KR940701447A - 캡시드를 형성하고 시스테인 변형된 키메라 ms-2 코트 단백질 - Google Patents
캡시드를 형성하고 시스테인 변형된 키메라 ms-2 코트 단백질Info
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- chimeric protein
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- 210000000234 capsid Anatomy 0.000 title claims abstract description 9
- 235000018417 cysteine Nutrition 0.000 title claims description 5
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 title claims description 5
- 235000018102 proteins Nutrition 0.000 title claims 6
- 102000004169 proteins and genes Human genes 0.000 title claims 6
- 108090000623 proteins and genes Proteins 0.000 title claims 6
- 230000015572 biosynthetic process Effects 0.000 title 1
- 101710132601 Capsid protein Proteins 0.000 claims abstract description 7
- 101710094648 Coat protein Proteins 0.000 claims abstract description 7
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 claims abstract description 7
- 101710125418 Major capsid protein Proteins 0.000 claims abstract description 7
- 101710141454 Nucleoprotein Proteins 0.000 claims abstract description 7
- 101710083689 Probable capsid protein Proteins 0.000 claims abstract description 7
- 102000037865 fusion proteins Human genes 0.000 claims abstract 15
- 108020001507 fusion proteins Proteins 0.000 claims abstract 15
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims abstract 11
- 230000037431 insertion Effects 0.000 claims abstract 2
- 238000003780 insertion Methods 0.000 claims abstract 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims 4
- 239000002299 complementary DNA Substances 0.000 claims 4
- 125000006850 spacer group Chemical group 0.000 claims 3
- 239000004471 Glycine Substances 0.000 claims 2
- 150000001413 amino acids Chemical group 0.000 claims 2
- 238000000034 method Methods 0.000 claims 2
- 239000000203 mixture Substances 0.000 claims 2
- 241000894007 species Species 0.000 claims 2
- 230000008685 targeting Effects 0.000 claims 2
- BQWBEDSJTMWJAE-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 4-[(2-iodoacetyl)amino]benzoate Chemical compound C1=CC(NC(=O)CI)=CC=C1C(=O)ON1C(=O)CCC1=O BQWBEDSJTMWJAE-UHFFFAOYSA-N 0.000 claims 1
- DIYPCWKHSODVAP-UHFFFAOYSA-N 1-[3-(2,5-dioxopyrrol-1-yl)benzoyl]oxy-2,5-dioxopyrrolidine-3-sulfonic acid Chemical group O=C1C(S(=O)(=O)O)CC(=O)N1OC(=O)C1=CC=CC(N2C(C=CC2=O)=O)=C1 DIYPCWKHSODVAP-UHFFFAOYSA-N 0.000 claims 1
- -1 2-pyridyldithio Chemical group 0.000 claims 1
- 241000588724 Escherichia coli Species 0.000 claims 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 claims 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims 1
- 230000000890 antigenic effect Effects 0.000 claims 1
- 230000001588 bifunctional effect Effects 0.000 claims 1
- 239000003431 cross linking reagent Substances 0.000 claims 1
- 239000013604 expression vector Substances 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 229930182830 galactose Natural products 0.000 claims 1
- 230000035772 mutation Effects 0.000 claims 1
- 229960005486 vaccine Drugs 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 abstract 2
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- 101100386050 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) cys-14 gene Proteins 0.000 description 2
- 238000000635 electron micrograph Methods 0.000 description 1
- 150000002256 galaktoses Chemical class 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
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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
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
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- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
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Abstract
본 발명에서는 캡시드 어셈블리의 일부분을 형성할 수 있고, 파지 MS-2의 코트 단백질의 아미노산 서열, 또는 그것의 보존적으로 변형된 변이체, 또는 캡시드 어셈블리를 형성하는 능력을 보유하기에 충분한 상기 서열 또는 변이체를 포함할 수 있으며, 상기 아미노산 서열은 코트 단백질의 N-말단에 돌출되어 있는 β-헤어핀의 외부에 존재하는 시스테인 잔기의 제거 및 N-말단의 돌출된 β-헤어핀에 상응하는 영역내에 시스테인 잔기의 삽입에 의해 변형되는 것을 특징으로 하는 키메라 단백질이 제공된다.
Description
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 면역친화성 정제된 cys-14 변형캡시드의 SDS-PAGE 결과를 도시한 도면,
제2도는 야생형 코트단백질에 의해 생성된 것과 유사한 어셈블된 입자들의 존재를 보여주는 면역친화성 정제된 cys-14변형 캡시드의 전자형미경 사진이다.
제3도는 유리시스테인과 DTNB와의 반응에 대한 대조곡선을 도시한다.
제4도는 시스테인(4mM)이 1 내지 10mM의 활성화 갈락토오스와 혼합되어 실온에서 1시간동안 방치된 다음 DTNB를 사용하여 100㎕의 알리컷으로 측정되었을때 얻어진 곡선을 나타낸다.
Claims (15)
- 캡시드 어셈블리의 일부분을 형성할 수 있고, 파지 MS-2의 코트 단백질의 아미노산 서열, 또는 그것의 보존적으로 변형된 변이체, 또는 캡시드 어셈블리를 형성하는 능력을 보유하기에 충분한 상기 서열 또는 변이체를 포함할 수 있으며, 상기 아미노산 서열은 코트 단백질의 N-말단에 돌출되어 있는 β-헤어핀의 외부에 존재하는 시스테인 잔기의 제거 및 N-말단의 돌출된 β-헤어핀에 상응하는 영역내에 시스테인 잔기의 삽입에 의해 변형되는 것을 특징으로 하는 키메라 단백질.
- 제1항에 있어서, 시스테인 잔기가 코트단백질의 글리신 13 및 14 잔기 영역에 삽입되는 것을 특징으로 하는 키메라 단백질.
- 제2항에 있어서, 시스테인 잔기가 글리신 14를 대체하기 위하여 삽입되는 것을 특징으로 하는 키메라 단백질.
- 제1항, 2항 또는 3항에 있어서, β-헤어핀 외부의 시스테인이 세린에 의해 대체되는 것을 특징으로 하는 키메라 단백질.
- 제1항, 2항, 3항 또는 4항에 있어서, 삽입된 시스테인 잔기가 직접 또는 간접적으로 외래분자종에 결합되어 있는 것을 특징으로 하는 키메라 단백질.
- 제5항에 있어서, 삽입된 시스테인 잔기가 스페이서 부분을 경유하여 외래분자종에 결합되어 있는 것을 특징으로 하는 키메라 단백질.
- 제6항에 있어서, 스페이서 부분이 m-말레이미도벤조일-N-히드록시 술포숙신이미드 에스테르, N-숙신이미딜-(4-요오드아세틸) 아미노벤조에이트 및 N-숙신이미딜-3-(2-피리딜디티오)프로피오네이트로부터 선택된 이중기능성 교차 결합 시약으로부터 유도된 것을 특징으로 하는 키메라 단백질.
- 제1항 내지 제7항 중 어느 한 항에 있어서, 시스테인 잔기가 직접 또는 간접적으로 항원단백질에 결합되어 있는 것을 특징으로 하는 키메라 단백질.
- 제1항 내지 7항 중 어느 한 항에 있어서, 시스테인 잔기가 직접 또는 간접적으로 표적화부분에 결합되어 있는 것을 특징으로 하는 키메라 단백질.
- 제9항에 있어서, 표적화 부분이 갈락토오스를 포함하는 것을 특징으로 하는 키메라 단백질.
- 제1항 내지 10항 중 어느 한 항의 키메라 단백질 또는 그러한 단백질들의 혼합물을 포함하는 캡시드.
- 선행항 중 어느 한 항의 키메라 단백질의 제조방법으로서, 시스테인 46 잔기를 제거하기 위한 돌연변이체와 시스테인 101 잔기를 제거하기 위한 돌연변이체의, 파지코트 단백질의 2개의 단일 돌연변이체를 제조하는 단계, 해당하는 cDNA를 소화시키고 재결찰시켜서 이중 돌연변이된 코트 단백질 cDNA를 제공하는 단계, 그리고 이중 돌연변이된 cDNA에 부위특정 돌연변이를 생성시켜서 시스테인 잔기를 N-말단에 돌출되어 있는 β-헤어핀에 상응하는 영역내에 삽입시키고 그 결과의 cDNA를 발현시켜서 키메라 단백질을 얻는 단계로 이루어지는 방법.
- 제12항에 있어서, 발현이 대장균에서 수행되는 것을 특징으로 하는 방법.
- 제1항 내지 4항 중 어느 한 항의 키메라 단백질을 코드하는 DNA를 포함하는 발현벡터.
- 제8항의 단백질 또는 그러한 단백질을 포함하는 캡시드를 포함하는 백신 제제.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB919114003A GB9114003D0 (en) | 1991-06-28 | 1991-06-28 | Chimaeric protein |
| GB9114003.8 | 1991-06-28 | ||
| PCT/GB1992/001159 WO1993000434A1 (en) | 1991-06-28 | 1992-06-26 | Capsid forming and cystein modified chimaeric ms2-coat protein |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR940701447A true KR940701447A (ko) | 1994-05-28 |
| KR100222163B1 KR100222163B1 (ko) | 1999-10-01 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1019930703986A KR100222163B1 (ko) | 1991-06-28 | 1992-06-26 | 캡시드를 형성하고 시스테인 변형된 키메라 ms-2 코트 단백질 |
Country Status (15)
| Country | Link |
|---|---|
| US (1) | US5698424A (ko) |
| EP (1) | EP0591369B1 (ko) |
| JP (1) | JP3514755B2 (ko) |
| KR (1) | KR100222163B1 (ko) |
| AT (1) | ATE205880T1 (ko) |
| AU (1) | AU657560B2 (ko) |
| CA (1) | CA2111683A1 (ko) |
| DE (1) | DE69232069T2 (ko) |
| FI (1) | FI110613B (ko) |
| GB (2) | GB9114003D0 (ko) |
| IE (1) | IE922085A1 (ko) |
| NO (1) | NO315050B1 (ko) |
| NZ (1) | NZ243330A (ko) |
| WO (1) | WO1993000434A1 (ko) |
| ZA (1) | ZA924774B (ko) |
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| GB9213601D0 (en) * | 1992-06-26 | 1992-08-12 | Mastico Robert A | Protein based delivery system |
| DE19618797C2 (de) * | 1996-05-10 | 2000-03-23 | Bertling Wolf | Vehikel zum Transport molekularer Substanz |
| DE69929232T2 (de) * | 1998-10-21 | 2006-08-31 | The United States Government As Represented By The Department Of Health And Human Services | Virusähnliche partikel zur induktion von autoantikörpern |
| BR9915771A (pt) * | 1998-11-30 | 2001-12-26 | Cytos Biotechnology Ag | Apresentação molecular ordenada de antìgenos,processo de preparação e utilização |
| DE19916224C1 (de) | 1999-04-10 | 2000-06-21 | November Ag Molekulare Medizin | Synthetisches biologisch aktives Molekül |
| IL142025A0 (en) * | 1999-07-20 | 2002-03-10 | Morphosys Ag | Novel methods for displaying (poly) peptides/proteins on bacteriophage particles via disulfide bonds |
| WO2001085208A2 (en) | 2000-05-05 | 2001-11-15 | Cytos Biotechnology Ag | Molecular antigen arrays and vaccines |
| US7264810B2 (en) * | 2001-01-19 | 2007-09-04 | Cytos Biotechnology Ag | Molecular antigen array |
| AU2007202761B2 (en) * | 2001-01-19 | 2010-01-21 | Kuros Biosciences Ag | Molecular antigen array |
| US7128911B2 (en) * | 2001-01-19 | 2006-10-31 | Cytos Biotechnology Ag | Antigen arrays for treatment of bone disease |
| US7094409B2 (en) * | 2001-01-19 | 2006-08-22 | Cytos Biotechnology Ag | Antigen arrays for treatment of allergic eosinophilic diseases |
| JP4516748B2 (ja) | 2001-09-14 | 2010-08-04 | サイトス バイオテクノロジー アーゲー | ウィルス様粒子中への免疫賦活物質のパッケージ化：調製法および使用法 |
| JP4360906B2 (ja) * | 2001-09-14 | 2009-11-11 | サイトス バイオテクノロジー アーゲー | ウィルス様粒子によって誘導される免疫応答を高めるための、抗原提示細胞のインビボでの活性化 |
| US7115266B2 (en) * | 2001-10-05 | 2006-10-03 | Cytos Biotechnology Ag | Angiotensin peptide-carrier conjugates and uses thereof |
| BR0213117A (pt) | 2001-10-05 | 2004-09-21 | Cytos Biotechnology Ag | Conjugados peptìdeo angiotensina-veìculo e seus usos |
| RU2322258C2 (ru) * | 2001-11-07 | 2008-04-20 | Цитос Байотекнолоджи Аг | Антигенные матрицы для лечения заболевания костей |
| US20030219459A1 (en) * | 2002-01-18 | 2003-11-27 | Cytos Biotechnology Ag | Prion protein carrier-conjugates |
| ATE489969T1 (de) * | 2002-06-20 | 2010-12-15 | Cytos Biotechnology Ag | Verpackte virusartige partikel in kombination mit cpg zur verwendung als adjuvantien mit allergenen. herstellungsverfahren und verwendung |
| EP1532167B1 (en) * | 2002-07-17 | 2012-01-25 | Cytos Biotechnology AG | Molecular antigen arrays using a virus like particle derived from the ap205 coat protein |
| NZ537003A (en) | 2002-07-18 | 2008-03-28 | Cytos Biotechnology Ag | Hapten-carrier conjugates comprising virus like particles and uses thereof |
| SI1524994T1 (sl) * | 2002-07-19 | 2011-08-31 | Cytos Biotechnology Ag | Sestavki cepiv, ki vsebujejo amiloidne beta 1-6 antigenske mreĹľe |
| US7537767B2 (en) * | 2003-03-26 | 2009-05-26 | Cytis Biotechnology Ag | Melan-A- carrier conjugates |
| CA2517675A1 (en) * | 2003-03-26 | 2004-10-07 | Cytos Biotechnology Ag | Packaging of immunostimulatory oligonucleotides into virus-like particles: method of preparation and use |
| US20060210588A1 (en) * | 2003-03-26 | 2006-09-21 | Cytos Biotechnology Ag | Hiv-peptide-carrier-conjugates |
| CA2553594A1 (en) * | 2004-01-20 | 2005-07-28 | Cytos Biotechnology Ag | Particle-induced ghrelin immune response |
| US20070248617A1 (en) * | 2004-06-02 | 2007-10-25 | Bachmann Martin F | Medical Uses of Carrier Conjugates of Non-Human Tnf -Peptides |
| AU2005291231B2 (en) | 2004-10-05 | 2010-12-23 | Cytos Biotechnology Ag | VLP-antigen conjugates and their uses as vaccines |
| CN101119749A (zh) * | 2004-10-25 | 2008-02-06 | 赛托斯生物技术公司 | 肠抑胃肽(gip)抗原阵列及其用途 |
| GB0424563D0 (en) * | 2004-11-05 | 2004-12-08 | Novartis Ag | Organic compounds |
| WO2006095345A2 (en) * | 2005-03-08 | 2006-09-14 | Ramot At Tel-Aviv University Ltd. | Targeted drug-carrying bacteriophages |
| US7767212B2 (en) * | 2005-03-18 | 2010-08-03 | Cytos Biotechnology Ag | CAT allergen conjugates and uses thereof |
| KR20080047564A (ko) | 2005-09-28 | 2008-05-29 | 사이토스 바이오테크놀로지 아게 | 인터루킨-1 컨쥬게이트 및 그의 용도 |
| US20070184068A1 (en) | 2005-12-14 | 2007-08-09 | Cytos Biotechnology Ag | Immunostimulatory nucleic acid packaged particles for the treatment of hypersensitivity |
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| DK2190987T3 (da) * | 2007-08-21 | 2013-02-18 | Morphosys Ag | Fremgangsmåder til dannelse af disulfidbindinger |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9101550D0 (en) * | 1991-01-24 | 1991-03-06 | Mastico Robert A | Antigen-presenting chimaeric protein |
-
1991
- 1991-06-28 GB GB919114003A patent/GB9114003D0/en active Pending
-
1992
- 1992-06-26 WO PCT/GB1992/001159 patent/WO1993000434A1/en active IP Right Grant
- 1992-06-26 CA CA002111683A patent/CA2111683A1/en not_active Abandoned
- 1992-06-26 NZ NZ243330A patent/NZ243330A/en unknown
- 1992-06-26 ZA ZA924774A patent/ZA924774B/xx unknown
- 1992-06-26 JP JP50123093A patent/JP3514755B2/ja not_active Expired - Fee Related
- 1992-06-26 GB GB9213644A patent/GB2257431B/en not_active Revoked
- 1992-06-26 AT AT92913894T patent/ATE205880T1/de not_active IP Right Cessation
- 1992-06-26 KR KR1019930703986A patent/KR100222163B1/ko not_active IP Right Cessation
- 1992-06-26 AU AU21955/92A patent/AU657560B2/en not_active Ceased
- 1992-06-26 US US08/167,982 patent/US5698424A/en not_active Expired - Fee Related
- 1992-06-26 DE DE69232069T patent/DE69232069T2/de not_active Expired - Fee Related
- 1992-06-26 EP EP92913894A patent/EP0591369B1/en not_active Expired - Lifetime
- 1992-07-01 IE IE208592A patent/IE922085A1/en not_active Application Discontinuation
-
1993
- 1993-12-27 NO NO19934842A patent/NO315050B1/no not_active IP Right Cessation
- 1993-12-27 FI FI935877A patent/FI110613B/fi active
Also Published As
| Publication number | Publication date |
|---|---|
| CA2111683A1 (en) | 1993-01-07 |
| GB2257431A (en) | 1993-01-13 |
| AU657560B2 (en) | 1995-03-16 |
| AU2195592A (en) | 1993-01-25 |
| EP0591369B1 (en) | 2001-09-19 |
| GB2257431B (en) | 1995-04-19 |
| GB9114003D0 (en) | 1991-08-14 |
| JP3514755B2 (ja) | 2004-03-31 |
| ATE205880T1 (de) | 2001-10-15 |
| KR100222163B1 (ko) | 1999-10-01 |
| ZA924774B (en) | 1993-04-22 |
| JPH06509223A (ja) | 1994-10-20 |
| NZ243330A (en) | 1994-06-27 |
| FI935877A (fi) | 1993-12-27 |
| NO315050B1 (no) | 2003-06-30 |
| EP0591369A1 (en) | 1994-04-13 |
| NO934842L (no) | 1994-02-28 |
| FI935877A0 (fi) | 1993-12-27 |
| DE69232069D1 (de) | 2001-10-25 |
| GB9213644D0 (en) | 1992-08-12 |
| IE922085A1 (en) | 1992-12-30 |
| FI110613B (fi) | 2003-02-28 |
| DE69232069T2 (de) | 2002-01-31 |
| US5698424A (en) | 1997-12-16 |
| WO1993000434A1 (en) | 1993-01-07 |
| NO934842D0 (no) | 1993-12-27 |
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