KR860700264A - 림포킨 생산 및 정제 - Google Patents
림포킨 생산 및 정제Info
- 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
- Authority
- KR
- South Korea
- Prior art keywords
- csf
- protein
- cdna
- activity
- amino acid
- Prior art date
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- C—CHEMISTRY; METALLURGY
- 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
- 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
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- C—CHEMISTRY; METALLURGY
- 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
- 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
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- 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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- C—CHEMISTRY; METALLURGY
- 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
- 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
-
- C—CHEMISTRY; METALLURGY
- 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
- 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
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- 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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- C—CHEMISTRY; METALLURGY
- 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)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Mycology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Immunology (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)
- Saccharide Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Steroid Compounds (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Debugging And Monitoring (AREA)
- Fats And Perfumes (AREA)
- Compounds Of Unknown Constitution (AREA)
- Detergent Compositions (AREA)
Abstract
내용 없음
Description
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)
- 재조합 CSF단백질.
- 골수 분석에서 1×107단위/㎎의 특이활성을 갖는 CSF단백질.
- 제2항에 있어서, 재조합 CSF단백질.
- 제1항에 있어서, 인체 과립구-대식기 CSF단백질.
- 제1항에 있어서, 제1도에서 CSF-Thr에 대하여 또는 CSF-ile에 대하여 또는 CSF-G에 대하여 나타낸 아미노산 서열을 가지는 CSF단백질.
- 제1항에 있어서, Ala, Pro…로 시작되는 제1도에 나타낸바의 아미노산 서열 또는 메티오닌 잔기가 선행된 Ala.Pro로 시작하는 아미노산 서열을 함유하는 CSF단백질.
- 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.
- 제1항에 있어서, 메티오닌 잔기가 선행되는 것을 제외하고는, 그 단백질이 천연 CSF의 아미노산 서열을 가지는 CSF단백질.
- CSF단백질을 암호하는 유전자가 삽입된 벡터로써 형질전환된 진핵 또는 원핵 숙주 세포로부터 표현된 CSF단백질을 분리하는 것을 특징으로 하는 CSF단백질의 생산방법.
- 제9항에 있어서, 그 표현이 E.coli CHO 또는 효모로부터 유도되는 것을 특징으로 하는 방법.
- 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.
- 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.
- 제1항의 CSF를 암호하는 cDNA 또는 표현 벡터.
- 제1항의 CSF를 함유하는 약학 조성물 또는 치료 목적의 제1항의 CSF.
- 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.
- 제12항의 방법으로 제조된 CSF를 암호하는 약학 조성물.
- 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.
- 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.
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US62834284A | 1984-07-06 | 1984-07-06 | |
US628342 | 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 |
Publications (2)
Publication Number | Publication Date |
---|---|
KR860700264A true KR860700264A (ko) | 1986-08-01 |
KR910001809B1 KR910001809B1 (ko) | 1991-03-26 |
Family
ID=27417446
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1019860700138A KR910001809B1 (ko) | 1984-07-06 | 1985-07-04 | Csf 단백질 생산방법 및 정제방법 |
Country Status (28)
Country | Link |
---|---|
EP (2) | EP0188479B1 (ko) |
JP (4) | JP2540509B2 (ko) |
KR (1) | KR910001809B1 (ko) |
AT (2) | ATE172494T1 (ko) |
AU (3) | AU599572B2 (ko) |
CA (2) | CA1341618C (ko) |
CY (1) | CY1629A (ko) |
DE (3) | DE3584089D1 (ko) |
DK (2) | DK168709B1 (ko) |
ES (1) | ES8701226A1 (ko) |
FI (1) | FI108796B (ko) |
GR (1) | GR851643B (ko) |
HK (2) | 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) | SK505985A3 (ko) |
UA (1) | UA39161C2 (ko) |
WO (1) | WO1986000639A1 (ko) |
Families Citing this family (58)
Publication number | Priority date | Publication date | Assignee | Title |
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AU594014B2 (en) * | 1984-03-21 | 1990-03-01 | Research Corporation Technologies, Inc. | Recombinant DNA molecules |
JPS61227526A (ja) * | 1984-07-25 | 1986-10-09 | Chugai Pharmaceut Co Ltd | 新規なコロニー刺激因子 |
AU588819B2 (en) * | 1984-10-29 | 1989-09-28 | Immunex Corporation | Cloning of human granulocyte-macrophage colony stimulating factor gene |
ZA856108B (en) * | 1984-10-29 | 1986-10-29 | Immunex Corp | Cloning of human granulocyte-macrophage colony simulating factor gene |
JP3038348B2 (ja) * | 1984-11-20 | 2000-05-08 | シェーリング コーポレイション | ヒト顆粒状、マクロファージおよび好酸球の細胞増殖因子活性を示すポリペプチドをコードするcDNAクローン |
EP0211899B1 (en) * | 1985-02-05 | 1992-09-23 | Cetus Oncology Corporation | Purification of native colony stimulating factor-1 |
DE3681551D1 (de) * | 1985-09-30 | 1991-10-24 | Chugai Pharmaceutical Co Ltd | Menschlicher granulozyten-colony stimulierender faktor. |
JPS63502795A (ja) * | 1985-10-03 | 1988-10-20 | バイオジェン インコーポレイテッド | 顆粒球‐マクロファージコロニー刺激因子‐様ポリペプチド、dna配列、組換えdna分子並びに微生物細胞中でヒト顆粒球−マクロファ−ジコロニ−刺激因子−様ポリペプチドを高収量で生産する方法 |
CA1295567C (en) * | 1988-07-25 | 1992-02-11 | Lawrence T. Malek | Expression system for the secretion of bioactive human granulocyte, macrophage-colony stimulating factor (gm-csf) and other heterologous proteins from streptomyces |
DE3545568A1 (de) * | 1985-12-21 | 1987-07-16 | Hoechst Ag | Gm-csf-protein, seine derivate, herstellung solcher proteine und ihre verwendung |
US5298603A (en) * | 1985-12-21 | 1994-03-29 | Hoechst Aktiengesellschaft | GM-CSF protein, its derivatives, the preparation of proteins of this type, and their use |
IL83003A (en) | 1986-07-01 | 1995-07-31 | Genetics Inst | Factors that soak bone formation |
GR871029B (en) | 1986-07-14 | 1987-11-02 | Genetics Inst | Novel osteoinductive factors |
US5162111A (en) * | 1986-07-30 | 1992-11-10 | Grabstein Kenneth H | Treatment of bacterial diseases with granulocyte-macrophage colony stimulating factor |
EP0256843A1 (en) * | 1986-08-11 | 1988-02-24 | Cetus Corporation | Expression of g-csf and muteins thereof and their use |
GB8624899D0 (en) * | 1986-10-17 | 1986-11-19 | Sandoz Ltd | Monoclonal antibodies |
JPH0618781B2 (ja) * | 1986-10-18 | 1994-03-16 | 中外製薬株式会社 | 感染症治療剤 |
EP0276846A3 (en) * | 1987-01-29 | 1989-07-26 | Zymogenetics, Inc. | Colony-stimulating factor derivatives |
JP2521094B2 (ja) * | 1987-04-28 | 1996-07-31 | アムジェン、インコーポレーテッド | ヒト顆粒球マクロファ―ジコロニ―刺激因子をコ―ドする合成dna、そのdnaを含むプラスミド、およびそのdnaで形質転換された大腸菌 |
DE3853590T2 (de) * | 1987-06-25 | 1996-01-04 | Immunex Corp | Rinder granulocyt-makrophagenkolonie stimulierender faktor. |
US5391706A (en) * | 1987-07-16 | 1995-02-21 | Schering Plough Corporation | Purification of GM-CSF |
ATE79884T1 (de) * | 1987-07-16 | 1992-09-15 | Schering Corp | Reinigung von gm-csf. |
AU626530B2 (en) * | 1987-07-17 | 1992-08-06 | Schering Biotech Corporation | Human granulocyte-macrophage colony stimulating factor and muteins thereof |
IE63514B1 (en) * | 1987-07-17 | 1995-05-03 | Schering Biotech Corp | Expression vectors for the production of human granulocyte-macrophage colony stimulation factor in a mammalian cell host |
AU2251488A (en) * | 1987-07-24 | 1989-03-01 | Cetus Corporation | Production of biologically active forms of csf using a baculovirus (acnpv)-insect cell expression system |
AU3834589A (en) * | 1988-05-31 | 1990-01-05 | Schering Biotech Corporation | Method of treating myeloid leukemias |
AU618283B2 (en) * | 1988-10-10 | 1991-12-19 | Instituto Nazionale Per Lo Studio E La Cura Dei Tumori | The use of GM-CSF in the treatment of a patient requiring high-dose chemo- or radiotherapy for cancer |
US5811523A (en) | 1988-11-10 | 1998-09-22 | Trinchieri; Giorgio | Antibodies to natural killer stimulatory factor |
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DE69326829D1 (de) | 1992-02-28 | 1999-11-25 | Univ Texas | Verwendung einer therapeutischen ZUSAMMENSETZUNGEN ZUR BEHANDLUNG VON VERBRENNUNGEN |
US6004807A (en) * | 1992-03-30 | 1999-12-21 | Schering Corporation | In vitro generation of human dendritic cells |
US5738849A (en) * | 1992-11-24 | 1998-04-14 | G. D. Searle & Co. | Interleukin-3 (IL-3) variant fusion proteins, their recombinant production, and therapeutic compositions comprising them |
US5772992A (en) * | 1992-11-24 | 1998-06-30 | G.D. Searle & Co. | Compositions for co-administration of interleukin-3 mutants and other cytokines and hematopoietic factors |
US6153183A (en) * | 1992-11-24 | 2000-11-28 | G. D. Searle & Company | Co-administration of interleukin-3 mutant polypeptides with CSF's or cytokines for multi-lineage hematopoietic cell production |
US6403076B1 (en) | 1992-11-24 | 2002-06-11 | S. Christopher Bauer | Compositions for increasing hematopoiesis with interleukin-3 mutants |
US6057133A (en) * | 1992-11-24 | 2000-05-02 | G. D. Searle | Multivariant human IL-3 fusion proteins and their recombinant production |
US6413509B1 (en) | 1992-11-24 | 2002-07-02 | S. Christopher Bauer | Methods of ex-vivo expansion of hematopoietic cells using interleukin-3 mutant polypeptides with other hematopoietic growth factors |
US7091319B1 (en) | 1992-11-24 | 2006-08-15 | Bauer S Christopher | IL-3 variant hematopoiesis fusion protein |
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JPH0751511B2 (ja) * | 1982-03-15 | 1995-06-05 | 味の素株式会社 | インターロイキン2を含有してなる癌治療剤 |
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