KR100663319B1 - 인간 17-1에이항원에 대해 특이성을 갖는 인간항체 및 그용도 - Google Patents
인간 17-1에이항원에 대해 특이성을 갖는 인간항체 및 그용도 Download PDFInfo
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- KR100663319B1 KR100663319B1 KR1020057008975A KR20057008975A KR100663319B1 KR 100663319 B1 KR100663319 B1 KR 100663319B1 KR 1020057008975 A KR1020057008975 A KR 1020057008975A KR 20057008975 A KR20057008975 A KR 20057008975A KR 100663319 B1 KR100663319 B1 KR 100663319B1
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Abstract
Description
Claims (20)
- 악성종양세포 표면에 발현된 것과 같은 인간 17-1A항원에 대해 특이성을 갖는 인간 항체에 있어서,상기 항체의 적어도 VH 영역이 인간 IgD 레퍼토리(repertoire)로부터 얻어지는 것을 특징으로 하는 인간 17-1A항원에 대해 특이성을 갖는 인간 항체.
- 제 1항에 있어서,상기 인간항체는 항인(anti-human) 항원 수용체를 제조하기 위한 방법에 따라 얻어질 수 있고,상기 방법은,기능적으로 재배열된 VH 및 VL 면역글로블린 사슬의 조합을 선택하는 단계(b)로서, 적어도 상기 VH 사슬은 초회항원 자극되지 않은 성숙한 인간 B-림프구(unprimed mature human B-lymphocytes)의 인간 IgD 레퍼토리로부터 얻어지고, 상기 VL 사슬은 자연적으로 발생하는 인간 B세포 레퍼토리로부터 얻어지며, 상기 사슬들은 재조합 벡터로부터 발현되는 상기 단계(b)와,인간항원에 결합하기 위하여 인 비트로 디스플레이 시스템(in vitro display system)을 이용하는 단계(c)로 구성되며,상기 선택단계(b)에 앞서 다른 종으로부터의 목표 항원 특이항체(target antigen-specific antibody)의 V 사슬과 함께 상기 항원에 결합하기 위해 상기 VH 사슬 또는 상기 VL 사슬을 선택하는 단계(a)가 수행되는 것을 특징으로 하는 인간항체.
- 제 1항에 있어서,상기 인간항체는 면역글로블린 또는 Fv-단편과 같은 면역글로블린 단편을 제조하기 위한 방법에 따라 얻어질 수 있고,상기 방법은,기능적으로 재배열된 VH 및 VL 면역글로블린 사슬의 조합을 선택하는 단계(b)로서, 적어도 상기 VH 사슬은 초회항원 자극되지 않은 성숙한 인간 B-림프구(unprimed mature human B-lymphocytes)의 인간 IgD 레퍼토리로부터 얻어지고, 상기 VL 사슬은 자연적으로 발생하는 인간 B세포 레퍼토리로부터 얻어지며, 상기 사슬들은 재조합 벡터로부터 발현되는 상기 단계(b)와,인간항원에 결합하기 위하여 인 비트로 디스플레이 시스템(in vitro display system)을 이용하는 단계(c)로 구성되며,상기 선택단계(b)에 앞서 다른 종으로부터의 목표 항원 특이항체(target antigen-specific antibody)의 V 사슬과 함께 상기 항원에 결합하기 위해 상기 VH 사슬 또는 상기 VL 사슬을 선택하는 단계(a)가 수행되는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제3항에 있어서,상기 VH와 상기 VL면역글로블린 사슬은 모두 인간 IgD 레퍼토리로부터 얻어지는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 인 비트로 디스플레이 시스템은 파지(phage) 디스플레이 시스템이 되는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 재배열된 사슬의 조합은 하나 이상의 다른 라이브러리로부터 발현되는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 인간 항원은 17-1A와 같은 종양 항원인 되는 것을 특징으로 하는 인간항체.
- 제 7항에 있어서,상기 VH 사슬은 도 7(뉴클레오티드 1 내지 381)과 도 8(뉴클레오티드 1 내지 339)에 도시된 두 개의 서열중 하나로 구성되고,상기 VL 사슬은 도 6(뉴클레오티드 1 내지 321)과 도 9(뉴클레오티드 1 내지 321)에 도시된 두 개의 서열중 하나로 구성되는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 선택단계(b)는,(ⅰ) 항원수용체를 발현하는 표시 비히클(display vehicle)을,(a) 고정된 목표 항원 또는 그 단편에,(b) 목표 항원 또는 그 단편을 발현하는 선택적으로 표지된 세포에 또는,(c) 가용성 목표 항원 또는 그 단편과 결합시키고,(ⅱ) 비특이적으로 결합한 표시 비히클(a와 b)을 세척한 후, 특이적으로 결합한 표시 비히클을 추출하거나,(ⅲ) 목표 항원 용액 또는 목표 항원을 발현하는 세포의 현탁액에서 목표항원과 결합한 표시 비히클(b와 c)의 포지티브를 증가시키므로,항원 수용체를 포함하는 분리된 표시 비히클은 선택적으로 복제에 의해 배가되며, 상기 (ⅰ) 내지 (ⅲ)에 기술된 인 비트로 선택(in vitro selection)을 수차례 추가로 반복되는 과정을 거치게 되는 것을 특징으로 하는 인간항체.
- 제 9항에 있어서,상기 가용성 목표 항원 또는 그 단편은 표지된 목표 항원 또는 그 단편인 것을 특징으로 하는 인간항체
- 제 2항 또는 제 3항에 있어서,상기 다른 종으로부터의 목표항원 특이항체의 사슬은 쥐의 VH 또는 VL 사슬이 되는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 적절한 조합의 선택은,(a) 상기 인간 항원에 결합하기 위해 서로 다른 다양한 VL 사슬과 함께 한 개의 동일한 VH 사슬을 테스트하는 단계 또는,(b) 상기 인간 항원에 결합하기 위해 서로 다른 다양한 VH 사슬과 함께 한 개의 동일한 VL 사슬을 테스트하는 단계를 포함하는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 방법은,선택단계 후, 인간 VH 사슬과 인간 VL 사슬 또는 그에 해당하는 핵산을 얻는 단계와,상기 사슬을 동일하거나 또는 다른 VH 사슬 또는 VL 사슬과 융합하여, 무겁거나(CH) 가벼운 사슬(CL) 또는 그 일부의 면역글로블린 불변영역 또는 비면역글로블린 사슬과 그에 해당하는 핵산으로 각각 만드는 단계를 추가로 포함하여 구성되는 것을 특징으로 하는 인간항체.
- 제 13항에 있어서,상기 불변영역의 사슬은 IgG1 또는 IgG3로부터 얻어지는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 방법은,선택단계 후에, 인간 VH 사슬과 VL 사슬을 얻는 단계와,상기 사슬을 되도록이면 방사성 동위원소 또는 화학치료에 사용되는 물질과 같은 낮은 분자량의 비단백질 약물과 물리적으로 연결하는 단계를 추가로 포함하여 구성되는 것을 특징으로 하는 인간항체.
- 제 2항 또는 제 3항에 있어서,상기 VH 사슬 또는 VL 사슬은 초회항원 자극되지 않은 성숙한 인간 B-림프구(unprimed mature human B-lymphocytes)의 인간 IgD 레퍼토리로부터 얻어지는 mRNA의 RT-PCR 증폭의 결과인 핵산서열로부터 발현되는 것을 특징으로 하는 인간항체.
- 제 1항 또는 제 2항에 있어서,상기 항체는 바람직하게는, 펩티드 번호 8, 11, 13, 14, 59, 60, 77 및 79의 적어도 하나의 아미노산 서열로 구성되는 17-1A 항원의 세포외 영역의 에피토프(epitope)를 인식하는 것을 특징으로 하는 인간항체.
- 제 1항 또는 제 2항에 있어서,상기 VH 사슬은 도 7(뉴클레오티드 1 내지 381) 및 도 8 (뉴클레오티드 1 내지 339)에 도시된 두 개의 서열 중 적어도 하나의 CDR로 구성되며, 그리고/또는 상기 VL 사슬은 도 6(뉴클레오티드 1 내지 321) 및 도 9(뉴클레오티드 1 내지 321)에 도시된 두 개의 서열 중 적어도 하나의 CDR로 구성되는 것을 특징으로 하는 인간항체.
- 제 1항 또는 제 2항 중 어느 한 항의 항체와 선택적으로 약학적으로 수용가능한 담체(carrier)를 포함하는 종양을 치료하거나 또는 예방하기 위한 약학적 조성물.
- 제 19항에 있어서,상기 종양은 상피에서 기원하는 것을 특징으로 하는 약학적 조성물.
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KR10-1999-7009425A KR100516133B1 (ko) | 1997-04-14 | 1998-04-14 | 항-인간항원 수용체의 신규한 제조방법 및 용도 |
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