ПОЛИПЕПТИДЫ

Настоящее изобретение относится к области биотехнологии, конкретно к сконструированным полипептидам со связывающей сывороточный альбумин аффинностью, и может быть использовано в медицине. Получают полипептид, связывающий сывороточный альбумин, включающий последовательность: LA[Е,S,Q,С]AK[Е,S,С][А,S]AN[A,S,R]ELD[A,S,С,K]YGVSDFYKRLI[D,Е]KAKTVEGVEALK[D,Е][А,Е]IL[А,K][A,S,E]LP, где L45 и P46 присутствует или отсутствует, гибридный белок и конъюгат с полипептидом с необходимой биологической активностью для увеличения времени полужизни в крови и/или снижения иммуногенности второго полипептида, а также полинуклеотиды, кодирующие полученный полипептид и гибридный белок, и композицию для повышения растворимости соединения в воде. Изобретение позволяет получить стабильный полипептид, связывающий сывороточный альбумин, с улучшенной аффинностью к человеческому сывороточному альбумину и сниженной иммуногенностью, способный увеличить время полужизни в крови и/или снизить иммуногенность полипептида с ...

СПОСОБ ПОЛУЧЕНИЯ ПОЛИПЕПТИДОВ В БЕСКЛЕТОЧНОЙ СИСТЕМЕ

Способ синтеза полипептидов в эукариотических и прокариотических бесклеточных системах основан на модификации способов синтеза с использованием клеточных лизата или экстракта в режимах непрерывного потока или непрерывного обмена, в которых наряду с поддержанием процесса синтеза за счет ввода в реакционную смесь компонентов, поддерживающих синтез, и вывода из реакционной смеси низкомолекулярных компонентов, ингибирующих синтез, осуществляют непрерывное изменение концентрации одного или нескольких компонентов, которые выбирают из группы Mg+2, К+, NTP, полиаминов или их комбинаций и которые определяют эффективность синтеза в заданном диапазоне изменения концентраций. Способ не требует применения дорогостоящего фермента Т7 экзогенной полимеразы, что обуславливает соответствующую экономичность получаемых продуктов синтеза. 6 з.п. ф-лы, 13 ил., 6 табл.

Ингибиторы фактора VIIa

Изобретение относится к группе новых соединений общей формулы I R1-A-B-D-En-R2 (I), в которой R1 представляет собой R12C(O), причем R12 выбран из группы, включающей алкенил, алкенилокси или алкениламино; А представляет собой группу А1-А2-А3, в которой A1 представляет собой NH, А2 представляет собой CHR93, в которой R93 представляет 4-амидинофенилметил; A3 представляет собой C(O); В представляет собой группу В1-В2-В3, в которой B1 представляет собой NH; B2 представляет собой CHR97, где R97 представляет этил, который замещен в положении 2 гидроксикарбонилом или алкилоксикарбонилом; В3 представляет собой C(O); D представляет группу D1-D2-D3, в которой D1 представляет собой NH, D2 представляет собой CR81R82, в которой R81 и R82 независимо выбраны из группы, состоящей из водорода и незамещенных или замещенных остатков алкила, арила, арилалкила, гетероарилалкила; D3 представляет собой С(О); Enпредставляет собой (E1-Е2-Е3)n, в которой n равно 0 или 1; E1 представляет NR70, где R70 представляет ...

СОЛЬ ПЕПТИДНОЙ ВАКЦИНЫ, СПОСОБ ЕЕ ПОЛУЧЕНИЯ И ФАРМАЦЕВТИЧЕСКИЙ ПРЕПАРАТ, СОДЕРЖАЩИЙ ЭТУ СОЛЬ

Изобретение относится к области фармацевтической технологии и относится к ацетату KIFGSLAFL в форме твердого аморфного вещества, способу его получения и фармацевтическому препарату пептидной вакцины, содержащему указанный ацетат. Ацетат KIFGSLAFL обладает хорошей чистотой, стабильностью и растворимостью в воде. 3 н. и 2 з.п. ф-лы, 3 ил., 13 пр.

СПОСОБ СИНТЕЗА ТЕРАПЕВТИЧЕСКИХ ПЕПТИДОВ

Изобретение относится к способу крупномасштабного синтеза терапевтических пептидов, представляющих собой аналоги грелина, методом поэтапной твердофазной Fmoc-химии с использованием амидной смолы Зибера. 13 з.п. ф-лы, 3 ил., 2 пр.

АВТОМАТИЗИРОВАННАЯ РЕАКТОРНАЯ СИСТЕМА ДЛЯ СИНТЕЗА С КОНТУРОМ РЕЦИРКУЛЯЦИИ

Настоящее изобретение относится к автоматизированной системе реакторов для проведения твердофазного пептидного синтеза, и более конкретно, к синтезатору пептидов в твердой фазе, автоматизированному посредством реактора с контуром рециркуляции жидкости, позволяющим в реальном времени измерять химические вещества в реакторе посредством измерительных ячеек. Реакторная система для проведения твердофазного пептидного синтеза содержит реактор (9) для сборки, входную трубу (1), предназначенную для введения смолы в реактор (9) для сборки, входную трубу (2), предназначенную для введения растворителя для синтеза и промывки в реактор (9) для сборки, входную трубу (3), предназначенную для введения средства снятия защиты подаваемой аминокислоты в реактор (9) для сборки, входную трубу (4), предназначенную для введения реагентов в реактор (9) для сборки, вход (6), предназначенный для введения в реактор (9) для сборки аминокислоты, которая предварительно активирована, контур (10) рециркуляции жидкости ...

ПЕПТИД ИЛИ ЕГО ОРГАНИЧЕСКИЕ ИЛИ НЕОРГАНИЧЕСКИЕ СОЛИ, СПОСОБ ПОЛУЧЕНИЯ, ФАРМАЦЕВТИЧЕСКАЯ КОМПОЗИЦИЯ, СПОСОБЫ СТИМУЛИРОВАНИЯ ВЫСВОБОЖДЕНИЯ ГОРМОНА РОСТА И УВЕЛИЧЕНИЯ ЕГО СОДЕРЖАНИЯ В КРОВИ

Изобретние относится к новым пептидам формулы 1, где А1, А2, А5, С1, С2, C3 представляют собой различные аминокислотные остатки или (низший алкил) замещенные остатки аминокарбоновых кислот, определенные в п.1 формулы изобретения. Раскрыты также их соли, способ получения, фармацевтическая композиция на основе пептидов, а также способ стимулирования высвобождения гормона роста и увеличения его содержания в крови. Соединения фopмулы 1 могут быть использованы в терапевтических целях для лечения заболеваний, где необходимо повышение уровней гормона роста, а также для увеличения надоев молока и повышения массы тела у животных. 5 с. и 14 з.п. ф-лы, 7 табл., 3 ил. А1 - А2 - С1 - С2 - C3 - А5 (1) ...

АНТАГОНИСТЫ LHRH, ИХ ПОЛУЧЕНИЕ, ФАРМАЦЕВТИЧЕСКАЯ КОМПОЗИЦИЯ И ЕЕ ПОЛУЧЕНИЕ, СПОСОБ ЛЕЧЕНИЯ ОПУХОЛЕЙ И БЕСПЛОДИЯ У МЛЕКОПИТАЮЩИХ

Изобретение относится к антагонистам LHRH - декапептидам общей формулы I: AXxx1-Xxx2-Xxx3-Xxx4-Xxx5-Xxx6-Xxx7-Xxx8-Xxx9-Xxx10-NH2, где А - ацетильная группа, Xxx1=D-Nal(2), Xxx2=D-Cpa, Xxx3=D-Pal(3), Xxx4=Ser, Xxx5=N-Me-Tyr, Xxx6 означает D-Cit, D-Hci, D-[ε-N1-4-(4-амидинофенил)амино-1,4-диоксобутил]-Lys [D-Lys(B)], Xxx7=Leu, Nle; Xxx8=Arg, Lys (iPr); Xxx9=Pro, Xxx10=D-Ala или Sar, при условии, что если Xxx6=D-Lys(B), то Xxx7 означает Nle, если Xxx6=D-Cit, то Xxx7=Nle и Xxx10=D-Ala, или если Xxx6 означает D-Hci, то Xxx7 означает Leu и Xxx10 означает D-Ala, а также их солям с фармацевтически приемлемыми кислотами; фармацевтической композиции, способу получения фармацевтической композиции, способу лечения гормонально зависимых опухолей у млекопитающих. Соединения по изобретению обладают повышенной растворимостью в воде. 5 с. и 11 з.п. ф-лы, 2 табл.

СПОСОБ ОТЩЕПЛЕНИЯ ПЕПТИДОВ, СВЯЗАННЫХ С ТВЕРДОЙ ФАЗОЙ, ОТ ТВЕРДОЙ ФАЗЫ

CПOCOБ ПOЛУЧEHИЯ TBEPДOФAЗHOГO PEAГEHTA

Способ получения циклического тетрадекапептида или его солей

ANTIGEN BINDENDE PEPTIDE (ABTIDES) AUS PEPTIDBIBLIOTHEKEN

Herstellung und Durchmusterung von hochgradig diversen Peptidbanken hinschtlich Bindungsaktivität

VERFAHREN ZUR SCHNELLEN SYNTHESE VON GROSSEN ZAHLEN TRAEGERGEBUNDENER ODER FREIER PEPTIDE

The invention concerns a method for the rapid synthesis of substrate-bound or free peptides, apparatus for carrying out the method, a flat material with peptides prepared by the method bound to it, and a use of this material.

IDENTIFIZIERUNG VON AMINOSÄUREN DURCH MASSENSPEKTROMETRIE

SYNTHESE ÄQUIMOLARER MISCHUNGEN VIELZÄHLIGER OLIGOMERE, SPEZIELL OLIGOPEPTIDMISCHUNGEN

PEPTIDE MIT AFFINITÄT ZU GP120, UND IHRE VERWENDUNGEN

PEPTIDE UND VERWANDTE MOLEKÜLE, DIE AN TALL-1 BINDEN

Peptidderivate, wertvoll als Antagonisten von Bombesin und Gastrin-freisetzendem Peptid

Schnellsynthese und Screening von Peptidmimetika

NEUES CADIODILATIN-FRAGMENT, PROZESS ZU DESSEN HERSTELLUNG UND DESSEN ANWENDUNG.

A peptide having the amino acid sequence 95-126 of ANF/CDD 1-126 (gamma-LANaP) having the designation urodilatin (ANF/CDD 95-126) according to formula (I) is claimed. In the formula, R1, R2 = further peptide fragments of ANF/CDD 1-126 (gamma-LANaP), more specifically R1 = Thr-Ala-Pro-Arg-Ser--Arg-Ser-Ser and R2 = Asn-Ser-Phe-Arg-Tyr.

NEUES CADIODILATIN-FRAGMENT, PROZESS ZU DESSEN HERSTELLUNG UND DESSEN ANWENDUNG.

POLYPEPTID UND VERFAHREN ZU SEINER HERSTELLUNG.

Verfahren zur Spaltung von Acylthiohydantoinen und Anwendung davon zur C-terminalen Peptidsequenzierung

VERFAHREN ZUR HERSTELLUNG UND AUSWAHL VON PEPTIDEN MIT SPEZIFISCHEN EIGENSCHAFTEN

Verfahren zur Herstellung von Tritylgruppen enthaltenden Polystyrolharzen

Vorrichtung zum Nachweis von fluoreszenten, der Bindung eines Liganden auf einem Substrat entsprechenden Merkmalen

Antikoagulierende Peptid-Alkohole.

Automatisierter Polypeptidsyntheseapparat.

LEITFÄHIGKEITSMESSUNG IN ORGANISCHEN LÖSUNGSMITTELN.

Ribonukleotidreduktase-Hemmer.

Zyklische Peptide mit aggregationshemmenden Eigenschaften

POLYÄTHYLENGLYKOLDERIVATE ZUR FESTPHASEANWENDUNGEN

ZUR PEPTIDSYNTHESE VERWENDBARE POLYOXETANE

NEUES VERFAHREN ZUR HERSTELLUNG VON PEPTIDEN

ETHYLENE DIAMINE FUNCTIONAL ACRYLIC RESINS AND PREPARATION THEREOF

VERFAHREN ZUR HERSTELLUNG VON AKTIVITAETSSTABILISIERTEN UND IN IHREN PHARMAKOLOGISCHEN EIGENSCHAFTEN MODIFIZIERTEN PEPTIDDERIVATEN

HAPTEN-MARKIERTE PEPTIDE

SCHUTZ- BZW. ANKERGRUPPEN UND DEREN VERWENDUNG (CARBAMIDE)

MODIFIED D-RETRO CYCLIC HEXAPEPTIDE SOMATOSTATIN ANALOGS, PROCESS FOR THEIR PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM

VERFAHREN ZUM AUFBAU VON PEPTIDKETTEN

GNRH ANTAGONISTS

Verfahren und Vorrichtungen zum Aufbringen von Substanzen auf einen Träger, insbesondere von monomeren für die kombinatorische Synthese von Molekülbibliotheken

SYNTHETISCHE ANTIBIOTIKA

MACROMONOMERE AUF DER BASIS VON PEG, DAMIT HERGESTELLTE INERTE POLYMERE UND DEREN VERWENDUNG FÜR ORGANISCHE SYNTHESEN UND ENZYM- REAKTIONEN

VERFAHREN ZUR BILDUNG EINES PROTEIN-MIKROARRAYSYSTEMS

Alpha-N-Methylierung von Aminosäuren

SOLUBILISABLE POLYMER SUPPORT SUITABLE FOR SOLID PHASE PEPTIDE SYNTHSIS & FOR INJECTION INTO EXPERIMENTAL ANIMALS

Process for preparing conjugates of metalloproteins and derivatives thereof

A process for obtaining biologically active metalloprotein-albumin conjugates or metalloprotein-albumin-antibody conjugates in high yield and without loss of biological activity, comprises cross-linking a metal-free apoprotein by means of a cross-linking agent in presence of human albumin and optionally an antibody fragment, purifying said metal-free conjugates and reconstituting the biological activity by adding the desired metals to said conjugates.

SYNTHESIS OF PEPTIDE ALCOHOLS

A process for the production of a peptide alcohol which at the C-terminal end of the peptide chain bears two alcohol groups or one alcohol group and one thiol group, by acidic hydrolysis of an acetal of the peptide alcohol and a resin containing formylphenyl residues. Thus a peptide alcohol (where Y is a peptide residue, X is 0 or S, R is H or methyl) may be prepared by hydrolysis of acetalised resin (where P is an insoluble resin and Z is a band or linking group). The acetalised resin may be prepared by reacting a polymer of formula with a compound of formula (e.g. the N-trifluoromethyl carbonyl-Threoninol acetal of P-formylphenoxyacetic acid).

PROCESS FOR THE MANUFACTURE OF PEPTIDES

... 1,234,982. Peptide synthesis. FARBWERKE HOECHST A.G. 17 June, 1968 [16 June, 1967], No. 28683/68. Heading C2C. [Also in Division C3] Peptides are prepared by synthesis on a solid synthetic surface that is coated with polystyrene and then chloromethylated, the polystyrene coating having been chemically bound to the solid synthetic surface by a graft reaction. Examples describe the preparation of H-leugly-OH.HBr and H-leu-gly-NH 2 .HCl by the above method.

PROCESS FOR BINDING AMINO ACIDS

... 1312920 Synthesizing polypeptides UCB SA 24 July 1970 [25 July 1969] 37542/69 Heading C3P A process for preparing bound amino acids suitable for use in the solid phase synthesis of polypeptides comprises reacting a halomethylated polymer with a salt of a protected amino acid in a solvent which swells the polymer without dissolving it, the salt of the protected amino acid being a salt of a quaternary ammonium hydroxide NR 1 R 2 R 3 R 4 +OH- where R 1 , R 2 , R 3 and R 4 are C 1-18 alkyl, C 5-12 cycloalkyl, phenyl, naphthyl, C 7-16 aralkyl or heterocyclic radicals or at least two thereof are joined to form a heterocyclic radical. Specified hydroxides are tetramethyl, tetraethyl, tetrabutyl, methyl trin-butyl, phenyl trimethyl and benzyl trimethyl ammonium hydroxides, and dodecyl pyridinium and methyl styryl pyridinium hydroxides. The halogen may be chlorine, bromine or iodine, and the preferred polymer is a chloromethylated styrene/divinyl benzene copolymer. Many solvents ...

Monitoring method for the synthesis of a linear of amino acid residues

PCT No. PCT/GB87/00078 Sec. 371 Date Feb. 8, 1988 Sec. 102(e) Date Feb. 8, 1988 PCT Filed Feb. 3, 1987 PCT Pub. No. WO87/04713 PCT Pub. Date Aug. 13, 1987.A monitoring method for application in solid phase peptide synthesis. In one aspect, the synthesis starts with an amino acid residue protected by an N-alpha-amino protecting group and involves the steps: (a) removing the N-alpha-amino protecting group to obtain an N-alpha-amino group, (b) adding an amino acid residue protected by an N-alpha-amino protecting group, via a peptide bond, to the N-alpha-amino group obtained in step (a) by use of a reactive protected amio acid derivative, and (c) repeating steps (a) and (b). The reaction system includes 3-hydroxy-1,2,3-benzotriazin-4(3H)-one or a derivative thereof; and the color of the reaction system or of a component thereof is monitored during the synthesis.

Protective group

The protection group has the following formula (I): Ar-L- where Ar represents a substantially flat system of fused rings containing at least 4 aromatic rings, and L represents a group containing at least one carbon atom capable of bonding to a group to be protected.

Process

TRIPEPTIDES OF THE GENERAL FORMULA

A VALVING SYSTEM

PROCESS FOR THE MANUFACTURE OF PEPTIDES

... 1310656 Peptide process F HOFFMANNLA ROCHE & CO AG 24 Sept 1971 [26 Sept 1970] 44701/71 Heading C2C [Also in Division C3] In a peptide process an amino acid which is linked via an ester or amide bond to a polymer which is soluble in water and/or an organic solvent is stepwise coupled in a homogeneous phase with further amino acids and the polymer removed from the peptide. Polymers have mol. wts. of up to 100,000 and are preferably polyethylene glycol optionally esterified with citric acid or a copolymerizate of vinyl pyrrolidone and acrylic acid #-hydroxyethylacrylate or of acrylamide and vinyl acetate in which the ester groups are partially saponified at least, or a copolymerizate of polyethylene glycol and succinic acid. In the process lower molecular coupling reagents are removed by dialysis, membrane filtration, column chromatography or countercurrent distribution and the polymer is removed from the peptide by treatment with dilute alkali hydroxide. Maleic acid di-(p-nitrophenyl) ester ...

REACTION SYSTEM FOR SEQUENTIALLY TRANSFERRING REAGENTS FROM A PLURALITY OF STORAGE VESSELS

... 1,235,726. Dispensing measured quantities of liquids. DANISH INSTITUTE OF PROTEIN CHEMISTRY. 17 Sept., 1968 [18 Sept., 1967], No. 44160/68. Heading B8N. [Also in Divisions B1-B2] A reaction system for use for example in the solid state synthesis of peptides comprises a plurality of first vessels 111a-111c for reagents, a second vessel 112 which may be a metering vessel, a plurality of conduits 115a- 115c free of valves and each connecting one first vessel to the second vessel, and an automatic control system for controlling the flow of reagents from the first vessels to the second vessel by sequentially selectively applying to the reagents in the first vessels 111a &c. by means of gas pressure in lines 122, 116a &c., a first pressure which is higher than the pressure in the second vessel 112 so that reagent flows into the second vessel, and a second pressure which is lower than the pressure in the second vessel so that reagent ceases to flow to the second vessel and flows back from the ...

PROCESSES FOR THE PRODUCTION OF PEPTIDES AND PROTEINS

... 1,218,459. Peptide derivatives of graft polymers. IMPERIAL CHEMICAL INDUSTRIES OF AUSTRALIA & NEW ZEALAND Ltd., and MONASH UNIVERSITY. 19 Sept., 1968 [21 Sept., 1967], No. 44616/68. Heading C3G. [Also in Division C2] A peptide or protein is prepared by (1) reacting a first protected amino acid which, optionally, may be attached to a peptide or part of a peptide, with a copolymer which is characterized in that it is a graft copolymer of a chemically inert polymeric backbone and grafted-on side chains comprising a multiplicity of mer units of formula wherein X is at least one amino acid reactive group forming a bond with the first amino acid sufficiently strong not to be cleaved during subsequent reaction of said first amino acid or peptide with a second amino acid or peptide and Y stands for one or more optional substituents which is non-reactive to amino acids, to form a stable linkage between the protected amino acid and the copolymer; (2) deprotecting the aminoacid- or peptide-copolymer ...

PROCESS FOR THE PREPARATION OF PEPTIDES

... 1464938 Synthesis of peptides on glycol polymers HOECHST AG 6 Dec 1974 [6 Dec 1973] 52987/74 Heading C3H A process for the preparation of a peptide comprises condensing a suitably protected amino acid or a peptide with a free amino group of an otherwise suitably protected amino acid or a peptide bound via an ester linkage formed from a carboxyl group of the amino acid or peptide to an unsubstituted, monoacylated or monoalkylated homopolymer of ethylene glycol or propylene glycol or copolymer of ethylene glycol and propylene glycol, said homopolymer or copolymer having a molecular weight of from 2000-40,000, separating the resulting polymerpeptide complex by crystallisation from an organic solvent or mixture of organic solvents, and splitting the peptide from the complex. The reaction may be effected stepwise, and the peptide-polymer complex purified after each step. In the Examples, H-Leu-Ala-Gly-Val-OH is obtained by stepwise reaction of BOC-protected amino acids with H-Val-PEG and its ...

Solid-phase N-terminal peptide capture and release

A method of storing or stabilizing a plurality of peptides, proteins, or combinations thereof, comprising using a plurality of supports comprising a plurality of capture moieties to capture said peptides, proteins, or combinations thereof, wherein a capture moiety of said plurality of capture moieties (i) is not an antibody or (ii) comprises an aromatic carboxaldehyde or a heteroaromatic carboxaldehyde. Preferably, said capture moiety of said plurality of capture moieties is a 2-pyridinecarboxaldehyde or a derivative thereof. The plurality of supports preferably comprises solid or semi-solid supports and/or comprises beads, gel beads, or resins. Preferably, said plurality of supports comprises a barcode, more preferably wherein said barcode is coupled to the support through a linker or is directly coupled to the support. Even more preferably, the barcode comprises DNA, RNA, PNA, or any combination thereof. Most preferably, plurality of supports comprises a unique barcode which is associated ...

2-Adrenergic receptor agonists

Process for the preparation of resin-bound cyclic peptides

Process for the preparation of resin-bound cyclic peptides.

This invention is directed to a process for the solid phase, fragment-based sythesis of resin bound cyclic peptide analogs of parathyroid hormones and analogs of parathyroid hormone-related proteins, which analogs contain at least one bridge between the side chains of two non-adjacent amino acid residues, and to peptide fragments useful therefor.

Process for the preparation of resin-bound cyclic peptides

2-Adrenergic receptor agonists

Methods for designing and preparing vaccines comprising directed sequence polymer compositions via the directed expansion of epitopes

METHODS FOR DESIGNING AND PREPARING VACCINES COMPRISING DIRECTED SEQUENCE POLYMER COMPOSITIONS VIA THE DIRECTED EXPANSION OF EPITOPES

b2- Adrenergic receptor agonists.

"Synthèse peptidique en phase solide".

Process for the preparation of resin-bound cyclic peptides.

2-Adrenergic receptor agonists

Process for the preparation of resin-bound cyclic peptides

Methods for designing and preparing vaccines comprising directed sequence polymer compositions via the directed expansion of epitopes

HEMOREGULATORY PEPTIDES

HEMOREGULATORY PEPTIDES

Methods for designing and preparing vaccines comprising directed sequence polymer compositions via the directed expansion of epitopes

PROCEDURE FOR THE PRODUCTION OF A SENSOR

PROCEDURE FOR THE FORMATION OF A PROTEIN OF MICRO ARRAY SYSTEM

SCREENING FROM COMBINATORIAL PEPTIDBIBLIOTHEKEN TO THE SELECTION FROM PEPTIDLIGANDEN TO THE USE IN THE AFFINITY CLEANING OF GOAL PROTEINS

PROCEDURE FOR THE PRODUCTION OF A NEW TETRA DECAPEPTIDS AND ITS ACID ADDITION SALTS

CYCLIC SOMATOSTATIN SIMILAR ONES WITH A MAIN CHAIN WITH A STRAINED ONE KONFORMATION

PROCEDURE FOR THE PRODUCTION OF POLYMERE ONES FIXED PHASE CARRIERS

PEPTIDE WITH AFFINITY FOR GP120, AND YOUR USES

TOPOLOGICALLY SEPARATIONS, CODING FIXED PHASE LIBRARIES

PROCEDURE FOR THE PROOF AND FOR THE ANALYSIS OF MACROMOLECULAR COMPLEXES

NUCLEIC ACID BINDING PROTEINS

REACTOR FOR THE FIXED PHASE SYNTHESIS

HYBRID PROCEDURE FOR THE PRODUCTION OF CARRIERS FOR THE ANALYTBESTIMMUNG

PROCEDURE FOR THE PRODUCTION OF NEW POLYPEPTIDE

PROCEDURE FOR THE BACK FOLDING OF CHEMICALLY MANUFACTURED POLYPEPTIDEN

INHIBITORS THE HUMAN ELASTASE OF NEUTROPHILEN

MORE IMPROVED EPITOP PRESENTING PHAGE

NACHSPALTIGE SULFURENTSCHÜTZUNG FOR CONVERGENT PROTEIN PRODUCTION WHEN PROCEEDING CHEMICAL LIGATION

PEPTID CYCLISIERUNG ON RESIN

POROUS RESIN PARTICLES WITH HYDROXY GROUP OR PRIMARY AMINO GROUP AND MANUFACTURING PROCESS FOR IT

STENCILS FROM MODIFIED GROWTH FACTORS TO THE STRUCTURE OF FABRIC

Whole proteome tiling microarrays

The present invention relates to a microarray comprising at least 50,000 oligopeptide features per cm 2 where the oligopeptide features represent at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% of the proteome of a virus or an organism. The present invention further relates to methods for the synthesis of such microarrays and methods of using microarrays comprising at least 50,000 oligopeptide features per cm 2 . In an embodiment of the invention, the oligopeptide features represent proteins expressed in the same species, wherein the oligopeptide features are presented in a tiling pattern representing at least about 5,000 to-at least about 25,000 proteins expressed in a species. In some embodiments, the oligopeptide microarray features represent proteins expressed in the same species, wherein the microarray features are present in a tiling pattern that represents at least about 5,000 to at least about 50,000 expressed proteins.

Arrays Of Biological Membranes And Methods And Use Thereof

The present invention overcomes the problems and disadvantages associated with prior art arrays by providing an array comprising a plurality of biological membrane microspots associated with a surface of a substrate that can be produced, used and stored, not in an aqueous environment, but in an environment exposed to air under ambient or controlled humidities. Preferably, the biological membrane microspots comprise a membrane bound protein. Most preferably, the membrane bound protein is a G-protein coupled receptor, an ion channel, a receptor serine/threonine kinase or a receptor tyrosine kinase.

NOVEL PROCESS FOR THE SYNTHESIS OF 37-MER PEPTIDE PRAMLINTIDE

A process for the production of pramlintide, a 37-mer peptide, is provided. The synthesis provides a high yield synthesis of the peptide in relatively pure form. Further purification can be achieved by preparative HPLC. 2. The process of claim 1 , wherein the acid-labile resin is selected from the group consisting of Rink amide AM resin claim 1 , 5-(4-N-Fmoc-aminomethyl-3 claim 1 ,5-dimethoxyphenoxy)valeryl-(PAL resin) claim 1 , Nova PEG-Rink amide claim 1 , NovaSyn TG R resin claim 1 , Rink amide MBHA resin claim 1 , Rink Amide NovaGel claim 1 , and Rink amide PEGA resin.3. The process of claim 2 , wherein the resin is Rink amide AM resin.4. The process of claim 1 , wherein the acid-labile resin has a substitution value ranging from 0.25 to 0.35 mmole/gm.5. The process of claim 1 , wherein the polar aprotic solvent used is dimethyl formamide (DMF).6. The process of claim 1 , wherein the (X) group is selected from the group consisting of tBu claim 1 , Trt claim 1 , and chlorotrityl claim 1 , and the (Y) group is selected from the group consisting of Trt claim 1 , Tmob claim 1 , Mtt claim 1 , and Xan.7. The process of claim 6 , wherein the (X) group is tBu.8. The process of claim 6 , wherein the (Y) group is Trt.9. The process of claim 1 , wherein Fmoc removal is carried out in a medium comprising 0.1M-0.5M-hydroxybenzotriazole (HOBt) in 20%-50% piperidine in DMF.10. The process of claim 1 , wherein the peptide of Formula (4) is subjected to air oxidation claim 1 , or hydrogen peroxide or copper sulphate or iodine oxidation claim 1 , to yield the peptide of Formula (1).11. The process of claim 1 , wherein step (g) contacting with acid comprises contacting with trifluoracetic acid.12. The process of claim 1 , further comprising converting the peptide of formula (1) to an acetate salt thereof.13. The process of claim 1 , wherein the peptide of formula (1) has a purity of 98% claim 1 , or of 99% claim 1 , or of 99.9% by weight.14. The peptide of formula (prepared by the ...

CHEMICAL SYNTHESIS USING UP-CONVERTING PHOSPHOR TECHNOLOGY AND HIGH SPEED FLOW CYTOMETRY

The invention offers the ability to rapidly synthesize multiple chemical compounds, particularly polymers of varying sequences, in parallel on the surfaces of carrier beads. Tinvention involves attaching up-converting phosphors (UCP's) to beads to create up-converting phosphor-loaded beads (UCP-loaded beads) with unique spectral characteristics. Using a dynamic sorting architecture each bead is cataloged based on its spectral characteristics, assigned a compound or polymer to be synthesized, and subjected to multiple rounds of sorting by a flow cytometer, wherein each round sorts the bead to an appropriate bin for a selected chemical reaction, such as the attachment of a monomeric subunit of the polymer sequence. 1. A carrier bead having a generally spherical shape and a layer of at least one up-converting phosphor particle on the bead's surface.2. A bead according to claim 1 , wherein the bead has a metallic layer between the bead surface and the up-converting phosphor particle layer.3. A bead according to claim 1 , wherein the bead is a ceramic bead.4. A bead according to claim 1 , having an external coating encapsulating the bead and up-converting phosphor particle layer.5. A bead according to claim 4 , wherein the external coating is a silica coating claim 4 , a glass coating claim 4 , or a ceramic coating.6. A bead according to claim 1 , wherein the up-converting phosphor particle layer comprises at least two up-converting phosphor particles having distinct emission wavelengths.7. A bead of claim 1 , wherein the diameter of the bead core is any diameter up to about 20 μm.8. A bead of claim 7 , wherein the up-converting phosphor particles have a diameter of at least 50 nm claim 7 , at least 75 nm claim 7 , at least 100 nm claim 7 , or at least 300 nm.9. A bead according to claim 8 , having an external coating encapsulating the bead and up-converting phosphor particle layer.10. A bead according to claim 9 , wherein the external coating is a silica coating claim 9 ...

Method For Synthesizing A Cyclic Multivalent Peptide Using A Thiol-Mediated Reaction

A method has been developed for the formation of multivalent cyclic peptides. This procedure exploits on-resin peptide cyclization using a photoinitiated thiol-ene click reaction and subsequent clustering using thiol-yne photochemistry. Both reactions utilize the sulfhydrl group on natural cysteine amino acids to participate in the thiol-mediated reactions. 1. A method of forming a cyclic peptide , said method comprising the steps of:(a) providing a linear peptide having at least one free thiol group and at least one unsaturated carbon-carbon bond; and(b) forming the cyclic peptide in a radical-mediated reaction wherein said at least one free thiol group reacts with said at least one unsaturated carbon-carbon bond.2. The method of claim 1 , wherein said linear peptide is attached to a solid support.3. The method of claim 1 , wherein said at least one unsaturated carbon-carbon bond comprises at least one carbon-carbon double bond.4. The method of claim 1 , wherein said radical-mediated reaction is a photoreaction.5. The method of claim 1 , further comprising the step of treating said linear peptide with a photoinitiator.6. The method of claim 2 , further comprising the step of cleaving the cyclized peptide from the solid support.7. The method of claim 1 , wherein said linear peptide comprises one or more cysteine residue.8. The method of claim 1 , wherein said linear peptide has a formula of:{'br': None, 'sub': x', 'y', 'z', 'x', 'y', 'z, '(Aaa)-Aaa(SH)-(Aaa)-Aaa(R)-(Aaa)or (Aaa)-Aaa(R)-(Aaa)-Aaa(SH)-(Aaa),'}wherein R is a side chain comprising an unsaturated carbon-carbon bond, and Aaa(SH)is an amino acid residue having at least one free thiol group, x and z each being an integer between 0 and 100, y being an integer between 2 and 100.9. The method of wherein R comprises a linear alkene or a cyclic alkene.10. The method of wherein R comprises a linear alkyne or a cyclic alkyne.11. The method of wherein R comprises an alkene selected from the group consisting of ...

MODIFIED PEPTIDE DISPLAY

The invention provides a replicable genetic package displaying a cyclic peptide having at least one intramolecular bond between amino acid side chains. Also provided are a method of preparing such a genetic package displaying cyclic peptides having at least one intramolecular bond. Further provided is a library of replicable genetic packages displaying cyclic peptides each having at least one intramolecular cyclic bond between amino acid side chains; and a method of producing such a library. 1. A replicable genetic package displaying a peptide having a plurality of amino acid side chains and at least one intramolecular cyclic bond between two of the amino acid side chains.2. The replicable genetic package according to claim 1 , which is selected from the group consisting of a phage particle claim 1 , a bacterium claim 1 , a yeast claim 1 , a fungus claim 1 , a spore of a microorganism and a ribosome.3. The replicable genetic package according to claim 1 , which is selected from the group consisting of M13 claim 1 , T4 claim 1 , T7 claim 1 , fd and lambda phages.4. The replicable genetic package according to claim 1 , wherein said peptide is a precursor peptide comprising a leader and a core peptide.5. The replicable genetic package according to claim 4 , wherein said core peptide is derived from a natural ribosomal peptide.6. The replicable genetic package according to claim 1 , wherein said cyclic bond is linking two atoms independently selected from the group consisting of C claim 1 , N claim 1 , O and S.7. The replicable genetic package according to claim 6 , wherein said cyclic bond is linking two different atoms.8. The replicable genetic package according to claim 1 , wherein said peptide comprises a polycyclic structure.9. The replicable genetic package according to claim 1 , wherein said peptide comprises a randomised amino acid sequence.10. A method of preparing a replicable genetic package displaying a peptide having a plurality of amino acid side chains ...

Stabilized Compounds Having Secondary Structure Motifs

The present invention provides novel stabilized crosslinked compounds having secondary structure motifs, libraries of these novel compounds, and methods for the synthesis of these compounds libraries thereof. The synthesis of these novel stabilized compounds involves (1) synthesizing a peptide from a selected number of natural or non-natural amino acids, wherein said peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and (2) contacting said peptide with a reagent to generate at least one crosslinker and to effect stabilization of a secondary structure motif. The present invention, in a preferred embodiment, provides stabilized p53 donor helical peptides. Additionally, the present invention provides methods for disrupting the p53/MDM2 binding interaction comprising (1) providing a crosslinked stabilized α-helical structure; and (2) contacting said crosslinked stabilized α-helical structure with MDM2.

TOTAL CHEMICAL SYNTHESIS OF UBIQUITIN, UBIQUITIN MUTANTS AND DERIVATIVES THEREOF

The present invention relates to the field of total chemical synthesis of ubiquitin and related peptides. More in particular, a method is provided of solid phase synthesis of ubiquitin, ubiquitin mutants and derivatives thereof. It was the object of the present invention to provide an approach for the total chemical synthesis of ubuiqitin, which allows for the chemical synthesis of virtually any Ub mutant and giving high overall efficiency and purity. The present inventors have surprisingly found that this object can be realized with a method relying on incorporation of special amino acid building blocks. This approach was found to allow for exceptionally high yields of up to 14% and to provide an synthetic entry into virtually any ubiquitin derivative. 117.-. (canceled)19. The method according to claim 18 , wherein each amino acid pair added as an amide protected building block is separated from any proline residue by at least 4 amino acids.20. The method according to claim 18 , wherein claim 18 , in step (a) claim 18 , at least five amino acid pairs are added during synthesis in the form of a building block claim 18 , wherein the amino acid pairs are separated from each other by at least two amino acids and are selected from the pairs at positions 6-7; 8-9; 11-12; 13-14; 21-22; 46-47; 52-53; 56-57; and 65-66 of ubiquitin sequence (SEQ ID no. 1) or from corresponding pairs of a ubiquitin mutant sequence.21. The method according to claim 18 , wherein claim 18 , in step (a) claim 18 , at least six amino acid pairs are added during synthesis in the form of a building block claim 18 , wherein the amino acid pairs are separated from each other by at least two amino acids and are selected from the pairs at positions 6-7; 8-9; 11-12; 13-14; 21-22; 46-47; 52-53; 56-57; and 65-66 of ubiquitin sequence (SEQ ID no. 1) or from corresponding pairs of a ubiquitin mutant sequence.22. The method according to claim 18 , wherein claim 18 , in step (a) claim 18 , amino acid pairs at ...

METHODS AND SYSTEMS FOR GENERATING, VALIDATING AND USING MONOCLONAL ANTIBODIES

Provided herein is a library of antibodies, wherein the library of antibodies can comprise a plurality of monoclonal, monospecific, or immunoprecipitating antibodies. Also provided herein is a method for producing and using the library of antibodies. 1. A library of antibodies comprising: a plurality of different antibodies , wherein at least 10% of the plurality is produced by the same platform , and wherein each antibody of the plurality of antibodies produced by the same platform(a) is a monospecific antibody,{'sup': '−7', 'sub': 'D', '(b) has a binding affinity of at least 10M (K) for its target protein,'}(c) is an immunoprecipitating antibody, or(d) binds a native form of its target protein.2. The library of claim 1 , wherein at least 20% claim 1 , 30% claim 1 , 40% claim 1 , 50% claim 1 , 60% claim 1 , 70% claim 1 , 80% claim 1 , 90% claim 1 , 95% claim 1 , 99% or 100% of the plurality are antibodies produced by the same platform.36.-. (canceled)7. The library of claim 1 , wherein the plurality comprises at least 50 claim 1 , 75 claim 1 , 100 claim 1 , 125 claim 1 , 150 claim 1 , 175 claim 1 , 200 claim 1 , 225 claim 1 , 250 claim 1 , 275 claim 1 , 300 claim 1 , 325 claim 1 , 350 claim 1 , 375 claim 1 , 400 claim 1 , 425 claim 1 , 450 claim 1 , 475 claim 1 , 500 claim 1 , 525 claim 1 , 550 claim 1 , 575 claim 1 , 600 claim 1 , 625 claim 1 , 650 claim 1 , 675 claim 1 , 700 claim 1 , 725 claim 1 , 750 claim 1 , 775 claim 1 , 800 claim 1 , 825 claim 1 , 850 claim 1 , 875 claim 1 , 900 claim 1 , or 1000 different antibodies.8. (canceled)9. The library of claim 1 , wherein the plurality binds at least 0.5% claim 1 , 1% claim 1 , 2% claim 1 , 3% claim 1 , 4% claim 1 , 5% claim 1 , 6% claim 1 , 7% claim 1 , 8% claim 1 , 9% claim 1 , 10% claim 1 , 11% claim 1 , 12% claim 1 , 13% claim 1 , 14% claim 1 , 15% claim 1 , 16% claim 1 , 17% claim 1 , 18% claim 1 , 19% claim 1 , 20% claim 1 , 25% claim 1 , 30% claim 1 , 35% claim 1 , 40% claim 1 , 45% claim 1 , 50% claim 1 , ...

METHODS OF PREPARING CYCLIC PEPTIDES AND USES THEREOF

This invention is directed to the discovery of improved methods of preparing cyclic peptides, cyclic peptide esters, cyclic peptide amidines, and libraries of these compounds. The invention also includes uses of these compounds and libraries for screens as drugs and binders of biologics. 1. A method for synthesizing a cyclic peptide ligand with selectivity and affinity for a biologic of interest which comprises:(a) synthesizing a solid-phase library of reversible cyclic heterodetic peptides;(b) selecting a reversible cyclic heterodetic peptide that shows selectivity and affinity for the biologic of interest;(c) linearizing and sequencing the selected reversible cyclic heterodetic peptide; and(d) solid-phase synthesizing a cyclic peptide ligand with a sequence corresponding to the selected reversible cyclic heterodetic peptide.2. The method of claim 1 , wherein the reversible cyclic heterodetic peptide is a cyclic depsipeptide.3. The method of claim 1 , wherein the reversible cyclic heterodetic peptide is a cyclic amidine-peptide.4. The method of claim 1 , wherein a plurality of cyclic peptide ligands are synthesized.5. A method for synthesizing a cyclic depsipeptide which comprises:(a) coupling a protected tri-functional molecule with a plurality of protecting groups onto a solid support under suitable conditions;(b) cleaving a protecting group from the protected tri-functional molecule to yield a deprotected tri-functional molecule coupled on the solid support;(c) reacting the deprotected tri-functional molecule coupled on the solid support under suitable conditions so as to link at least one protected amino acid or peptide to the tri-functional molecule;(d) cleaving a protecting group from either (i) the protected amino acid or peptide, or (ii) the tri-functional molecule so as to form a deprotected amino acid or peptide, or a deprotected tri-functional molecule coupled on the solid support;(e) coupling a protected cleavable linker with either (iii) the ...

BOC AND FMOC SOLID PHASE PEPTIDE SYNTHESIS

A solid phase method for synthesizing a peptide containing three or more amino acid residues utilizing both Boc and Fmoc protected amino acids and a chloromethylated polystyrene resin. 1. A method for preparing a peptide comprising three or more amino acid residues having an N-terminal amino acid , a next to last amino acid adjacent to the N-terminal amino acid and a C-terminal amino acid , wherein said method comprises the steps of:(a) attaching a first amino acid to a solid support resin through an ester bond to form a first-coupled-product, which comprises (i) reacting an aqueous solution of cesium carbonate with an alcohol solution of the first amino acid to form a cesium salt of the first amino acid, (ii) obtaining a solvent free cesium salt of the first amino acid, (iii) reacting the solid support resin with the cesium salt of the first amino acid in a dry polar aprotic solvent to form a first-coupled product;wherein the first amino acid corresponds to the C-terminal amino acid of the peptide, the first amino acid's non-side chain amino group is blocked by a Boc and the first amino acid does not have a side chain functionality requiring protection, and the solid support resin is a chloromethylated polystyrene resin;(b) deblocking the Boc from the first-coupled-product with an acid to form a first-deblocked-coupled-product;(c) optionally coupling a next-amino-acid to the first-deblocked-coupled-product, which comprises reacting the next-amino-acid with the first-deblocked-coupled-product in an organic solvent comprising a peptide coupling reagent to form a next-blocked-coupled-product, wherein the next-amino-acid has a non-side chain amino group blocked by Boc and if the next-amino-acid has one or more side chain functionalities then the side chain functionalities do not require protection or the side chain functionalities have protecting groups that are stable to acid and base reagents used to deblock Boc and Fmoc, respectively;(d) deblocking the Boc from the ...

USE OF TRIFLUOROACETAMIDE FOR N-TERMINAL PROTECTION

This disclosure teaches the novel use of trifluoroacetamide for N-terminal protection. The disclosure also teaches novel compositions and chemical structures associated therewith. These methods and compositions are useful for site-specific methylation of peptide backbone amides, performed, for example, to modulate the pharmacokinetic properties of peptide drugs. 1. A method for solid phase peptide synthesis (SPPS) comprising N-terminal protection of an amino acid , the method comprising binding of trifluoroacetamide (Tfac) to the N-terminal of the amino acid.2. The method of further comprising deprotection of the amino acid by the removal of the Tfac group claim 1 , by treatment with excess sodium borohydride in a mixed solvent system.3. The novel use of trifluoroacetamide (Tfac) for N-terminal protection of amino acids claim 1 , dipeptides claim 1 , peptides claim 1 , and ploypeptides during solid phase peptide synthesis.4. A Method for the site specific N-terminal methylation of a resin-bound peptide containing an N-terminal trifluoroacetamide claim 1 , the method comprising treatment of the peptide with an excess of methanol and triphenylphosphine in tetrahydrofuran claim 1 , followed by the addition of excess diisopropyl azadicarboxylate therby yielding >99% N-methylation product within 30 minutes.5. The method of claim 4 , wherein the method employs Mitsunobu methylation of resin-bound Tfac-protected peptides.6. The method of claim 5 , the method comprising the steps of: (i) drying Methanol and DIAD prior to use claim 5 , (ii) rinsing the resin with anhydrous THF claim 5 , (iii) dissolving Triphenylphosphine in anhydrous THF claim 5 , (iv) adding anhydrous methanol and (v) transferring to a synthesis vial containing the resin-bound Tfa-protected peptide claim 5 , and (vi) adding DIAD dropwise with agitation claim 5 , then after the DIAD addition is complete claim 5 , (vii) shaking for about 30 min reaction time.7. The method of further comprising repeating the ...

METHOD FOR PREPARING SERMAGLUTIDE

A method for preparing Sermaglutide. Amino acid protected by Fmoc-Lys(Alloc)-OH is used as a raw material, and protection is carried out by selecting Pd(PPh3)4. In one aspect, the operation process is simple, one or two times of elimination reactions are required only, each time lasts 10 min to 30 min, and no side reaction occurs, and the operation process is safe, so that the preparation method is suitable for expanding production. The risk of His racemization can be reduced to the greatest extent in the process by using Boc-His(Boc)-OH.DCHA and Boc-His(Trt)-OH as raw materials. The synthesis efficiency is improved by performing coupling by using special segments. 1. A preparation method of Sermaglutide , comprising the steps of:step 1: coupling Gly to a resin by solid phase synthesis to obtain Gly-resin; andstep 2: successively coupling the Gly-resin prepared in step 1 to an amino acid according to the sequence of Sermaglutide by sequential coupling; cleaving, and purifying;{'sup': '26', 'wherein Fmoc-Lys(Alloc)-OH is used as a raw material of Lys;'}{'sup': '7', 'H-Hisis synthesized using Boc-His(Trt)-OH or Boc-His(Boc)-OH.DCHA as a raw material; and'}{'sup': '26', 'the Lysside chain is prepared by sequential coupling or fragmental synthesis.'}2. The preparation method according to claim 1 , wherein a fully-protected fragment of Fmoc-PEG-PEG-γ-Glu-octadecanedioic acid used in the fragmental synthesis of the Lysside chain is prepared by successively coupling a chloride resin to Fmoc-PEG-OH claim 1 , Fmoc-PEG-OH claim 1 , Fmoc-Glu(OH)-OtBu and octadecanedioic acid mono-tert-butyl ester claim 1 , cleaving claim 1 , and recrystallizing to obtain PEG-PEG-γ-Glu-Octadecanedioic Acid mono-tert-butyl ester.3. The preparation method according to claim 1 , wherein the resin in step 1 is selected from wang resin or 2-CTC resin;wherein the wang resin has a degree of substitution of 0.1-0.5 mmol/g;the 2-CTC resin has a degree of substitution of 0.2-0.6 mmol/g;the wang resin is ...

SELECTIVE PHOTOACTIVATION OF AMINO ACIDS FOR SINGLE STEP PEPTIDE COUPLING

Disclosed herein are formulations, substrates, and arrays for amino acid and peptide synthesis on microarrays. In certain embodiments, methods for manufacturing and using the formulations, substrates, and arrays including one-step coupling, e.g., for synthesis of peptides in a C→N orientation are disclosed. In some embodiments, disclosed herein are formulations and methods for high efficiency coupling of biomolecules to a substrate. 113.-. (canceled)14. A method of attaching a coupling molecule to a substrate , comprising:obtaining a substrate comprising a plurality of amine groups for linking to a coupling molecule;contacting said substrate with a carboxylic acid activating formulation comprising: a carboxylic acid activating compound, wherein said compound is 1-(3-(diethylamino)-propyl)-4-(2-methoxyphenyl)-1,4-dihydro-5H-tetrazole-5-thione, a coupling molecule, and a solvent;selectively exposing said photoactive coupling formulation to light, thereby activating a carboxylic group of said coupling molecule at a selectively exposed area;coupling the activated carboxylic group of said coupling molecule to at least one of said plurality of amine groups at said selectively exposed area, wherein said coupling is performed multiple times at different selectively exposed areas on said substrate and wherein said coupling step has a coupling efficiency of at least 98.5%; andoptionally repeating said method to produce a desired polymer at said at least one carboxylic acid group.15. The method of claim 14 , wherein said coupling molecule is an amino acid.16. The method of claim 15 , wherein said amino acid has a protecting group attached to an amine group.17. The method of claim 16 , wherein said protecting group is Fmoc.18. A method of attaching a coupling molecule to a substrate claim 16 , comprising:obtaining a substrate comprising a plurality of amine groups for linking to a coupling molecule;contacting said substrate with a carboxylic acid activating formulation ...

PARALLEL ORGANIC SYNTHESIS ON PATTERNED PAPER USING A SOLVENT-REPELLING MATERIAL

The present application is directed to a porous support for parallel organic synthesis comprising: a solvophilic area for spotting an organic solvent comprising a reagent for synthesizing an organic compound. and a solvophobic area that repels the organic solvent. Methods of synthesizing the support and compounds thereon are also provided. 1. A method of fabricating a porous support for performing organic synthesis or biochemical assays in an organic solvent. comprising the steps of:(a) applying a pattern of a hydrophobic material onto the porous support, the pattern defining unmodified areas separated from modified area;(b) protecting the unmodified areas with an aqueous solution;(c) applying a solvophobic material to the modified area;(d) removing the protective aqueous solution to yield a porous support with unmodified areas separated from modified solvophobic area.2. The method of wherein the protective aqueous solution creates a convex droplet covering an unmodifed area.3. The method of claim 1 , wherein the protective aqueous solution is a sucrose solution.4. The method of wherein the solvophobic material is a perfluorinated polymer.5. The method of wherein the hydrophobic material pattern is performed by wax printing.6. The method of wherein the protective aqueous material is an aqueous gel.7. The method of wherein the aqueous gel is agarose.8. The method of wherein the solvophobic material is applied in a solvent. which is removed by evaporation under conditions which prevent evaporation of water.9. The method of wherein the protective aqueous solution is removed by rinsing with water.10. The method of wherein at least one of the patterning step claim 1 , protection step and the solvphobic material application step is automated with a robotic spotter.11. A method of synthesizing one or more compounds on a porous support produced by the method of claim 1 , comprising;(a) applying one or more solvents comprising one or more reagents in an unmodified. area, ...

METHODS AND SYSTEMS FOR GENERATING, VALIDATING AND USING MONOCLONAL ANTIBODIES

Provided herein is a library of antibodies, wherein the library of antibodies can comprise a plurality of monoclonal, monospecific, or immunoprecipitating antibodies. Also provided herein is a method for producing and using the library of antibodies. 152.-. (canceled)53. A method comprising:(a) contacting a plurality of monoclonal antibodies with an array comprising at least 11,000 distinct human proteins, wherein the plurality of monoclonal antibodies are isolated from antibody-generating cells or hybridoma cells generated therefrom, wherein the antibody-generating cells are from a non-human animal immunized with a plurality of native human antigens, wherein the at least 11,000 distinct human proteins are listed in Table 5;(b) measuring an antibody affinity signal intensity for a human protein on the array, wherein the antibody is from the plurality of monoclonal antibodies; and (i) has the affinity signal intensity for the human protein on the array that is greater than 6 standard deviations above a mean affinity signal intensity of the antibody for the at least 11,000 distinct human proteins on the array;', '(ii) has a highest affinity signal intensity for the human protein on the array that is at least 3 times greater than a second-highest signal intensity of the antibody for a human protein on the array; and', {'sup': '−7', 'sub': 'D', 'claim-text': (b) is an immunoprecipitating antibody, or', '(c) binds a native form of the human protein., '(iii) (a) has a binding affinity of 10M (K) or less for the human protein,'}], '(c) selecting the antibody, wherein the antibody54. The method of claim 53 , further comprising pre-screening the plurality of monoclonal antibodies prior to the contacting.55. The method of claim 54 , wherein the pre-screening is by performing immunocytochemistry.56. The method of claim 54 , wherein the pre-screening is by determining binding of antibodies from the antibody-generating cells with a mixture comprising one or more target antigens. ...

METHOD AND COMPOSITION FOR DETECTION OF PEPTIDE CYCLIZATION USING PROTEIN TAGS

This invention relates to peptide microarrays, methods of generating peptide microarrays, and methods of identifying peptide binders using microarrays. More specifically, this invention relates to peptide microarrays, methods of generating peptide microarrays, and methods of identifying peptide binders using microarrays wherein the microarrays comprise cyclic peptides. The invention also relates to methods and compositions for detecting the formation of cyclized peptides from linear peptides on a microarray by contacting the microarray with a detectable protein. The cyclized peptides include tags that are activated upon cyclization, facilitating the detection of successful cyclization reactions. In additional aspects, the invention relates to developing fragmented peptide tags that, upon cyclization, bind to detectable proteins. Additionally, the invention relates to methods of generating linear and cyclic peptides subarrays on a microarray. 1. A method of detecting peptide cyclization comprisinga) generating a peptide microarray comprising at least one linear peptide coupled to a solid support, wherein the at least one linear peptide comprises a peptide tag sequence fragmented across a first end portion and a second end portion of the at least one linear peptide;b) cyclizing the at least one linear peptide to form at least one cyclic peptide by combining the first end portion and the second end portion, wherein a binding interaction of the peptide tag sequence with a detectable target protein increases after the first end portion and the second end portion combine to cyclize the at least one linear peptide;c) contacting the at least one cyclic peptide with the detectable target protein; andd) detecting the presence of the detectable target protein that is bound to the at least one cyclic peptide.2. The peptide microarray of claim 1 , wherein the detectable target protein is an antibody against the peptide tag sequence.3. The peptide microarray of claim 1 , wherein ...

Hplc free purification of peptides by the use of new capping and capture reagents

The present disclosure relates to the use of a capping and capture reagent in solid phase peptide synthesis. The present disclosure further relates to a method of solid phase peptide synthesis, wherein a capping and capture reagent according to the present disclosure is used. The present disclosure further relates to a method for purification of a (full-length) synthetic peptide via use of a capping and capture reagent according to the present disclosure. The present disclosure also relates to a kit comprising a capping and capture reagent according to the present disclosure and an amino oxy resin or a hydrazine resin and the use of the kit.

TRANSFER RNA LIGAND ADDUCT LIBRARIES

The present invention is drawn to, among other things, compositions of matter and methods for producing an aminoacyl-tRNA analogue comprising an adaptor tRNA and modified amino acid for ribosome-directed translation in vitro. 2. An acyl-tRNA analogue capable of ribosome-directed translation having a structure:{'br': None, 'tRNA-A-z-L'}wherein:tRNA has a 3′ terminus to which at least one hydroxyacyl or aminoacyl group may be transferred;A is an aminoacyl or α-hydroxyacyl group selected from the group consisting of canonical amino acids, α-hydroxyl acids, non-canonical amino acids and α-hydroxyl acids, each with an orthogonally reactive moiety y;L is a ligand with a reactive moiety x andz is a covalent linker formed by a reaction of tRNA-A-y with x-L.3. The acyl-tRNA analogue according to claim 2 , where the aminoacyl group comprises at least one non-canonical amino acid with an orthogonally reactive moiety y.4. The acyl-tRNA analogue according to claim 2 , wherein the 3′ cytosine C75 is not 2′-deoxycytosine.5. A method of reacting a starting aminoacyl-tRNA compound represented by a structural formula:{'br': None, 'tRNA-A-y'}wherein A is a non-canonical amino acid with an orthogonally reactive moiety y,with a ligand, x-L, containing a reactive moiety x, under conditions suitable for a reaction, the method comprising forming a covalent linker of tRNA-A-y with x-L, the covalent linker forming a covalently linked product aminoacyl-tRNA analogue tRNA-A-z-L in greater than 50% yield, relative to the starting tRNA-A-y and wherein the product aminoacyl-tRNA analogue is capable of ribosome-directed translation.6. The method according to claim 5 , wherein the conditions suitable for a reaction comprise an acidic pH.7. The method according to claim 6 , wherein the pH is approximately between about 1 and about 5.8. The method according to claim 7 , wherein then pH is about 5.9. The method according to claim 5 , wherein the starting aminoacyl-tRNA is produced substantially pure ...

STABILIZED IMMUNOGLOBULIN DOMAINS

The invention relates autonomous VH domains (aVH) with cysteines in positions 52a and 71 or in positions 33 and 52 in order to stabilize the autonomous VH domains. Said cysteines are capable of forming a disulfide bond and/or form a disulfide bond under suitable conditions. The invention further relates to aVH libraries. 1. An autonomous VH domain with cysteines in positions (i) 52a and 71 or (ii) 33 and 52 according to Kabat numbering , wherein said cysteines are capable of forming a disulfide bond and/or form a disulfide bond under suitable conditions.2. An autonomous VH domain of comprising a substitution selected from the group consisting of 44E claim 1 , 45E and (101-1)Y according to Kabat numbering.3. An autonomous VH domain of comprising the substitutions 44E claim 2 , 45E claim 2 , and (101-1)Y according to Kabat numbering.4. An autonomous VH domain of comprising a substitution selected from the group consisting of G44E claim 1 , T45E and F(101-1)Y according to Kabat numbering.5. An autonomous VH domain of comprising the substitutions G44E claim 4 , T45E claim 4 , and F(101-1)Y according to Kabat numbering.6. An autonomous VH domain of any of to comprising a heavy chain variable domain framework comprising a(a) FR1 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 207,(b) FR2 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 208,(c) FR3 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 209, and(d) FR4 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 210;or(a) FR1 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 211,(b) FR2 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 208,(c) FR3 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 209, and(d) FR4 comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: ...

METHODS FOR GENERATING AND SCREENING COMPARTMENTALISED PEPTIDE LIBRARIES

A method for co-compartmentalising a cyclic polypeptide with a polynucleotide encoding the cyclic polypeptide, comprising the steps of a) forming a compartment containing a polynucleotide encoding the cyclic polypeptide, b) expressing a polypeptide from the polynucleotide, and c) cyclising the polypeptide. Co-compartmentalised cyclic polypeptides and encoding polynucleotides. Libraries of co-compartmentalised cyclic polypeptide and encoding polynucleotide. Methods for screening libraries of co-compartmentalised cyclic polypeptide and encoding polynucleotide. Incorporation of non-canonical nucleic acids into such libraries. 1. A method for co-compartmentalising a cyclic polypeptide with a polynucleotide encoding the cyclic polypeptide , comprising the steps of:a) forming a compartment containing a polynucleotide encoding the cyclic polypeptide;b) expressing a polypeptide from the polynucleotide; andc) cyclising the polypeptide.2. The method according to and further comprising the steps of:d) screening the cyclic polypeptide for activity; ande) selecting the cyclic polypeptide exhibiting a desired activity.3. The method according to claim 1 , further comprising the step of amplifying the polynucleotide.4. The method according to claim 3 , wherein the compartment further comprises a gel-forming agent claim 3 , wherein the gel-forming agent is solidified into a gel bead after the polynucleotide has been amplified.5. The method according to claim 4 , wherein the compartment is disrupted after the gel-forming agent has been solidified into a gel bead.6. The method according to claim 5 , wherein the gel bead is exposed to conditions for expressing the cyclic polypeptide.7. The method according to claim 5 , wherein new compartment is formed around the gel bead.8. The method according to claim 1 , wherein the polynucleotide comprises a sequence encoding an N-terminal intein fragment claim 1 , followed by a sequence encoding the cyclic polypeptide claim 1 , followed by a ...

PHAGE DISPLAY USING COTRANSLATIONAL TRANSLOCATION OF FUSION POLYPEPTIDES

The present invention relates to a filamentous phage display method wherein the polypeptides of interest displayed on the phage particle are cotranslationally translocated across the cytoplasmic membrane of Gram-negative bacteria based on the signal recognition particle pathway. This method is particularly suitable for polypeptides, which are known to be difficult to display on phages, and for proteins of cDNA libraries and other combinatorial libraries, in particular when derived from very fast folding, stable protein scaffolds. The invention further relates to phage or phagemid vectors useful in the method comprising a gene construct coding for a fusion polypeptide comprising the polypeptide to be displayed on the phage particle and an N-terminal signal sequence promoting cotranslational translocation. 115-. (canceled)16. A filamentous phage display method wherein a polypeptide of interest displayed on the phage particle is cotranslationally translocated across the cytoplasmic membrane of Gram negative bacteria using a signal sequence selected from the group consisting of signal sequences of TorT , SfmC , FocC , CcmH , YraI , TolB , NikA , FlgI , and DsbA , and homologs thereof.17. The method of wherein the signal sequence is selected from the group consisting of signal sequences of TorT claim 16 , SfmC claim 16 , TolB and DsbA.18. A phage or phagemid vector comprising a gene construct coding for a fusion protein comprising the polypeptide of interest to be displayed on the phage particle and a signal sequence selected form the group consisting of signal sequences of TorT claim 16 , SfmC claim 16 , FocC claim 16 , CcmH claim 16 , YralI claim 16 , TolB claim 16 , NikA claim 16 , FlgI claim 16 , and Dsba claim 16 , and homologs thereof.19. The vector according to wherein the signal sequence is selected from the group consisting of signal sequences TorT claim 18 , SfmC claim 18 , TolB claim 18 , and Dsba.20. A library of phage or phagemid vectors comprising gene ...

Process for the preparation of guanylate cyclase 2c agonist

The present invention relates to an improved process for the preparation of Linaclotide of Formula I. The process disclosed in the present invention is simple, economical and eco-friendly with reduced reaction times.

SOLID PHASE PEPTIDE SYNTHESIS PROCESSES AND ASSOCIATED SYSTEMS

Systems and processes for performing solid phase peptide synthesis are generally described. Solid phase peptide synthesis is a known process in which amino acid residues are added to peptides that have been immobilized on a solid support. In certain embodiments, the inventive systems and methods can be used to perform solid phase peptide synthesis quickly while maintaining high yields. Certain embodiments relate to processes and systems that may be used to heat, transport, and/or mix reagents in ways that reduce the amount of time required to perform solid phase peptide synthesis. 1. A process for adding amino acid residues to peptides , comprising:providing a plurality of peptides comprising protection groups, each peptide immobilized on a solid support;exposing a deprotection reagent to the immobilized peptides to remove the protection groups from at least a portion of the immobilized peptides;removing at least a portion of the deprotection reagent;exposing activated amino acids to the immobilized peptides such that at least a portion of the activated amino acids are bonded to the immobilized peptides to form newly-bonded amino acid residues; andremoving at least a portion of activated amino acids that do not bond to the immobilized peptides;wherein an amino acid residue is added to at least about 99% of the immobilized peptides during the amino acids exposing step; and the total amount of time taken to perform the combination of all of the deprotection reagent exposing step, the deprotection reagent removal step, the activated amino acid exposing step, and the activated amino acid removal step is about 10 minutes or less and the protection groups comprise fluorenylmethyloxycarbonyl protection groups and/or', 'the total amount of time taken to perform the combination of all of the deprotection reagent exposing step, the deprotection reagent removal step, the activated amino acid exposing step, and the activated amino acid removal step is about 5 minutes or less., ...

Use of Excess Carbodiimide for Peptide Synthesis at Elevated Temperatures

An improved method of coupling amino acids into peptides or peptidomimetics is disclosed in which the activation and coupling are carried out in the same vessel, in the presence of a carbodiimide in an amount greater than 1 equivalent as compared to the amino acid, in the presence of an activator additive, and at a temperature greater than 30° C. 1. In a method of coupling amino acids into peptides or peptidomimetics , the improvement comprising:carrying out activation and coupling in the same vessel;incorporating a carbodiimide in an amount greater than 1 equivalent as compared to the amino acid;in the presence of an activator additive; andat a temperature greater than 30° C.2. A solid phase peptide synthesis (SPPS) method according to in which the activated acid is coupled to at least one other amino acid that is linked to a solid phase resin.3. A solid phase peptide synthesis (SPPS) method according to in which at least one of the added acids is initially Fmoc-protected.4. A method according to wherein the carbodiimide is present in an amount between 1.5 and 4 equivalents compared to the amino acid.5. A method according to carried out a temperature greater than 70° C.6. A method according to limited to a total coupling time less than 10 minutes.7. A method according to limited to a total coupling time less than 15 minutes.8. A method according to wherein the activator additive is present in an amount between 1 and 1.5 equivalents compared to the amino acid.9. A method according to wherein the carbodiimide is present in an amount to between 1.5 and 4 equivalents compared to the amino acid.10. A composition for coupling amino acids into peptides or peptidomimetics claim 8 , the composition comprising:an amino acid;an organic solvent;a carbodiimide in an amount greater than 1 equivalent as compared to said amino acid; andan activator additive.11. The composition of claim 10 , wherein said composition further comprisesbetween 1 and 1.5 equivalents of said activator ...

Biopolymer synthesis system and method

The present invention provides improved automated systems and methods for synthesis of biopolymers including DNA and RNA. The automated systems and methods represent a number of improvements over existing systems for multiplex synthesis of biopolymers in a combinatorial fashion.

METHOD OF IMPROVING CHARACTERISTICS OF PROTEINS

The invention provides efficient methods for combining single-substitution libraries of nucleic acids that span and encode proteins of interest and for selecting resultant mutant proteins after expression which have improved properties or characteristics. Specifically, the methods comprising synthesizing a single substitution library for each of a plurality of domains of a protein; expressing separately each member of each single substitution library as a pre-candidate protein; selecting members of each single substitution library which encode pre-candidate proteins which exhibit an improvement in the one or more predetermined characteristics to form a selected library; shuffling members or the selected libraries in a PCR to produce a combinatorial shuffled library; expressing members of the shuffled library as candidate proteins; and selecting mutant proteins which have improved properties or characteristics. 1. A method of improving one or more predetermined characteristics of a protein , the method comprising the steps of:synthesizing a single substitution library for each of a plurality of domains of a protein, each member of a single substitution library having a nucleotide sequence that overlaps a nucleotide sequence of at least one member of a different single substitution library;expressing separately each member of each single substitution library as a pre-candidate protein;selecting members of each single substitution library which encode pre-candidate proteins which exhibit an improvement in the one or more predetermined characteristics to form a selected library for each domain of the protein;shuffling members of the selected libraries in a PCR to produce a combinatorial shuffled library;expressing members of the shuffled library as candidate proteins; andselecting members of the shuffled library which encode candidate proteins which exhibit an improvement in at least one of the one or more predetermined characteristics.2. The method of wherein said ...

Stabilized Compounds Having Secondary Structure Motifs

The present invention provides novel stabilized crosslinked compounds having secondary structure motifs, libraries of these novel compounds, and methods for the synthesis of these compounds libraries thereof. The synthesis of these novel stabilized compounds involves (1) synthesizing a peptide from a selected number of natural or non-natural amino acids, wherein said peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and (2) contacting said peptide with a reagent to generate at least one crosslinker and to effect stabilization of a secondary structure motif. The present invention, in a preferred embodiment, provides stabilized p53 donor helical peptides. Additionally, the present invention provides methods for disrupting the p53/MDM2 binding interaction comprising (1) providing a crosslinked stabilized α-helical structure; and (2) contacting said crosslinked stabilized α-helical structure with MDM2.

CONDITIONED SURFACES FOR IN SITU MOLECULAR ARRAY SYNTHESIS

Described herein are in situ synthesized arrays and methods of making them, wherein array signal sensitivity and robustness is enhanced by carrying out conditioning steps and/or generating linkers during synthesis. An array comprises a surface with a collection of features, wherein the features comprise molecules or polymers attached to the surface. In certain embodiments of the invention, carrying out conditioning steps during array synthesis can yield arrays with improved signal. In other embodiments, linkers are synthesized on the array surface prior to synthesis of functional molecules, wherein increasing linker length can correspond to an improvement in the signal generated by the array. 1a. performing a conditioning step on a surface of the array in the absence of monomers;b. repeating step (a) at least once;c. performing a synthesis step upon the surface to add at least one monomer; andd. repeating step (c) at least once to form a sequence, wherein the conditioning step performed prior to synthesizing the sequence enhances attachment of the sequence to the surface of the array.. A method for making an array comprising: This application is a continuation of U.S. patent application Ser. No. 15/264,426, filed Sep. 13, 2016 with claims the benefit of U.S. Provisional Application No. 62/218,418, filed Sep. 14, 2015, each of which is incorporated herein by reference in its entirety.This invention was made with government support under MCB-1243082 awarded by the National Science Foundation. The government has certain rights in the invention.The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Nov. 28, 2016, is named 42206-708_201_SL.txt and is 3,456 bytes in size.Array technologies allow for large-scale, quantitative analyses of biological samples. However, the density and robustness of such technologies should be improved to ...

STABILIZED COMPOUNDS HAVING SECONDARY STRUCTURE MOTIFS

The present invention provides novel stabilized crosslinked compounds having secondary structure motifs, libraries of these novel compounds, and methods for the synthesis of these compounds libraries thereof. The synthesis of these novel stabilized compounds involves (1) synthesizing a peptide from a selected number of natural or non-natural amino acids, wherein said peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and (2) contacting said peptide with a reagent to generate at least one crosslinker and to effect stabilization of a secondary structure motif. The present invention, in a preferred embodiment, provides stabilized p53 donor helical peptides. Additionally, the present invention provides methods for disrupting the p53/MDM2 binding interaction comprising (1) providing a crosslinked stabilized α-helical structure; and (2) contacting said crosslinked stabilized α-helical structure with MDM2. 111-. (canceled)12. A method for synthesizing a peptide comprising a cross-link , the method comprising:(a) synthesizing an amino acid sequence, wherein the amino acid sequence comprises a first amino acid comprising a first moiety and a second amino acid comprising a second moiety, and wherein the first and second moieties are reactive toward one another in the presence of a catalyst; and(b) reacting the amino acid sequence in conditions sufficient to promote a reaction between the first and second moieties toward each other in the presence of the catalyst, thereby resulting in formation of the cross-link in the peptide;wherein the peptide disrupts binding of p53 to MDM2.13. The method of claim 12 , wherein the peptide is stabilized in comparison to a corresponding uncross-linked peptide.14. The method of claim 12 , wherein the peptide comprises an MDM2 binding site of a p53 donor helix.15. The method of claim 12 , wherein the cross-link comprises a double bond.16. The method of claim 15 , wherein the double ...

METHODS OF GENERATING NANOARRAYS AND MICROARRAYS

The methods described herein provide a means of producing an array of spatially separated proteins. The method relies on covalently attaching each protein of the plurality of proteins to a structured nucleic acid particle (SNAP), and attaching the SNAPs to a solid support. 1. A method of producing an array of spatially separated proteins , the method comprising:obtaining a solid support with an array of attachment sites;obtaining a sample comprising a plurality of proteins;obtaining seeds, wherein at least one or more seeds are labeled with a functional group respectively;growing seeds to a plurality of structured nucleic acid particles (SNAPs), wherein said one or more SNAPs comprise substantial internal complementarity and wherein one or more proteins of said plurality of proteins are configured to attach to one or more seeds via said functional group to form a plurality of attached seeds, wherein each attached seed comprises a single protein and a single seed; andattaching said one or more SNAPs to attachment sites of the array of attachment sites, thereby producing an array of spatially separated proteins.2. The method of claim 1 , wherein said solid support comprises glass or tin oxide.3. The method of claim 1 , wherein the distance separating said attachment site from any other attachment site on said array is greater than the radius of said SNAP.4. The method of claim 1 , wherein said solid support is optically opaque.5. The method of claim 1 , wherein said solid support is optically clear.6. The method of claim 1 , wherein said solid support comprises a negatively charged surface.7. The method of claim 1 , wherein said attachment sites on said array are positively charged.8. The method of claim 1 , wherein said solid support comprises functional groups configured to bind said SNAPs.9. The method of claim 1 , wherein said SNAPs of said plurality of SNAPs have a diameter between 10 nm and 50 μm.10. The method of claim 1 , wherein said plurality of SNAPs are ...

COMBINATORIAL SYNTHESIS AND BIOMARKER DEVELOPMENT

Provided herein are methods, kits, and compositions for use in the diagnosis and treatment of diseases. Peptoids recognized by Alzheimer s disease specific antibodies are identified. 1. A method comprisinga. contacting a sample with one or more peptoid or pharmaceutically acceptable salt thereof, wherein said one or more peptoid or pharmaceutically acceptable salt thereof is capable of binding at least two antibody subtypes or fragments thereof; andb. using a computer system to detect whether or not a molecule is bound to said one or more peptoid or pharmaceutically acceptable salt thereof.2. The method of , wherein said one or more peptoid or pharmaceutically acceptable salt thereof comprises said peptoid as described in .3. The method of or , wherein said at least two antibody subtypes comprises at least one of an IgG , IgM , IgD , IgE or an IgA.5. The method of claim 4 , wherein claim 4 , in said one or more peptoid or pharmaceutically acceptable salt thereof of formula I claim 4 , Ris selected from the group consisting of deuterium and hydrogen.6. The method of or claim 4 , wherein claim 4 , in said one or more peptoid or pharmaceutically acceptable salt thereof of formula I claim 4 , Ris selected from the group consisting of piperonyl; cyclopropyl; dimethoxybenzyl; morpholyl; aminobutyl; and pyridyl.7. The method of any one of - claim 4 , wherein claim 4 , in said one or more peptoid or pharmaceutically acceptable salt thereof of formula I claim 4 , Ris selected from the group consisting of allyl; methylbenzyl; cyclopropyl; diphenylethyl; benzyl; dimethoxybenzyl; and methyl.8. The method of any one of - claim 4 , wherein claim 4 , in said one or more peptoid or pharmaceutically acceptable salt thereof of formula I claim 4 , Ris selected from the group consisting of benzyl; isobutyl; cyclopropyl; piperonyl; and aminobutyl.9. The method of any one of - claim 4 , wherein claim 4 , in said one or more peptoid or pharmaceutically acceptable salt thereof of formula I ...

Peptide Synthesis Apparatus and Methods Using Infrared Energy

Apparatus and methods utilizing infrared energy for heating reactions associated with peptide synthesis, such as activation, deprotection, coupling, and cleavage. Thorough agitation of the contents of reaction vessels during heating and real-time monitoring and adjustment of temperature and/or reaction duration are also described. Existing peptide synthesizers may be retrofitted to include an infrared energy source. 1. A process for the solid-phase synthesis of peptides , comprising the step of:applying infrared energy to a reaction vessel during said solid-phase synthesis of peptides.2. The process of claim 1 , wherein said step of applying infrared energy to a reaction vessel comprises heating one or more of a deprotection reaction claim 1 , an activation reaction claim 1 , a cleavage reaction claim 1 , or a coupling reaction.3. The process according to claim 1 , wherein the contents of the reaction vessel are agitated during application of the infrared energy.4. The process according to claim 1 , wherein temperature in said reaction vessel is monitored in real time and the output of the infrared energy is adjusted to a predetermined point.5. The process according to claim 2 , wherein the deprotection reaction is monitored in real time and the duration or repetitions of one or more steps of deprotection or coupling is adjusted to a predetermined point.6. A process for the solid phase synthesis of peptides claim 2 , comprising:performing a deprotection step while irradiating a reaction vessel containing an amino acid and deprotection reagents with infrared energy;activating and coupling a second amino acid to a deprotected amine of said amino acid while irradiating said reaction vessel with infrared energy;agitating said reaction vessel; andperforming successive deprotection and coupling steps while irradiating said reaction vessel until a desired peptide is synthesized.7. The peptide synthesis process according to claim 6 , wherein said agitating of the reaction ...

SELECTIVE PHOTOACTIVATION OF AMINO ACIDS FOR SINGLE STEP PEPTIDE COUPLING

Disclosed herein are formulations, substrates, and arrays for amino acid and peptide synthesis on microarrays. In certain embodiments, methods for manufacturing and using the formulations, substrates, and arrays including one-step coupling, e.g., for synthesis of peptides in a C→N orientation are disclosed. In some embodiments, disclosed herein are formulations and methods for high efficiency coupling of biomolecules to a substrate. 1. A carboxylic acid activating formulation , comprising: a carboxylic acid activating compound , a coupling molecule and a solvent.2. The formulation of claim 1 , wherein said carboxylic acid activating compound is a carbodiimide precursor.3. The formulation of claim 2 , wherein said carbodiimide precursor converts to carbodiimide upon exposure to electromagnetic radiation at a defined wavelength.4. The formulation of claim 4 , wherein said defined wavelength is 248 nm.5. The formulation of claim 3 , wherein said carbodiimide precursor is a thione.6. The formulation of claim 5 , wherein said thione is 1-(3-(diethylamino)-propyl)-4-(2-methoxyphenyl)-1 claim 5 ,4-dihydro-5H-tetrazole-5-thione.7. A method of attaching a coupling molecule to a substrate claim 5 , comprising:obtaining a substrate comprising a plurality of amine groups for linking to a coupling molecule;{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'contacting said substrate with the carboxylic acid activating formulation of ;'}selectively exposing said photoactive coupling formulation to light, thereby activating a carboxylic group of said coupling molecule at a selectively exposed area;coupling the activated carboxylic group of said coupling molecule to at least one of said plurality of amine groups at said selectively exposed area; andoptionally repeating said method to produce a desired polymer at said at least one carboxylic acid group.8. The method of claim 7 , wherein said coupling step is performed multiple times at different selectively exposed areas on said ...

BETA-HAIRPIN PEPTIDOMIMETICS AS CXC4 ANTAGONISTS

β-Hairpin peptidomimetics of the general formula cyclo(-Tyr-His-Xaa-Cys-Ser-Ala-Xaa-Xaa-Arg-Tyr-Cys-Tyr-Xaa-XaaPro-Pro-), disulfide bond between Cysand Cys, and pharmaceutically acceptable salts thereof, with Xaa, Xaa, Xaa, Xaaand Xaabeing amino acid residues of certain types which are defined in the description and the claims, have favorable pharmacological properties and can be used for preventing HIV infections in healthy individuals or for slowing and halting viral progression in infected patients; or where cancer is mediated or resulting from CXCR4 receptor activity; or where immunological diseases are mediated or resulting from CXCR4 receptor activity; or for treating immunosuppression; or during apheresis collections of peripheral blood stem cells and/or as agents to induce mobilization of stem cells to regulate tissue repair. These peptidomimetics can be manufactured by a process which is based on a mixed solid- and solution phase synthetic strategy. 1. A backbone cyclized peptidic compound , built up from 16 amino acid residues , of the formula{'br': None, 'sup': 1', '2', '3', '4', '5', '6', '7', '8', '9', '10', '11', '12', '13', '14', 'D', '15', '16, 'cyclo(-Tyr-His-Xaa-Cys-Ser-Ala-Xaa-Xaa-Arg-Tyr-Cys-Tyr-Xaa-Xaa-Pro-Pro-)\u2003\u2003(I),'}in which{'sup': '3', 'Xaais Ala; Tyr; or Tyr(Me),'}Tyr(Me) is (2S)-2-amino-(4-methoxyphenyl)-3-propionic acid,{'sup': 7', 'D', 'D', 'D, 'Xaais Tyr; Tyr(Me); or Pro,'}{'sup': 'D', 'Tyr(Me) is (2R)-2-amino-(4-methoxyphenyl)-3-propionic acid, or,'}{'sup': '8', 'Xaais Dab; or Orn(iPr),'}Dab is (2S)-2,4-diaminobutyric acid,{'sup': 'ω', 'Orn(iPr) is (2S)—N-isopropyl-2,5-diaminopentanoic acid,'}{'sup': '13', 'Xaais Gln; or Glu,'}{'sup': '14', 'Xaais Lys(iPr),'}{'sup': 'ω', 'Lys(iPr) is (2S)—N-isopropyl-2,6-diaminohexanoic acid,'}all of the amino acid residues, which are not explicitly designated as D-amino acid residues, are L-amino acid residues, and{'sup': 4', '11, 'the two —SH groups in the two L-cysteine residues Cysand ...

SOLID PHASE PEPTIDE SYNTHESIS PROCESSES AND ASSOCIATED SYSTEMS

Systems and processes for performing solid phase peptide synthesis are generally described. Solid phase peptide synthesis is a known process in which amino acid residues are added to peptides that have been immobilized on a solid support. In certain embodiments, the inventive systems and methods can be used to perform solid phase peptide synthesis quickly while maintaining high yields. Certain embodiments relate to processes and systems that may be used to heat, transport, and/or mix reagents in ways that reduce the amount of time required to perform solid phase peptide synthesis. 177-. (canceled)78. A process for adding amino acid residues to peptides , comprising:merging a first stream comprising first amino acids that comprise first protecting groups with a second stream comprising a first amino acid activating agent such that the first and second streams form a first mixed fluid comprising first activated amino acids;exposing the first mixed fluid to a plurality of peptides immobilized on a solid support such that first amino acid residues are added to the plurality of peptides;flowing a deprotection reagent to remove at least a portion of the first protecting groups;merging a third stream comprising second amino acids that comprise second protecting groups and a fourth stream comprising a second amino acid activating agent to form a second mixed fluid comprising second activated amino acids; andexposing the second mixed fluid to the plurality of peptides immobilized on the solid support such that second amino acid residues are added to the plurality of peptides;wherein there is no washing step between the first merging step and the second merging step.79. The process of claim 78 , further comprising washing the solid support with a washing solvent after the addition of the second amino acid residues to the plurality of peptides.80. The process of claim 78 , further comprising flowing a second deprotection reagent to remove at least a portion the second ...

METHOD OF IMPROVING CHARACTERISTICS OF PROTEINS

The invention provides efficient methods for combining single-substitution libraries of nucleic acids that span and encode proteins of interest and for selecting resultant mutant proteins after expression which have improved properties or characteristics. 18-. (canceled)9. A method of improving a protein binding site , the method comprising the steps of:synthesizing a single substitution library for each of a plurality of domains of a protein binding site, wherein each member of a single substitution library has a nucleotide sequence that encodes amino acid changes at a single amino acid position of its associated domain and that overlaps a nucleotide sequence of at least one member of a different single substitution library, and wherein the domains of the protein binding site are singly substituted at from 1 to 250 amino acid positions;expressing separately each member of each single substitution library as a pre-candidate protein;incubating in a reaction mixture under binding conditions the pre-candidate proteins of each single substitution library with target molecules;washing the target molecules and from pr-candidate proteins remaining bound form a selected library for each domain of the protein;shuffling members of the selected libraries in a PCR to produce a combinatorial shuffled library;expressing members of the shuffled library as candidate proteins;incubating in a reaction mixture under binding conditions the candidate proteins with target molecules;washing the target molecules so that a fraction of candidate proteins remain bound; andselecting members of the shuffled library which encode candidate proteins which remain bound.10. The method of wherein said binding site is that of an antibody or an antibody fragment expressed by a protein display system.11. The method of wherein said protein display system is a yeast display system claim 10 , a mammalian display system claim 10 , a bacterial display system claim 10 , an insect cell display system or a ...

COMBINATORIAL SYNTHESIS AND BIOMARKER DEVELOPMENT

Provided herein are methods, kits, and compositions for use in the diagnosis and treatment of diseases. Peptoids recognized by Alzheimer's disease specific antibodies are identified. 1109-. (canceled)110. A method comprising:a. contacting a sample with a non-random peptoid library, wherein said non-random peptoid library comprises one or more peptoid or pharmaceutically acceptable salt thereof having an affinity to an antibody; andb. detecting whether said antibody is bound to said one or more peptoid or pharmaceutically acceptable salt thereof.111. The method of claim 110 , wherein said antibody is bound to said one or more peptoid or pharmaceutically acceptable salt thereof.112. The method of claim 111 , wherein said antibody comprises an IgA or a fragment thereof.113. The method of claim 111 , wherein said antibody comprises an IgM or a fragment thereof.114. The method of claim 111 , wherein said detecting comprise identifying a binding between said one or more peptoid or pharmaceutically acceptable salt thereof and at least two antibody isotypes.115. The method of claim 114 , wherein said at least two antibody isotypes comprises at least one of an IgG claim 114 , IgM claim 114 , IgD claim 114 , IgE or an IgA.116. The method of claim 111 , wherein the detecting utilizes a method comprising radio immunoassay (“RIA”) claim 111 , fluorescence immunoassay (“FIA”) claim 111 , enzyme-linked immunosorbent assay (“ELISA”) claim 111 , western blot claim 111 , flow cytometry claim 111 , Forster resonance energy transfer (“FRET”) claim 111 , surface plasmon resonance claim 111 , or any combination thereof.117. The method of claim 110 , wherein said sample is tissue claim 110 , cell claim 110 , urine claim 110 , serum claim 110 , whole blood claim 110 , cerebrospinal fluid claim 110 , sputum claim 110 , saliva claim 110 , or semen.118. The method of claim 117 , wherein said sample is serum.119. The method of claim 110 , wherein said sample is from a subject suspected of ...

Artificial Transcription Factors and Uses Thereof

The present invention relates to artificial transcription factors (ATFs) that alter gene expression, including inducing pluripotency in cells or promoting the conversion of cells to specific cell fates. In particular, provided herein is a zinc-finger based ATF library that can be screened in cells by looking for expression of a specific gene (e.g., reporter expression), monitoring for cell surface markers or morphology, or via functional assays.

PROTEIN RETROSPLICING ENABLED BY A DOUBLE LIGATION REACTION

Proteins containing a C-terminal thioester are important intermediates in semi-synthesis. Currently there is one main method for the synthesis of protein thioesters that relies upon the use of engineered inteins. The invention involves, in some aspects a method, utilizing Sortase A, for preparation of recombinant proteins containing a C-terminal thioester. This new method for double ligatation is useful for synthesizing new or naturally occurring molecules such as a protein thioester. 16-. (canceled)7. A method comprisingperforming a ligation reaction of a N-terminal protein domain with a peptide thioester in the presence of a cysteine transpeptidase enzyme to produce a N-terminal protein domain-COSR product,reacting the N-terminal protein domain-COSR product with a C-terminal protein domain, wherein the C-terminal protein domain has a cysteine at the N-termini, to produce a modified protein having a chemical entity linking the N-terminal protein domain and the C-terminal protein domain.8. The method of claim 7 , wherein the cysteine transpeptidase enzyme is a SrtA enzyme.9. The method of claim 8 , wherein SrtA is SrtA*.10. The method of claim 7 , wherein the ligation reaction is performed in a sortase buffer.11. The method of claim 7 , whereinprior to the ligation reaction the N-terminal protein or N-terminal protein domain is recombinantly expressed as a SUMO-protein having a SUMO tag.12. The method of claim 11 , wherein the SUMO tag is removed using SUMO protease.13. The method of claim 7 , wherein the peptide thioester is G-Xaa-COSR (SEQ ID NOs:1-3) claim 7 , wherein n is 1-6 and wherein Xaa is an amino acid.14. The method of claim 7 , wherein the peptide thioester is G-Xaa-COSR (SEQ ID NOs:4-5) claim 7 , wherein n is 3-5 and claim 7 , wherein Xaa is Gly claim 7 , Phe claim 7 , Ser or Leu.15. The method of claim 7 , wherein the peptide thioester is GGGGG-Xaa-COSR (SEQ ID NOs:5) claim 7 , wherein Xaa is Gly claim 7 , Phe claim 7 , Ser or Leu.16. The method of ...

Single Molecule Arrays for Genetic and Chemical Analysis

Random arrays of single molecules are provided for carrying out large scale analyses, particularly of biomolecules, such as genomic DNA, cDNAs, proteins, and the like. In one aspect, arrays of the invention comprise concatemers of DNA fragments that are randomly disposed on a regular array of discrete spaced apart regions, such that substantially all such regions contain no more than a single concatemer. Preferably, such regions have areas substantially less than 1 μmand have nearest neighbor distances that permit optical resolution of on the order of 10single molecules per cm. Many analytical chemistries can be applied to random arrays of the invention, including sequencing by hybridization chemistries, sequencing by synthesis chemistries, SNP detection chemistries, and the like, to greatly expand the scale and potential applications of such techniques. 1. An array of single molecules comprising:{'sup': '2', 'claim-text': [ (i) a macromolecular structure and', '(ii) at least one analyte having an attachment moiety,, 'a plurality of single molecules attached to the surface, wherein each single molecule comprises'}, 'such that each macromolecular structure comprises a unique functionality and a plurality of attachment functionalities that are capable of forming linkages with the reactive functionalities of the discrete spaced apart regions,', 'and such that the analyte is attached to the macromolecular structure by a linkage between the unique functionality and the attachment moiety of the analyte;', 'wherein each of said discrete spaced apart regions is surrounded by an inter-regional space that is substantially free of said macromolecular structures; and', 'wherein the plurality of single molecules are randomly disposed on the discrete spaced apart regions such that at least a majority of the discrete spaced apart regions contain only one single molecule comprising a macromolecular structure and at least one analyte., 'a support having a planar surface having a ...

SOLID PHASE PEPTIDE SYNTHESIS PROCESSES AND ASSOCIATED SYSTEMS

Systems and processes for performing solid phase peptide synthesis are generally described. Solid phase peptide synthesis is a known process in which amino acid residues are added to peptides that have been immobilized on a solid support. In certain embodiments, the inventive systems and methods can be used to perform solid phase peptide synthesis quickly while maintaining high yields. Certain embodiments relate to processes and systems that may be used to heat, transport, and/or mix reagents in ways that reduce the amount of time required to perform solid phase peptide synthesis. 171-. (canceled)72. A process for adding amino acid residues to peptides , comprising:flowing a first stream comprising amino acids and a second stream comprising an amino acid activating agent substantially simultaneously such that the first stream and the second stream are merged to form a mixed fluid stream comprising activated amino acids; andflowing the mixed fluid stream into a reactor containing solid support on which peptides have been immobilized such that an activated amino acid is added to at least 99% of the immobilized peptides.73. The process of claim 72 , further comprising performing at least 10 amino acid addition cycles subsequent to flowing the mixed fluid.74. The process of claim 72 , further comprising performing at least 50 amino acid addition cycles subsequent to flowing the mixed fluid.75. The process of claim 72 , further comprising performing at least 100 amino acid addition cycles subsequent to flowing the mixed fluid.76. The process of claim 72 , wherein the flowing is performed such that peptides comprising two or more amino acid residues are added to at least 99% of the immobilized peptides.77. The process of claim 72 , further comprising exposing a deprotection reagent to the immobilized peptides to remove one or more protection groups from at least a portion of the immobilized peptides.78. The process of claim 77 , wherein the one or more protection groups ...

SOLID PHASE PEPTIDE SYNTHESIS PROCESSES AND ASSOCIATED SYSTEMS

Systems and processes for performing solid phase peptide synthesis are generally described. Solid phase peptide synthesis is a known process in which amino acid residues are added to peptides that have been immobilized on a solid support. In certain embodiments, the inventive systems and methods can be used to perform solid phase peptide synthesis quickly while maintaining high yields. Certain embodiments relate to processes and systems that may be used to heat, transport, and/or mix reagents in ways that reduce the amount of time required to perform solid phase peptide synthesis. 177-. (canceled)78. A peptide synthesizer reactor , comprising:a tubular wall having an outer dimension and an inner dimension;an upstream tubular insert having a first portion positioned within and in contact with the tubular wall and a second portion positioned outside of the tubular wall, wherein the first portion comprises a first wall having a first outer dimension and a first inner dimension, the second portion comprises a second wall having second outer dimension and a second inner dimension, and the first outer dimension is greater than second outer dimension; anda downstream insert having a third portion positioned within and in contact with the tubular wall and a fourth portion positioned outside of the tubular wall, wherein the third portion comprises a third wall having a third outer dimension and a third inner dimension, the fourth portion comprises a fourth wall having a fourth outer dimension and a fourth inner dimension, and the third outer dimension is greater than fourth outer dimension.79. The peptide synthesizer reactor of claim 78 , wherein the first inner dimension is substantially the same as the second inner dimension.80. The peptide synthesizer reactor of claim 78 , wherein the third inner dimension is substantially the same as the fourth inner dimension.81. The peptide synthesizer reactor of claim 78 , further comprising a first tube claim 78 , wherein at least a ...

Modified peptide display

The invention provides a replicable genetic package displaying a cyclic peptide having at least one intramolecular bond between amino acid side chains. Also provided are a method of preparing such a genetic package displaying cyclic peptides having at least one intramolecular bond. Further provided is a library of replicable genetic packages displaying cyclic peptides each having at least one intramolecular cyclic bond between amino acid side chains; and a method of producing such a library.

SOLID-PHASE POLYMER SYNTHESIS ON REUSABLE SUBSTRATES

Substrates for solid-phase synthesis are reused by freeing synthesized polymers without removing the linkers that hold the polymers to the substrate. The linkers may be made of oligonucleotides or polypeptides. In an implementation, the polymers are released by cleavage of the linkers and then the truncated linkers are regenerated by adding back the portion that was removed. In an implementation, molecular bonds between the linkers and the polymers are cleaved releasing the polymers while leaving the linkers available for reuse without regeneration. In an implementation, single-stranded oligonucleotide linkers are hybridized to complementary strands that hold the polymers to the substrate with double-stranded oligonucleotide complexes. The double-stranded oligonucleotide complexes are denatured releasing the polymers while leaving the original linkers attached to the substrate. The polymers that are synthesized with these techniques may be the same or different type of molecules than the linkers. 1. A method for solid-phase synthesis of polymers , the method comprising:generating polymer strands by adding monomers to free ends of linkers attached to a substrate;contacting the linkers with a linker cleavage agent that cleaves the linkers at a recognition site thereby releasing the polymer strands from the substrate and generating truncated linkers; andregenerating the linkers from the truncated linkers.2. The method of claim 1 , wherein the linkers comprise single-stranded oligonucleotide linkers and the linker cleavage agent comprises a restriction endonuclease that cleaves the linkers within the recognition site.3. The method of claim 2 , further comprising claim 2 , prior to contacting the linkers with the linker cleavage agent claim 2 , contacting the linkers with linker complement strands under conditions that cause the linker complement strands to hybridize with the linkers claim 2 , wherein the recognition site is a double-stranded oligonucleotide sequence ...

DIRECTED SEQUENCE POLYMER COMPOSITIONS AND ANTIBODIES THEREOF FOR THE TREATMENT OF PROTEIN CONFORMATIONAL DISORDERS

The instant invention comprises a process for the solid phase synthesis of directed epitope peptide mixtures useful in the treatment and diagnosis of protein conformational disorders, such process defined by a set of rules regarding the identity and the frequency of occurrence of amino acids that substitute a base or native amino acid of a known epitope. The resulting composition is a mixture of related peptides for therapeutic use. The invention also pertains to the process of generating antibodies using the directed epitope peptide mixtures as the antigens, and antibodies generated by such process, useful in the treatment and diagnostics of the said protein conformational disorder. 134-. (canceled)35. A process for generating antibodies comprising the steps of:{'sup': '2', 'claim-text': (1) selecting a base peptide sequence, wherein the base peptide sequence is derived from an epitope of an antigen associated with a protein conformational disorder,', wherein, for at least one amino acid position of the first cassette of the DSPs, an amino acid is added, said amino acid randomly selected from a mixture of amino acids consisting of the original amino acid found at the corresponding amino acid position of the base peptide sequence, optionally alanine (A), and, optionally, at least one conserved substitution,', 'wherein the amino acids in the mixture are present in a fixed molar input ratio relative to each other, determined prior to starting synthesis,', 'wherein the relative molar amount of A is more than 10% and less than 90% of the total amino acid concentration of the DSPs;, '(2) synthesizing by solid phase peptide synthesis a first cassette of the DSPs, the cassette having a sequence of amino acid positions corresponding to each amino acid of the base peptide sequence,'}, (a) repeating step (2) for 1 to 15 cycles and elongating the DSPs under the same condition including the input ratio of amino acids in the mixture;', '(b) repeating step (2) for 1 to 15 cycles ...

Methods of Constructing and Screening Libraries of Peptide Structures

The present invention provides the means for producing libraries of peptide structures for drug screening applications that are capable of folding or assuming their native conformations independently of artificial scaffolds or flanking sequences in the proteins from which they are derived. The libraries can be highly diverse such that they are representative of the repertoire of protein structures existing in nature. The libraries can also be non-redundant or normalized such that the bias towards specific structures existing in source data sets and/or in nature is/are removed. In a particularly preferred embodiment, the present invention provides 30,000 independent fold structures produced by this method. The present invention also provides computer-readable media and systems comprising structural data in relation to the peptide libraries, and methods for displaying and screening the libraries. 1. A method for producing a peptide library , said method comprising:(i) obtaining a plurality of amino acid sequences capable of independently-forming secondary structures and/or assemblies of secondary structures and/or folds:(ii) producing peptides having the amino acid sequences obtained at (i); and(iii) displaying the peptides at (ii) such that said peptides form secondary structures and/or assemblies of secondary structures and/or folds.27-. (canceled)8. The method of wherein the peptides mimic tertiary structures produced by interaction of non-contiguous portions of native proteins.9. The method of further comprising size-selecting sequences at (i) to thereby identify a sub-set of sequences having the average length of an independent protein fold.10. The method of further comprising identifying claim 1 , redundant sequences and removing or deleting redundant sequences to thereby leave a non-redundant or normalized plurality of amino acid sequences.1112-. (canceled)13. The method of further comprising mutating peptides that are predicted to form a secondary structure or ...

SOLID PHASE PEPTIDE SYNTHESIS PROCESSES AND ASSOCIATED SYSTEMS

Systems and processes for performing solid phase peptide synthesis are generally described. Solid phase peptide synthesis is a known process in which amino acid residues are added to peptides that have been immobilized on a solid support. In certain embodiments, the inventive systems and methods can be used to perform solid phase peptide synthesis quickly while maintaining high yields. Certain embodiments relate to processes and systems that may be used to heat, transport, and/or mix reagents in ways that reduce the amount of time required to perform solid phase peptide synthesis. 1. A process for adding amino acid residues to peptides , comprising:providing a plurality of peptides comprising protection groups, each peptide immobilized on a solid support;exposing a deprotection reagent to the immobilized peptides to remove the protection groups from at least a portion of the immobilized peptides;removing at least a portion of the deprotection reagent;exposing activated amino acids to the immobilized peptides such that at least a portion of the activated amino acids are bonded to the immobilized peptides to form newly-bonded amino acid residues; andremoving at least a portion of activated amino acids that do not bond to the immobilized peptides;wherein an amino acid residue is added to at least about 99% of the immobilized peptides during the amino acids exposing step; and the total amount of time taken to perform the combination of all of the deprotection reagent exposing step, the deprotection reagent removal step, the activated amino acid exposing step, and the activated amino acid removal step is about 10 minutes or less and the protection groups comprise fluorenylmethyloxycarbonyl protection groups and/or', 'the total amount of time taken to perform the combination of all of the deprotection reagent exposing step, the deprotection reagent removal step, the activated amino acid exposing step, and the activated amino acid removal step is about 5 minutes or less., ...

RAPID AND SIMPLIFIED DEVELOPING ANTIBODIES AND FABs OR OTHER MOLECULES CONSISTING OF AMINO ACIDS RECOGNIZING NATURAL ANTIGENS OR NON-NATURAL MOLECULAR TARGETS USING SPECIAL BLOCKING AGENT

A rapid and simplified method of developing antibodies or other monoclonal antigen-recognizing polypeptides comprised of amino acids (polypeptides capable of forming complexes with targeted antigens, comprised of natural or non-natural molecular targets), using a Bioparticle Display Library approach, with each bioparticle of the Bioparticle Display Library containing gene coding polypeptide and capable of multiplying in the presence of some particular multiplication limiting factor. A polypeptide-carrying bioparticle insulated from a complex mixture of similar bioparticles of the Bioparticle Display Library using a first Blocking Agent, said Blocking Agent consisting of a bioparticle strain which lacks resistant to this multiplication limiting factor (for example for Phage Display Library multiplication limiting factor can be some antibiotic, and blocking agent would be phage strain which is lacks resistance to this particular antibiotic). 1. A rapid and simplified method of developing antibodies or other monoclonal antigen-recognizing polypeptides , comprising:amino acids comprising polypeptides capable of recognizing and forming complexes with targeted antigens, comprised of natural or artificial molecular targets, using a Bioparticle Display Library approach, with each bioparticle of the Bioparticle Display Library containing genes composed of nucleic acids and capable of multiplying in the presence of a multiplication limiting factor;a polypeptide-carrying bioparticle identified and separated from a complex mixture of similar bioparticles of the Bioparticle Display Library using a first blocking agent, said first blocking agent consisting of a bioparticle, carrying only one individual type of polypeptide or not carrying any type of polypeptide, and resistant to different ones of the Bioparticle Display Library antibiotic or some other multiplication-limiting factor, or lacks resistance to any antibiotic, or to other multiplication-limiting agents; andperforming ...

CONOTOXIN PEPTIDE K-CPTX-BTL05, PREPARATION METHOD THEREFOR, AND USES THEREOF

Provided are a conotoxin peptide κ-CPTx-btl05 and a derivative polypeptide. The amino acid sequence of the conotoxin peptide is indicated by SEQ ID NO: 1. Also provided are a conotoxin peptide preparation method and uses of the conotoxin peptide in the treatment of diseases related to a calcium ion channel. 1. A conotoxin peptide κ-CPTx-btl05 , which is:(a) a polypeptide having the amino acid sequence shown in SEQ ID NO: 1; or(b) a polypeptide derived from polypeptide (a) by substitution, addition and deletion of one or more amino acids in the amino acid sequence of SEQ ID NO: 1 of polypeptide (a) and having the function of polypeptide (a); wherein the amino acids to be substituted, added or deleted do not include cysteine;the amino acid sequences of polypeptides (a) and (b) contain two pairs of disulfide bonds.2. The conotoxin peptide κ-CPTx-btl05 according to claim 1 , wherein the substitution of the one or more amino acids is selected from the group consisting of:(i) substitution of the isoleucine at position 2 with leucine or valine;(ii) substitution of the valine at position 5 with leucine or isoleucine;(iii) substitution of the threonine at position 8 or 10 with serine;(iv) substitution of the leucine at position 16 with isoleucine or valine.3. A polynucleotide encoding the conotoxin peptide κ-CPTx-btl05 according to .4. A nucleic acid construct comprising the polynucleotide of claim 3 , and one or more control sequences operably linked thereto and being able to direct the production of the polypeptide in an expression host.5. An expression vector comprising the nucleic acid construct of .6. A transformed cell into which the nucleic acid construct of .7. A method for inhibiting a calcium ion channel comprising administration of the conotoxin peptide κ-CPTx-btl05 according to .8. (canceled)9. (canceled)10. A method for producing the conotoxin peptide κ-CPTx-btl05 according to claim 1 , which comprises:(1) synthesizing the linear peptide of the conotoxin peptide ...

PEPTIDE LIBRARY AND USE THEREOF

Disclosed are compositions and method related to variants of SPINK2 that bind to targets other than an endogenous target of SPINK2. In one embodiment, a peptide is provided that comprises the amino acid sequence SEQ ID NO: 1. In further embodiments, an amino acid sequences encoded by nucleotide positions 4 to 42 and/or nucleotide positions 94 to 189 in the nucleotide sequence of SEQ ID NO: 14 flank the amino terminus and the carboxyl terminus, respectively, of the amino acid sequence. In another embodiment, a peptide is provided that comprises an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 1 in which a conservative substitution, deletion, addition and/or insertion of 1 to 5 (inclusive) amino acids has occurred at amino acids other than the 1st Xaa to the 12th Xaa counting from the amino terminus. 1. A peptide library comprising a peptide comprising an amino acid sequence encoded by a nucleotide sequence derived from SEQ ID NO: 14 wherein the nucleotide sequence consisting of the 43rd base thymine to the 93rd base thymine is replaced with a nucleotide sequence encoding the amino acid sequence represented by SEQ ID NO: 1 , wherein the peptide library has a diversity of at least 1×10.2. The peptide library according to claim 1 , wherein the library has a diversity of at least 1×10.3. The peptide library according to claim 1 , wherein each of the 1st Xaa to the 5th Xaa claim 1 , the 7th Xaa claim 1 , the 9th Xaa claim 1 , and the 10th Xaa counting from the amino terminus of SEQ ID NO: 1 is any amino acid other than cysteine and proline.4. The peptide library according to claim 1 , wherein each of the 6th Xaa and the 8th Xaa counting from the amino terminus of SEQ ID NO: 1 is any amino acid other than cysteine.5. The peptide library according to claim 1 , wherein the 11th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of tyrosine claim 1 , serine claim 1 , phenylalanine claim 1 , leucine ...

SELECTIVELY CONTROLLABLE CLEAVABLE LINKERS

Selectively controllable cleavable linkers include electrochemically-cleavable linkers, photolabile linkers, thermolabile linkers, chemically-labile linkers, and enzymatically-cleavable linkers. Selective cleavage of individual linkers may be controlled by changing local conditions. Local conditions may be changed by activating electrodes in proximity to the linkers, exposing the linkers to light, heating the linkers, or applying chemicals. Selective cleaving of enzymatically-cleavable linkers may be controlled by designing the sequences of different sets of the individual linkers to respond to different enzymes. Cleavable linkers may be used to attach polymers to a solid substrate. Selective cleavage of the linkers enables release of specific polymers from the solid substrate. Cleavable linkers may also be used to attach protecting groups to the ends of growing polymers. The protecting groups may be selectively removed by cleavage of the linkers to enable growth of specific polymers. 1. A method of site-specific cleavage of attached polymers from a substrate , the method comprising:preparing the substrate with selectively-cleavable linkers;growing polymers on the selectively-cleavable linkers; andselectively cleaving a portion of the selectively-cleavable linkers by changing local conditions on a portion of the substrate thereby releasing polymers attached to the portion of the substrate.2. The method of claim 1 , wherein the selectively-cleavable linkers comprise electrochemically-cleavable linkers and the substrate comprises spatially-addressable electrodes.3. The method of claim 1 , wherein the selectively-cleavable linkers comprise photolabile linkers and the local conditions comprise light contacting the photolabile linkers.4. The method of claim 1 , wherein the selectively-cleavable linkers comprise thermolabile linkers and the substrate comprises spatially-addressable resistors.5. The method of claim 1 , wherein the selectively-cleavable linkers comprise ...

METHOD FOR PEPTIDE SYNTHESIS AND APPARATUS FOR CARRYING OUT A METHOD FOR SOLID PHASE SYNTHESIS OF PEPTIDES

The invention relates to a method for peptide synthesis, wherein said method comprises the steps of reacting a first amino acid or a first peptide with an α-amine protected second amino acid in a solvent selected from the group consisting of water, alcohol, and a mixture of water and alcohol, and removing the α-amine protecting group with a deprotecting solution. The invention further relates to protective agents, their use and an apparatus for carrying out a method for solid phase synthesis of peptides. 1. A method of forming protecting groups on functional groups during water based peptide synthesis , the method comprising using a protective agent ,wherein the protective agent comprisesa) a backbone structure,b) at least one water-solubility enhancing functional group andc) at least one reactive group,wherein the backbone structure comprises a moiety selected from the group consisting of 9-methylfluorene, t-butane and/or mono-, di or triphenylmethane,{'sub': 3', '3', '3', '2', '3', '3', '3', '3, 'sup': −', '2−', '+', '−', '2−, 'wherein the water-solubility enhancing functional group is selected from the group consisting of SO, PO, N(CH), N(CH), CN, OSO ester, OPO ester and combinations thereof, and'}wherein the water-solubility enhancing functional group and the reactive group are attached to the backbone structure via at least one covalent bond,for protecting a functional group in a chemical reaction, wherein the agent is used for forming protecting groups on functional groups during water based peptide synthesis.2. The method according to claim 1 , wherein the agent is used for forming protecting groups on functional groups on a peptide and/or amino acid during water based peptide synthesis.3. The method according to claim 2 , wherein the functional group to be protected is selected from amine claim 2 , alcohol claim 2 , thiol and carboxyl groups.4. The method according to claim 3 , wherein the reactive group is selected from the group consisting of oxycarbonyl ...

SYNTHESIS METHOD FOR LIRAGLUTIDE WITH LOW RACEMATE IMPURITY

A synthesis method for low-racemization impurity liraglutide comprises the following steps: performing synthesis to obtain a propeptide, coupling 2 to 5 peptides comprising Thr-Phe on the propeptide by using a solid-phase synthesis method; further, performing solid-phase synthesis to obtain a liraglutide resin; the liraglutide resin is cracked after modification, or the liraglutide resin is directly cracked, purified and frozen dry, so as to obtain the liraglutide. The provided liraglutide synthesis method effectively restrains or reduces the generation of racemization impurity D-Thrhighly similar to a product property, which facilitates the purification of the coarse liraglutide, and the high yield of the liraglutide is ensured, thereby greatly reducing production costs; during the synthesis of the liraglutide, the syntheses between dipeptide fragments, tripeptide fragments, the tetrapeptide fragments and pentapeptide fragments and the Gly-resin or the syntheses between the combination of the dipeptide fragments, the tripeptide fragments, the tetrapeptide fragments and pentapeptide fragments and the Gly resin can be carried out at the same time, and accordingly the synthesis time is shortened to some extent. 1. A process for synthesizing liraglutide with a low racemate impurity comprising:synthesizing to obtain a propeptide, and then coupling a 2˜5 amino acid residue-containing peptide with Thr-Phe to the propeptide by using solid-phase synthesis,further performing a solid-phase synthesis to obtain a liraglutide resin, cleaving the liraglutide resin after side chain modification, or directly cleaving the liraglutide resin, purifying, and lyophilizing to give liraglutide.2. The process according to claim 1 , wherein the 2˜5 amino acid residue-containing peptide with Thr-Phe is selected from the group consisting of Thr-Phe claim 1 , Gly-Thr-Phe claim 1 , Thr-Phe-Thr claim 1 , Gly-Thr-Phe-Thr claim 1 , Glu-Gly-Thr-Phe claim 1 , Thr-Phe-Thr-Ser claim 1 , Ala-Glu-Gly- ...

MODIFIED PEPTIDE DISPLAY

The invention provides a replicable genetic package displaying a cyclic peptide having at least one intramolecular bond between amino acid side chains. Also provided are a method of preparing such a genetic package displaying cyclic peptides having at least one intramolecular bond. Further provided is a library of replicable genetic packages displaying cyclic peptides each having at least one intramolecular cyclic bond between amino acid side chains; and a method of producing such a library. 1. A replicable genetic package displaying a peptide comprising a leader portion operationally linked to a core peptide portion having a plurality of amino acid side chains and at least one intramolecular cyclic bond between two atoms of the amino acid side chains , wherein the replicable genetic package is a phage , wherein the cyclic bond is between C—N , C—O , or C—S.2. The replicable genetic package of claim 1 , wherein the replicable genetic package is selected from the group consisting of M13 claim 1 , T4 claim 1 , T7 claim 1 , fd and lambda phages.3. The replicable genetic package of claim 1 , wherein the peptide is a precursor peptide comprising a leader and a core peptide.4. The replicable genetic package of claim 3 , wherein the core peptide is derived from a natural ribosomal peptide.5. The replicable genetic package of claim 1 , wherein the peptide comprises a polycyclic structure.6. The replicable genetic package of claim 1 , wherein the peptide comprises a randomized amino acid sequence.7. The replicable genetic package of claim 1 , wherein the cyclic bond is between C—O or C—N.8. A replicable genetic package displaying a peptide having a plurality of amino acid side chains and at least one intramolecular cyclic bond between two atoms of the amino acid side chains claim 1 , wherein the cyclic bond is between C—N claim 1 , C—O claim 1 , or C—S claim 1 , and wherein the replicable genetic package is a phage particle and is produced by a method comprising the ...

METHOD FOR PREPARING EXENATIDE

Disclosed in the present invention is a method for preparing Exenatide. Serine resin is obtained through a first coupling of serine and resin and successively with amino acids through a second coupling to obtain a peptide resin with a sequence as shown by SEQ ID No. 1; Exenatide resin is obtained through a third coupling of histidine containing a protecting group or salts thereof and the peptide resin with a sequence as shown by SEQ ID No. 1, then it is cracked and purified to obtain purified Exenatide peptide. The method for preparing Exenatide of the present invention inhibits the formation of D-His-Exenatide, and thereby improves the yield and purity of Exenatide. 1. A method for the preparation of exenatide comprising:(a) obtaining a serine-resin by a first coupling between serine and resin;(b) obtaining a peptide-resin of SEQ ID NO: 1 by a secondary coupling of said serine-resin sequentially with amino acids; and(c) obtaining a peptide-resin of SEQ ID NO: 2 by a third coupling between the protective group-containing histidine or a salt thereof with said peptide-resin of SEQ ID No. 1, and exenatide is finally obtained after cleavage and purification.2. The preparation method according to claim 1 , wherein the protective group-containing histidine or a salt thereof in step 3 has the structure as shown in formula II:{'br': None, 'X-His(Y)—OH\u2003\u2003(Formula II)'}wherein X is selected from the group consisting of Fmoc, Boc and Trt; and Y is selected from the group consisting of Trt, Bum, Boc, mtt and mmt.3. The preparation method according to claim 1 , wherein the protective group-containing histidine or a salt thereof in step 3 has the structure as shown in formula III:{'br': None, 'X-His(Boc)-OH\u2003\u2003(Formula III)'}wherein X is selected from the group consisting of Fmoc, Boc and Trt.4. The preparation method according to claim 1 , wherein the protective group-containing histidine or a salt thereof in step 3 has the structure as shown in formula IV:{'br ...

Ultra high-density oligomer arrays and method of production thereof

The present invention relates to a method of producing an oligomer array. The invention comprises the steps of: providing a substrate with a multitude of recesses; introducing a first particle with a first molecule into a recess; releasing the first molecule from the first particle; binding the first molecule to a second molecule to form an oligomer while immobilizing the second molecule in the recess; optionally repeating the steps, wherein at least one of the first particles and/or the first molecules comprises a detectable marker.

Reaction cell for asynchroniousmultipeptide instrument

A reaction cell for an automated peptide synthesizer consists of a body having adjacent first and second reaction wells for simultaneous reactions. The first reaction well is in fluid communication with the second reaction well for reagent pre-activation simultaneously with an amino acid addition for solid state peptide production.

Methods and Products for Fusion Protein Synthesis

The present invention provides a method of producing a fusion protein, said method comprising: a) contacting a first protein with a second protein under conditions that enable the formation of an isopeptide bond between said proteins, wherein said first protein and said second protein each comprise a peptide linker, wherein said peptide linkers are a pair of peptide linkers which react to form an isopeptide bond that links said first protein to said second protein to form a linked protein; and b) contacting the linked protein from (a) with a third protein under conditions that enable the formation of an isopeptide bond between said third protein and said linked protein, wherein said third protein comprises a peptide linker which reacts with a further peptide linker in the linked protein from (a), and wherein said peptide linkers are a pair of peptide linkers that react to form an isopeptide bond that links said third protein to said linked protein to form a fusion protein, wherein said pair of peptide linkers used in (a) are orthogonal to the pair of peptide linkers used in (b). Peptide linkers and the use of orthogonal pairs of said linkers in the synthesis of fusion proteins are also provided. Recombinant proteins comprising said linkers, nucleic acid molecules encoding said proteins and linkers, vectors comprising said nucleic acid molecules and host cells comprising said vectors and nucleic acid molecules are also contemplated. 1. A method of producing a fusion protein , said method comprising:a) contacting a first protein with a second protein under conditions that enable the formation of an isopeptide bond between said proteins, wherein said first protein and said second protein each comprise a peptide linker, wherein said peptide linkers are a pair of peptide linkers which react to form an isopeptide bond that links said first protein to said second protein to form a linked protein; andb) contacting the linked protein from (a) with a third protein under ...

SYSTEM AND METHOD FOR CHARACTERIZING PROTEIN DIMERIZATION

Systems and methods to characterize dimerization interfaces at the subdomain level of a protein are provided. An exemplary method includes digesting a protein dimer sample into subdomains, labeling the digested protein sample, isolating labeled dimeric and monomeric subdomain fragments, and peptide mapping the labeled sample to determine where the dimer fragments are labeled and where the dimer fragments are not labeled. Regions that show decreased labeling extents in the dimer fraction than that in the monomer fraction are likely involved or in close proximity to the dimerization interface. 1. A method for identifying noncovalent interaction sites or dimerization interfaces in a protein drug product , comprising:employing limited digestion of a protein drug product under native conditions to form a protein drug dimer sample mixture comprising monomers and noncovalently linked dimers;introducing detectable modifications into the dimers and monomers of the protein drug dimer sample mixture to produce modified dimers and modified monomers;separating the modified dimers and modified monomers using native size exclusion chromatography into modified dimer and modified monomer fractions;digesting the modified dimer and modified monomer fractions to form peptide samples;separating the peptide samples using liquid chromatography/mass spectrometry (LC-MS) to obtain mass spectrometry data of the peptide samples;determining modification sites in each peptide in the peptide samples using the mass spectrometry data of the peptides compared to known mass data of the protein drug product by calculating the mass of each peptide and modified peptide of such protein drug product.2. The method of claim 1 , wherein native limited digestion conditions retain disulfide bonds between polypeptides and noncovalent interactions between dimers.3. The method of claim 1 , wherein the dimers and monomers are modified by fast photochemical oxidation to form detectable oxidative modifications into ...

Synthesis of Hydantoin Containing Peptide Products

The present invention relates to a method of synthesizing a peptide product comprising at least one hydantoin group. The peptide product may be used as a reference material for the quality control of pharmaceutical peptides, particularly for the quality control of exendin peptides. Further, the invention relates to hydantoin building blocks, a method for manufacturing such building blocks and their use for the synthesis of peptide products. 2. The method of claim 1 , wherein R and/or R are amino acid side chains protected with an acid-labile protecting group claim 1 , a base-labile protecting group or another protecting group.3. The method of claim 1 , wherein R and/or R are protected Glu claim 1 , Gln claim 1 , Asp claim 1 , Asn or Ser side chains.6. Use of a peptide product of any one of - as a reference material for the quality control of pharmaceutical exendin peptide products.7. A reagent kit for determining the amount of impurities in a lixisenatide (AVE0010) product composition comprising at least one stock preparation of the peptide product of or .8. A method for the quality control of a composition comprising a pharmaceutical exendin peptide product claim 4 , more particularly a lixisenatide (AVE0010) product claim 4 , comprising quantitatively determining the amount of a peptide product with a hydantoin group of formula (I) or a salt or solvate thereof claim 4 , according to claim 4 , in said composition.10. Use of a compound of as a building block for the synthesis of peptides claim 9 , particularly in the manufacture of a reference material for the quality control of peptide products.12. The method of wherein R and/or R are amino acid side chains protected with an acid-labile protecting group such as trityl claim 11 , t-butyl or butoxy carbonyl. The present invention relates to a method of synthesizing a peptide product comprising at least one hydantoin group. The peptide product may be used as a reference material for the quality control of ...

LIPOPROTEIN COMPLEXES AND MANUFACTURING AND USES THEREOF

The present disclosure relates to lipoprotein complexes and lipoprotein populations and their use in the treatment and/or prevention of dyslipidemic diseases, disorders, and/or conditions. The disclosure further relates to recombinant expression of apolipoproteins, purification of apolipoproteins, and production of lipoprotein complexes using thermal cycling-based methods. 1. A population of lipoprotein complexes , each comprising a lipid fraction and an apolipoprotein fraction consisting essentially of an Apolipoprotein A-I (“ApoA-I”) , wherein the lipoprotein complexes are at least 80% , at least 85% , at least 90% or at least 95% homogeneous , as reflected by a single peak in gel permeation chromatography.2. The composition of claim 1 , further characterized by one claim 1 , two claim 1 , three claim 1 , or four claim 1 , of the following characteristics:(a) at least 80%, at least 85%, at least 90%, at least 95%, at least 99% of the lipoprotein complexes are in the form of particles of 4 nm to 15 nm in size, 6 nm to 15 nm in size, 4 nm to 12 nm in size, 6 nm to 12 nm in size, or 8 nm to 12 in size as measured by gel permeation chromatography (“GPC”) or dynamic light scattering (“DLS”);(b) at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% by weight over ApoA-I in said population is in mature form;(c) no more than 25%, no more than 20%, no more than 15%, no more than 10% or no more than 5% by weight of ApoA-I in said population is in immature form; and(d) no more than 25%, no more than 20%, no more than 15%, no more than 10% or no more than 5% by weight of ApoA-I in the population is in truncated form.3. The composition of or claim 1 , further characterized by one claim 1 , two claim 1 , three claim 1 , four claim 1 , or five of the following characteristics:(a) no more than 25%, no more than 20%, no more than 15%, no more than 10%, no more than 5%, no more than 3%, no more than 2% or no more than 1% of each of methionine 112 and methionine ...

STABILIZED COMPOUNDS HAVING SECONDARY STRUCTURE MOTIFS

The present invention provides novel stabilized crosslinked compounds having secondary structure motifs, libraries of these novel compounds, and methods for the synthesis of these compounds libraries thereof. The synthesis of these novel stabilized compounds involves (1) synthesizing a peptide from a selected number of natural or non-natural amino acids, wherein said peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and (2) contacting said peptide with a reagent to generate at least one crosslinker and to effect stabilization of a secondary structure motif. The present invention, in a preferred embodiment, provides stabilized p53 donor helical peptides. Additionally, the present invention provides methods for disrupting the p53/MDM2 binding interaction comprising (1) providing a crosslinked stabilized α-helical structure; and (2) contacting said crosslinked stabilized α-helical structure with MDM2. 111-. (canceled)12. A method for disrupting the binding of a first protein to a second protein , the method comprising contacting a peptide with the second protein , wherein the second protein comprises a helix acceptor; the first protein binds the helix acceptor of the second protein; the peptide binds the helix acceptor of the second protein; and the peptide comprises at least two amino acids connected by a hydrocarbon covalent cross-link.13. The method of claim 12 , wherein the hydrocarbon covalent cross-link stabilizes an alpha-helix of the peptide.14. The method of claim 12 , wherein the second protein is a natural protein.15. The method of claim 12 , wherein the peptide is a synthetic peptide.16. The method of claim 12 , wherein the peptide comprises a binding site of a donor helix of the first protein.17. The method of claim 12 , wherein the cross-link comprises a carbon-carbon double bond.18. The method of claim 12 , wherein the at least two amino acids in the peptide comprise olefins.19. The method of ...

METHODS OF PREPARING PEPTIDES

The present invention provides methods of preparing a peptide having an N-terminal histidine, and compositions comprising a plurality of peptides prepared by the methods. The methods disclosed herein reduce racemization of the N-terminal histidine in the peptides during the synthesis process, thereby improving the yield and purity of the peptide compositions. Exemplary peptides that can be manufactured with the methods include Exenatide, Lixisenatide, and Liraglutide. 1. A method of preparing a peptide having an N-terminal histidine , comprising:(a) contacting a resin-bound peptide intermediate having the N-terminal histidine with an acidic cleavage solution to provide a resin-free peptide intermediate, wherein the N-terminus of the resin-bound peptide intermediate is protected by an Fmoc group; and(b) contacting the resin-free peptide intermediate with a basic deblock solution to remove the Fmoc group from the N-terminus of the resin-free peptide intermediate to provide the peptide having the N-terminal histidine.2. The method of claim 1 , further comprising synthesizing the resin-bound peptide intermediate on a resin using Fmoc-protected amino acids according to the sequence of the peptide having the N-terminal histidine.3. The method of claim 1 , wherein the side chain of the N-terminal histidine of the resin-bound peptide intermediate is protected by a group selected from trityl (Trt) claim 1 , 4-methyltrityl (Mtt) claim 1 , and p-methoxytrityl (Mmt).4. The method of claim 1 , wherein the acidic cleavage solution comprises trifluoroacetic acid (TFA).5. The method of claim 1 , wherein the basic deblock solution comprises piperidine.6. The method of claim 5 , wherein the concentration of the piperidine in the deblock solution is about 15% to about 25% by volume.7. The method of claim 5 , wherein the resin-free peptide intermediate is contacted with the basic deblock solution for about 15 minutes to about 30 minutes.8. The method of claim 1 , wherein step (a) ...

LIPOSOMAL METHODS FOR PURIFICATION OF PROTEINS

A general method is described for purifying any protein from a complex mixture by anchoring the protein of interest to liposomes that can be easily isolated from the complex mixture, then separated from the liposome and purified if desired. 1. A method for the purification of a protein comprising the steps of:a) providing the protein in the presence of liposomes, wherein the protein comprises means for binding to the liposomes and the liposomes comprise means for binding to the protein, and wherein the protein binds to the liposomes forming protein-decorated liposomes, and wherein the binding of the protein to the liposomes enables the separability of the protein-decorated liposomes;b) isolating the protein-decorated liposomes;c) separating the protein from the protein-decorated liposomes; andd) purifying the protein from the protein-decorated liposomes.2. The method of wherein the means for binding of the liposomes to the protein comprises a surface moiety on the liposomes that binds to the protein.3. The method of wherein the means for protein binding to the liposomes comprises a moiety on the protein that binds to the liposomes.4. The method of wherein the moiety is added to the protein or the moiety is expressed with the protein.5. The method of wherein the binding is reversible claim 1 , irreversible claim 1 , non-covalent or covalent.6. The method of wherein the non-covalent means is selected from nickel-NTA-6×His tag binding claim 5 , streptavidin/biotin binding claim 5 , conjugate antibody binding claim 5 , binding between complementary protein fragments claim 5 , hybridization of complementary DNAs or RNA and binding to DNA or RNA aptamers.7. The method of wherein the protein comprises a His tag and the liposomes comprise a nickel-NTA moiety.8. The method of wherein the covalent means is selected from click chemistry and spytag/spycatcher chemistry.9. The method of wherein the separability is selected from a change in density of the liposomes associated ...

PROCESSES FOR PREPARING PHARMACEUTICALLY RELEVANT PEPTIDES

The present technology provides methods of generating the peptides, and pharmaceutically acceptable salts of the peptides and intermediates thereof. In some embodiments, the peptide is D-Arg-2′6′-Dmt-Lys-Phe-NH. 114.-. (canceled)15. A compound selected from Boc-D-Arg-DMT-OBn and Cbz-Lys(Boc)-Phe-NH. This application claims priority to U.S. Provisional Application No. 62/129,575, filed on Mar. 6, 2015, the contents of which are incorporated by reference in its entirety.The present technology relates general methods of generating pharmaceutically relevant peptides and intermediates thereof.In an aspect, a process is provided for the preparation of a compound of formula VIII or a salt thereof:the method comprising reacting a compound of formula I-A or a salt thereof,with a compound of formula I-B or a salt thereof,to form a compound of formula I-C or a salt thereof:converting the compound of formula I-C to a compound of formula I-D or a salt thereof:reacting the compound of formula I-D or a salt thereof with a compound of formula I-E or a salt thereof to form the compound of formula VIII or a salt thereof:whereinIn an aspect, a process is provided for the preparation of a compound of formula VIII or a salt thereof:the method comprising reacting a compound of formula II-D or a salt thereof with a compound of formula I-B or a salt thereof to form the compound of formula VIII or a salt thereof:whereinRand Rare each independently selected fromIn some embodiments, the compound of formula II-D or a salt thereof is prepared by a method comprising converting the compound of formula II-C or a salt thereof to a compound of formula II-D or a salt thereofwherein Wis a substituted or unsubstituted alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl, or heterocyclylalkyl group, and the other variables are as defined in formula II-D.In some embodiments, the compound of formula II-C or a salt thereof is prepared by a method comprising ...

System and Method for Characterizing Protein Dimerization

Systems and methods to characterize dimerization interfaces at the subdomain level of a protein are provided. An exemplary method includes digesting a protein dimer sample into subdomains, labeling the digested protein sample, isolating labeled dimeric and monomeric subdomain fragments, and peptide mapping the labeled sample to determine where the dimer fragments are labeled and where the dimer fragments are not labeled. Regions that show decreased labeling extents in the dimer fraction than that in the monomer fraction are likely involved or in close proximity to the dimerization interface. 1. A method for identifying noncovalent interaction sites or dimerization interfaces in a protein drug product , comprising:employing limited digestion of a protein drug product under native conditions to form a protein drug dimer sample mixture comprising monomers and noncovalently linked dimers;introducing detectable modifications into the dimers and monomers of the protein drug dimer sample mixture to produce modified dimers and modified monomers;separating the modified dimers and modified monomers using native size exclusion chromatography into modified dimer and modified monomer fractions;digesting the modified dimer and modified monomer fractions to form peptide samples;separating the peptide samples using liquid chromatography/mass spectrometry (LC-MS) to obtain mass spectrometry data of the peptide samples;determining modification sites in each peptide in the peptide samples using the mass spectrometry data of the peptides compared to known mass data of the protein drug product by calculating the mass of each peptide and modified peptide of such protein drug product.2. The method of claim 1 , wherein native limited digestion conditions retain disulfide bonds between polypeptides and noncovalent interactions between dimers.3. The method of claim 1 , wherein the dimers and monomers are modified by fast photochemical oxidation to form detectable oxidative modifications into ...

COUPLING METHOD FOR PEPTIDE SYNTHESIS AT ELEVATED TEMPERATURES

An improved method for coupling amino acids into peptides or peptidomimetics is disclosed that includes the steps of combining an amino acid, a carbodiimide, an activator additive, and a base at less than 1 equivalent compared to the amino acid to be activated; and carrying out the activation and coupling at a temperature greater than 30° C. 1. In a method for coupling amino acids into peptides or peptidomimetics , the improvement comprising:combining an amino acid, a carbodiimide, an activator additive, and a base, with the base in an amount of less than 1 equivalent compared to the amino acid to be activated; andcarrying out the activation and coupling at a temperature greater than 30° C.2. A method according to carried out within a total coupling time less than 10 minutes.3. A method according to carried out within a total coupling time less than 4 minutes.4. A method according to carried out within a total coupling time less than 2 minutes.5. A solid phase peptide synthesis method according to .6. A method according to wherein the base is selected from the group consisting of DIEA claim 5 , NMM claim 5 , TMP claim 5 , TEA and combinations thereof.7. A method according to wherein the amount of base is no more than 0.2 equivalent based on the amount of amino acid present.8. A method according to wherein the amount of base is no more than 0.1 equivalent based on the amount of amino acid present.9. A method according to wherein the carbodiimide is selected from the group consisting of DCC claim 5 , DIC claim 5 , EDC claim 5 , and mixtures thereof.10. A method according to wherein the activator additive is selected from the group consisting of HOBt claim 9 , HOAt claim 9 , 6-Cl-HOBt claim 9 , Oxyma claim 9 , NHS and mixtures thereof.11. A method according to wherein the activation and coupling are carried out at temperature of between about 30° C. and 110° C.12. A method according to wherein the activation and coupling are carried out at temperature of at least about ...

Fluorogenic peptide substrates for in solution and solid phase factor Xa measurements

The measurement of Factor Xa (FXa) enzymatic activity using novel fluorogenic peptide substrates that have a C-terminus cleavable fluorophore and optionally the ability to attach to a solid support. Fluorogenic measurements increase sensitivity and flexibility of measurements of enzymatic reactions over traditional absorbance-based approaches. The measurement of FXa generation is applicable to a range of biological reactions. 1. A fluorogenic peptide substrate having the formula{'br': None, 'peptide-fluorophore-(linker)n-(X)m'}whereinX is an attachment group;the peptide comprises a C-terminus; andthe fluorophore is cleavable at the C-terminus.2. The fluorogenic peptide substrate of wherein the fluorophore is selected from the group consisting of ACC claim 1 , AMC claim 1 , and AFC.3. The fluorogenic peptide substrate of wherein the fluorophore is excitable by at least one of a UV-light source and a violet light source.4. The fluorogenic peptide substrate of wherein the fluorophore comprises an amine group capable of being functionalized and conjugated with the at least one linker and the at least one attachment group claim 1 , X.5. The fluorogenic peptide substrate of wherein:n is an integer from 0 to 4; andm is an integer from 0 to 4.6. The fluorogenic peptide substrate of wherein the C-terminus is Arg.7. The fluorogenic peptide substrate of wherein the peptide comprises the sequence DArg-Gly-Arg.8. The fluorogenic peptide substrate of wherein the peptide comprises the sequence Ile-Glu(gamma-pip)-Gly-Arg.9. The fluorogenic peptide substrate of wherein:the peptide comprises the sequence Ile-Glu(gamma-OR);{'sub': '3', 'and R is selected from the group consisting of H and CH.'}10. The fluorogenic peptide substrate of wherein the linker is selected from the group comprising PEG and C—C.11. The fluorogenic peptide substrate of wherein the linker is spherical PEG synthesized generating microfluidic droplets.12. The fluorogenic peptide substrate of wherein the linker is ...

WHOLE PROTEOME TILING MICROARRAYS

The present invention relates to a microarray comprising at least 50,000 oligopeptide features per cmwhere the oligopeptide features represent at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% of the proteome of a virus or an organism. The present invention further relates to methods for the synthesis of such microarrays and methods of using microarrays comprising at least 50,000 oligopeptide features per cm. In an embodiment of the invention, the oligopeptide features represent proteins expressed in the same species, wherein the oligopeptide features are presented in a tiling pattern representing at least about 5,000, at least about 10,000, at least about 15,000, at least about 20,000, or at least about 25,000 proteins expressed in a species. In some embodiments, the oligopeptide microarray features represent proteins expressed in the same species, wherein the microarray features are present in a tiling pattern that represents between about 5,000 and 50,000 expressed proteins, between about 10,000 and 50,000 expressed proteins, between about 15,000 and 50,000 expressed proteins, between about 20,000 and 50,000 expressed proteins, or between about 25,000 and 50,000 expressed proteins. 1. A microarray comprising at least 50 ,000 oligopeptide features per cmwherein the features represent between about 90% and 100% of a target proteome , the target selected from a virus and an organism.2. The microarray of claim 1 , comprising at least 100 claim 1 ,000 oligopeptide features per cm.3. The microarray of claim 1 , comprising at least 200 claim 1 ,000 oligopeptide features per cm.4. The microarray of claim 1 , wherein the organism is human.5. The microarray of claim 1 , wherein substantially all of the oligopeptides are the same length.6. The microarray of claim 1 , wherein substantially all of the oligopeptides are 9 to 18 amino acid residues in length.7. The microarray of claim 6 , wherein substantially all of the ...

Elongate Solid Phase Body

This invention relates to an elongate solid phase body suitable for performing solid phase synthesis. The solid phase body comprises a plurality of enclosures formed of a material comprising a chemically inert mesh and, within each enclosure, a plurality of solid phase beads.

Compounds for use in synthesis of peptidomimetics

Synthesis of O-benzotriazole and O-imidazole synthons are described. Uses of synthons in synthesis of azapeptides and other peptidomimetics, azapeptides and other peptidomimetics synthesized from the synthons and uses of azapeptides and other peptidomimetics are also described.

In-situ Solvent Recycling Process for Solid Phase Peptide Synthesis at Elevated Temperatures

An improvement in of deprotection in solid phase peptide synthesis is disclosed. The method includes the steps of adding the deprotection composition in high concentration and small volume to the mixture of the coupling solution, the growing peptide chain, and any excess activated amino acid from the preceding coupling cycle; and without any draining step between the coupling step of the previous cycle and the addition of the deprotection composition for the successive cycle; and with the coupling solution at a temperature of at least 30° C. 1. A method of deprotection in solid phase peptide synthesis in which the improvement comprises:adding the deprotection composition in high concentration and small volume to the mixture of the coupling solution, the growing peptide chain, and any excess activated amino acid from the preceding coupling cycle; andwithout any draining step between the coupling step of the previous cycle and the addition of the deprotection composition for the successive cycle; andwith the coupling solution at least 30° C.2. A method according to wherein the deprotection composition is an organic base3. A method according to using Fmoc solid phase peptide chemistry4. A method according to with the deprotection solution having a concentration of organic base of at least 50% by volume5. A method according to where the deprotection composition is added in an amount that is less than ⅓ of the volume of the coupling solution.6. A method of deprotection in solid phase peptide synthesis in which the improvement comprises:adding the deprotection composition in high concentration and small volume to the mixture of the coupling solution, the growing peptide chain, and any excess activated amino acid from the preceding coupling cycle; andwithout any draining step between the coupling step of the previous cycle and the addition of the deprotection composition for the successive cycle which removes at least 50% of the volume of the previous cycle coupling ...

System and method for longitudinal analysis of peptide synthesis

The present invention provides a system and method for assessing a synthetic peptide population including interrogating a population of peptide features in the presence of a receptor having an affinity for a binder sequence. The population of peptide features is synthesized over a plurality of synthesis periods and includes a plurality of control peptide features synthesized to have an amino acid sequence including the binder sequence. The control peptide features include a first feature synthesized beginning with a first one of the synthesis periods, and a second feature synthesized beginning after the first one of the synthesis periods such that synthesis of the second control peptide feature is delayed by at least one synthesis period. The method further includes detecting a signal output characteristic of an interaction of the receptor with the control peptide features, the signal output indicative of the fidelity of synthesis of the population of peptide features.

Method for Preparing Peptides

The invention relates to a method for preparing peptides comprising the step of forming a peptide bond wherein the carboxyl group of a first amino acid or first peptide is activated and an amino group of the first activated amino acid or first peptide is protected by a protecting group having a water-solubility enhancing group and the activated carboxyl group of the first amino acid or first peptide is reacted with an amino group of a second amino acid or second peptide wherein said carboxyl group of the first amino acid or first peptide is activated in the absence of the second amino acid or second peptide. The invention further relates to peptides comprising a protecting group having a water-solubility enhancing group being bound to the amino group and an activated or free carboxyl group.

Solvent System for Solid Phase Peptide Synthesis

A solvent system for solid phase peptide synthesis is disclosed that combines a morpholine-based compound and an alkoxybenzene-based compound to form a solvent that can be compatible with both polystyrene and/or PEG resins and that can produce purity yields at least comparable to conventional solvents such as DMF, DMA, and/or NMP. 1. A solid phase peptide synthesis (SPPS) solvent system comprising a solvent mixture including N-formylmorpholine and an alkoxybenzene-based compound , wherein the solvent mixture has a boiling point of about 140° C. or greater suitable for use with elevated SPPS temperatures.2. The solvent system of claim 1 , comprising about 20 wt % to about 50 wt % N-formylmorpholine and about 50 wt % to about 80 wt % of the alkoxybenzene-based compound.3. The solvent system of claim 1 , wherein the solvent mixture has a flash point of about 93° C. or higher.5. The solvent system of claim 4 , wherein m is 2.6. The solvent system of claim 5 , wherein each R is methyl.7. The solvent system of claim 5 , wherein each Ris hydrogen.8. The solvent system of claim 5 , wherein one or more of Ris C-Calkyl.10. The solvent system of claim 9 , comprising about 20 wt % to about 50 wt % N-formylmorpholine and about 50 wt % to about 80 wt % 1 claim 9 ,3-dimethoxybenzene.11. The solvent system of claim 9 , comprising about 25 wt % to about 50 wt % N-formylmorpholine and about 50 wt % to about 75 wt % 1 claim 9 ,3-dimethoxybenzene.12. The solvent system of claim 4 , wherein m is 1.13. The solvent system of claim 12 , wherein R is methyl.14. The solvent system of claim 12 , wherein each Ris H.15. The solvent system of claim 12 , wherein one or more of Ris C-Calkyl.16. The solvent system of claim 12 , wherein one Ris C-Calkyl and the remaining Rare hydrogen.17. The solvent system of claim 16 , wherein one Ris methyl and the remaining Rare hydrogen.20. The solvent system of claim 1 , wherein the solvent mixture has a boiling point suitable for use at a SPPS reaction ...

PROTEIN-TARGETED DRUG COMPOUND IDENTIFICATION

Methods and systems are provided for identifying drug compounds for targeting proteins in tissue cells. Such a method includes providing a neural network model which comprises an attention-based protein encoder and a molecular decoder. The protein encoder is pretrained in an autoencoder architecture to encode an input protein sequence into an output vector in a latent space representing proteins. The molecular decoder is pretrained in an autoencoder architecture to generate compound data, defining a compound molecule, from an input vector in a latent space representing molecules. The protein encoder and molecular decoder are coupled such that the input vector of the molecular decoder is dependent on the output vector of the protein encoder for an input protein sequence. 1. A method for identifying drug compounds for targeting proteins in tissue cells , the method comprising:providing a neural network model which comprises an attention-based protein encoder, pretrained in an autoencoder architecture to encode an input protein sequence into an output vector in a latent space representing proteins, and a molecular decoder which is pretrained in an autoencoder architecture to generate compound data, defining a compound molecule, from an input vector in a latent space representing molecules, wherein the protein encoder and molecular decoder are coupled such that the input vector of the molecular decoder is dependent on the output vector of the protein encoder for an input protein sequence;training the model in a reinforcement learning architecture in which reward values dependent on affinity of compound molecules, defined by compound data generated by the molecular decoder for respective protein sequences encoded by the protein encoder, to proteins corresponding to those sequences are used to progressively train the model to optimize the reward value for compound data generated thereby; andafter training the model, supplying a protein sequence for a target protein to the ...

Peptide libraries

The invention relates to novel libraries of linear and cyclic peptides, and methods of generating and screening such libraries for biological, pharmaceutical and other uses.

Fluid delivery system

A fluid delivery system is provided to deliver precise volumes of a plurality of fluids in sequence to a treatment reservoir. Fluid channels are formed in a plate structure to provide a means for delivering fluids from a plurality of fluid reservoirs to one or a plurality of treatment reservoirs. Each fluid is delivered by a system comprising a check valve in fluid communication with a fluid reservoir, a positive displacement pump in fluid communication with the check valve and an injector in fluid communication with the positive displacement pump. A partition valve is provided to direct fluid to the desired treatment reservoirs.

Nucleic acid binding proteins

The invention provides a method for preparing a nucleic acid binding protein of the Cys2-His2 zinc finger class capable of binding to a nucleic acid quadruplet in a target nucleic acid sequence, wherein binding to base 4 of the quadruplet by an α-helical zinc finger nucleic acid binding motif in the protein is determined as follows: if base 4 in the quadruplet is A, then position +6 in the α-helix is Gln and position ++2 is not Asp; and if base 4 in the quadruplet is C, then position +6 in the α-helix may be any residue, as long as position ++2 in the α-helix is not Asp.

Identification of compound-protein interactions using libraries of protein-nucleic acid fusion molecules

Disclosed herein is a method for detecting a compound-protein interaction, involving: (a) providing a compound library in which each member of the compound library is immobilized on a solid support; (b) contacting each member of the immobilized compound library in a single reaction chamber with each member of a protein-nucleic acid fusion library under conditions which allow the formation of compound-fusion complexes; (c) isolating the immobilized compound-fusion complexes; and (d) detecting a compound-fusion complex as an indication that the protein of the fusion interacts with the compound. In preferred embodiments, the protein is identified by reading the nucleic acid portion of the fusion, and the compound is identified by reading a detectable tag bound to either the compound or the solid support.

Screening of drugs from chemical combinatorial libraries employing transponders

Materials and methods are disclosed for identifying chemical compounds having desired binding properties towards a binding partner of pharmaceutical interest. The method employs transponders associated with the solid phase material used in the assay and a scanner to encode and decode data stored electronically on the transponder. The data stored on the transponder identifies the monomeric building blocks added during the synthesis. The structural identification of synthesized compounds bound to the solid phase is done by decoding the transponder.

Labeled complex, process for producing the same and process for utilizing the same

A labeled complex which makes it possible, as a multimolecular marking in combinatorial chemistry, to stably and clearly distinguish various fine substances of several thousand or several ten thousand order or more at a high sensitivity and a high accuracy and to simultaneously satisfy the requirements for improving the ability to capture targets, enhancing the distinguishing sensitivity and elevating the number of substances to be distinguished. The above labeled substance is composed of a fine particle, a number of target-carrying substances each bonding at one end to the fine particle, and a label bonding to each target-carrying substance at another end, wherein each of the target-carrying substances carries one or more targets or is capable of carrying the same, and the whole labeled substance is constituted so that definite targets are contained therein at a definite ratio and distributed to the support and all of the target-carrying substances bonded thereto.

Spatially-addressable immobilization of oligonucleotides and other biological polymers on surfaces

Substrates with surfaces comprising compounds with thiol functional groups protected with a photoremovable protecting group can be used to construct arrays of immobilized anti-ligands, such as oligonucleotide probes or other biological polymers. The arrays can be used in assays to detect the presence of complementary nucleic acids in a sample. Spatially addressed irradiation of predefined regions on the surface permits immobilization of oligonucleotides and other biological polymers at the activated regions on the surface. Cycles of irradiation on different regions of the surface and immobilization of different anti-ligands allow formation of an immobilized matrix of anti-ligands at defined sites on the surface. The immobilized matrix of anti-ligands permits simultaneous screenings of a liquid sample for ligands having high affinities for certain anti-ligands of the matrix.

Apparatus for multiple simultaneous synthesis

An apparatus and method which provides a suitable location for multiple, simultaneous synthesis of compounds. The apparatus consists of: a reservoir block having a plurality of wells; a plurality of reaction tubes, usually gas dispersion tubes, having filters on their lower ends; a holder block, having a plurality of apertures; and a manifold, which may have ports to allow introduction/maintenance of a controlled environment. The manifold top wall has apertures and a detachable plate with identical apertures. The apparatus is constructed from materials which will accommodate heating, cooling, agitation, or corrosive reagents. Gaskets are placed between the components. Rods or clamps are provided for fastening the components together. Apparatus operation involves placing the filters on the lower ends of the reaction tubes in the reservoir block wells, and the upper ends passing through the holder block apertures and into the manifold. The apparatus provides in excess of 1 mg of each product with structural knowledge and control over each compound. Using the apparatus a series of building blocks are covalently attached to a solid support. These building blocks are then modified by covalently adding additional different building blocks or chemically modifying some existing functionality until the penultimate structure is achieved. This is then cleaved from the solid support by another chemical reaction into the solution within the well yielding an array of newly synthesized individual compounds, which after postreaction modification, if necessary, are suitable for testing for activity.

Apparatus for multiple simultaneous synthesis

Combinatorial libraries of monomer domains

Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Novel proteins with targeted binding

Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Combinatorial libraries of monomer domains

Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains (as depicted in figure 5) are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Combinatorial libraries of monomer domains

Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Protein scaffolds and uses thereof

Specific monomer domains and multimers comprising the monomer domains are provided. Methods, compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Combinatorial libraries of monomer domains

Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Combinatorial libraries of monomer domains

Methods for identifying discrete monomer domains and immuno-domains with a desired property are provided. Methods for generating multimers from two or more selected discrete monomer domains are also provided, along with methods for identifying multimers possessing a desired property. Presentation systems are also provided which present the discrete monomer and/or immuno-domains, selected monomer and/or immuno-domains, multimers and/or selected multimers to allow their selection. Compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Random peptides that bind to gastro-intestinal tract (GIT) transport receptors and related methods

This invention relates to proteins (e.g., peptides) that are capable of facilitating transport of an active agent through a human or animal gastro-intestinal tissue, and derivatives (e.g., fragments) and analogs thereof, and nucleotide sequences coding for said proteins and derivatives. The proteins of the invention have use in facilitating transport of active agents from the lumenal side of the GIT into the systemic blood system, and/or in targeting active agents to the GIT. Thus, for example, by binding (covalently or noncovalently) a protein of the invention to an orally administered drug, the drug can be targeted to specific receptor sites or transport pathways which are known to operate in the human gastrointestinal tract, thus facilitating its absorption into the systemic system.

PROTEIN SYNTHESIS BY NATIVE CHEMICAL LIGATION (07/01/97)

Process for the production of peptides by solid phase synthesis

스페이서를 아실화 가능형태로 전환시키고, 전환된 스페이서를 적합한 포름산 유도체와 반응시킨 후 적당한 아미노 하이드라자이드와 반응시키고, 적당한 경우, 보호그룹을 염기-불안정성이거나 약산에 대해 불안정성인 보호그룹으로 전환시키고, 스페이서를 수지에 결합(coupling)시키고, 목적하는 펩티드를 C-종결 말단으로부터 단계적으로 합성하고, 이어서 수지로부터 펩티드를 분리하고, 적당한 경우, 분리된 펩티드를 이의 생리학적으로 허용되는 염으로 전환시킴을 특징으로 하여, 하기 일반식(I)의 펩티드 및 이의 생리학적으로 허용되는 염을 제조하는 방법이 기술되어 있다. Convert the spacer into an acylizable form, react the converted spacer with a suitable formic acid derivative and then with the appropriate amino hydrazide, if appropriate, convert the protecting group to a base-labile or weakly labile protecting group Coupling the spacer to the resin, synthesizing the desired peptide step by step from the C-terminated end, and then separating the peptide from the resin and, where appropriate, converting the isolated peptide into its physiologically acceptable salt. Characterized in that, a method for preparing a peptide of formula (I) and physiologically acceptable salts thereof is described. (X)n-A-NH 2 (Ⅰ) (X) nA-NH 2 (I) 상기식에서, X는 천연 또는 비천연 아미노산, 아자-아미노산 또는 이미노산이고 ; n은 1 내지 50, 바람직하게는 1 내지 30의 정수이며 ; A는 아자-아미노산이다. Wherein X is a natural or unnatural amino acid, aza-amino acid or imino acid; n is an integer of 1 to 50, preferably 1 to 30; A is an aza-amino acid.

Protein scaffolds and uses thereof

Specific monomer domains and multimers comprising the monomer domains are provided. Methods, compositions, libraries and cells that express one or more library member, along with kits and integrated systems, are also included in the present invention.

Method of synthesis of peptideamide derivatives or their physiologically compatible acetates or hydrochlorides

FIELD: peptide chemistry. SUBSTANCE: product: derivatives of peptideamides of the formula (I) (X) n -A-NH 2 where X - Ac-L (or D)-(Hal), p-Cl-L (or D)-Phe, L (or D)-Ser, L (or D)-Tyr, L (or D)-Trp, L (or D)-Ser(α-L-Phe), L (or D)-Leu, L (or D)-Arg, L (or D)-Pro, p-Slu, His, D-Ser(t-Bu); n = 3-10; A - azaglycine, or their biologically compatible acetates or hydrochlorides. Reagent 1: compound of the formula (II) where Y 1 -Y 5 - hydrogen; C 1 -C 4 -alkoxy-group; -(CH 2 ) m -C(O)-OH [radicals are similar or different but at least one of their is (CH 2 ) n -C(O)-OH]; m = 1-3; R 1 - hydrogen; 4-methoxyphenyl. Reagent 2: silyling agent of the group: tert.-butyldimethylsilyl chloride, tert.-butylphenylsilyl chloride, trimethylchlorosilane, bis-trimethylsilylacetamide. Reaction conditions: process is carried out in the medium of solvent following by treatment of silylated compound with chloroformic acid derivative. Compounds of the formula (III) R 2 were prepared where Y 1 -Y 5 and R 1 - as indicated above; R 2 - halogeno (or nitro)-phenyl. These compounds were treated with hydrazide hydrochloride R 3 -protected with corresponding α-amino acid (X meaning) where R 3 , Fmoc Bpoc, BOC - are protective groups, in the medium of solvent with formation of compound of the formula (IV) . Protective group -BOC- is removed by hydrogenation on the Pt-catalyst. Before the further reaction protective group is conversed to the Fmoc- or BOC-protective group. Then compounds of the formula (IV) (at meaning R 3 - Fmoc, Bpoc-urethane-protective group) were copulated using usual peptide reagent (through -(CH) n -C(O)-OH-grouping on the resin), protective group R 3 is split, and then step by step Fmoc- or Bpoc-protected α-amino acids (in some cases in the form of activated derivatives) were joined. After synthesis termination of peptide of the formula (I) the latter is released from resin by treatment with moderate force acid. Simultaneously or using conventional procedures temporarily added ...

Systems and methods for characterizing protein dimerization

단백질의 하위도메인 수준에서 이합체화 계면을 특징규명하기 위한 시스템 및 방법이 제공된다. 예시적인 방법은 단백질 이합체 샘플을 하위도메인으로 소화시키는 단계, 소화된 단백질 샘플을 표지하는 단계, 표지된 이합체성 하위도메인 단편 및 단량체성 하위도메인 단편을 단리하는 단계, 및 표지된 샘플을 펩타이드 맵핑하여 이합체 단편이 어디서 표지되는지 및 이합체 단편이 어디서 표지되지 않는지를 결정하는 단계를 포함한다. 단량체 분획에서보다 이합체 분획에서 감소된 표지 정도를 나타내는 영역은 이합체화 계면에 관여되고 이와 근접할 것으로 예상된다. Systems and methods are provided for characterizing dimerization interfaces at the subdomain level of proteins. An exemplary method includes digesting a protein dimer sample into subdomains, labeling the digested protein sample, isolating labeled dimeric subdomain fragments and monomeric subdomain fragments, and mapping the labeled sample to peptides to determining where the dimer fragments are labeled and where the dimer fragments are unlabeled. Regions showing a reduced degree of labeling in the dimer fraction than in the monomer fraction are expected to be involved in and close to the dimerization interface.

Methods for Improving Target Affinity of Peptides

본 발명은 단백질 타겟에 결합하는 펩타이드의 타겟 친화도를 증가시키는 방법에 관한 것이다. 본 발명은 공지 펩타이드의 타겟 친화도를 획기적으로 향상시킬 수 있는 방법으로서, 매우 효율적이면서도 간단하다. 본 발명에 의하여, 공지 펩타이드의 타겟 친화도를 예컨대 1000배 증가킬 수 있다. 본 발명의 KPI-바이포달 펩타이드 바인더 기술은 공지의 펩타이드의 응용성을 크게 향상시킬 수 있다. The present invention relates to a method for increasing the target affinity of a peptide that binds to a protein target. The present invention is a method which can dramatically improve the target affinity of a known peptide, and is very efficient and simple. According to the present invention, the target affinity of the known peptide can be increased, for example, 1000 times. The KPI-Bifadal peptide binder technique of the present invention can greatly improve the applicability of known peptides.