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KR860700264A - 림포킨 생산 및 정제 - Google Patents

림포킨 생산 및 정제

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Publication number
KR860700264A
KR860700264A KR1019860700138A KR860700138A KR860700264A KR 860700264 A KR860700264 A KR 860700264A KR 1019860700138 A KR1019860700138 A KR 1019860700138A KR 860700138 A KR860700138 A KR 860700138A KR 860700264 A KR860700264 A KR 860700264A
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csf
protein
amino acid
activity
acid sequence
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KR910001809B1 (ko
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씨. 클라크 스티븐
제이. 카우프만 란달
웡 고르돈 지
에이. 왕 엘리자베스
Original Assignee
예안 크라메르,한스 루돌프 하우스
산도즈 아크티엔 게젤샤프트
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Application filed by 예안 크라메르,한스 루돌프 하우스, 산도즈 아크티엔 게젤샤프트 filed Critical 예안 크라메르,한스 루돌프 하우스
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    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • C07K14/53—Colony-stimulating factor [CSF]
    • C07K14/535—Granulocyte CSF; Granulocyte-macrophage CSF
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2840/00—Vectors comprising a special translation-regulating system
    • C12N2840/44—Vectors comprising a special translation-regulating system being a specific part of the splice mechanism, e.g. donor, acceptor

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  • Health & Medical Sciences (AREA)
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  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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Abstract

내용 없음

Description

림포킨 생산 및 정제
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 본 발명에 의한 CPS단백질을 암호하는 DNA서열을 나타낸다. DNA서열은 인체 CSF의 하나의 변이(CSF-Thr)에 대한 코드를 전부 나타낸다. 다른 대립 유전자는 100으로 표시된 위치에서 Thr이 Ile로 대치된다(CSF-Ile)는 것을 제외하고는 동일한 생산물을 암호한다. 인체서열에 대해 상술한 변화는 긴팔원숭이(gibbon) CSF (CSF-G)를 암호하는 DNA서열에서와는 다르다. 연역한 아미노산 서열이 또한 표시되어 있다.

Claims (18)

  1. 재조합 CSF단백질.
  2. 골수 분석에서 1×107단위/㎎의 특이활성을 갖는 CSF단백질.
  3. 제2항에 있어서, 재조합 CSF단백질.
  4. 제1항에 있어서, 인체 과립구-대식기 CSF단백질.
  5. 제1항에 있어서, 제1도에서 CSF-Thr에 대하여 또는 CSF-ile에 대하여 또는 CSF-G에 대하여 나타낸 아미노산 서열을 가지는 CSF단백질.
  6. 제1항에 있어서, Ala, Pro…로 시작되는 제1도에 나타낸바의 아미노산 서열 또는 메티오닌 잔기가 선행된 Ala.Pro로 시작하는 아미노산 서열을 함유하는 CSF단백질.
  7. 제1항에 있어서, 천연 CSF의 생물학적 활성에 거의 영향을 미치지 않고서 하나이상의 아미노산이부가, 치환 또는 제거되는 것을 제외하고는 그 아미노산 서열에 있어서 CSF단백질이 천연 발생한 CSF에 해당하는 CSF단백질.
  8. 제1항에 있어서, 메티오닌 잔기가 선행되는 것을 제외하고는, 그 단백질이 천연 CSF의 아미노산 서열을 가지는 CSF단백질.
  9. CSF단백질을 암호하는 유전자가 삽입된 벡터로써 형질전환된 진핵 또는 원핵 숙주 세포로부터 표현된 CSF단백질을 분리하는 것을 특징으로 하는 CSF단백질의 생산방법.
  10. 제9항에 있어서, 그 표현이 E.coli CHO 또는 효모로부터 유도되는 것을 특징으로 하는 방법.
  11. CSF를 생산하는 세포로부터 RNA를 제조하고; 그 RNA로부터 폴리아데닐화 mRNA를 제조하여 그 mRNA로부터 단일쇄 cDNA를 제조한 후 일본쇄 cDNA를 이본쇄 cDNA로 전환시키고; 이본쇄 cDNA를 형질전환 벡터에 삽입, 그 벡터로써 박테리아를 형질전환시켜 콜로니를 형성시키고; 200 내지 500콜로니들의 각각의 푸울을 선발하여 각 푸울로부터 플라스미드 DNA를 분리; 그 플라스미드 DNA를 CSF단백질을 포현하는 적합한 숙주 세포에 트랜스펙숀하며; 트랜스펙숀된(이입)셀을 배양하여 CSF 활성에 대해 그 상청액을 분석하고; CSF 양성 푸울을 선발, 푸울제조에 사용된 콜로니를 스크리닝하여 CSF 활성을 갖는 콜로니를 동정하는 것을 특징으로 하는, CSF/cDNA 함유의 형질전환 벡터의 제조 및 분리방법.
  12. 암모늄 설페이트로써 단백질을 80% 포화 침전시켜서 VCSF단백질이 함유된 펠릿을 형성시키고; 그 펠릿을 약 6 내지 8의 pH에서 완충 제용액에 재현탁시키고; CSF 함유의 완충 용액을 크로마토그래프 컬럼에 적용시키고, 염화나트륨 함유의 완충 용액으로 CSF 활성을 용출시켜 CSF 활성을 갖는 분획을 수거하며; 그 활성 분획을 혼주하고, 그 혼주된 분획들을 C4역상 컬럼에 적용시킨 후 0 내지 90% 아세토니트릴 그래디언트로 CSF 활성을 용출시켜 CSF 활성이 함유된 분획을 수거하는 것을 특징으로 하는 수용성, 배지에 현탁된 단백질 혼합물로부터 CSF단백질을 정제하는 방법.
  13. 제1항의 CSF를 암호하는 cDNA 또는 표현 벡터.
  14. 제1항의 CSF를 함유하는 약학 조성물 또는 치료 목적의 제1항의 CSF.
  15. 치료가 필요한 동물에 제1항의 CSF를 투여하는 것을 특징으로 하는 동물의 감염 또는 과립구 감소증의 치료 또는 호중구의 활성화 방법. 또는 그 방법을 위한 약학 조성물 제조에 사용되는 제1항의 CSF의 용도.
  16. 제12항의 방법으로 제조된 CSF를 암호하는 약학 조성물.
  17. 치료가 필요한 동물에 제12항의 CSF를 투여하여 포유동물의 감염 또는 과립구 감소증의 치료 또는 호중구 활성화 방법 또는 그 방법을 위한 약학 조성물 제조에 사용되는 제12항의 CSF의 용도.
  18. ATCC 제39754호로 ATCC에 기탁된 E.coli MC1061 또는 그에 기탁된 플라스미드 p91023 (B)-CSF의 127 아미노산 CSF 암호 뉴클레오티드 서열을 표현함으로써 얻을 수 있는 CSF단백질.
    ※참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019860700138A 1984-07-06 1985-07-04 Csf 단백질 생산방법 및 정제방법 Expired KR910001809B1 (ko)

Applications Claiming Priority (7)

Application Number Priority Date Filing Date Title
US62834284A 1984-07-06 1984-07-06
US65244784A 1984-09-19 1984-09-19
US65274284A 1984-09-19 1984-09-19
US652742 1984-09-19
US652447 1984-09-19
PCT/EP1985/000326 WO1986000639A1 (en) 1984-07-06 1985-07-04 Lymphokine production and purification
US628342 1990-12-14

Publications (2)

Publication Number Publication Date
KR860700264A true KR860700264A (ko) 1986-08-01
KR910001809B1 KR910001809B1 (ko) 1991-03-26

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KR1019860700138A Expired KR910001809B1 (ko) 1984-07-06 1985-07-04 Csf 단백질 생산방법 및 정제방법

Country Status (28)

Country Link
EP (2) EP0337359B1 (ko)
JP (4) JP2540509B2 (ko)
KR (1) KR910001809B1 (ko)
AT (2) ATE67244T1 (ko)
AU (3) AU599572B2 (ko)
CA (2) CA1341618C (ko)
CY (1) CY1629A (ko)
DE (3) DE3588199T2 (ko)
DK (2) DK168709B1 (ko)
ES (1) ES8701226A1 (ko)
FI (1) FI108796B (ko)
GR (1) GR851643B (ko)
HK (1) HK111594A (ko)
HU (1) HU208711B (ko)
IE (2) IE930961L (ko)
IL (2) IL75725A (ko)
LU (1) LU88337I2 (ko)
MY (1) MY102902A (ko)
NL (1) NL930089I2 (ko)
NO (4) NO179455C (ko)
NZ (2) NZ212645A (ko)
PL (1) PL153139B1 (ko)
PT (1) PT80776B (ko)
SA (1) SA94150015A (ko)
SG (1) SG108991G (ko)
SK (1) SK280265B6 (ko)
UA (1) UA39161C2 (ko)
WO (1) WO1986000639A1 (ko)

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