KR960704052A - 인간 에리스로포이에틴 수용체 단편 및 그에 대한 항체(Human ery thropoietin receptor fragment and antibodies thereto) - Google Patents
인간 에리스로포이에틴 수용체 단편 및 그에 대한 항체(Human ery thropoietin receptor fragment and antibodies thereto)Info
- Publication number
- KR960704052A KR960704052A KR1019960700177A KR19960700177A KR960704052A KR 960704052 A KR960704052 A KR 960704052A KR 1019960700177 A KR1019960700177 A KR 1019960700177A KR 19960700177 A KR19960700177 A KR 19960700177A KR 960704052 A KR960704052 A KR 960704052A
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- Prior art keywords
- polypeptide
- human erythropoietin
- erythropoietin receptor
- solid phase
- fusion protein
- Prior art date
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- 101000852145 Homo sapiens Erythropoietin receptor Proteins 0.000 title claims abstract description 12
- 239000012634 fragment Substances 0.000 title abstract 3
- 238000003776 cleavage reaction Methods 0.000 claims abstract description 7
- 230000007017 scission Effects 0.000 claims abstract description 7
- 230000027455 binding Effects 0.000 claims abstract description 6
- 102000003951 Erythropoietin Human genes 0.000 claims abstract description 4
- 108090000394 Erythropoietin Proteins 0.000 claims abstract description 4
- 229940105423 erythropoietin Drugs 0.000 claims abstract description 4
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229920001184 polypeptide Polymers 0.000 claims abstract 14
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract 14
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract 14
- 108020001507 fusion proteins Proteins 0.000 claims abstract 8
- 102000037865 fusion proteins Human genes 0.000 claims abstract 8
- 239000000203 mixture Substances 0.000 claims abstract 7
- 108020003175 receptors Proteins 0.000 claims abstract 2
- 108090000190 Thrombin Proteins 0.000 claims description 6
- 229960004072 thrombin Drugs 0.000 claims description 6
- 108091026890 Coding region Proteins 0.000 claims description 2
- 239000002299 complementary DNA Substances 0.000 claims description 2
- 239000007790 solid phase Substances 0.000 claims 8
- 239000003153 chemical reaction reagent Substances 0.000 claims 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 210000004899 c-terminal region Anatomy 0.000 claims 3
- 239000002773 nucleotide Substances 0.000 claims 3
- 125000003729 nucleotide group Chemical group 0.000 claims 3
- 101000987586 Homo sapiens Eosinophil peroxidase Proteins 0.000 claims 2
- 101000920686 Homo sapiens Erythropoietin Proteins 0.000 claims 2
- 102000044890 human EPO Human genes 0.000 claims 2
- 150000001413 amino acids Chemical class 0.000 claims 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims 1
- 230000015556 catabolic process Effects 0.000 claims 1
- 230000007547 defect Effects 0.000 claims 1
- 238000006731 degradation reaction Methods 0.000 claims 1
- 238000003018 immunoassay Methods 0.000 claims 1
- 239000000463 material Substances 0.000 claims 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- 102000005962 receptors Human genes 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
- 241000588724 Escherichia coli Species 0.000 abstract description 2
- 239000013612 plasmid Substances 0.000 abstract description 2
- 108010075944 Erythropoietin Receptors Proteins 0.000 abstract 3
- 102100036509 Erythropoietin receptor Human genes 0.000 abstract 3
- 239000003446 ligand Substances 0.000 abstract 1
- 238000000034 method Methods 0.000 abstract 1
- 230000019491 signal transduction Effects 0.000 abstract 1
- 230000009870 specific binding Effects 0.000 abstract 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
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- C07—ORGANIC CHEMISTRY
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- C07K14/3156—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci from Streptococcus pneumoniae (Pneumococcus)
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
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- C12N15/62—DNA sequences coding for fusion proteins
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
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Abstract
본 발명은 인간 에리스로포이에틴 수용체의 세포외 영역을 함유하는 융합단백질 발현용 대장균 재조합 플라스미드, 일한 벡터로부터 생성되어 정제된 것으로서, 에리스로포이에틴 수용체의 세포외 영역을 융합단백질의 나머지 부분으로부터 분리하기에 적합한 분해부위를 갖는 융합단백질, 정제된 세포외 영역 폴리펩타이드에 대한 특이적 결합 친화성을 갖는 항제, 에리스로포이에틴에 결합하는 정제된 인간 에리스로포이에틴 수용체 단편 폴리펩타이드에 관한 것이다. 본 발명에 따른 물질, 조성물 및 방법들은 에리스로포이에틴 수용체에 대한 리간드 결합을 연구하고 에리스로포이에틴 수용체를 정량하는 것 뿐 아니라 수용체의 구조 및 신호전달체계를 이해하는 데도 유용하다.
Description
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 발현벡터 pGEX-2T 내로 삽입된 인간 에리스로포이에틴 수용체 cDNA의 5′ 코드화 서열중 약 678bp를 함유하는 플라스미드 pJYL26을 도식적으로 나타낸 것이다. 제1도는 또한, pJYL 26으로부터 발현되는 재조합 융합단백질 EpoRex-th을 나타내고 있다.
제2a도는 EpoRex-th를 발현하는 대장균 세포 추출액을 글루타치온 친화성 칼럼상에서 정제하여 회수한 분획의 280㎚에서의 흡광도(A280)를 나타낸다.
제2b도는 트롬빈 처리된 EpoRex-th를 에리스로포이에틴 친화성 크로마토그래피에 적용한 결과 회수된 Epo-bp를 함유하는 분획의 A280을 나타낸다.
제3도는 트롬빈에 의한 EpoRex-th의 절단을 보여주는 쿠마시 블루 염색된 폴리아크릴아미드 겔 사진이다.
제4도는 Epo-bp에 대한 양의 체내에서 생성된 항-Epo-bp 항체의 결합을 보여주는 웨스턴블롯이다.
제5도는 표지되지 않은 Epo의 존재 및 부재하에, Epo-bp에 대한 다양한 농도의 인간125I-Epo의 결합을 나타낸다.
Claims (7)
- (a) 카르복시 말단에 트롬빈 분해부위를 갖는 폴리펩타이드를 발현시킬 수 있는 첫 번째 뉴클레오티드 서열, 및; (b) 전체 길이 인간 에리스로포이에틴 수용체의 cDNA 코딩 서열중 뉴클레오티드 73 내지 750을 필수적으로 포함하며, 상기 트롬빈 분해부위의 3′에 위치하고 상기 첫 번째 서열에 해독적으로(translationally)연결되어 있는 두 번째 뉴클레오티드 서열을 함유하는 발현벡터.
- (a)아미노 말단 및 카르복시 말단을 가지며, 카르복시 말단에 트롬빈 분해부위를 포함하는 첫 번째 폴리펩타이드 세그먼트; 및 (b)전체 길이 인간 에리스로포이에틴 수용체 단백질중 아미노산 서열 약25 내지 약250을 필수적으로 포함하며, 상기 첫 번째 폴리펩타이드 세그먼트의 카르복시 말단에 공유적으로 결합되어 있는 두 번째 폴리펩타이드 세그먼트를 필수적으로 포함하는 정제된 융합 단백질.
- 인간 에리스로포이에틴과 결합할 수 있고 전체 길이 인간 에리스로포이에틴 수용체 단백질중 아미노산 서열 약 25내지 약 250을 필수적으로 포함하는 정제된 인간 에리스로포이에틴 수용체 폴리펩타이드.
- 인간 에리스로포이에틴과 결합할 수 있고 전체 길이 인간 에리스로포이에틴 수용체 단백질중 아미노산 서열 약 25 내지 약 250을 필수적으로 포함하는 정제된 인간 에리스로포이에틴 수용체 폴리펩타이드에 대한 특이적 결함 친화성을 갖는 정제된 항체.
- (a) 고체상 면역분석 시약; 및 (b) 상기 시약에 작동적으로 결합된 제3항의 단백질을 함유하는 면역분석 조성물.
- (a) 고체상 시약; 및 (b) 상기 시약에 작동적으로 결합된 제4항의 항체를 함유하는 면역분석 조성물.
- (a) 제2항의 정제된 융합단백질을 준비하고;(b) 융합단백질로부터 인간 에리스로포이에틴 수용체 폴리펩타이드를 절단할 수 있는 조건하에 상기 융합 단백질을 트롬빈으로 처리하여 분해혼합물을 형성하고,; (c) 상기 폴리펩타이드로 하여금 고체상 시약과 결합하도록 하는 조건하에 에리스로포이에틴이 결합되어 있는 고체상 시약에 상기 분해 혼합물을 첨가하여 폴리펩타이드-고체상 조성물을 형성하고; (d) 상기 폴리펩타이드-고체상 조성물을 세척하여 결합되지 않은 물질을 제거하고;(e) 상기 폴리펩타이드-고체상 조성물로부터 폴리펩타이드를 용출시키는 과정을 포함하여, 에리스로포이에틴과 결합할 수 있고 전체 길이 인간 에리스로포이에틴 수용체 단백질중 아미노산 서열 약 25내지 약 250을 필수적으로 포함하는 실질적으로 순수한 인간 에리스로포이에틴 수용체 폴리펩타이드를 수득하는 방법.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US10681593A | 1993-08-16 | 1993-08-16 | |
| US08/106815 | 1993-08-16 | ||
| PCT/US1994/009298 WO1995005469A1 (en) | 1993-08-16 | 1994-08-15 | Human erythropoietin receptor fragment and antibodies thereto |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR960704052A true KR960704052A (ko) | 1996-08-31 |
| KR100355159B1 KR100355159B1 (ko) | 2002-12-26 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1019960700177A KR100355159B1 (ko) | 1993-08-16 | 1994-08-15 | 인간 에리스로포이에틴 수용체 단편 및 그에 대한 항체 |
Country Status (8)
| Country | Link |
|---|---|
| US (2) | US5843726A (ko) |
| EP (1) | EP0776370B1 (ko) |
| JP (1) | JPH09501833A (ko) |
| KR (1) | KR100355159B1 (ko) |
| AT (1) | ATE323771T1 (ko) |
| CA (1) | CA2169629C (ko) |
| DE (1) | DE69434704T2 (ko) |
| WO (1) | WO1995005469A1 (ko) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030064480A1 (en) * | 1990-06-28 | 2003-04-03 | Leander Lauffer | Fusion proteins with immunoglobulin portions, the preparation and use thereof |
| US6142857A (en) * | 1998-01-06 | 2000-11-07 | Speedfam-Ipec Corporation | Wafer polishing with improved backing arrangement |
| US7345019B1 (en) * | 1999-04-13 | 2008-03-18 | The Kenneth S. Warren Institute, Inc. | Modulation of excitable tissue function by peripherally administered erythropoietin |
| JP2002544123A (ja) * | 1999-04-14 | 2002-12-24 | スミスクライン・ビーチャム・コーポレイション | エリトロポイエチン受容体抗体 |
| US6998124B1 (en) | 1999-04-14 | 2006-02-14 | Smithkline Beecham Corporation | Erythropoietin receptor antibodies |
| AU780935B2 (en) | 1999-09-13 | 2005-04-28 | Equitech Laboratories, Inc. | Materials and methods for the determination of an analyte |
| JP2004533596A (ja) * | 2000-04-05 | 2004-11-04 | リー，ジョン・ワイ | エリスロポエチンおよびエリスロポエチン受容体の検出 |
| US20030072737A1 (en) * | 2000-12-29 | 2003-04-17 | Michael Brines | Tissue protective cytokines for the protection, restoration, and enhancement of responsive cells, tissues and organs |
| US7767643B2 (en) | 2000-12-29 | 2010-08-03 | The Kenneth S. Warren Institute, Inc. | Protection, restoration, and enhancement of erythropoietin-responsive cells, tissues and organs |
| US7045546B2 (en) * | 2001-07-26 | 2006-05-16 | Tagra Biotechnologies Ltd. | Stabilized derivatives of ascorbic acid-3-phosphate |
| US7396913B2 (en) | 2002-10-14 | 2008-07-08 | Abbott Laboratories | Erythropoietin receptor binding antibodies |
| US20040178714A1 (en) * | 2003-03-11 | 2004-09-16 | Pan Wun Fang | Decorative bulb of a series connected light string |
| US7718363B2 (en) | 2003-04-25 | 2010-05-18 | The Kenneth S. Warren Institute, Inc. | Tissue protective cytokine receptor complex and assays for identifying tissue protective compounds |
| US20060216757A1 (en) * | 2003-04-25 | 2006-09-28 | The Kenneth S. Warren Institute, Inc. | Tissue protective cytokine receptor complex, assays for identifying tissue protective compounds and uses thereof |
| CN1897959A (zh) * | 2003-09-29 | 2007-01-17 | 沃伦药品公司 | 用于治疗和预防脓毒病和粘连形成的组织保护细胞因子 |
| US20050255112A1 (en) * | 2004-05-17 | 2005-11-17 | Lee Jong Y | Compositions and methods for preventing erythropoietin-associated hypertension |
| CN102131829A (zh) * | 2008-08-28 | 2011-07-20 | 弗·哈夫曼-拉罗切有限公司 | 抗人epo受体抗体 |
| ES2528219T3 (es) | 2009-01-15 | 2015-02-05 | F. Hoffmann-La Roche Ag | Anticuerpos contra el receptor de la EPO humana |
| WO2012171996A1 (en) | 2011-06-15 | 2012-12-20 | F. Hoffmann-La Roche Ag | Anti-human epo receptor antibodies and methods of use |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4558005A (en) * | 1982-09-13 | 1985-12-10 | University Patents, Inc. | Monoclonal anti-erythropoietin |
| WO1990008822A1 (en) * | 1989-02-03 | 1990-08-09 | Genetics Institute, Inc. | Erythropoietin receptor |
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1994
- 1994-08-15 AT AT94927925T patent/ATE323771T1/de not_active IP Right Cessation
- 1994-08-15 JP JP7507168A patent/JPH09501833A/ja active Pending
- 1994-08-15 CA CA002169629A patent/CA2169629C/en not_active Expired - Lifetime
- 1994-08-15 EP EP94927925A patent/EP0776370B1/en not_active Expired - Lifetime
- 1994-08-15 KR KR1019960700177A patent/KR100355159B1/ko not_active IP Right Cessation
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| EP0776370A4 (en) | 2002-04-17 |
| EP0776370A1 (en) | 1997-06-04 |
| DE69434704D1 (de) | 2006-05-24 |
| EP0776370B1 (en) | 2006-04-19 |
| KR100355159B1 (ko) | 2002-12-26 |
| ATE323771T1 (de) | 2006-05-15 |
| CA2169629C (en) | 2002-06-11 |
| US20020031806A1 (en) | 2002-03-14 |
| CA2169629A1 (en) | 1995-02-23 |
| WO1995005469A1 (en) | 1995-02-23 |
| US7053184B2 (en) | 2006-05-30 |
| JPH09501833A (ja) | 1997-02-25 |
| US5843726A (en) | 1998-12-01 |
| DE69434704T2 (de) | 2006-10-19 |
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