KR890701759A - 에스케리키아 콜라이내에서 발현된 사람 인터루킨-4의 정제방법 - Google Patents

에스케리키아 콜라이내에서 발현된 사람 인터루킨-4의 정제방법

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Publication number
KR890701759A
KR890701759A KR1019890700539A KR890700539A KR890701759A KR 890701759 A KR890701759 A KR 890701759A KR 1019890700539 A KR1019890700539 A KR 1019890700539A KR 890700539 A KR890700539 A KR 890700539A KR 890701759 A KR890701759 A KR 890701759A
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South Korea
Prior art keywords
human interleukin
aqueous solution
reduced
reducing agent
concentration
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KR1019890700539A
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English (en)
Inventor
에이. 카스텔레인 로버트
엠.에이. 반 킴메나드 애니타
Original Assignee
스테이너 브이. 칸스태드
쉐링 바이오텍 코포레이션
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Application filed by 스테이너 브이. 칸스태드, 쉐링 바이오텍 코포레이션 filed Critical 스테이너 브이. 칸스태드
Publication of KR890701759A publication Critical patent/KR890701759A/ko

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—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
    • 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
    • C07K1/1133—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 by redox-reactions involving cystein/cystin side chains
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/54—Interleukins [IL]
    • C07K14/5406—IL-4

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

내용 없음

Description

에스케리키아 콜라이내에서 발현된 사람 인터루킨-4의 정제방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 사람 IL-4 cDNA 삽입물을 함유한 이, 콜라이 발현 벡터의 도면이다. 제2도는 제폴딩된 사람 IL-4의 겔 여과 용출 프로우필의 그래프이다. 제3도는 다양한 방법으로 변성시키고, 겔 전기영동으로 분리 한 IL-4 단백질의 웨스턴 블브로트을 나타낸다.

Claims (10)

  1. 세균의 세포막을 붕해시키고, 불용성 응집체를 붕해질로부터 분리하며, 불용성 응집체를 카오트로픽제(chaotropic agent) 및 환원제가 함유된 제1수용액으로 용해시켜 환원되고 변성된 사람 인터루킨-4를 형성하고, 제1수용액중의 환원되고 변성된 사람 인터루킨-4의 농축물을 환원제가 함유된 제2수용액으로 동일한 인터루킨-4분자의 상이한 부분간의 상호작용을 감소시키며, 카오트로픽제 및 환원제의 농도를 낮추어 환원되고 변성된 사람 인터루킨-4를 재생시키고 산화시켜 생물학적으로 활성인 사람 인터루킨-4를 형성하고, 생물학적으로 활성인 사람 인터루킨-4를 정제하는 단계를 함유함을 특징으로 하여, 세균내의 불용성 응집체로부터 생물학적으로 활성인 사람 인터루킨-4 분자를 회수하는 방법.
  2. 제1항에 있어서, 카오트로픽제가 구아니딘 HCI인 방법.
  3. 제1항에 있어서, 환원제가 글루타티온 또는 디티오 트레이톨과 같은 티올 환원제인 방법.
  4. 제3항에 있어서, 환원제가 환원형의 글루타티온 1 내지 3mM 및 산화형의 글루타티온 0.1 내지 0.6mM로 구성된 방법.
  5. 제2항 내지 4항중 어느 한 항에 있어서, 용해 단계가 불용성 응집체의 용액을 약 2.5㎎/ml 미만의 농도로 제1수용액중에 형성함을 포함하는 방법.
  6. 제2항 내지 4항중 어느 한 항에 있어서, 제2수용액으로의 희석단계에 의해 환원되고 변성된 사람 인터루킨-4 분자간의 평균 분자간 간격이 1.5 내지 3배로 확장되는 방법.
  7. 제6항에 있어서, 수용액이 약 50 내지 200mM 이상의 농도로 염을 함유하는 방법.
  8. 제7항에 있어서, 제2수용액으로 희석한후 카오트로픽제 및 환원제의 농도를 투석으로 약 1000배까지 낮추는 방법.
  9. 제7항에 있어서, 정제 단계를 겔 여과 크로마토 그래피로 수행하는 방법.
  10. 제1항에 있어서, 정제 단계로 적어도 5×107단위/㎎의 특이활성을 갖는 사람 인터루킨-4를 수득하는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019890700539A 1987-07-29 1988-07-27 에스케리키아 콜라이내에서 발현된 사람 인터루킨-4의 정제방법 KR890701759A (ko)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US7966687A 1987-07-29 1987-07-29
US?079,666? 1987-07-29
PCT/US1988/002472 WO1989001046A1 (en) 1987-07-29 1988-07-27 Purification of human interleukin-4 expressed in escherichia coli

Publications (1)

Publication Number Publication Date
KR890701759A true KR890701759A (ko) 1989-12-21

Family

ID=22152026

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1019890700539A KR890701759A (ko) 1987-07-29 1988-07-27 에스케리키아 콜라이내에서 발현된 사람 인터루킨-4의 정제방법

Country Status (6)

Country Link
EP (1) EP0301835A1 (ko)
KR (1) KR890701759A (ko)
AU (1) AU2257488A (ko)
IL (1) IL87270A0 (ko)
WO (1) WO1989001046A1 (ko)
ZA (1) ZA885540B (ko)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU620537B2 (en) * 1986-12-19 1992-02-20 Immunex Corporation Human interleukin-4 muteins
US5260202A (en) * 1988-09-07 1993-11-09 Delta Biotechnology Limited Fermentation method
DE69129747T2 (de) * 1990-09-05 1998-11-12 Southern Cross Biotech Pty Ltd In lösung bringen von proteinen in aktiver form
US5494662A (en) * 1992-04-27 1996-02-27 Ono Pharmaceutical Co., Ltd. Stimulator for bone formation
PL306858A1 (en) * 1992-07-02 1995-04-18 Pitman Moore Inc Method of isolating recombination protein in biologically active form from a solution containing non-active protein
US6737513B1 (en) 1996-06-07 2004-05-18 Icos Corporation Macrophage derived chemokine (MDC) and chemokine analogs and assay to identify modulators of MDC activity, and therapeutic uses for same
US6498015B1 (en) 1995-06-07 2002-12-24 Icos Corporation Methods of identifying agents that modulate the binding between MDC and an MDC receptor
US7018627B1 (en) 1995-06-07 2006-03-28 Icos Corporation Macrophage derived chemokine (MDC), MDC analogs, MDC inhibitor substances, and uses thereof
JP3288720B2 (ja) 1996-06-11 2002-06-04 ロシュ ダイアグノスティクス ゲゼルシャフト ミット ベシュレンクテル ハフツング 変性タンパク質の活性化方法
EP1314739A1 (en) * 2001-11-22 2003-05-28 Bayer Ag Process for renaturation of recombinant, disulfide containing proteins at high protein concentrations in the presence of amines

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2129810B (en) * 1982-06-07 1985-09-18 Celltech Ltd A process for the preparation of chymosin
US4569790A (en) * 1984-03-28 1986-02-11 Cetus Corporation Process for recovering microbially produced interleukin-2 and purified recombinant interleukin-2 compositions
PT83761B (pt) * 1985-11-19 1989-06-30 Schering Biotech Corp Metodo para a producao de interleuquina-4 de mamifero

Also Published As

Publication number Publication date
WO1989001046A1 (en) 1989-02-09
IL87270A0 (en) 1988-12-30
AU2257488A (en) 1989-03-01
EP0301835A1 (en) 1989-02-01
ZA885540B (en) 1989-04-26

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