID NO:25, a CDRH2 of SEQ ID NO:26 and a CDRH3 of SEQ ID NO:27 and a light chain variable region comprising as CDRs a CDRL1 of SEQ ID NO:28, a CDRL2 of SEQ ID NO:29, and a CDRL3 of SEQ ID NO:30.7. The bispecific antibody of any one of the preceding claims claim ...

LONG GERMLINE DH GENES AND LONG HCDR3 ANTIBODIES

The invention relates to a long DH (LDH) cassette comprising a recombinant DH construct comprising at least two DH gene segments encoding at least 10 amino acids of the HCDR3 amino acid sequence, wherein at least one of the DH gene segments is a heterologous DH gene segment; an immunoglobulin heavy chain locus and a transgenic non-human animal comprising the same; and their use in producing an immunoglobulin library with long HCDR3 regions. 1. A long DH (LDH) cassette comprising a recombinant DH construct comprising at least two DH gene segments encoding at least 10 amino acids of the HCDR3 amino acid sequence , wherein at least one of the DH gene segments is a heterologous DH gene segment.2. The expression cassette of claim 1 , wherein the heterologous DH gene segment is any of:a) a DH gene segment of a different species origin; or i) the fusion of naturally-occurring DH gene segments and/or', 'ii) a 23 (+/−1) bps recombination signal sequence (RSS) spacer; and/or', 'iii) the deletion of one or more stop codons; and/or', 'iv) the fusion to at least one intergenic region which is artificial or of different species origin., 'b) an artificial DH segment comprising'}3. The expression cassette of claim 1 , wherein the heterologous DH gene segment is a DH gene segment of the same species.4. The expression cassette of claim 3 , wherein all DH gene segments are of the same species origin.5. The expression cassette of claim 4 , wherein all DH gene segments are of human origin.6. The expression cassette of claim 1 , wherein the recombinant DH construct is a chimeric DH construct comprising at least one human DH gene segment and at least one heterologous DH gene segment claim 1 , which is of a different species origin.7. The expression cassette of claim 6 , wherein said different species origin is bovine origin.8. The expression cassette of claim 1 , wherein the heterologous DH gene segment is a bovine DH-2 gene segment claim 1 , or a bovine DH-2 gene segment which is ...

MICE THAT MAKE VL BINDING PROTEINS

Genetically modified mice and methods for making an using them are provided, wherein the mice comprise a replacement of all or substantially all immunoglobulin heavy chain V gene segments, D gene segments, and J gene segments with at least one light chain V gene segment and at least one light chain J gene segment. Mice that make binding proteins that comprise a light chain variable domain operably linked to a heavy chain constant region are provided. Binding proteins that contain an immunoglobulin light chain variable domain, including a somatically hypermutated light chain variable domain, fused with a heavy chain constant region, are provided. Modified cells, embryos, and mice that encode sequences for making the binding proteins are provided. 1. A mouse , comprising in its germline an unrearranged light chain V segment and an unrearranged J segment operably linked with a heavy chain constant region nucleic acid sequence.2. The mouse of claim 1 , wherein the unrearranged light chain V segment is selected from a human κ segment claim 1 , a human λ segment claim 1 , and a combination thereof.3. The mouse of claim 1 , wherein the heavy chain constant region nucleic acid sequence is selected from the group consisting of a C1 sequence claim 1 , a hinge sequence claim 1 , a C2 sequence claim 1 , a C3 sequence claim 1 , and a combination thereof.4. The mouse of claim 1 , wherein the unrearranged light chain V segment and the unrearranged J segment replace an endogenous mouse heavy chain V segment and an endogenous mouse heavy chain J segment at the endogenous mouse heavy chain locus.5. The mouse of claim 4 , wherein the unrearranged light chain V segment replaces all or substantially all functional mouse heavy chain V segments of the endogenous mouse heavy chain locus.6. The mouse of claim 4 , wherein the unrearranged J segment comprises a light chain J segment claim 4 , and the light chain J segment replaces all or substantially all functional mouse heavy chain J ...

NON-HUMAN ANIMALS HAVING A HUMANIZED SIGNAL-REGULATORY PROTEIN GENE

Genetically modified non-human animals and methods and compositions for making and using the same are provided, wherein the genetic modification comprises a humanization of an endogenous signal-regulatory protein gene, in particular a humanization of a SIRPα gene. Genetically modified mice are described, including mice that express a human or humanized SIRPα protein from an endogenous SIRPα locus. 1. A mouse expressing a SIRPα polypeptide comprising an extracellular portion of a human SIRPα protein and an intracellular portion of a mouse SIRPα protein.2. The mouse of claim 1 , wherein the extracellular portion of a human SIRPα protein comprises amino acids corresponding to residues 28-362 of a human SIRPα protein that appears in Table 3.3. The mouse of or claim 1 , wherein the extracellular portion of a human SIRPα protein shares a percent identity of at least 50% with a corresponding extracellular portion of a human SIRPα protein that appears in Table 3.4. The mouse of or claim 1 , wherein the shared percent identity is at least 60% with a corresponding extracellular portion of a human SIRPα protein that appears in Table 3.5. The mouse of or claim 1 , wherein the shared percent identity is at least 70% with a corresponding extracellular portion of a human SIRPα protein that appears in Table 3.6. The mouse of or claim 1 , wherein the shared percent identity is at least 80% with a corresponding extracellular portion of a human SIRPα protein that appears in Table 3.7. The mouse of or claim 1 , wherein the extracellular portion of a human SIRPα protein has a sequence that is at least 90% identical to an extracellular portion of a human SIRPα protein that appears in Table 3.8. The mouse of or claim 1 , wherein the extracellular portion of a human SIRPα protein has a sequence that is at least 95% identical to an extracellular portion of a human SIRPα protein that appears in Table 3.9. The mouse of or claim 1 , wherein the extracellular portion of a human SIRPα protein ...

ANTI-RHO GTPASE CONFORMATIONAL SINGLE DOMAIN ANTIBODIES AND USES THEREOF

The present invention relates to active form specific anti-Rho GTPase conformational single domain antibodies and their uses in particular in the therapeutic and diagnostic fields. In particular, the present invention relates to a single domain antibody wherein the amino acid sequences of CDR1-IMGT, CDR2-IMGT and CDR3-IMGT have at least 90% of identity with the amino acid sequences of the CDR1-IMGT, CDR2-IMGT and CDR3-IMGT of the H12, B6, 4P75, 4SP1, 4SNP36, 4SNP61, 5SP10, 5SP11, 5SP58, 5SNP47, 5SNP48, 5SNP65, B20, B15, B5, B71, E3, A6, G12, NB61, 212B, 111B or 404F (hs2dAb) single domain antibody which are defined in Table B. 111-. (canceled)12. A nucleic acid molecule which encodes a single domain antibody wherein the amino acid sequences of CDR1-1MGT , CDR2-IMGT and CDR3-IMGT of the single domain antibody are set forth as SEQ ID NOS: 5-7.13. A vector which includes the nucleic acid molecule of .14. A host cell transformed with the nucleic acid molecule of .1516-. (canceled)17. A method for treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the nucleic acid molecule of claim 12 , or the single domain antibody encoded by the nucleic acid molecule. The present invention relates to anti-Rho GTPase conformational single domain antibodies and their uses thereof in particular in the therapeutic and diagnostic fields.Rho GTPases belong to the family of the 20 small GTPases homologous to Ras which are largely considered as undrugable proteins. Their physiologic activity resides more in a conformational change of the switch I and switch II highly conserved domains than in their guanine nucleotide triphosphate hydrolase very slow catalytic activity. Small G proteins are therefore molecular switch that cycle between an inactive GDP bound state an active GTP bound conformation. The Rho subfamily, which contains RhoA, RhoB and RhoC that share more than 85% sequence identities are pleiotropic proteins ...

NATIVE PROTEIN PURIFICATION TECHNOLOGY

The present invention relates to an isolated polypeptide comprising (a) a protein of interest; (b) a first member of a pair of binding partners; (c) an affinity tag for immobilizing the polypeptide on a solid support; and (d) a modified endoprotease recognition site, wherein the modified endoprotease recognition site is located directly adjacent to the N-terminal amino acid of the protein of interest and comprises or only consists of the amino acid sequence N-terminal of the cleavage site of the native endoprotease recognition site. The present invention also relates to a nucleic acid encoding the above polypeptide and a host cell thereof, a method for isolating a protein of interest using the above polypeptide as a fusion partner and a protease fusion protein with the second member of the pair of binding partners and kits thereof. In addition, a method of degrading a target protein, a method of treatment and use of a fusion protease protein comprising a protease and a target protein binding element are also disclosed. 1. Isolated polypeptide comprising(a) a protein of interest;(b) a first member of a pair of binding partners;(c) an affinity tag for immobilizing the polypeptide on a solid support; and 'wherein the modified endoprotease site is located directly adjacent to the N-terminal amino acid of the protein of interest and comprises or only consists of the amino acid sequence N-terminal of the cleavage site of the native endoprotease recognition site.', '(d) a modified endoprotease recognition site,'}2. The isolated polypeptide according to claim 1 , wherein the first member of the pair of binding partners is located N-terminal to the modified protease recognition site.3. The isolated polypeptide according to or claim 1 , wherein the affinity tag is located on the N- or C-terminus of the polypeptide claim 1 , preferably the N-terminus.4. The isolated polypeptide according to any one of - claim 1 , wherein the polypeptide has in N- to C-terminal orientation the ...

TRANSGENIC ANIMAL FOR PRODUCTION OF ANTIBODIES HAVING MINIMAL CDRS

A transgenic animal is provided. In certain embodiments, the transgenic animal comprises a genome comprising: an immunoglobulin light chain locus comprising: a) a functional immunoglobulin light chain gene comprising a transcribed variable region encoding: i. light chain CDR1, CDR2 and CDR3 regions that are composed of 2 to 5 different amino acids; and ii. a light chain framework; and, operably linked to the functional immunoglobulin light chain gene: b) a plurality of pseudogene light chain variable regions each encoding: i. light chain CDR1, CDR2 and CDR3 regions that are composed of the same 2 to 5 different amino acids as the CDRs of the functional gene; and ii. a light chain framework that is identical in amino acid sequence to the light chain framework of the transcribed variable region. 120-. (canceled)21. A transgenic chicken that comprises a genome comprising a recombinant immunoglobulin heavy chain (IgH) locus comprising: (i) heavy chain complementarity determining (CDR) regions; and', '(ii) a heavy chain framework; and,, '(a) a functional IgH gene comprising a nucleic acid encoding a heavy chain variable region comprising (i) heavy chain CDR regions; and', '(ii) a heavy chain framework region that is identical in amino acid sequence to the heavy chain framework of a) (ii);, '(b) a plurality of pseudogenes that encode heavy chain variable regions each comprisingwherein said nucleic acid of (a) and pseudogene of (b), are exogenous to the genome of said transgenic chicken,wherein said plurality of pseudogenes are operably linked to said functional IgH gene and donate nucleotide sequences to the nucleic acid of (a) by gene conversion in said transgenic chicken; andwherein said transgenic chicken expresses a polyclonal antisera comprising a diversified IgH variable region.22. The transgenic chicken of claim 21 , wherein said heavy chain framework is identical to a human framework.23. The transgenic chicken of claim 21 , wherein said heavy chain framework is at ...

METHODS OF PRODUCING NOR-OPIOID AND NAL-OPIOID BENZYLISOQUINOLINE ALKALOIDS

A method of demethylizing an opioid to a nor-opioid is provided. The method comprises contacting an opioid with at least one enzyme. Contacting the opioid with the at least one enzyme converts the opioid to a nor-opioid. A method of converting a nor-opioid to a nal-opioid is provided. The method comprises contacting a nor-opioid with at least one enzyme. Contacting the nor-opioid with the at least one enzyme converts the nor-opioid to a nal-opioid. 13.-. (canceled)4. A method of altering an opioid to a nal-opioid , comprising:contacting the opioid with at least a first enzyme, wherein contacting the opioid with the at least a first enzyme converts the opioid to a nor-opioid through removal of a N-linked methyl group from the opioid; andcontacting the nor-opioid with at least a second enzyme, wherein contacting the nor-opioid with the at least a second enzyme in the presence of a cofactor converts the nor-opioid to the nal-opioid through transfer of a sidechain from the cofactor.5. A method of altering an opioid to a nal-opioid , comprising:contacting a first opioid with at least one enzyme, wherein contacting the first opioid with the at least one enzyme converts the first opioid to a second opioid through loss of an O-linked methyl group;contacting the second opioid with at least a second enzyme, wherein contacting the opioid with the at least a second enzyme converts the second opioid to the nor-opioid through loss of an N-linked methyl group; andcontacting the nor-opioid with at least a third enzyme, wherein contacting the nor-opioid with the at least a third enzyme in the presence of a cofactor converts the nor-opioid to a nal-opioid through transfer of a sidechain from the cofactor.68.-. (canceled)9. The method of claim 4 , wherein the nal-opioid is produced by culturing an engineered cell comprising a coding sequence for encoding the first enzyme and the second enzyme.10. The method of claim 5 , wherein the nor-opioid is produced by culturing an engineered ...

Human Monoclonal Antibodies to Human Endogenous Retrovirus K Envelope (HERV-K) and Use Thereof

The present invention features anti-HERV-K monoclonal antibodies or antigen-binding portions thereof. The present invention also features uses of the antibodies for treating HIV infection or HIV-associated conditions or diseases. 1. An isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to human HERV-K envelope polypeptide comprising:a. a heavy chain variable region comprising a HCDR1 comprising an amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising an amino acid sequence of SEQ ID NO: 5 and a HCDR3 comprising an amino acid sequence of SEQ ID NO: 6; and a light chain variable region comprising a LCDR1 comprising an amino acid sequence of SEQ ID NO: 7, a LCDR2 comprising an amino acid sequence of SEQ ID NO: 8 and a LCDR3 comprising an amino acid sequence of SEQ ID NO: 9; orb. a heavy chain variable region comprising a HCDR1 comprising an amino acid sequence of SEQ ID NO: 10, a HCDR2 comprising an amino acid sequence of SEQ ID NO: 11 and a HCDR3 comprising an amino acid sequence of SEQ ID NO: 12; and a light chain variable region comprising a LCDR1 comprising an amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising an amino acid sequence of SEQ ID NO: 14 and a LCDR3 comprising an amino acid sequence of SEQ ID NO: 15.2. The isolated antibody or the antigen-binding fragment thereof of claim 1 , wherein the heavy chain variable region comprises an amino acid sequence of SEQ ID NO: 23 and light chain variable region comprises an amino acid sequence of SEQ ID NO: 22.3. The isolated antibody or the antigen-binding fragment thereof of claim 1 , wherein the heavy chain variable region comprises an amino acid sequence of SEQ ID NO: 25 and light chain variable region comprises an amino acid sequence of SEQ ID NO: 24.4. The isolated antibody or the antigen-binding fragment thereof of claim 1 , further comprising a variant Fc constant region.5. The isolated antibody or the antigen-binding fragment thereof of claim 1 , wherein the ...

NOVEL T CELL RECEPTORS AND IMMUNE THERAPY USING THE SAME

The present invention pertains to antigen recognizing constructs against tumor associated antigens (TAA), in particular the TAA Serine protease inhibitor Kazal-type 2 (SPINK2). The invention in particular provides novel T cell receptor (TCR) based molecules which are selective and specific for the tumor expressed antigen of the invention. The TCR of the invention, and SPINK2 binding fragments derived therefrom, are of use for the diagnosis, treatment and prevention of SPINK2 expressing cancerous diseases. Further provided are nucleic acids encoding the antigen recognizing constructs of the invention, vectors comprising these nucleic acids, recombinant cells expressing the antigen recognizing constructs and pharmaceutical compositions comprising the compounds of the invention. 1. A method of treating a patient who has cancer , comprising administering to the patient a population of transformed T cells expressing at least one vector encoding a T cell receptor (TCR) , SEQ ID NOs: 1, 3, 7, and 9, or', 'SEQ ID NOs: 13, 15, 19, and 21, or', 'SEQ ID NOs: 25, 27, 31, and 33, or', 'SEQ ID NOs: 37, 39, 43, and 45, or', 'SEQ ID NOs: 49, 51, 55, and 57, or', 'SEQ ID NOs: 61, 63, 67, and 69, or', 'SEQ ID NOs: 73, 75, 79, and 81, or', 'SEQ ID NOs: 85, 87, 91, and 93, or', 'SEQ ID NOs: 97, 99, 103, and 105, or', 'SEQ ID NOs: 109, 111, 115, and 117,, 'wherein the TCR comprises'}wherein each of SEQ ID NOs: 1, 3, 7, 9, 13, 15, 19, 21, 25, 27, 31, 33, 37, 39, 43, 45, 49, 51, 55, 57, 61, 63, 67, 69, 73, 75, 79, 81, 85, 87, 91, 93, 97, 99, 103, 105, 109, 111, 115, and 117 comprises at most one conservative amino acid substitution,wherein the TCR binds to a peptide consisting of the amino acid sequence of ALSVLRLAL (SEQ ID NO: 133) in a complex with an MHC class I molecule, andwherein the cancer is selected from acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal, or anorectum, cancer of the ...

COMPOSITIONS AND METHODS FOR TREATMENT OF CANCER

Disclosed herein are transmembrane proteins comprising at least two chains, each chain comprising an endodomain, a transmembrane domain, and an ectodomain; wherein the ectodomains of the at least two chains of the transmembrane protein interact together to bind to a drug or soluble protein; wherein the endodomains of the at least two chains of the transmembrane protein together comprise a structure functionally similar to that of an IL-2 receptor endodomain, an IL-7 receptor endodomain, or an IL-15 receptor endodomain; and wherein binding of the drug or soluble protein to the ectodomain activates IL-2, IL-7, or IL-15 signaling in CAR-bearing immune effector cells. 1. A composition comprising a population of CAR-bearing immune effector cells , the cells comprising a transmembrane protein , the transmembrane protein comprising at least two recombinant chains , each recombinant chain comprising an endodomain , a transmembrane domain , and an ectodomain;wherein the ectodomains of the at least two chains of the transmembrane protein interact together to bind to a drug or soluble protein;wherein the endodomains of the at least two chains of the transmembrane protein together comprise a structure functionally similar to that of an IL-2 receptor endodomain, an IL-7 receptor endodomain, or an IL-15 receptor endodomain; andwherein binding of the drug or soluble protein to the ectodomain activates IL-2, IL-7, or IL-15 signaling in CAR-bearing immune effector cells.2. The composition of claim 1 , wherein the ectodomains of the at least two chains of the transmembrane protein each comprise an antibody or binding portion thereof.3. The composition of claim 1 , wherein the binding portion of each antibody is chosen from a Fab claim 1 , scFv claim 1 , F(ab′) claim 1 , minibody claim 1 , or other binding arrangement of Vand Vchains or CDR-containing proteins.4. The composition of claim 1 , wherein the ectodomains of the at least two chains of the transmembrane protein each comprise ...

ANTI-C5A ANTIBODIES AND USES THEREOF

This invention relates to inhibition of the complement signaling using an anti-C5a antibody. Specifically, the invention relates to methods of treating a complement-mediated disease or complement-mediated disorder in an individual by contacting the individual with an anti-C5a antibody. 1. An antibody that specifically binds to human C5a.2. The antibody of claim 1 , wherein the antibody is a chimeric antibody.3. The antibody of claim 1 , wherein the antibody comprises at least one of the CDRs selected from the group consisting of: VH-CDR1: SEQ ID NO:3; VH-CDR2: SEQ ID NO:4; VH-CDR3: SEQ ID NO:5; VL-CDR1: SEQ ID NO:8; VL-CDR2: SEQ ID NO:9; and VL-CDR3: SEQ ID NO:10 claim 1 , or a variant of variants thereof.4. The antibody of claim 1 , wherein the antibody comprises the CDRs: VH-CDR1: SEQ ID NO:3; VH-CDR2: SEQ ID NO:4; VH-CDR3: SEQ ID NO:5; VL-CDR1: SEQ ID NO:8; VL-CDR2: SEQ ID NO:9; and VL-CDR3: SEQ ID NO:10 claim 1 , or a variant of variants thereof.5. The antibody of claim 1 , wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:2 claim 1 , or a variant thereof.6. The antibody of claim 1 , wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:7 claim 1 , or a variant thereof.7. The antibody of claim 1 , wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:2 and a light chain comprising the amino acid sequence of SEQ ID NO:7 claim 1 , or a variant or variants thereof.8. A method of treating a complement pathway-mediated disease or disorder in an individual claim 1 , comprising the step of administering to said individual the anti-C5a antibody of .9. The method of claim 8 , wherein the disease or disorder is at least selected from the group consisting of: macular degeneration (MD) claim 8 , age-related macular degeneration (AMD) claim 8 , ischemia reperfusion injury claim 8 , arthritis claim 8 , rheumatoid arthritis claim 8 , lupus claim 8 , ...

ANTI-PD-L1 NANOBODY, CODING SEQUENCE AND USE THEREOF

Provided in the present invention are a type of anti-human PD-L1 specific nanobodies and VHH chains thereof, coding sequences of the foregoing nanobodies or VHH chains thereof, corresponding expression vectors and host cells, and a method for producing antibodies. 1. A VHH chain of an anti-PD-L1 nanobody , wherein the amino acid sequence of the VHH chain is shown in any one of SEQ ID NOs.: 2-8 , 10-11 , 13-17 , 19 , 21-44 , 46-49 , 51-62 , 64-69 , 71-73 , 75-150.2. The VHH chain of an anti-PD-L1 nanobody according to claim 1 , wherein the amino acid sequence of the VHH chain is shown in SEQ ID NOs.: 2-8 claim 1 , 10 or 11.3. The VHH chain of an anti-PD-L1 nanobody according to claim 1 , wherein the amino acid sequence of the VHA chain is shown in any one of SEQ ID NOs.: 3-5 claim 1 , 7 claim 1 , 13-17 claim 1 , 19 claim 1 , 21-23 claim 1 , 25-26 claim 1 , 28-33. 36-39 claim 1 , 41-44 claim 1 , 46-49 claim 1 , 51-57 claim 1 , 59-60 claim 1 , 62 claim 1 , 64 claim 1 , 66 claim 1 , 68-69 claim 1 , 75-76 claim 1 , 78 claim 1 , 80--81 claim 1 , 83 claim 1 , 85-86 claim 1 , 88-90 claim 1 , 92-95 claim 1 , 97-99 claim 1 , 101-104 claim 1 , 108 claim 1 , 113 claim 1 , 117-119 claim 1 , and 123.4. The VHH chain of an anti-PD-L1 nanobody according to claim 1 , wherein the amino acid sequence of the VHH chain is shown in any one of SEQ ID NOs.: 58 and 61.5. The VHH chain of an anti-PD-L1 nanobody according to claim 1 , which has a very high specificity or selectivity for PD-L1 (relative to PD-L2) and has a selectivity ratio (such as a ratio of OD value)(PD-L1/PD-L2) as high as preferably 20-40 claim 1 , or 21-35.6. An immunoconjugate claim 1 , wherein the immunoconjugate comprises:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) the VHH chain of the anti-PD-L1 nanobody according to ; and'}(b) a conjugating part selected from the group consisting of a detectable marker, drug, toxin, cytokine, radionuclide, and enzyme.7. A pharmaceutical composition comprising:{'claim-ref ...

EXPRESSION FROM TRANSPOSON-BASED VECTORS AND USES

Recombinant expression vectors are disclosed that include a control sequence for recombinant expression of proteins of interest; the control sequence combines a mCMV enhancer sequence with a rat EF-1alpha intron sequence. Some of the vectors are useful for tetracycline-inducible expression. Some of the vectors contain a 5′ PiggyBac ITR and a 3′ PiggyBac ITR to promote genomic integration into a host cell chromosome. A method of selecting a stable production cell line for manufacturing a protein of interest is also disclosed. Also disclosed are mammalian host cells comprising the inventive recombinant expression vectors and a method of producing a protein of interest, in vitro, involving the mammalian host cell. 19-. (canceled)10: A recombinant expression vector , comprising: [ (1) a murine cytomegalovirus (mCMV) enhancer sequence, comprising a mCMV enhancer element (mCMV-E) and a CMV promoter (CMV-P) sequence at its 3′ end, operably linked 5′ to a rat EF-1alpha intron sequence;', '(2) an intervening first leader sequence operably linked, 3′ to the CMV-P sequence of the mCMV enhancer sequence, and 5′ to the rat EF-1alpha intron sequence; and', '(3) a second leader sequence operably linked 3′ to the rat EF-1alpha intron sequence;, '(i) a control sequence comprising, '(ii) an open reading frame encoding a first immunoglobulin subunit operably linked to the control sequence; and', '(iii) a first polyadenylation site operably linked 3′ to the open reading frame;, '(a) a first expression cassette, comprising (i) a control sequence comprising a promoter;', '(i) an open reading frame encoding a second immunoglobulin subunit operably linked to the promoter; and', '(iii) a second polyadenylation site operably linked 3′ to the open reading frame; and, '(b) a second expression cassette 3′ to the first expression cassette, comprising(c) a transcription termination sequence 3′ to the first expression cassette and 5′ to the second expression cassette.11: The vector of claim 10 , ...

Human Lambda Light Chain Mice

Genetically modified mice are provided that express human λ variable (hVλ) sequences, including mice that express hVλ sequences from an endogenous mouse λ light chain locus, mice that express hVλ sequences from an endogenous mouse κ light chain locus, and mice that express hVλ sequences from a transgene or an episome wherein the hVλ sequence is linked to a mouse constant sequence. Mice are provided that are a source of somatically mutated human λ variable sequences useful for making antigen-binding proteins. Compositions and methods for making antigen-binding proteins that comprise human λ variable sequences, including human antibodies, are provided.

METHODS FOR PRODUCING AND USING IN VIVO PSEUDOTYPED RETROVIRUSES

The present invention provides novel pseudotyped retroviral vectors that can transduce human and other cells. Vectors are provided that are packaged efficiently in packaging cells and cell lines to generate high titer recombinant virus stocks expressing novel envelope glycoproteins. The present invention further relates to compositions for gene therapy. 1. A method of delivering a transgene to an airway epithelia cell comprising contacting the cell with a pseudotyped retrovirus virion comprising a baculovirus envelope glycoprotein and a nucleic acid encoding a promoter and the transgene.2. The method of claim 1 , wherein the cell is contacted with the virion on the apical surface of the cell.3. The method of claim 1 , wherein the envelope glycoprotein is glycoprotein-64 (GP64).4Autographa californica. The method of claim 3 , wherein the envelope glycoprotein is an multinuclear polyhedrosis virus (AcMNPV) glycoprotein.5. The method of claim 1 , wherein the retrovirus is feline immunodeficiency virus (FIV).6. The method of claim 1 , wherein the transgene is a remedial gene.7. The method of claim 6 , wherein the transgene is a nucleic acid encoding a cystic fibrosis transmembrane regulator protein (CFTR).8. The method of claim 1 , wherein the cell is in vitro.9. The method of claim 1 , wherein the cell is in vivo.10. The method of claim 1 , wherein the promoter is a tissue-specific promoter.11. The method of claim 1 , wherein the envelope glycoprotein is a type D influenzae virus glycoprotein claim 1 , an F protein for an insect virus glycoprotein claim 1 , or a metaviridae envelope protein.12. The method of claim 11 , wherein the glycoprotein is derived from a type D influenzae virus is a glycoprotein-75 (GP75) protein.13. The method of claim 12 , wherein the influenze D virus is a thogoto virus or a dhori virus. This application is a Continuation of U.S. application Ser. No. 11/943,871, filed Nov. 21, 2007, which is a Continuation of U.S. application Ser. No. 10/964, ...

ANTI-GALACTAN II MONOCLONAL ANTIBODIES TARGETING KLEBSIELLA PNEUMONIAE

A cross-neutralizing monoclonal antibody that specifically recognizes a cross-reactive epitope of the lipopolysaccharide (LPS) antigen structure of , which is an O3b epitope, cross-reacting with an O3a epitope and an O3 epitope, wherein the antibody is characterized by specific CDR sequences or VH and VL sequences. 115-. (canceled)16Klebsiella pneumoniae. A monoclonal antibody that specifically recognizes serotype O1 , which is capable of neutralizing the LPS endotoxin activity , which antibody is selected from any ofa) an antibody comprising the CDR1-CDR6 sequences of any one of the antibodies listed in Table 1a or Table 1b; or{'figref': {'@idref': 'DRAWINGS', 'i': 'b', 'FIG. 2'}, 'b) an antibody comprising the the VH and VL sequences of any one of the antibodies depicted in ; or'}{'i': 'Klebsiella pneumoniae', 'c) an antibody which is a functionally active variant of a parent antibody that is any one of the antibodies of a) or b), which functionally active variant specifically recognizes serotype O1 and is capable of neutralizing the LPS endotoxin activity, and comprises at least one point mutation in any of the CDR, wherein the number of point mutations is either 0, 1, 2, or 3 point mutations in each of the CDR sequences, wherein the sequence identity in each of the CDR sequences is at least 60% compared to the respective CDR sequences of the parent antibody.'}17. The antibody of claim 16 , which isA)selected from the group consisting of group members i) to ii), whereini) a) CDR1 consisting of the amino acid sequence SEQ ID 9;', 'b) CDR2 consisting of the amino acid sequence SEQ ID 10;', 'c) CDR3 consisting of the amino acid sequence of SEQ ID 11;', 'd) CDR4 consisting of the amino acid sequence SEQ ID 15;', 'e) CDR5 consisting of the amino acid sequence SEQ ID 16; and', 'f) CDR6 consisting of the amino acid sequence of SEQ ID 17;, 'is an antibody which comprises'}ii) a) CDR1 consisting of the amino acid sequence SEQ ID 12;', 'b) CDR2 consisting of the amino acid ...

ANTI-HUMAN INTERLEUKIN-17A MONOCLONAL ANTIBODY, AND PREPARATION METHOD AND USE THEREOF

An anti-human IL-17A monoclonal antibody, the preparation method, and the use thereof are provided. The anti-human IL-17A monoclonal antibody can specifically bind to human IL-17A and has a good effect for inhibiting the secretion of inflammatory cytokines, such as IL-6 and the like by means of various cell lines induced by IL-17A, and can be used for preparing drugs for treating diseases involving immune-medicated inflammatory responses. 1. An anti-human IL-17A monoclonal antibody , comprising:(1) a heavy chain complementarity determining region CDR1, CDR2, CDR3, wherein the CDR1 has the amino acid sequence as shown in SEQ ID NO: 5, the CDR2 has the amino acid sequence as shown in SEQ ID NO: 6, and the CDR3 has the amino acid sequence as shown in SEQ ID NO: 7; and(2) a light chain complementarity determining region CDR1′, CDR2′, CDR3′, wherein the CDR1′ has the amino acid sequence as shown in SEQ ID NO: 8, the CDR2′ has the amino acid sequence as shown in SEQ ID NO: 9, and the CDR3′ has the amino acid sequence as shown in SEQ ID NO: 10.2. The anti-human IL-17A monoclonal antibody of claim 1 , characterized in that the anti-human IL-17A monoclonal antibody comprises a heavy chain variable region having the amino acid sequence as shown in SEQ ID NO: 2 claim 1 , and a light chain variable region having the amino acid sequence as shown in SEQ ID NO: 4.3. The anti-human IL-17A monoclonal antibody of claim 1 , characterized in that the anti-human IL-17A monoclonal antibody is a humanized anti-human IL-17A monoclonal antibody claim 1 , and the humanized anti-human IL-17A monoclonal antibody comprises a heavy chain variable region having the amino acid sequence as shown in SEQ ID NO: 12 claim 1 , and a light chain variable region having the amino acid sequence as shown in SEQ ID NO: 14.4. The anti-human IL-17A monoclonal antibody of claim 3 , characterized in that the humanized anti-human IL-17A monoclonal antibody comprises a heavy chain having the amino acid sequence as ...

Bispecific Antibodies Directed Against OX40 and a Tumor-Associated Antigen

The invention provides bispecific polypeptides comprising a first binding domain, designated B1, which is capable of binding specifically to OX40, and a second binding domain, designated B2, which is capable of specifically binding to a tumour cell-associated antigen. Also provided are pharmaceutical compositions of such bispecific polypeptides and uses of the same in medicine. 1. A bispecific polypeptide comprising a first binding domain , designated B1 , which is capable of binding specifically to OX40 , and a second binding domain , designated B2 , which is capable of specifically binding to a tumour cell-associated antigen.2. A polypeptide according to claim 1 , wherein the first and/or second binding domains are/is selected from the group consisting of antibodies and antigen-binding fragments thereof.3. A polypeptide according to wherein the antigen-binding fragment is selected from the group consisting of: Fv fragments (such as a single chain Fv fragment claim 2 , or a disulphide-bonded Fv fragment) claim 2 , Fab-like fragments (such as a Fab fragment; a Fab′ fragment or a F(ab)fragment) and domain antibodies.4. A polypeptide according to any one of the preceding claims wherein the polypeptide is a bispecific antibody.5. A polypeptide according to wherein:(a) binding domain B1 and/or binding domain B2 is an intact IgG antibody;(b) binding domain B1 and/or binding domain B2 is an Fv fragment;(c) binding domain B1 and/or binding domain B2 is a Fab fragment; and/or(d) binding domain B1 and/or binding domain B2 is a single domain antibody.6. A polypeptide according to or wherein the bispecific antibody comprises a human Fc region or a variant of a said region claim 4 , where the region is an IgG1 claim 4 , IgG2 claim 4 , IgG3 or IgG4 region claim 4 , preferably an IgG1 or IgG4 region.7. A polypeptide according to wherein the Fc exhibits no or very low affinity for FcgR.8. A polypeptide according to or wherein the Fc region is a variant of a human IgG1 Fc region ...

COMPOSITIONS AND METHODS RELATED TO THERAPEUTIC CELL SYSTEMS EXPRESSING EXOGENOUS RNA

The invention includes compositions and methods related to erythroid cells comprising exogenous RNA encoding a protein. The exogenous RNA can comprise a heterologous untranslated region comprising a regulatory element. Alternatively or in combination, the exogenous RNA can comprise chemical modifications. 1. A method of making an erythroid cell comprising an mRNA encoding an exogenous protein , comprising:a) providing an erythroid cell in maturation phase, andb) contacting the erythroid cell with an mRNA encoding the exogenous protein, under conditions that allow uptake of the mRNA by the erythroid cell,thereby making an erythroid cell comprising an mRNA encoding an exogenous protein.2. The method of claim 1 , wherein the erythroid cell takes up the mRNA encoding the exogenous protein.3. The method of claim 1 , comprising providing a population of erythroid cells in maturation phase and contacting a plurality of cells of the population of erythroid cells with the mRNA encoding the exogenous protein.4. The method of claim 3 , wherein the plurality of cells of the population of erythroid cells each takes up the mRNA encoding the exogenous protein.5. The method of any of - claim 3 , wherein after uptake of the mRNA encoding the exogenous protein claim 3 , the cell or the plurality of cells express the exogenous protein.6. The method of claim 5 , wherein the cell or the plurality of cells comprise the exogenous protein.7. The method of any of - claim 5 , wherein the population of erythroid cells in maturation phase is a population of cells expanded in a maturation medium for 3-7 days claim 5 , e.g. claim 5 , 4-5 or 4-6 days.9. The method of claim 8 , wherein the further culturing comprises fewer than 3 claim 8 , 2 claim 8 , or 1 population doubling.10. The method of any of - claim 8 , wherein the population of erythroid cells or the population of differentiating erythroid cells is a population of erythroid cells comprising one or more (e.g. claim 8 , 2 claim 8 , 3 claim ...

ANTI-ROBO2 ANTIBODIES, COMPOSITIONS, METHODS AND USES THEREOF

The invention provides antibodies, and antigen-binding fragments thereof, that specifically bind to ROBO2, as well as uses, and methods thereof. 1. An isolated antibody , or antigen-binding fragment thereof , that binds an epitope in Ig Domain 1 , or Domains 1 and 2 , of Roundabout Receptor 2 (ROBO2) , wherein said epitope comprises amino acid residue arginine 100 (R100) , according to the numbering of SEQ ID NO: 1.2. The antibody claim 1 , or antigen-binding fragment thereof claim 1 , of claim 1 , wherein said epitope further comprises one or more residues selected from the group consisting of: V96 claim 1 , G98 claim 1 , R99 claim 1 , and S101 claim 1 , according to the numbering of SEQ ID NO: 1.3. The antibody claim 1 , or antigen-binding fragment thereof claim 1 , of claim 1 , wherein said epitope further comprises one or more residues selected from the group consisting of: E69 claim 1 , E72 claim 1 , R79 claim 1 , H81 claim 1 , R82 claim 1 , R94 claim 1 , and P103 claim 1 , according to the numbering of SEQ ID NO: 1.4. The antibody claim 1 , or antigen-binding fragment thereof claim 1 , of claim 1 , wherein said epitope further comprises one or more residues selected from the group consisting of: K66 claim 1 , D67 claim 1 , R70 claim 1 , V71 claim 1 , T73 claim 1 , D74 claim 1 , D77 claim 1 , P78 claim 1 , H97 claim 1 , and K102 claim 1 , according to the numbering of SEQ ID NO: 1.5. The antibody claim 1 , or antigen-binding fragment thereof claim 1 , of any one of claim 1 , wherein said antibody claim 1 , or antigen-binding fragment thereof claim 1 , binds said epitope with a binding affinity (K) value that is at least 100-fold less than its Kvalue for an epitope wherein R100 is replaced with K.6. The antibody claim 1 , or antigen-binding fragment thereof claim 1 , of claim 1 , wherein said antibody claim 1 , or antigen-binding fragment thereof claim 1 , does not substantially bind to Roundabout Receptor 1 (ROBO1).7. The antibody claim 6 , or antigen-binding ...

ANTI-CD3 ANTIBODIES AND METHODS OF MAKING AND USING THEREOF

The applications provides the anti-CD3 monoclonal antibodies, antigen-binding portions thereof, therapeutic compositions thereof and/or nucleic acid encoding the same, and their use to active CD3+ T-cells to enhance cell-mediated immune responses in the treatment of cancer and other T-cell dysfunctional disorders. 1. An isolated monoclonal antibody (mAb) or antigen-binding fragment thereof having a binding specificity to human CD3 , wherein the isolated mAb or antigen-binding fragment comprises an amino acid sequence having an identity with a sequence selected from SEQ ID NO:4 , SEQ ID NO:8 , SEQ ID NO:12 , SEQ ID NO:16 , SEQ ID NO:20 , SEQ ID NO:24 , SEQ ID NO:28 , SEQ ID NO:32 , SEQ ID NO:36 , SEQ ID NO:40 , SEQ ID NO:44 , SEQ ID NO:48 , SEQ ID NO:52 , SEQ ID NO:56 , SEQ ID NO:60 , SEQ ID NO:64 , SEQ ID NO:68 , or SEQ ID NO:72 , wherein the identity is not less than at least 98%.2. The isolated monoclonal antibody (mAb) or antigen-binding fragment according to claim 1 , wherein the identity is not less than 98%.3. The isolated mAb or antigen-binding fragment according to claim 1 , having a binding affinity to human CD3 with a Kd not greater than 70 nM.4. The isolated mAb or antigen-binding fragment according to claim 1 , wherein the isolated mAb or antigen-binding fragment is capable of high affinity binding to CD3 claim 1 , enhancing T cell activation claim 1 , stimulating antibody response claim 1 , or reversing the suppressive function of an immunosuppressive cell.5. The isolated mAb or antigen-binding fragment according to claim 4 , wherein the enhancing T-cell activation comprises T-cell proliferation claim 4 , IFN-γ and/or IL-2 secretion claim 4 , or a combination thereof.6. The isolated mAb or antigen-binding fragment according to claim 4 , wherein the immunosuppressive cell comprises a T regulatory cell.7. The isolated mAb or an antigen-binding fragment thereof according to claim 1 , wherein the isolated mAb is a humanized antibody claim 1 , a chimeric ...

Anti-ALK Antibodies and Methods for Use Thereof

Provided herein are compositions, methods and uses involving antibodies that specifically bind to ALK, a receptor tyrosine kinase, and modulate the expression and/or activity of ALK. Also provided are uses and methods for treating, e.g., managing, disorders, such as cancer. 1. An antibody or an antigen-binding fragment thereof , which specifically binds to an anaplastic lymphoma kinase (ALK) extracellular domain (ECD) , comprising: (a) a VL complementarity determining region (CDR) 1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13);', '(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and', {'sub': 1', '2', '1', '2, '(c) a VL CDR3 comprising the amino acid sequence of QXYNSYPYMXT (SEQ ID NO:468), wherein Xis Q or R and Xis Y or F; and'}], '(i) a light chain variable region (VL) comprising [{'sub': 3', '3, '(a) a VH CDR1 comprising the amino acid sequence XYWMH (SEQ ID NO:469), wherein Xis N or S;'}, {'sub': 4', '4, '(b) a VH CDR2 comprising the amino acid sequence of YIXPSSGYTKYNQKFKD (SEQ ID NO:470), wherein Xis N or K; and'}, '(c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18)., '(ii) a heavy chain variable region (VH) comprising2. The antibody or antigen-binding fragment of claim 1 , comprising a VL comprising:(a) a VL CDR1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13);(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and(c) a VL CDR3 comprising the amino acid sequence of QQYNSYPYMYT (SEQ ID NO:15).3. The antibody or antigen-binding fragment of claim 1 , comprising a VL comprising:(a) a VL CDR1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13);(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and(c) a VL CDR3 comprising the amino acid sequence of QRYNSYPYMFT (SEQ ID NO:41).4. The antibody or antigen-binding fragment of any one of - claim 1 , comprising a VH comprising:(a) a VH CDR1 comprising the amino acid ...

Non-human animals expressing exogenous terminal deoxynucleotidyltransferase

Provided herein are methods and compositions related to non-human animals that express exogenous Terminal Deoxynucleotidyltransferase (TdT).

Anti-ryk antibodies and methods of using the same

Methods for inhibiting degeneration of a neuron, methods of treating a neurological/neurodegenerative disease, methods of modulating the directional growth of a neuron, and methods of interfering with the interaction of Wnt and Ryk are provided herein. Also provided are isolated anti-Ryk antibodies and antibody fragments that specifically bind to a binding domain of Wnt.

Circular RNA For Translation In Eukaryotic Cells

Disclosed are methods and constructs for engineering circular RNA. Disclosed is a vector for making circular RNA, said vector comprising the following elements operably connected to each other and arranged in the following sequence: 1. A vector for making circular RNA , said vector comprising the following elements operably connected to each other and arranged in the following sequence:a) a 5′ homology arm,b) a 3′ Group I intron fragment containing a 3′ splice site dinucleotide,c) a 5′ spacer sequence,d) an internal ribosome entry site (IRES),e) a protein coding region,f) a 3′ spacer sequence,g) a 5′ Group I intron fragment containing a 5′ splice site dinucleotide, andh) a 3′ homology arm,said vector allowing production of a circular RNA that is translatable or biologically active inside eukaryotic cells.2. The vector of claim 1 , wherein the protein coding region encodes a therapeutic protein claim 1 , a chimeric antigen receptor claim 1 , an antibody or a bispecific antibody.3. A composition comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) vector for making circular RNA of , and'}b) a delivery vehicle for delivering the vector to a cell, said delivery vehicle comprising a nanocarrier selected from the group consisting of a lipid, a polymer and a lipo-polymeric hybrid.4. A circular RNA produced by the vector of claim 1 , wherein the vector is used to transcribe circular RNA with a size of at least 500 nucleotides.5. A eukaryotic cell comprising the circular RNA of .6. A cell population comprising the circular RNA of .7. The circular RNA of claim 4 , wherein the circular RNA comprises in the following order claim 4 , a portion of a 3′ group I intron fragment claim 4 , an Internal Ribosome Entry Site (IRES) claim 4 , a protein coding region claim 4 , and a portion of a 5′ group I intron fragment.8. A composition comprising an effective amount of the circular RNA of and a pharmaceutically acceptable carrier.9. A composition comprising:{'claim-ref': {'@ ...

Compositions and methods for enhancing gene expression

The present disclosure generally relates to nucleic acid molecules for use in regulating gene expression. Disclosed herein include nucleic acid molecules containing one or more structural elements of the viral capsid enhancer operably linked to a coding sequence of a gene of interest. In some embodiments, the viral capsid enhancer comprises a Downstream Loop (DLP) from a viral capsid protein, or a variant of the DLP.

COMPOSITIONS AND METHODS FOR INHIBITING ENDOGENOUS IMMUNOGLOBULIN GENES AND PRODUCING TRANSGENIC HUMAN IDIOTYPE ANTIBODIES

The invention relates to transgenic animals lacking endogenous Ig and capable of producing transgenic antibodies, as well as methods of making the same. The invention further relates to Methods for producing transgenic antibodies in such animals, and transgenic antibodies so produced. 134-. (canceled)35. A chimeric monoclonal antibody comprising a non-human IgG constant region , a light chain and/or heavy chain , and a human idiotype , and wherein the non-human constant region comprises a rat IgG CH2 and CH3 domain.36. The chimeric monoclonal antibody of claim 35 , wherein the non-human constant region further comprises a human IgG CH1 domain.37. The chimeric monoclonal antibody of claim 35 , wherein the monoclonal antibody comprises a heavy chain and a light chain.38. The chimeric monoclonal antibody of claim 35 , wherein the monoclonal antibody is a heavy chain only antibody.39. The chimeric monoclonal antibody of claim 35 , wherein the non-human constant region comprises an IgG CH2 and CH3 domain and lacks a CH1 domain.40. A polyclonal antiserum comprising a plurality of chimeric antibodies claim 35 , wherein the individual chimeric antibodies comprise a non-human IgG constant region claim 35 , a heavy chain and/or light chain claim 35 , and a human idiotype claim 35 , and wherein the non-human constant region comprises a rat IgG CH2 and CH3 domain.41. The polyclonal antiserum of claim 40 , wherein the non-human constant region further comprises a human IgG CH1 domain.42. The polyclonal antiserum of claim 40 , wherein the individual chimeric antibodies comprise a heavy chain and a light chain.43. The polyclonal antiserum of claim 40 , wherein the individual chimeric antibodies are heavy chain only antibodies.44. The polyclonal antiserum of claim 40 , wherein the non-human constant region comprises an IgG CH2 and CH3 domain and lacks a CH1 domain.45. The polyclonal antiserum of claim 40 , wherein the antiserum is:a. a bird antiserum;b. a chicken antiserum;c. a ...

MODIFIED ANTI-TENASCIN ANTIBODIES AND METHODS OF USE

The present invention relates to improved antibodies specific for Tenascin-C (TnC), in particular domain specific anti-TnC antibodies with improved cross-species reactivity. In addition, the invention relates to polynuleotides encoding such antibodies, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the antibodies and methods of using them in the treatment of disease. 1. An antibody that specifically binds to Tenascin-C (TnC) , wherein said antibody comprises:(i) a heavy chain variable region comprising a heavy chain CDR1 of SEQ ID NO: 49, a heavy chain CDR2 of SEQ ID NO: 50 and a heavy chain CDR3 of SEQ ID NO:51; and a light chain variable region comprising a light chain CDR1 of SEQ ID NO: 37, a light chain CDR2 of SEQ ID NO: 38 and a light chain CDR3 of SEQ ID NO: 39; or(ii) a heavy chain variable region comprising a heavy chain CDR1 of SEQ ID NO: 52, a heavy chain CDR2 of SEQ ID NO: 53 and a heavy chain CDR3 of SEQ ID NO: 54; and a light chain variable region comprising a light chain CDR1 of SEQ ID NO: 40, a light chain CDR2 of SEQ ID NO:41 and a light chain CDR3 of SEQ ID NO: 42; wherein said antibody has cross-species reactivity.2. The antibody of claim 1 , wherein said antibody comprises a heavy chain variable region comprising(a) the heavy chain CDR1 of SEQ ID NO: 49;(b) the heavy chain CDR2 of SEQ ID NO: 50;(c) the heavy chain CDR3 of SEQ ID NO: 51,and a light chain variable region comprising(a) the light chain CDR1 of SEQ ID NO: 37;(b) the light chain CDR2 of SEQ ID NO: 38, and(c) the light chain CDR3 of SEQ ID NO: 39.3. The antibody of claim 1 , wherein said antibody comprises a heavy chain variable region comprising(a) the heavy chain CDR1 of SEQ ID NO: 52;(b) the heavy chain CDR2 of SEQ ID NO: 53;(c) the heavy chain CDR3 of SEQ ID NO: 54,and a light chain variable region comprising(a) the light chain CDR1 of SEQ ID NO: 40;(b) the light chain CDR2 of SEQ ID NO: 41, and(c) the light chain ...