KR100897938B1 - 면역글로불린 융합 단백질 - Google Patents
면역글로불린 융합 단백질 Download PDFInfo
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- KR100897938B1 KR100897938B1 KR1020087018012A KR20087018012A KR100897938B1 KR 100897938 B1 KR100897938 B1 KR 100897938B1 KR 1020087018012 A KR1020087018012 A KR 1020087018012A KR 20087018012 A KR20087018012 A KR 20087018012A KR 100897938 B1 KR100897938 B1 KR 100897938B1
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Abstract
Description
hFc 도메인 | Ig 단편의 허용가능한 범위 | N-말단에서 C-말단 방향으로, 허용가능한 범위내의 가장 긴 단편의 서열 | 서열번호 | 서열번호의 위치 | EU 색인의 위치 * |
CH1 (Z1) | IgG1 CH1의 5-9 C-말단 아미노산 잔기 | SNTKVDKRV** | 11 | 90-98 | 207-215 |
IgD CH1의 5-9 C-말단 아미노산 잔기 | ASKSKKEIF | 14 | 90-98 | Not available | |
Hinge (Y) | IgG1 힌지 영역의 5-15 C-말단 아미노산 잔기 | EPKSCDKTHTCPPCP | 11 | 99-113 | 216-230 |
IgD 힌지 영역의 5-64 C-말단 아미노산 잔기 | RWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECP | 14 | 99-162 | Not available | |
CH2, N-말단 쪽 (Z2) | IgG2 CH2의 4-37 N-말단 아미노산 잔기 | APPVAGPSVFLFPPKPKDTLMISRTPEVTWVVVDVSH | 12 | 111-147 | 231-267 |
IgD CH2 도메인의 4-37 N-말단 아미노산 잔기 | SHTQPLGVYLLTPAVQDLWLRDKATFTCFVVGSDLKD | 14 | 163-199 | Not available | |
CH2, C-말단 쪽 (Z3) + CH3(Z4) | IgG4 CH2의 71-106 C-말단 아미노산 잔기 + IgG4 CH3의 80-107 N-말단 아미노산 잔기 | LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHN AKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK + GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ EGNVFSCSVMHEALHNHYTQKSLSLSLGK | 13 | 115-220 + 221-327 | 235-340 + 341-447 |
IgG3 CH2의 71-106 C-말단 아미노산 잔기 + IgG3 CH3 도메인의 80-107 N-말단 아미노산 잔기 | LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFKWYVDGVEVHNAKTKPREEQYNSTFRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKTK + GQPREPQVYTLPPSREEMTKNQVSLTCPVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQ QGNIFSCSVMHEALHNRFTQKSLSLSPGK | 24 | 165-270 + 271-377 | 235-340 + 341-447 | |
IgG2 CH2의 71-106 C-말단 아미노산 잔기 + IgG2 CH3 도메인의 80-107 N-말단 아미노산 잔기 | VAGPSVFLFPPKPKDTLMISRTPEVTWVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFCVVSVLTVVHQD WLNGKEYKCKVSNKGLPAPIEKTISKTK + GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK | 12 | 114-219 + 220-326 | 234-340 + 341-447 | |
IgG1 CH2의 71-106 C-말단 아미노산 잔기 + 80-107 N-말단 아미노산 잔기 | LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVL TVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAK + GQPREPQVYTLPPSRDELTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | 11 | 118-223 + 224-330 | 234-340 + 341+447 |
Claims (90)
- 하기 식으로 표현되는 폴리펩타이드:N'-(Z1)p-Y-Z2-Z3-Z4-C'상기 N'은 폴리펩타이드의 N-말단이고 C'는 폴리펩타이드의 C-말단이고;p는 0 또는 1인 정수이고; 및(i) Z1은 서열번호 11의 90 내지 98 위치의 아미노산 잔기를 가지는 아미노산 서열이고,Y는 서열번호 11의 99 내지 113 위치의 아미노산 잔기를 가지는 아미노산 서열이며,Z2는 서열번호 12의 111 내지 116 위치의 아미노산 잔기를 가지는 아미노산 서열이고,Z3-Z4는 서열번호 13의 118 내지 220 위치의 아미노산 잔기 및 서열번호 13의 221 내지 327 위치의 아미노산 잔기를 가지는 연속된 아미노산 서열로 구성된 아미노산 서열이거나, 또는(ii) Z1은 서열번호 14의 90 내지 98 위치의 아미노산 잔기를 가지는 아미노산 서열이고,Y는 서열번호 14의 158 내지 162 위치의 아미노산 잔기, 서열번호 14의 153 내지 162 위치의 아미노산 잔기, 서열번호 14의 148 내지 162 위치의 아미노산 잔기, 서열번호 14의 143 내지 162 위치의 아미노산 잔기, 서열번호 14의 133 내지 162 위치의 아미노산 잔기, 또는 서열번호 14의 99 내지 162 위치의 아미노산 잔기를 가지는 아미노산 서열이고,Z2는 서열번호 14의 163 내지 170 위치의 아미노산 잔기를 가지는 아미노산 서열이고,Z3-Z4는 서열번호 13의 121 내지 220 위치의 아미노산 잔기 및 서열번호 13의 221 내지 327 위치의 아미노산 잔기를 가지는 연속된 아미노산 서열로 구성된 아미노산 서열이다.
- 제1항에 있어서, 서열번호 1, 서열번호 2, 서열번호 3, 서열번호 4, 서열번호 5, 서열번호 6, 서열번호 26 및 서열번호 27로 구성된 군으로부터 선택된 뉴클레오타이드 서열에 의해 코딩되는 폴리펩타이드.
- 제1항에 있어서, 서열번호 18, 서열번호 19, 서열번호 20, 서열번호 21, 서열번호 22, 서열번호 23, 서열번호 28 및 서열번호 29로 구성된 군으로부터 선택된 아미노산 서열을 갖는 폴리펩타이드.
- 제1항의 폴리펩타이드 및 생물학적 활성 분자를 포함하는 키메릭 폴리펩타이드에 있어서, 상기 생물학적 활성 분자는 제1항의 폴리펩타이드의 N-말단 또는 C-말단에 융합된 것이고, 상기 생물학적 활성 분자는 그 생물학적 활성 분자의 천연형의 순환 반감기에 비해 증가된 순환 반감기를 나타내는 것인 키메릭 폴리펩타이드.
- 제4항에 있어서, 상기 생물학적 활성 분자는 가용성 단백질인 키메릭 폴리펩타이드.
- 제5항에 있어서, 상기 생물학적 활성 분자는 호르몬, 사이토카인, 성장인자, 공동-자극 분자, 호르몬 수용체, 사이토카인 수용체, 성장인자 수용체, 또는 짧은 펩타이드인 키메릭 폴리펩타이드.
- 제5항에 있어서, 상기 생물학적 활성 분자는 EPO 또는 그의 변이체/단편인 키메릭 폴리펩타이드.
- 제5항에 있어서, 상기 생물학적 활성 분자는 p40, TNF 수용체 또는 이의 변이체/단편인 키메릭 폴리펩타이드.
- 제8항에 있어서, 상기 생물학적 활성 분자는 Asn303Gln 치환을 포함하는 p40 변이체인 키메릭 폴리펩타이드.
- 제5항에 있어서, 상기 생물학적 활성 분자는 G-CSF 또는 그의 변이체/단편인 키메릭 폴리펩타이드.
- 제5항에 있어서, 상기 생물학적 활성 분자는 GM-CSF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, 인간 성장 호르몬, BMP-1(bone morphogenetic protein-1), 인터페론 α, IL-10, FSH, 혈액 인자 VII, CTLA4, IFN-β, PD-1, IL-10R, CXCL-11, TGF-β수용체, TGF-β, IL-17R, GLP-1, BTC, OPG, RANK 또는 그들의 변이체/단편인 키메릭 폴리펩타이드.
- 제4항에 있어서, 상기 생물학적 활성 분자는 분비 단백질인 키메릭 폴리펩타이드.
- 제12항에 있어서, 상기 생물학적 활성 분자는 분비 단백질의 성숙형(mature form)인 키메릭 폴리펩타이드.
- 제4항에 있어서, 상기 생물학적 활성 분자는 항체의 Fab 영역인 키메릭 폴리펩타이드.
- 제4항에 있어서, 상기 폴리펩타이드 및 상기 생물학적 활성 분자는 링커를 통해 서로 연결된 것인 키메릭 폴리펩타이드.
- 제15항에 있어서, 상기 링커는 알부민 링커 또는 합성 링커인 키메릭 폴리펩타이드.
- 제16항에 있어서, 상기 링커는 알부민 링커인 키메릭 폴리펩타이드.
- 제17항에 있어서, 상기 알부민 링커는 서열번호 25의 아미노산 서열 321 내지 323, 318 내지 325, 316 내지 328, 313 내지 330, 311 내지 333, 또는 306 내지 338을 포함하는 것인 키메릭 폴리펩타이드.
- 제16항에 있어서, 상기 합성 링커는 Gly 및 Ser 잔기로 구성된 10 내지 20개의 아미노산 잔기의 펩타이드인 키메릭 폴리펩타이드.
- 제16항에 있어서, 상기 합성 링커는 GGGGSGGGGSGGGSG (서열번호 32)인 키메릭 폴리펩타이드.
- (i) 포유류 숙주 세포에 폴리펩타이드를 코딩하는 DNA 분자를 도입하는 단계; (ii) 성장 배지에서 폴리펩타이드가 발현될 수 있는 조건하에서 세포를 키우는 단계; 및 (iii) 발현된 폴리펩타이드를 수득하는 단계를 포함하는, 제1항에 따른 폴리펩타이드의 생산 방법.
- 제21항에 있어서, 상기 포유류 숙주세포는 CHO, COS 또는 BHK 세포인 방법.
- 제4항의 키메릭 폴리펩타이드를 코딩하는 단리된 핵산 분자.
- 제23항에 있어서, 상기 핵산 분자는 서열번호 18, 서열번호 19, 서열번호 20, 서열번호 21, 서열번호 22, 서열번호 23, 서열번호 28 및 서열번호 29로 이루어진 그룹으로부터 선택되는 아미노산 서열을 갖는 키메릭 폴리펩타이드를 코딩하는 핵산 분자.
- 제24항에 있어서, 상기 핵산 분자는 서열번호 1, 서열번호 2, 서열번호 3, 서열번호 4, 서열번호 5, 서열번호 6, 서열번호 26 및 서열번호 27로 구성된 그룹으로부터 선택되는 어느 하나인 핵산 분자.
- 제23항 내지 제25항 중 어느 한 항에 있어서, 상기 핵산 분자는 신호 서열 또는 리더 서열을 더 포함하는 것인 핵산 분자.
- 제26항에 있어서, 상기 신호 서열은 tPA 신호서열인 핵산 분자.
- 제23항에 따른 핵산 분자를 포함하는 발현 벡터.
- 제28항에 있어서, 상기 발현 벡터는 pAD11의 EPO-hFc-1, pAD11 G-SCF-hFc-1, pAD11 p40N303Q-hFc-1, pAD11 EPO-hFc-6, pAD11 G-SCF-hFc-6, pAD11 p40N303Q-hFc-6, pAD11 EPO-hFc-5, pAD11 G-CSF-hFc-5, pAD11 p40N303Q-hFc-5 및 pAD11 TNFR-hFc-5로 구성된 그룹으로부터 선택되는 것인 발현 벡터.
- 제28항의 발현 벡터를 포함하는 숙주 세포.
- 제29항의 발현 벡터를 포함하는 숙주 세포.
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