KR900006515A - 유전자 공학적으로 생성되고 원핵생물내 발현된 항체의 활성화 방법 - Google Patents

유전자 공학적으로 생성되고 원핵생물내 발현된 항체의 활성화 방법 Download PDF

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KR900006515A
KR900006515A KR1019890014951A KR890014951A KR900006515A KR 900006515 A KR900006515 A KR 900006515A KR 1019890014951 A KR1019890014951 A KR 1019890014951A KR 890014951 A KR890014951 A KR 890014951A KR 900006515 A KR900006515 A KR 900006515A
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concentration
protein
guanidine hydrochloride
mole
reactivation
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루돌프 라이너
부크너 조한네스
렌즈 헬무트
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데에르. 다움·데에르.포우크에트
뵈링거 만하임 게엠베하
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    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
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    • C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
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Abstract

내용 없음

Description

유전자 공학적으로 생성되고 원핵생물내 발현된 항체의 활성화 방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 MAB 33의 카파 사슬의 발현에 대한 플라즈미드 pBT Ⅲ의 서열을 표시한다.
제2도는 MAB 33의 Fd사슬의 발현에 대한 플라즈미드 p10169의 서열을 표시한다.

Claims (24)

1-1000g/ml의 단백질 농도로 방법을 행하고 가용화와 재활성화 사이에 4-8mole/ℓ구아니딘 하이드로클로라이드나 6-10mole/ℓ우레아를 함유하는 pH 1-4의 완충액에 대해 투석을 행하는 것으로 특징되며, 세포 분해후 변성 및 환원 조건하에서 가용화하고 연속해서 산화 및 재생 조건하에서 재활성화함에 의해 유전자 공학적으로 생성되고 원핵생물내 발현된 생물학적 활성 단백질을 활성화하는 방법.
제1항에 있어서, 투석 완충액이 5-7mole/ℓ의 구아니딘 하이드로클로라이드를 함유하는 방법.
제1항 또는 제2항에 있어서, 재활성화가 변성시키지 않는 농도의 변성화제와 함께 사이에 0.1-20mole/ℓ의 GSH농도, 0.01-13mmole/ℓ의 GSSG농도 및 pH 9-12에서 1-300시간 동안 행해지는 방법.
제3항에 있어서, GSH농도, 0.2-10mmole/ℓ이고 및/또는 GSSG농도가 0.05-1mmole/ℓ인 방법.
제1항 또는 제2항에 있어서, 재활성화가 단계에서, 변성 조건하에서 GSSG를 부가하므로써 단백질의 티올 그룹이 우선 글루타티온과 단백질의 혼합된 이황화물로 전환되고 구아니딘 하이드로클로라이드나 우레아를 함유하는 완충액에 대해 다시 투석된 다음 병성시키지않는 농도의 변성화제와 함께 0.1-5mmole/ℓ의 GSH농도 및 pH 6-10에서 1-300시간 동안 재활성화되는 방법.
전기한 항의 어느 한 항에 있어서, 재활성화 단계에서, 변성화제로서 아르기닌, 구아니딘 하이드로클로라이드 및/또는 최소한 하나의 다음 구조식 화합물이 사용되는 방법.
R2-CO-NRR1(I)
식중, R 및 R1은 수소원자 또는 C4까지의 알킬기이며 R2는 수소원자 또는 -NRR1그룹 또는 C3까지의 알킬기임.
제6항에 있어서, 아르기닌 및/또는 구아니딘 하이드로클로라이드의 농도가 0 .1-1.0mmole/ℓ인 방법.
제7항에 있어서, 아르기닌 및/또는 구아니딘 하이드로클로라이드의 농도가 0 .25-1.8mmole/ℓ인 방법.
제6항에 있어서, 구조식 I 화합물의 농도가 0.5-4mole/ℓ인 방법.
제9항에 있어서, 구조식 I 화합물의 농도가 1-3.5ole/ℓ인 방법.
전기한 항의 어느 한 항에 있어서, 재할성화 단계에서, 단백질 분해적으로 활성이 아닌 단백질의 존재에서 행해지는 방법.
제11항에 있어서, 단백질 분해적으로 활성이 아닌 단백질이 소의 혈청 알부민인 방법.
전기한 어느 한 항에 있어서, 재활성화 단계에서, 1-10mmole/ℓEDTA의 존재에서 행해지는 방법.
전기한 항의 어느 한 항에 있어서, 세포 분해가 초음파, 고압 분산 또는 리소자임에 의해 행해지는 방법.
제14항에 있어서, 분해가 중성 내지 약산성 pH값의 묽은 수성 완충용액에서 행해지는 방법.
제15항에 있어서, 묽은 수성 완충액이 0.1mmole/ℓ트리스인 방법.
전기한 항의 어느 한 항에 있어서, 세포 분해후에, 불용성 성분이 분리되는 방법.
전기한 항의 어느 한 항에 있어서, 가용화단계에 있어, 머캅탄 그룹을 지닌 환원제 및 변성화제의 존재내 알칼리성 pH값에서 행해지는 방법.
제15항에 있어서, 변성화제로서 구아니딘 하이드로클로라이드 및/또는 구조식 I 화합물의 존재에서 행해지는 방법.
제19항에 있어서, 구아니딘 하이드로클로라이드의 농도가 6mole/ℓ이고 구조식 I 화합물의 농도가 8mole/ℓ인 방법.
제18항 내지 제20항의 어느 한 항에 있어서, DTE, β-머캅토에탄올, 시스테인 또는 GSH의 존재에서 행해지는 방법.
전기한 항의 어느 한 항에 있어서, 항체나 항체 단편이 재활성화되는 방법.
제1항 내지 제21항의 어느 한 항에 있어서, tPA 또는 tPA같이 작용하는 단백질이 재활성화되는 방법.
제1항 내지 제23항의 어느 한 항의 방법에 의해 활성화된, 유전자 공학적으로생성되고 원핵생물내 발현된 생물학적 활성 단백질.
※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019890014951A 1988-10-17 1989-10-17 유전자 공학적으로 생성되고 원핵생물내 발현된 항체의 활성화 방법 KR920010873B1 (ko)

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DE3835350A DE3835350A1 (de) 1988-10-17 1988-10-17 Aktivierung von gentechnologisch hergestellten, in prokaryonten exprimierten antikoerpern
DEP3835350.4 1988-10-17

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US (1) US5077392A (ko)
EP (1) EP0364926B1 (ko)
JP (1) JPH0793879B2 (ko)
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AT (1) ATE101650T1 (ko)
AU (1) AU609645B2 (ko)
CA (1) CA2000604C (ko)
DD (1) DD285613A5 (ko)
DE (2) DE3835350A1 (ko)
DK (1) DK175439B1 (ko)
ES (1) ES2061876T3 (ko)
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Families Citing this family (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5453363A (en) * 1985-10-23 1995-09-26 Boehringer Mannheim Gmbh Process for the activation of t-PA or Ing after genetic expression in prokaryotes
US5676947A (en) * 1989-02-07 1997-10-14 Boehringer Manneheim Gmbh Method for treating thromboembolic conditions using thrombolytically active proteins
DE4037196A1 (de) * 1990-11-22 1992-05-27 Boehringer Mannheim Gmbh Verfahren zur reaktivierung von denaturiertem protein
DE4105480A1 (de) * 1991-02-21 1992-08-27 Boehringer Mannheim Gmbh Verbesserte aktivierung von rekombinanten proteinen
EP0586667A1 (en) * 1992-03-24 1994-03-16 Synergen, Inc. Refolding and purification of insulin-like growth factor i
NZ261571A (en) * 1993-02-04 1997-03-24 Denzyme Aps Producing correctly folded proteins using multiple denaturation and renaturation cycles
EP0943690B1 (en) * 1995-12-21 2006-11-29 Ajinomoto Co., Inc. Method for refolding human activin a
US6027888A (en) * 1996-04-05 2000-02-22 Board Of Regents, The University Of Texas System Methods for producing soluble, biologically-active disulfide-bond containing eukaryotic proteins in bacterial cells
US6083715A (en) * 1997-06-09 2000-07-04 Board Of Regents, The University Of Texas System Methods for producing heterologous disulfide bond-containing polypeptides in bacterial cells
WO1999008707A1 (fr) 1997-08-15 1999-02-25 Chugai Seiyaku Kabushiki Kaisha Preventifs et/ou medicaments contre le lupus erythemateux systemique, contenant un anticorps anti-recepteur d'il-6 comme ingredient actif
DK1095056T3 (da) * 1998-07-09 2007-04-23 Barofold Inc Höjtryksrefoldning af proteinaggregater og inklusionslegemer
EP1077263A1 (de) * 1999-07-29 2001-02-21 F.Hoffmann-La Roche Ag Verfahren zur Herstellung von natürlich gefalteten und sekretierten Proteinen durch Co-Sekretion von Chaperonen
AU7053500A (en) * 1999-09-15 2001-04-17 Eli Lilly And Company Chymotrypsin-free trypsin
US6808902B1 (en) * 1999-11-12 2004-10-26 Amgen Inc. Process for correction of a disulfide misfold in IL-1Ra Fc fusion molecules
MXPA03003867A (es) 2000-10-31 2004-10-15 Univ Colorado Regents Desagregacion mejorada de proteina y replegamiento utilizando alta presion.
DE10105912A1 (de) * 2001-02-09 2002-08-14 Roche Diagnostics Gmbh Rekombinante Proteinase K
DE102005033250A1 (de) 2005-07-15 2007-01-18 Bioceuticals Arzneimittel Ag Verfahren zur Reinigung von G-CSF
US20080032343A1 (en) * 2005-12-22 2008-02-07 Genentech, Inc. Recombinant Production of Heparin Binding Proteins
DE202006020194U1 (de) 2006-03-01 2007-12-06 Bioceuticals Arzneimittel Ag G-CSF-Flüssigformulierung
RU2439076C2 (ru) 2006-07-14 2012-01-10 Дженентек, Инк. Способ выделения повторно свернутого рекомбинантного белка из культуры прокариотических клеток (варианты)
EP2059523A2 (en) * 2006-09-15 2009-05-20 Barofold, Inc. High pressure treatment of proteins for reduced immunogenicity
CN102892780B (zh) 2010-03-17 2015-07-29 通益制药有限公司 获得生物活性的重组人g-csf的方法
HUP1200171A1 (hu) 2012-03-19 2013-09-30 Richter Gedeon Nyrt Módszerek polipeptidek elõállítására
US20160060291A1 (en) * 2012-03-30 2016-03-03 Biogenomics Limited Process for renaturation of polypeptides
US20150376228A1 (en) * 2013-02-22 2015-12-31 Biogenomics Limited Process for high efficiency refolding of recombinant proteins
WO2014167574A1 (en) * 2013-03-22 2014-10-16 Biogenomics Limited Process for isolation and stabilisation of key intermediates for high efficiency refolding of recombinant proteins
US10457716B2 (en) 2014-08-06 2019-10-29 University Of Notre Dame Du Lac Protein folding and methods of using same
CN110643595A (zh) * 2019-10-21 2020-01-03 黑龙江精益检测有限公司 一种对化学修饰后磁性固定化pla1再活化的方法

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GR79124B (ko) * 1982-12-22 1984-10-02 Genentech Inc
US4620948A (en) * 1982-12-22 1986-11-04 Genentech, Inc. Purification and activity assurance of precipitated heterologous proteins
US4816567A (en) * 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4634586A (en) * 1984-05-21 1987-01-06 The Board Of Trustees Of The Leland Stanford Junior University Reagent and method for radioimaging leukocytes
ES8800957A1 (es) * 1985-02-22 1987-12-01 Monsanto Co Un metodo para la solubilizacion y renaturalizacion de proteina somatotropina
US4929444A (en) * 1985-05-28 1990-05-29 Burroughs Wellcome Co. Low pH pharmaceutical formulation containing t-PA
ZW14486A1 (en) * 1985-07-29 1986-10-22 Smithkline Beckman Corp Pharmaceutical dosage unit
DE3537708A1 (de) * 1985-10-23 1987-04-23 Boehringer Mannheim Gmbh Verfahren zur aktivierung von t-pa nach expression in prokaryonten
US4766205A (en) * 1985-11-13 1988-08-23 Beatrice Companies, Inc. Method for isolation of recombinant polypeptides in biologically active forms
FR2594845B1 (fr) * 1986-02-21 1989-12-01 Genetica Preparation par voie microbiologique de l'activateur tissulaire humain du plasminogene (t-pa) et conversion de l'enzyme ainsi obtenue en sa forme active
DE3611817A1 (de) * 1986-04-08 1987-10-15 Boehringer Mannheim Gmbh Verfahren zur renaturierung von proteinen

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US5077392A (en) 1991-12-31
DK175439B1 (da) 2004-10-18
CA2000604C (en) 1999-07-06
DK515989A (da) 1990-04-18
AU4293189A (en) 1990-04-26
EP0364926A2 (de) 1990-04-25
JPH0793879B2 (ja) 1995-10-11
DK515989D0 (da) 1989-10-17
DE58906984D1 (de) 1994-03-24
AU609645B2 (en) 1991-05-02
DD285613A5 (de) 1990-12-19
EP0364926A3 (de) 1991-05-02
IL92012A (en) 1994-12-29
KR920010873B1 (ko) 1992-12-19
EP0364926B1 (de) 1994-02-16
CA2000604A1 (en) 1990-04-17
ES2061876T3 (es) 1994-12-16
DE3835350A1 (de) 1990-04-19
IL92012A0 (en) 1990-07-12
ATE101650T1 (de) 1994-03-15
JPH02227090A (ja) 1990-09-10

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