KR20130086036A - 인간 사이토메갈로바이러스 gB 단백질에 대한 고친화도 인간 항체 - Google Patents
인간 사이토메갈로바이러스 gB 단백질에 대한 고친화도 인간 항체 Download PDFInfo
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- KR20130086036A KR20130086036A KR1020137001220A KR20137001220A KR20130086036A KR 20130086036 A KR20130086036 A KR 20130086036A KR 1020137001220 A KR1020137001220 A KR 1020137001220A KR 20137001220 A KR20137001220 A KR 20137001220A KR 20130086036 A KR20130086036 A KR 20130086036A
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Abstract
인간 B 세포로부터 인간 사이토메갈로바이러스(CMV) gB 단백질에 대한 항체를 분리해냈다. 이 항체들의 친화도는 종전에 보고된 최고의 항체들보다도 더 높은 것이다. 높은 친화도가 태반을 통과하는 바이러스의 전달을 방지하는데 결정적이기 때문에, 본 발명의 항체는 임신 중의 CMV 감염이 태아에 미치는 영향을 예방하거나 또는 완화하기 위한 약학적 또는 예방적 의약으로써 유용하다.
Description
관련출원
[0001] 본원은 2010년 6월 16일에 출원된 미국 가출원 번호 제61/355,499호에 대하여 우선권을 주장하며, 그 기재 내용은 참조에 의하여 전체로서 본 명세서에 삽입된다.
EFS
-웹을 통하여 제출된 서열 목록에 대한 언급
[0002] MPEP §730 II.B.2(a)(C)에서 위임 및 규정한 바에 따른 미국특허상표청 EFS-웹 서버를 통한 다음과 같은 서열 목록의 전자 제출의 전체 내용은, 참조에 의하여 전체로서 모든 목적을 위하여 본 명세서에 삽입된다. 서열 목록은 전자적으로 제출된 다음의 텍스트 파일에 의하여 특정된다.
기술분야
[0003] 본 발명은 임신기간 동안 태아에 대한 CMV 감염의 효과를 예방하거나 완화하고, 이식 환자를 포함하여 면역력이 약화된 환자의 CMV 감염을 치료하기 위한, 치료 및 예방적 용도의 CMV gB 단백질에 대한 인간 단일클론 항체(mAbs)와 관련된다.
[0004] 사이토메갈로바이러스(CMV)는 이식 환자, 다른 면역력이 약화된 환자 및 신생아에게 질병을 유발하는 주요 인자이다. 미국에서 매년 약 40,000명의 신생아가 CMV를 흩뿌리며 태어난다. 이중 8,000명이 증상 및/또는 심각한 장애를 가지고 태어나고, 최대 8,000명 이상까지 추후 진행성 청력 상실이 발생할 것이다. 임신한 모체의 약 절반이 자연적으로 적절한 면역성을 가지고 있다. 따라서, 효과적인 mAb가 인간 혈액 중에 존재하는 것으로 알려져 있다. 또한, 이는 정맥 투여된 감마 글로불린(IVIG)에 의한 성공적인 면역성의 수동 전달에 의해서도 나타나는데, 이는 태아 보호에 매우 높은 효능을 나타내어 왔다. 이는 이식 환경에서 IVIG에 대하여 관찰된 제한된 효능과 반대되는 것인데, 이식 환경에서는 세포성 면역이 체액성 면역보다 명백하게 더 중요하다.
[0005] CMV에 대한 자연적 반응은 그 대부분이 gB 단백질로부터 기인하는 것이다(Park, J. W., et al ., J. Korean Med . Sci . (2000) 15:133-138). 타운(Towne) 백신은 인 비트로에서 강력하게 다루어 약하게 살아있는 바이러스 백신인데, 이는 섬유모세포 감염을 중화하지만 내피세포 감염은 중화하지 못하는 항체를 생산한다. 이 백신은 안전한 것으로 알려져 있고, 20년 동안 연구되어 왔다(Adler, S. P., et al ., Pediatr . Infect . Dis . J. (1998) 17:200-206). 이하에서 기술되는 바와 같이, gB에 대한 항체 분리에 유용한 혈액 기증자들은 이전에 CMV에 노출된 적이 있는 혈청 양성 반응자 및 타운 백신으로 백신화되기 전 및 후의 혈청 음성 반응자를 포함한다.
[0006] CMV의 gB 단백질에 대한 항체들이 제조되어 왔다(Nozawa N., et al ., J. Clin. Virol . (2009) [Epub ahead of print], Nakajima, N., et al . (US2009/0004198 A1), Lanzavecchia, A, et al . (US2009/0004198 A1), Ohlin, M., et al. (J Virol (1993) 67:703-710). gB의 AD-2 도메인에 대한 중화된 항체인 ITC88가 보고된바 있다(Lantto, Virology (2003) 305:201-209). 그러나 CMV에 대한 인간 항체를 복제하고자 한 기존의 노력은 성공적이었던 반면에 그 범위가 제한되었으며, 고도의 친화도(서브-나노몰)의 항체는 기술된 바 없다. CMV에 대한 약한 친화도의 항체가 인간 태반(Nozawa, supra)을 가로지르는 전달을 촉진하기 때문에,고도의 친화도는 핵심변수인데, 이는 설치류에서는 관찰되지 않는 병리학의 한 모습이다. 인간 CMV은 약 236 kb의 이중사 DNA 게놈을 가지며, β-헤르페스 바이러스 패밀리의 기본형 멤버이다. 게놈이 갖는 고도의 복잡성은 관심대상인 수많은 잠재적 항체가 존재함을 의미한다. 중화 항체들 및 그들과 관련된 에피토프를 밝혀내려는 노력에 의하여 효과적인 중화 반응을 이끌어내는 당단백질 B(gB)에 기반한 서브유닛 백신이 얻어졌지만, 혈청 음성인 여성의 코호트에 시험한 경우 단지 50%의 효능을 가질뿐이다. 이는 CMV 백신 중 최고로 높은 효능으로 보여진다. 백신은 전형적으로 친화도 범위를 가지는 항체를 생산하기 때문에, 현재까지 시험된 백신들의 실망스러운 효능으로부터 높은 친화도의 항체에 대한 요구가 있는데, 이는 직접적인 예방적 전략으로서 높은 친화도의 mAb 공급에 대한 선호를 논증한다.
[0007] 에피토프를 특이적으로 중화하는 면역 반응에 집중하지 못한 것이, 열악한 효능의 원인으로 여겨진다(Marshall, B. C., et al ., Viral Immunol . (2003) 16:491-500). 항-CMV 백신 개발에 있어 의심되는 또 다른 기술적 문제점은, 바이러스 병리학을 이해하기 위하여 다른 종류의 세포 감염이 더욱더 주안점이 되고 있음에도 불구하고, 그들이 오직 섬유 모세포 감염을 중화하는 항체를 생산하는 능력에 대해서만 평가되었다는 것이다. 이러한 편견은 인 비트로에서의 바이러스 성장에 관한 기술적 장애를 반영한다. 섬유 모세포에 대한 반복된 바이러스 경로는 내피 및 상피 세포에 대한 굴성을 잃은 많은 실험실 균주를 야기하였다고 여겨진다. 지난 수년 동안, 이러한 결함은 바이러스 표면에 대한 gH/gL/UL131-UL128 당단백질 복합체의 1 이상의 성분의 상실과 관련되어 왔다.
[0008] 더 효능있는 항-CMV 예방적 전략에 관한 요구가 명백히 존재한다.
[0009] 종전의 항체들(인간 또는 뮤린 항체)과 비교하여 개선된 친화도를 가지고 중화능력을 구비한, CMV gB 단백질에 대한 특이적인 면역반응성이 있는 인간 항체가 제조되었다. 체액성 면역 시스템은 수만개의 잘 분화된 결합능력을 구비한 항체 구조 수백만개를 생산할 수 있지만, 보호성 항체는 이들의 매우 작은 부분에 불과하다. 본 발명자들은 셀 스팟(CellSpot)TM 기술(Harriman, W. D., et al ., J. Immunol. Methods (2009) 34:135-145, Collarini, E. J., et al ., J. Immunol . (2009) 183:6338-6345, 및 미국특허 제7,413,868호, 모두 참조에 의하여 본 명세서에 삽입됨)을 채택하여, CMV에 높은 역가를 가지는 것으로 확인된 기증자의 혈액으로부터 유래한 mAbs 패널을 생산하였다.
[0010] 그러므로 하나의 관점에서, 본 발명은 gB 단백질의 에피토프에 결합하는 인간 단일클론 항체 또는 그의 면역반응성 단편, 바람직한 구현예로는 gB 단백질 내에 보존된 서열에 결합하는 것에 관한 것이다. 이들 항체들은, CMV의 중화에 대한 표준 플라그 형성 분석에서 중화능력을 나타내고, 그러한 분석법에서 <500 ng/ml, 바람직하게는 <200 ng/ml, 더 바람직하게는 <100 ng/ml의 EC50 을 나타낸다. 본 발명의 항체는 또한 CMV 스트레인 AD169 의 gB 단백질에 대하여 <10 nM 또는 <5 nM 또는 <1 nM의 친화도를 갖는다.
[0011] CMV 감염을 치료하거나 CMV에 대한 저항성을 증진시키기 위한 본 발명의 방법에 사용하기 위하여, 본 발명의 단일클론 항체 또는 단편은 다수의 CMV 스트레인에 대해 면역반응성일 수 있고, 1종의 단일클론 항체로도 원하는 효과를 가지기에 충분할 수 있다. 또는 동일하거나 다른 CMV 단백질에 결합하는 1종 초과의 단일클론 항체를 치료하고자 하거나 저항성 있게 만들려는 대상체에게 투여할 수도 있다.
[0012] 본 발명은 또한 본 발명의 항체 또는 면역성 단편 1종, 또는 본 발명의 항체 또는 단편 2종 이상을 활성 약제로서 포함하는, 예방 및 치료에 유용한 약제학적 조성물을 포함한다.
[0013] 본 발명의 다른 관점은 인간 대상체 중의 CMV를 치료하기 위하여 또는 인간 대상체 내 감염에 대한 저항성을 유도하기 위하여 항체를 사용하는 방법을 포함한다.
[0014] 본 발명의 단일클론 항체는 재조합적으로 생산될 수 있으며, 따라서 본 발명은 또한 그러한 항체의 생산을 위한 세포주 또는 불멸화된 세포 및 비인간 다세포 기관 또는 그의 세포, 또는 미생물 세포뿐만 아니라, 그러한 생산을 위한 재조합 물질들을 포함한다. 일 구현예에서, 인간 대상체로부터 취득한 세포는 "불멸화"된 형태로 생산되는데, 여기서 그들은 그들을 특징짓는 관련 인코딩 서열이 클론화된 항체를 충분한 시간 동안 분비하도록 변형된다.
[0015] 도 1A 및 1B는 4A2 및 19B10의 gB 단백질 및 그것의 보존된 영역에의 결합을 나타낸다.
[0016] 도 2A 및 B는 HUVEC 및 HFF 세포 내에서 mAbs 4A2, 310, 313, 338 및 345에 의한 VR1814 중화를 나타낸다.
[0017] 도 3은 HUVECs 내에서 mAbs 4A2, 310, 313, 338 및 345에 의한 VR1814 중화를 나타낸다.
[0018] 도 4는 HFF 세포 내에서 mAbs 4A2, 310, 313, 338 및 345에 의한 VR1814 중화를 나타낸다.
[0016] 도 2A 및 B는 HUVEC 및 HFF 세포 내에서 mAbs 4A2, 310, 313, 338 및 345에 의한 VR1814 중화를 나타낸다.
[0017] 도 3은 HUVECs 내에서 mAbs 4A2, 310, 313, 338 및 345에 의한 VR1814 중화를 나타낸다.
[0018] 도 4는 HFF 세포 내에서 mAbs 4A2, 310, 313, 338 및 345에 의한 VR1814 중화를 나타낸다.
[0019] 본 명세서에서 사용되는 용어 "치료"란 이미 CMV에 감염된 대상체 내 바이러스 부담을 경감시키거나 또는 그러한 대상체의 질병의 증상을 완화시키는 것을 의미한다. 그러한 증상은 망막염 및 간염을 포함한다.
[0020] 용어 "저항성을 부여하다"란 도전중인 RSV에 의한 바이러스 감염이 적어도 그 극심의 정도가 감소하는 예방적 효과를 의미한다.
[0021] "불멸화된 세포"란 변형되지 않은 일차적으로 분리된 세포보다 현저하게 많은 시간을 생존할 수 있는 세포를 의미한다. 본 발명의 문맥에서 사용되는 봐와 같이 "불멸화된"이란 세포가 매우 오린 기간에 걸쳐 항체를 계속적으로 분비하는 것을 필수적으로 의미하는 것이 아니며, 단지 그들이 일차 세포 배양과 비교하여 더 오래 생존하는 것을 의미한다. 항체 분비가 일어나는 시간은 그것을 동정하고, 인코딩하고 있는 핵산 서열을 수득하기에 충분한 시간이면 족하다.
[0022] 본 명세서에서 인간 세포로부터 분리한 것과 같은 인간 항체는 강한 면역 반응을 유도하지 않는다. 인간 항체는 심지어 인간으로부터 분리된 항체들의 경우에도, 처리된 인간의 5-10%에서 면역반응을 유도하는 것으로 알려져 있는데, 이는 유도되는 면역 반응 중에는 일정 수준의 배경 "노이즈"가 있기 때문이다. 면역반응은 체액성 또는 세포성이거나 또는 둘 모두일 수 있다. 특히 이러한 개인 퍼센트에서 사이토카인 수준이 상승되는 것이 발견될 수 있다.
[0023] CMV의 gB 단백질은 130 kDa의 전구체 단백질로서 합성되는데, 이는 공유적으로 연결된 채 남아 있는 116 kDa (N-말단) 및 58 kDa (C-말단)의 단편으로 절단될 수 있다. 관찰되는 분자량은 글리코실화 상태에 따라 달라질 수 있다. AD-2 항원결정부위란 gp116의 67-82 잔기를 의미한다. CMV에 대한 천연 면역성의 중요한 부분이 AD-2에의 결합으로 설명된다(즉, 이 영역을 덮는 펩타이드에 의하여 차단될 수 있다, 상술한 Ohlin 참조).
[0024] 본 발명의 항체는, 실시예 1에서 기재한 바와 같이 참조에 의하여 모두 삽입되는 미국특허 제7,413,868호, PCT 공개 WO 2005/045396 및 WO 2008/008858에 기술된 바와 같은, 독점적인 셀 스팟(CellSpot)TM 기법을 사용하여 CMV에 노출된 인간 기증자로부터 수득한 것이다.
[0025] 본 발명의 인간 또는 인간화된 항체의 생산은 중국 햄스터 난세포 또는 곤충 세포와 같은 다른 유핵 세포주 내에서의 생산과 같은 전통적인 재조합 기법을 사용하여 달성한다. 또는 항체를 포함하는 재조합 물질들을 식물 및 형질 전환 동물 중에서, 예컨대, 보바인유 중에서, 또는 미생물 또는 식물 또는 곤충 유래 단일 세포 시스템에서, 또는 그와 같은 세포들의 무세포 추출물중에서 생산하는 방법이 알려져 있다.
[0026] 또한, 항체를 인코딩하는 뉴클레오타이드 서열이 이용가능하기 때문에, 동일한 에피토프에 결합하는 관련 단편들, 예컨대 Fab, F(ab')2 또는 Fv 단편들을 재조합적으로(또는 단백질 그 자체를 단백질분해 처리하는 것에 의하여) 생산할 수 있고, 항체는 단일쇄 형태로 생산될 수 있다. 재조합 항체 생산을 조작하는 다양한 기법들이 당업계에 알려져 있다.
[0027] 키메라, 인간화 및 인간 항체 모두가 본 발명의 범위 내에 속하며, 피브로넥틴, 트랜스페린 또는 리포칼린과 같은 다른 단백질 스캐폴드에 기초한 항체 모조품 또한 그러하다. 유사하게, 분리된 2개의 항체로부터 유래한 항원 특이적 도메인이 결합된 단일의 항체 유사 분자를 만들기 위한 다수의 기술이 현재 존재한다. 적절한 기술은 문헌들(Macrogenics (Rockville, MD), Micromet (Bethesda, MD) 및 Merrimac (Cambridge, MA). 예컨대, Orcutt KD, Ackerman ME, Cieslewicz M, Quiroz E, Slusarczyk AL, Frangioni JV, Wittrup KD 참조. A modular IgG-scFv bispecific antibody topology. Protein Eng Des Sel . (2010) 23:221-228; Fitzgerald J, Lugovskoy A. Rational engineering of antibody therapeutics targeting multiple oncogene pathways. MAbs . (2011) 1;3(3); Baeuerle PA, Reinhardt C. Bispecific T-cell engaging antibodies for cancer therapy. Cancer Res. (2009) 69:4941-4944 참조)에 기술되어 있다.
[0028] 그러므로, 다른 CMV 에피토프에 대한 반응성을 갖는 개개의 항체들의 Fab 도메인을 사용하여, 매우 넓은 스트레인 반응성을 가지는 단일의 항체를 구성할 수 있는데, 예컨대 이중-특이적 항체는 gB 및 gH 복합체 모두에 대한 활동성을 갖거나, 또는 동일하거나 또는 다른 스트레인으로부터 유래한 gB 단백질에 대하여 반응성일 수 있다. 고도의 친화도 gH 항체들이 문헌(예컨대, Macagno A, Bernasconi NL, Vanzetta F, Dander E, Sarasini A, Revello MG, Gerna G, Sallusto F, Lanzavecchia A, gH/gL/UL128-131A 복합체 상의 상이한 에피토프들을 표적화함에 의하여 인간 사이토메갈로바이러스 감염을 잠재적으로 중화하는 인간 단일클론 항체의 분리, J Virol . (2010) 84:1005-1013)에 기술된 바 있다. 다른 스트레인으로부터 유래한 gH 단백질들에 대한 고도의 친화도 항체들이 또한 발생되고 사용될 수 있다.
[0029] 치료에 사용하기 위하여, 재조합적으로 생산된 항체 또는 단편들은 적절한 부형제를 사용하여 약학적 조성물로 제제화될 수 있고, 표준 프로토콜에 따라 투여될 수 있다. 약학적 조성물은 본 발명의 단일클론 항체 또는 그의 단편, 특히 모든 CMV 스트레인의 gB 단백질에 교차 반응성인 단일클론 항체 또는 단편을 오직 단일의 활성 성분으로서 포함할 수 있다. 또는, 2종의 단일클론 항체가 단일의 활성 성분일 수도 있는데, 여기서 1종은 하나의 스트레인 gB 단백질과 더 강하게 반응하고, 다른 것은 또 다른 스트레인 gB 단백질과 더 강하게 반응한다. 이 모든 경우에 있어서, 다른 CMV 단백질에 결합하는 것들을 포함하여, 추가적인 약학적 제제가 존재할 수 있다. 또한, 화합물들은 비타민 또는 항체가 아닌 다른 유익한 화합물과 같은 영양 물질을 포함할 수 있다.
[0030] 일 구현예에서, 투여를 위한 제제가 감염에 대한 저항성을 증가시키기 위하여 사용되는 경우, 보체-포함 Fc 영역을 포함하는 완전 항체가 채택될 수 있다. 전형적으로 항체들은 인간 대상체에게 0.01-20 mg/kg 의 복용 수준으로 투여되거나, 또는 0.01-5 mg/kg 범위 내 투여량 또는 그 범위 내 중간량으로 투여된다. 일 구현예에서 0.1-1.0 mg/kg 범위 내 양이 채택된다. 수일 또는 수주 또는 수개월에 의하여 분리된 반복된 투여가 이로울 수 있다.
[0031] 또 다른 구현예에서, 바이러스 로드를 감소시키기 위한 약학적 효과를 위하여 보체-포함 Fc 영역을 포함하는 완전항체가 또한 채택된다. 이러한 프로토콜에서 투여되는 양은 0.001-50 mg/kg의 처방 또는 0.01, 1 또는 10 mg/kg과 같은 그 범위 내 중간값이 채택된다. 반복되는 투여 또한 사용할 수 있다. 비록 수일 내 투여가 또한 본 발명의 범위 내에 속하지만, 감염 진단 이후 되도록 빨리 약학적 치료법을 투약한다. 반복되는 투여 또한 채택될 수 있다. 폐에서의 염증 반응을 감소시키기 위하여, 오직 항체의 면역 특이적 단편만이 채택될 필요가 있다. 투여 레벨은 전항체를 위한 것과 유사하다. 면역 특이적 단편과 전항체 혼합물으 투여 또한 본 발명의 범위 내에 포함된다.
[0032] 본 발명의 항체 조성물의 투여는 전형적으로 주사, 일반적으로 정맥 주사에 의한다. 따라서, 비경구적 투여가 선호된다. 그러나, 유전자 치료(인 비보에서 재조합 항체를 생산하는 것)를 포함하는 그 어떠한 실행 가능한 투여의 방식이 포함된다.
[0033] 제제는 항체 조성물을 투여하는 기술 분야에서 일반적으로 알려진 방법에 따라 제조된다. 적합한 제제는 참조에 의하여 본 명세서에 삽입되는 레밍톤 약제학 과학(Remington's Pharmaceutical Sciences , latest edition, Mack Publishing Co., Easton, PA)와 같은 표준 처방서 중에서 발견될 수 있다. 제제는 비경구 투여에 전형적으로 적합한 것들로서, 리포좀, 마이셀 및 나노입자와 같은 전달 담체를 포함하는 에멀전 뿐만 아니라, 완충액, 산화제 등을 포함하는 등장 용액을 포함한다.
[0034] 바람직한 프로토콜 및 제제는 대상체의 특정 컨디션뿐만 아니라, 수행하고 있는 실무자의 판단에 의존한다. 투여 수준은 적절하다면 대상체의 연령, 일반적인 건강 및 감염의 심각성에 의존할 수 있다.
[0035] 다음의 실시예들은 설명을 위하여 제공되며, 발명을 제한하기 위한 것이 아니다.
살시예
1
CMV
gB
에 대한 항체를 분비하는 인간 B 세포의 분리
[0036] CMV에 대한 확인된 역가를 가지는 50명의 성인으로부터 유래한 말초 혈액 단핵세포에 대하여, 항-바이러스 항체를 생산하는 인간 B 세포를 조사하였다. 특이적 mAbs를 클로닝하기 위하여 CMV gB 단백질에 대한 원하는 항체를 가진 대상체를 사용하였다. 조사 결과, ~10%의 대상체가 50,000 중 1보다 더 큰 빈도로 원하는 세포를 가지는 것으로 조사되었다.
[0037] 희귀한 유망 세포의 조사 및 회복을 달성하기 위하여, 본 발명자들은 종전에 기술된 셀스팟TM 기술(U.S. 7,413,868, incorporated herein by reference)을 사용하였다. 셀스팟TM 분석법은 세포 근처의 풋 프린트로서 단일 세포로부터 분비된 IgG를 포착함으로써 거의 단일 세포의 용적의 실질적인 웰까지 ELISA 등가 분석을 축소하여, 수백만개의 세포가 쉽게 분석되도록 한다. 더 나아가, 현미경 복합 시약(참조로서 본 명세서에 삽입된 미국특허 제6,642,062호의 조합된 형광색 라텍스 마이크로스피어)을 사용함에 의하여, 다중 생화학적 프로브를 사용하여 각각의 클론이 분비한 항체 풋 프린트가 그 특이성 및/또는 친화도에 대하여 상세하게 밝혀질 수 있다. 정량적 분석의 신뢰도는 기존의 감정 분석법과 일치하는 표현형을 나타내는 클론된 발현 세포를 가지고, 조사 군집으로부터 극히 희귀한 유망 세포의 포착을 가능하게 할 정도로 충분하다.
[0038] 스크리닝 표준은 바이러스 용리액뿐만 아니라, 정제된 gB 단백질에도 결합하는 것이었다. gB 단백질은 CMV의 AD169 스트레인으로 감염시킨 293개 세포로부터 정제하였다. 비드 상의 항원을 혈청 알부민으로 희석시키어, 클론의 친화도 정렬 순서를 결정한다. 이는 분비된 IgG 풋 프린트("친화도 효과")에 다 돌기성으로 결합하는 기회를 감소시키어, 더 높은 내재적 친화성을 선택한다.
[0039] 표준 자기 분리법을 사용하여, 인간 기증자의 혈장 중 PBMCs로부터 B 세포가 아닌 것을 고갈시켰다. 1e6/ml에서 IMDM/20% HI-FCS 중에 세포를 재현탁시키고, EBV(감염된 B95-8 세포의 상등액으로부터의 직접 알갱이화된 것)로 불멸화시켰다. 1:100 희석으로 EBV를 첨가하였고, 37℃에서 2시간 동안 세포를 배양하였다. 과량의 EBV는 세척해내었고, 세포는
(1) 오직 조사만을 위하여, IMDM, 20%의 HI-FCS, 20%의 거대 세포 종양 조절 배지, 2 mg/ml의 CpG (ODN2006), 및 10 ng/ml의 IL-10중에서, 2e6/ml으로 배양되거나, 또는
(2) 자기(magnetic) 양성 선택을 사용하여 표면 IgG를 더 선택하였다.
[0040] 세포는 IMDM, 20%의 HI-FCS, 20%의 거대 세포 종양 조절 배지, 2 mg/ml의 CpG (ODN2006), 및 10 ng/ml IL-10 중에서, 조사된 인간 폐 세포(MRC-5, 5,000 cells/well) 상의 200-300 세포/웰에서 배양되었다. 매 2~3일마다 배지를 보충하였다. 6일째 날에, 웰에 있는 성분의 절반을 셀스팟TM 중에서 분석하였다. 그 다음, 조사 분석에 의할 때 양성인 소수의 웰에 남아 있는 세포를 동일한 피더 세포 및 배양 조건으로 10, 5, 1 및 0.5 세포/웰까지 희석시켰다. 4~5일 후에, 이러한 제한적인 희석 접시를 ELISA 또는 셀스팟TM으로 다시 분석하였다.
[0041] 원하는 항체를 스크리닝하기 위하여, 셀스팟TM 나노 입자를 바이러스 용리액 또는 정제된 gB 단백질과 컨주게이션시켰다. CMV AD169 바이러스로 감염된 세포로부터 생산된 용출액은 Virusys (cat # CV046)로부터 구입하였다. (용리액은 NHDF(Normal Human Dermal Fibroblast ) 세포주로부터 생산된다.) 재조합 CMV gB 항원은 293개 세포 중 His-태그된 융합 단백질로서 생산되었고, 니켈 킬레이션 컬럼을 사용하여 정제되었다. 정제된 gB 단백질을 ELISA 및 셀스팟을 위하여 사용하였다. 상기 기술한 바와 같이, AD169 용리액 및 gB 정제된 단백질의 조합물을 나노 입자와 각각 컨주게이션시켰다(참고: Harriman et al(상술함) 및 Collarini et al(상술함)).
[0042] 그 다음으로, 항체의 중쇄 및 경쇄를 인코딩하는 mRNA를 수득하기 위하여 역전사-PCR을 사용하여 제한된 희석에서의 양성적 웰의 성분을 처리하였다. RT-PCR을 통하여 인코딩 mRNA 서열을 회복하기 위하여 PBMC를 해동하는 총 시간은 10~12일이었다.
실시예
2
CMV
gB
에 대한 인간 항체의
클로닝
[0043] 세미-네스티드 PCR(semi-nested polymerase chain reaction)을 사용하여 양성적 ELISA 웰로부터 유래한 재배열된 Ig 중쇄 및 Ig 경쇄의 증폭을 수행하였다. 종전에 알려진 V-유전자 재배열의 증폭을 위하여 패밀리-특이적인 V-유전자 프라이머들의 수집을 구성하였는데, 이는 인간 Ig 유전자 자리에 있어서 거의 모든 V-유전자 세그먼트를 인식한다. Cg, Ck 및 Cl 유전자 세그먼트에 대하여 특이적인 프라이머 혼합물과 함께 5' 프라이머를 사용하였다. 제한된 희석 CMV-gB 특이적 B 세포의 클론화도는 구별되는 자손 세포로부터 유래한 V-유전자 증폭물의 서열 비교에 의하여 명백하게 결정되었고, 증폭된 전장 V-유전자 재배열은 IgG 발현 벡터로 클론되었다.
[0044] 구체적으로, 상업적으로 이용가능한 RNA 정제 키트 (RNeasyTM Qiagen (독일))를 사용하여, 분리된 인간 B 세포로부터 유래한 총 mRNA를 추출하였다. 총 RNA 준비물 및 프라이머로서 올리고뉴클레오타이드를 사용하여 역 전사-PCR을 행하였다. 세개의 PCR 반응을 하나는 경쇄 카파(k), 하나는 경쇄 람다(l), 그리고 하나는 감마 중쇄(g)인 각각의 시료에 대하여 수행하였다. 증폭을 위하여 QIAGEN® 원스텝 RT-PCR 키트를 사용하였다(Qiagen 카탈로그 No. 210212). 커플된 RT-PCR 반응에서, C-k, C-l 또는 Cg 유전자의 CH1 영역의 교감에 해당하는 특이적 안티센스 프라이머 서열을 사용한 RT 효소의 독특한 혼합물(OmniscriptTM 및 SensiscriptTM)을 사용하여 cDNA를 합성하고, RT는 50℃에서 1시간 동안 수행되며, 그 다음으로 고도의 특이성 및 민감성을 위하여 HotStarTaq DNA 폴리머라제에 의하여 cDNA의 PCR 증폭을 수행한다. 각각의 PCR 반응은 5' 센스 프라이머의 혼합물을 사용하였다. 프라이머 서열은 VH, VK 및 VL의 리더 서열에 기초하였다. PCR 반응은 초기 열시점 95℃에서 15분간 수행하고, 그 다음으로 95℃에서 30초 (변성화), 60℃에서 45초(어닐링) 및 72℃에서 1분간(신장)의 20 사이클을 수행하였다.
[0045] 가변 Ig 단편의 탐지 및 발현 벡터로의 클로닝을 위한 네스티드 PCR . 두번째 라운드에, 첫번째 증폭 반응의 5 ml 등분량을 적용하였다. 사용한 프라이머는 5' BglII 및 3' XbaI 제한 부위를 수반한다. 30회의 PCR 사이클을 수행하였다. 증폭의 첫번째 및 두번째 라운드에 대하여 동일한 조건을 사용하였다. 각 반응의 5 마이크로리터를 올리고, 1% 아가로스 겔에서 분리한 다음, 에티디움 브로마이드로 염색하였다. V-C PCR 생산물이 재배열된 VH 및 VL 단편을 각각 500 및 450 bp로 증폭할 것으로 예측된다. 대략 500 bp의 분자 크기를 갖는 PCR 밴드가 양성적 결과를 나타내었다. PCR 산물을 정제하고(Qiagen gel purification kit catalog number 28704), 불변 영역 특이적 프라이머를 사용하여 추출한 PCR 산물을 직접 시퀀싱하였다. 클론화된 단편의 서열은 재조합 산물을 위하여 준비한 플라스미드 시퀀싱으로 확인하였다.
[0046] 포유류 세포 중에서 인 비트로 항체 생산을 위하여, 상술한 PCR 단편을 소화시키고, 인간 감마 1 또는 인간 카파 또는 람다의 불변 영역을 수반하는 개별 발현 벡터 속으로 클로닝하였다. 중쇄 및 경쇄를 코딩하는 발현 벡터를 293 (인간 신장) 세포주(Invitrogen) 속으로 공동 감염시켰다. 양이온성 지질계 감염 시약(293fectinTM Invitrogen)을 사용하여, 발현 플라스미드를 도입하였다. 각각의 감염 반응을 위하여, 정제된 플라스미드 20 ㎍ 및 293fectinTM 40 ㎕를 Opti-MEM®(Invitrogen) 1 mL와 혼합하고, 상온에서 20분 동안 조합되어 복합체가 형성되는 것을 허여하기 전에, 상온에서 5분간 배양하였다. DNA-293fectin 복합체를 90 mm 페트리 접시에 접종된 3×106개의 세포에 첨가하고, 37℃, 8% CO2 조건에서 배양하였다. 최종 절차에서, 감염 72시간 후에 원심분리(3,000 g, 4℃에서 15분)로 상등액을 수확하여 분비된 항체들을 수득하였다.
[0047] ~2백만개의 림프구 세포로부터 AD169 용리액에 결합하는 45개 클론이 분리되었다. 이중에서 대다수는 재조합 gB 단백질에도 또한 결합하였다.
[0048] AD169 및 gB 둘 모두에 결합하는 mAbs 중 2가지(4A2 및 19B10) 는 중화능력을 가졌다. 이들 중 하나(4A2)는 AD-2 펩타이드에 결합하는데, 이는 gB 단백질 상에서 보존된 부위이다. 추가적인 mAb(5C5)는 AD-2에 결합하지만 바이러스를 중화하지는 않는다.
[0049] 가변영역, D 및 J 결합 영역, 프레임워크(FR) 및 CDR(complementarity determining regions) 영역을 포함하는 4A2 및 19B10의 중쇄 및 경쇄의 아미노산 서열은 아래에 보이는 바 같다. 중쇄의 분비 신호 서열은 이탤릭체로, CDR 1-3은 밑줄로 표시하였다.
[0050] 형광 비드 상에 고밀도 또는 저밀도에서 전장 gB 단백질 또는 Ad-2 펩타이드로 코팅된 셀 스팟 프로브를 제조하는 것에 의하여 고 친화도 항체를 생산하였다. 이론에 의한 제한 없이, 저밀도가 다돌기성 친화성 효과를 감소시키기 때문에, 내재적으로 고도의 친화성을 갖는 항체를 선호하는 것으로 연구가 편향된다. 다수의 고도의 친화도 항체를 분리하고 시퀀싱하였다. CMV에 반응성이 있는 다른 단일클론 항체들의 서열을 결정하였는데, 이는 SEQ ID NOs:9-36 및 38-66로 보여지는 바와 같다. 편견이 있다. 인간 IgG1 중쇄의 불변 영역의 뉴클레오타이드 서열을 SEQ ID NO:37로 표시하였다.
[0051] 본 발명의 항체에서, 중쇄는 GFTFNRHG (SEQ ID NO:67) 또는 GSISSEDFC (SEQ ID NO:68)의 CDR1; 및/또는 SSDGANQ (SEQ ID NO:69) 또는 ICYTGD (SEQ ID NO:70)의 CDR2 영역; 및/또는 ARDGRCEGERCYSGVTDF (SEQ ID NO:71) 또는 AREDRRQLHSRPYFYYGLDV (SEQ ID NO:72)의 CDR3 영역을 가질 수 있다. 다른 구현에에서, 경쇄는 QNIGGY (SEQ ID NO:73) 또는 QSLLHSNGHNY (SEQ ID NO:74)의 CDR1 영역; 및/또는 DAS (SEQ ID NO:75) 또는 LG (SEQ ID NO:76)의 CDR2 영역; 및/또는 QQRNSWPPLT (SEQ ID NO:77) 또는 QALQTPNT (SEQ ID NO:78)의 CDR3 영역을 가질 수 있다.
실시예
3
친화도 측정
[0052] 본 발명의 항체의 친화도를 포르테바이오®(Menlo Park, CA) 바이오센서 분석법으로 측정하였다. 이 방법에서, 아민 반응성 바이오센서 상의 카복시산 기는 EDC/NHS으로 활성화되었다. pH 5에서 MES 완충액 중에 희석시킨 항체를 프로브의 활성화된 표면에 부착시키고, 남아 있는 활성화된 카복시화 그룹은 에탄올아민으로 차단하였다. gB 단백질을 Ab-코팅된 프로브와 함께 배양하고, Ab- 코팅된 프로브로의 결합율 및 그로부터의 해리율을 포르테바이오® 기기로 측정하였다.
[0053] 하나의 실험에서, 표 1의 ㎍/ml 에서 대응하는 IC50으로 보여지는 바와 같이, 4A2은 168 pM의 친화도을 갖는 것으로, 19B10는 697 pM의 친화도를 갖는 것으로 밝혀졌다. 이러한 친화도 상수는, 공지의 gB에 대한 단일클론 항체보다 실질적으로 우수한 것이다.
표 1
mAbs의 중화 잠재력 비교.
[0054] gB의 AD-2 에피토프에 대한 mAbs 310, 313, 345 및 4A2의 결합 친화도를 측정하였다. gB 결합 동태를 표 2에 나타내었다. mAbs 310, 313 및 338은 mAb 4A2보다 약 10x 높은 잠재력을 갖는다. mAbs 323, 316 및 338는 또한 mAb 4A2보다 더 높은 잠재력을 갖는다(데이터 미표시). 남아있는 mAbs의 결합 친화도 또한 측정하였으며, 공개된 gB에 대한 단일클론 항체들보다 또한 우수하다.
표 2
gB 결합 동태
실시예
4
Elisa 결합 분석 및
에피토프
매핑
[0055] mAbs 4A2 및 19B10의 정제된 gB 단백질에 대한 결합 및 AD-2로 명명된 보존된 펩타이드(NETIYNTTLKYGDV (SEQ ID NO:79))에 대한 결합을 측정하였다. 도 1에서 보여지는 바와 같이, 4A2는 전장 길이 단백질 및 펩타이드 AD-2 모두에 잘 결합한 반면에, 19B10는 오직 단백질에만 결합한다.
실시예
5
바이러스 중화 분석
[0056] mAbs 4A2 및 19B10는 MRC5 1차 섬유모세포 내에서 CMV의 AD169 스트레인을 중화시켰다. 항체의 점진적 희석액을 동일한 부피의 AD169 (웰 당 2000개의 감염된 세포를 산출하도록 희석된108/ml 스톡)와 혼합하고, 96-웰 마이크로플레이트 안의 표적 세포 단일층에 첨가하기 전에 상온에서 1시간 동안 배양하였다. 24시간 후에 세포를 고정하고, 투과화하고, HRP에 컨주게이션된 IE1 (중간 초기 단백질 1, UL123로도 알려짐, 복제 중인 바이러스의 마커임)에 대한 단일클론 항체로 염색하였다. HRP 기질의 첨가 후에 따라 감염된 세포를 탐지하였다. 감염된 세포의 수를 항체의 농도에 대하여 도시하였다.
[0057] 또한, VR1814 스트레인(Revello, et al ., J. Gen . Virol . (2001) 82:1429-1438)을 사용하여 바이러스 중화를 산정하였다. mAbs 4A2, 310, 313, 338 및 345은 인간 제대 내피세포(HUVEC)와 인간 포피 섬유모세포(HFF) 모두에서 VR1814 스트레인을 중화하였다. 도 2A 및 2B는 mAbs 4A2, 310, 313, 338 및 345 각각에 대한 IC50 및 IC90 값을 나타낸다. 도 3 및 4는 mAbs 4A2, 310, 313, 338 및 345 각각에 의한 HUVEC 및 HFF 세포에 대한 중화를 각각 나타낸다. 결과는 2번 시험하였다.
[0058] AD169 및 VR1814 스트레인을 중화하는 다른 mAbs 또한 시험하였다.
서열목록
인간
IgG1
HC
불변영역의 아미노산 서열 (
SEQ
ID
NO
:9)
ASTKGPSVFPLVPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
MAB297
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:10)
QVQLVQSGGGVVQPGRSLRLSCSASGFTFSNYNMHWVRQAPGKGPEWVAVISKDGNEKHYAESAKGRFTISRDNSKNTLYMEMHSLTPEDTAMYYCTRDGRTDGTGYSGILDIWGQGTKVIVS
MAB309
및 318
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:11)
QVQLVQSGGGVVQPGTSLRLSCAASGFMFNTYNMHWVRQAPGKGLEWVAVISNDGTYKHFADSLKGRFSISRDDSKNTLYLHMNSLRPDDTAIYYCARDGRSVGGFSGILDPWGQGTLVTVSS
MAB310
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:12)
QVQLVQSGGGVVQPGTSLRLSCAASGFMFNTYNMHWVRQAPGKGLEWVAVISNDGTYKYSADSLKGRFSISRDNSKNTLYLHMNSLRPDDTAVYYCARDGRSVGGFSGILDPWGQGTLVTVSS
MAB313
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:13)
QVQLVQSGGGVIQPGRSLTLSCAASGFTFSAYSLHWVRQAPGKGLQWVAVISFDGNFKHFADSLRGRFTISRDNSKNRFYLQMNGLRGEDTAVYYCARDGRAVDGFSGILDFWGQGTLVSVSS
MAB314
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:14)
QVQLQESGGGLVQPGGSLKLSCAVSGFSFGGSAMHWVRQASGKGLEWIGHIRSGANNFETAYAPSLDGRFTISRDDSKNTAYLHMNSLKTDDTAMYFCTTGLIASGDANFDYWGQGTQVTVSS
MAB316
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:15)
QVQLVQSGGGVVQPGRSLTLSCAASGFTFSGFSLHWVRQAPGKGLQWVAVISFDGNHKHFADSLKGRFTISRDNSKNTLYLQINDLRGEDTAVYYCARDGRAVDGFSGILDFWGQGTLVSVSS
MAB319
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:16)
QVQLVESGGGVVQPGRSLRLSCSASGFTFSDYNLHWVRQAPGKGLEWVAVISIDGSDKHHADSVKGRFTVSRDNSKNTVSLQMDSLRPEDTAVYYCARDGRSVGGYSGILDPWGQGTLVTVSS
MAB321
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:17)
EVQLVESGAEVKKPGESLKISCQGSGYRFTNYWIAWVRQMPGKGLEWMGIIYPGDSDTRYHPSFQGQVTISSDKSLNTAYLQWSSLKPSDTAVYYCARHHCLSTNCQTAVAGYNDYWGQGNPGRRLLS
MAB322
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:18)
QVQLVQSGGGVVQPGRSLRLSCSASGFTFTNYNMHWVRQAPGKGLEWVAVTSKDGNEKHFADSVKGRFTISRDNSKNTLYLEMNTLTAEDTAIYYCTRDGRTDGTGYSGILDIWGQGTKVTVSS
MAB323
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:19)
QVQLVQSGGGVVQPGRSLRLSCAASGFTFSNFAMHWVRQAPGKGLEWVAVISNAGRETHYADSVKGRFTVSRDNSKNMLSLQMNSLRGEDTAVYYCARDGRTDGSGYSGVLDIWAQGTLVTVSS
MAB338
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:20)
QVQLVESGGGVVQPGRSLRLSCSGSGFTFSDYNLHWVRQAPGKGLEWVAVISIDGTNKHHADSVKGRFTISRDNSKNTVNLEMSRLKAEDTAVYYCVRDGRSIGGYSGIFDPWGQGTLVTVSS
MAB343
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:21)
QVQLQESGGGVVQPGRSLRLSCAASGFTFNTYNMHWVRQAPGKGLEWVAVISNDGTYKYSADSVKGRFSISRGNSKNTLYLQMNSLRPDDTAVYYCARDGRSVGGFSGILDPWGQGTLATVSS
MAB345
HC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:22)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSDYNMHWVRQAPGKGLEWVAVISIDGTYKYSADSVAGRFSLSRDNSKNTLYLQMNSLRPDDTAIYYCARDGRSVGGFSGILDPWGQGTLVTVSS
인간
LC
불변
카파
영역의 아미노산 서열 (
SEQ
ID
NO
:23)
TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
MAB297
및
MAB322
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:24)
EIVMTQSPATLSLSPGERATLSCRASQSVGGYLAWYQQKPDQAPRLLIYDVSNRAAGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRNTWPPLTFGGGTKVEIKR
MAB309
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:25)
EIVLTQSPATLSLSPGDRATLSCRASQTVGRYLAWYQQKPGQAPRLLIYDASDRATGISARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSSWPPLTFGGGTKVEIKR
MAB310
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:26)
EIVLTQSPATLSLSPGDRATLSCRASQTVGRYLAWYQQKPGQAPRLLIYDASDRATGISARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPLTFGGGTKVEIKR
MAB313
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:27)
EIVMTQSPATLSLSPGERATLSCRASQSVGRYLTWFQQKPGQAPRLLIYDASERATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRANWPPLTFGGGTKVEIK
MAB314
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:28)
EIVMTQSPGTLSLFPGERATLSCRASQTVRNGYLAWYQQKPGQAPRLLIYGASIRATGIPDRFSGSGSETDFTLSITRVEPEDFAVYYCQQYGRLSSTFGQGTKLDLK
MAB316
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:29)
EIVMTQSPATLSLSPGERATLSCRASQSVGRYLTWFQQKPGQAPRLLIYDASERATGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPLTFGGGTKVEIK
MAB318
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:30)
EVVLTQSPATLSLSPGDRATLSCRASQTVGRYLAWYQQKPGQAPRLLIYDASDRATGISARFSGSGSGTDFTLTIGSLEPEDFAVYYCQQRSSWPPLTFGGGTKVEIK
MAB319
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:31)
EIVLTQSPATLSLSPGERATLSCRASQSVGSYLAWYQQKPGQAPRLLIYDASERATGIPARFSGSGSGTDFTLTISSLEPEDVAVYYCQQRNNWPPLTFGGGTKVEIK
MAB321
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:32)
EIVMTQSPDSLAVSLGERATINCKSSQSILFSSKNQNHLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYNIPHTFGGGTKVEIK
MAB323
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:33)
EIVLTQSPATLSLSPGERATLSCRASQSVNRYLAWFQHRPGQPPRLLIYDASKRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIK
MAB338
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:34)
EIVLTQSPATLSLSPGERATLSCRASQSVDRYLAWYQQKPGQAPRLLIYDASQRATGIPARFSGSGSGTDFTLAISSLEPEDVAVYYCQQRSNWPPLTFGGGTKIEIK
MAB343
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:35)
EIVMTQSPATLSLSPGDRATLSCRASQSVGSYLAWYQQKPGQAPRLLIYDASDRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPLTFGGGTKVEIK
MAB345
LC
가변 도메인 아미노산 서열 (
SEQ
ID
NO
:36)
EIVMTQSPATLSLSPGDRATLSCRASQSVGSYLAWYQQKPGQAPRLLMYDSSVRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRNNWPPLTFGGGTKVEIK
인간
IgG1
HC
불변 영역의
뉴클레오타이드
서열 (
인트론은
밑줄침) (
SEQ
ID
NO
:37)
GCCTCCACCAAGGGCCCATCAGTCTTCCCCCTGGCACCCTCTACCAAGAGCACCTCTGGGGGCACAACGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGGTGAGAGGCCAGCACAGGGAGGGAGGGTGTCTGCTGGAAGCCAGGCTCAGCGCTCCTGCCTGGACGCATCCCGGCTATGCAGTCCCAGTCCAGGGCAGCAAGGCAGGCCCCGTCTGCCTCTTCACCCGGAGGCCTCTGCCCGCCCCACTCATGCTCAGGGAGAGGGTCTTCTGGCTTTTTCCCCAGGCTCTGGGCAGGCACAGGCTAGGTGCCCCTAACCCAGGCCCTGCACACAAAGGGGCAGGTGCTGGGCTCAGACCTGCCAAGAGCCATATCCGGGAGGACCCTGCCCCTGACCTAAGCCCACCCCAAAGGCCAAACTCTCCACTCCCTCAGCTCGGACACCTTCTCTCCTCCCAGATTCCAGTAACTCCCAATCTTCTCTCTGCAGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGGTAAGCCAGCCCAGGCCTCGCCCTCCAGCTCAAGGCGGGACAGGTGCCCTAGAGTAGCCTGCATCCAGGGACAGGCCCCAGCCGGGTGCTGACACGTCCACCTCCATCTCTTCCTCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGTGGGACCCGTGGGGTGCGAGGGCCACATGGACAGAGGCCGGCTCGGCCCACCCTCTGCCCTGAGAGTGACCGCTGTACCAACCTCTGTCCCTACAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCCCCGGGTAAATGA
MAB297
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:38)
CAGGTGCAACTGGTGCAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTTCAGCCTCTGGATTCACCTTCAGCAACTATAATATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCCGGAGTGGGTGGCAGTTATATCAAAAGATGGAAACGAAAAACACTATGCAGAGTCTGCGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATATGGAAATGCACAGCCTGACACCTGAGGACACGGCTATGTATTACTGTACGAGAGATGGGCGAACCGATGGTACTGGGTACTCCGGTATTCTTGATATCTGGGGCCAAGGGACAAAGGTCATCGTCTCT
MAB309
및 318
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:39)
CAGGTGCAGCTGGTGCAGTCTGGGGGAGGCGTGGTCCAGCCTGGGACGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCATGTTCAATACCTATAATATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCAAATGATGGAACCTATAAGCATTTCGCTGACTCCCTGAAGGGCCGATTCAGCATCTCCAGAGACGATTCCAAGAACACGCTGTATCTGCACATGAACAGCCTGAGACCTGACGACACGGCTATATATTACTGTGCGAGAGATGGCCGTAGTGTTGGCGGGTTTAGTGGGATCCTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG
MAB310
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:40)
CAGGTGCAGCTGGTGCAGTCTGGGGGAGGCGTGGTCCAGCCTGGGACGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCATGTTCAATACCTACAATATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCAAATGATGGAACCTATAAGTACTCCGCTGACTCCCTGAAGGGCCGATTCAGCATCTCCAGAGACAATTCCAAGAACACGTTGTATCTGCACATGAACAGCCTGAGACCTGACGACACGGCTGTATATTACTGTGCGAGAGATGGCCGTAGTGTTGGCGGGTTTAGTGGGATCCTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG
MAB313
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:41)
CAGGTGCAGCTGGTGCAGTCTGGGGGAGGCGTGATCCAGCCTGGGAGGTCCCTGACACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTGCCTATTCTCTACACTGGGTCCGCCAGGCTCCAGGCAAAGGGCTACAGTGGGTGGCGGTTATCTCATTTGATGGGAATTTTAAACACTTCGCAGACTCCCTGAGGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACAGATTCTATTTGCAAATGAATGGCCTGAGAGGTGAGGACACGGCTGTATATTACTGTGCGAGAGATGGACGTGCTGTTGACGGGTTTAGTGGGATCCTCGACTTCTGGGGCCAGGGAACCCTAGTCAGCGTCTCCTCAG
MAB314
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:42)
CAGGTGCAGCTGCAGGAGTCGGGGGGAGGCTTGGTCCAGCCGGGGGGGTCCCTGAAACTCTCCTGTGCAGTCTCTGGATTCTCCTTCGGTGGCTCTGCAATGCACTGGGTCCGCCAGGCTTCCGGGAAAGGGCTGGAGTGGATTGGCCATATTAGAAGCGGAGCTAATAATTTCGAGACAGCATATGCTCCGTCGCTGGATGGCAGGTTCACCATCTCCAGAGACGATTCAAAGAACACGGCGTATCTGCACATGAACAGCCTGAAAACCGATGACACGGCCATGTATTTCTGCACTACCGGACTTATAGCGTCAGGTGATGCAAATTTTGACTACTGGGGCCAGGGAACCCAGGTCACCGTCTCCTCGG
MAB316
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:43)
CAGGTGCAGCTGGTGCAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGACACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTGGCTTTTCTCTACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTACAGTGGGTGGCGGTTATCTCATTTGATGGGAACCATAAACACTTCGCAGACTCCCTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACATTGTATTTGCAAATTAATGACCTGAGAGGTGAGGACACGGCTGTATATTACTGTGCGAGAGATGGACGTGCTGTTGACGGGTTTAGTGGGATTCTCGACTTCTGGGGCCAGGGAACCCTGGTCAGCGTCTCCTCAG
MAB319
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:44)
CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTTCAGCCTCAGGATTCACCTTCAGTGACTATAATCTACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTCATCTCAATTGATGGAAGCGATAAACACCACGCAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAACACAGTGAGTCTACAAATGGACAGCCTGAGACCTGAAGACACGGCTGTATATTACTGTGCGAGAGATGGCCGTAGTGTGGGCGGCTACAGTGGGATCCTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG
MAB321
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:45)
GAGGTGCAGCTGGTGGAGTCCGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAAGATCTCCTGTCAGGGTTCTGGATACAGGTTTACCAATTACTGGATCGCCTGGGTGCGCCAGATGCCCGGGAAAGGCCTGGAGTGGATGGGGATCATCTATCCTGGTGACTCTGATACCAGATATCACCCGTCCTTCCAAGGCCAGGTCACCATCTCATCCGACAAATCCCTCAACACCGCCTACCTGCAGTGGAGCAGCCTGAAGCCCTCGGACACCGCCGTGTATTACTGTGCGAGACACCACTGCCTTAGTACCAACTGCCAAACCGCAGTGGCTGGATATAATGACTACTGGGGCCAGGGAAACCCTGGTCGCCGTCTCCTCAG
MAB322
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:46)
CAGGTGCAGCTGGTGGAGTCCGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTTTCCTGTTCAGCCTCTGGATTCACCTTCACCAACTATAACATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTACGTCAAAAGATGGAAACGAAAAACACTTTGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGGAAATGAACACCCTGACAGCTGAGGACACGGCGATATATTACTGTACGAGAGATGGGCGAACCGATGGTACTGGGTACTCCGGTATTCTTGATATCTGGGGCCAAGGGACAAAGGTCACCGTCTCCTCA
MAB323
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:47)
CAGGTGCAGCTGGTGCAGTCTGGGGGAGGGGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAACTTTGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCAAATGCTGGAAGGGAAACACACTACGCAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAATATGTTGTCTCTGCAAATGAACAGCCTGAGAGGTGAGGACACGGCTGTGTATTACTGTGCGAGAGATGGGCGAACCGATGGTAGTGGCTATTCCGGTGTTCTTGATATCTGGGCCCAAGGGACACTGGTCACTGTCTCCTCA
MAB338
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:48)
CAGGTGCAGCTGGTGGAGTCCGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTTCAGGCTCTGGATTCACCTTCAGTGACTATAATCTACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAATGGGTGGCAGTCATTTCAATTGATGGAACTAATAAACACCACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAACTCCAAGAATACAGTGAATCTGGAAATGAGTCGGCTGAAAGCAGAAGACACGGCTGTATATTACTGTGTGAGAGATGGGCGAAGTATTGGCGGCTACAGTGGAATCTTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA
MAB343
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:49)
CAGGTGCAGCTGCAGGAGTCAGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAATACCTACAATATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCAAATGATGGAACCTATAAATACTCCGCTGACTCCGTGAAGGGCCGATTCAGCATCTCCAGAGGCAATTCCAAGAACACGTTGTATCTGCAGATGAACAGCCTGAGACCTGACGACACGGCTGTATATTACTGTGCGAGAGATGGGCGTAGTGTTGGCGGGTTTAGTGGGATCCTCGACCCCTGGGGCCAGGGAACCCTGGCCACCGTCTCCTCA
MAB345
HC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:50)
CAGGTGCAGCTGGTGGAGTCCGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTGACTACAATATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATTTCAATTGATGGAACGTATAAATACTCCGCTGACTCCGTGGCGGGCCGATTCAGTCTCTCCAGAGACAATTCCAAGAACACGTTGTATTTGCAGATGAATAGTCTGAGACCTGACGACACGGCTATATATTATTGCGCGAGAGATGGGCGTAGTGTTGGCGGGTTTAGTGGGATCCTCGACCCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG
인간
LC
불변
카파
영역의
뉴클레오타이드
서열 (
SEQ
ID
NO
:51)
ACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCTAGCGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG
MAB297
및
MAB322
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:52)
GAAATTGTAATGACGCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCGGCTACTTAGCCTGGTACCAACAGAAACCTGACCAGGCTCCCAGGCTCCTCATCTATGATGTTTCCAATAGGGCCGCTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGGAACACCTGGCCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGA
MAB309
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:53)
GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGATAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACTGTTGGCAGGTACTTAGCCTGGTACCAACAAAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCTTCCGACAGGGCCACTGGCATCTCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGGAGCCTGAAGATTTTGCAGTCTATTACTGTCAGCAGCGGAGCAGCTGGCCGCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGA
MAB310
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:54)
GAAATTGTGTTGACTCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGATAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACTGTTGGCAGGTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCTTCCGACAGGGCCACTGGCATCTCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTCTATTACTGTCAGCAGCGGAGCAACTGGCCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGA
MAB313
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:55)
GAAATTGTGATGACTCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCAGATACTTAACTTGGTTCCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCTTCCGAGAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAACAGCGTGCTAACTGGCCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGA
MAB314
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:56)
GAAATTGTGATGACCCAGTCTCCAGGCACCCTGTCCTTGTTTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACTGTTAGGAACGGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCTTCCATCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGGTCTGAGACAGACTTCACCCTCAGCATCACCAGAGTGGAGCCTGAAGATTTTGCAGTTTATTACTGTCAACAGTATGGAAGGTTATCGTCCACTTTTGGCCAGGGGACCAAGCTGGACCTCAAACGA
MAB316
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:57)
GAAATTGTGATGACCCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCAGATACTTAACTTGGTTCCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCTTCCGAGAGGGCCACTGGCGTCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAACAGCGTAGTAACTGGCCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAAC
MAB318
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:58)
GAAGTTGTGCTGACGCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGATAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACTGTTGGCAGGTACTTAGCCTGGTACCAACAAAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCTTCCGACAGGGCCACTGGCATCTCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGATTTCACTCTCACCATCGGCAGCCTGGAGCCTGAAGATTTTGCAGTCTATTACTGTCAGCAGCGGAGCAGCTGGCCGCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAAC
MAB319
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:59)
GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGGGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCAGCTACTTAGCCTGGTATCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCATCCGAGAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATGTTGCAGTTTATTACTGTCAGCAGCGTAACAACTGGCCTCCGCTCACCTTCGGCGGAGGGACCAAGGTGGAGATCAAAC
MAB321
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:60)
GAAATTGTGATGACCCAGTCTCCAGACTCCCTTGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGTCAGAGTATTTTATTCAGCTCCAAGAATCAGAACCACTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTGATTTACTGGGCATCTACCCGGGAATCCGGGGTCCCCGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTCCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGCAATATTATAATATTCCTCACACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
MAB323
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:61)
GAAATTGTGTTGACTCAGTCTCCAGCCACCTTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCCGGGCCAGTCAGAGTGTTAACCGCTACTTAGCCTGGTTCCAACACAGACCTGGCCAGCCTCCCAGGCTCCTCATCTATGATGCGTCCAAGAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAG
MAB338
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:62)
GAAATTGTGTTGACCCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGACAGGTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGACTCCTCATCTATGATGCATCCCAGAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCCGGGACAGACTTCACTCTCGCCATCAGCAGCCTGGAGCCTGAAGATGTTGCAGTTTATTACTGTCAGCAGCGTAGTAACTGGCCTCCGCTCACCTTCGGCGGAGGGACCAAAATAGAGATCAAA
MAB343
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:63)
GAAATCGTGATGACCCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGATAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCTTCCGACAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAAC
MAB345
LC
가변 도메인
뉴클레오타이드
서열 (
SEQ
ID
NO
:64)
GAAATTGTGATGACCCAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGATAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATGTATGATTCTTCCGTCAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAACAACTGGCCTCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
인간
LC
불변
카파
영역의
뉴클레오타이드
서열 (
SEQ
ID
NO
:65)
ACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCTAGCGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG
인간
LC
불변
람다
영역의
뉴클레오타이드
서열 (
SEQ
ID
NO
:66)
GGTCAGCCCAAGGCTGCCCCCTCTGTCACTCTGTTCCCGCCCTCTAGCGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGTCCCTGCAGAATGCTCT
SEQUENCE LISTING
<110> TRELLIS BIOSCIENCE, INC.
KAUVAR, Lawrence M.
ELLSWORTH, Stote
USINGER, William
MCCUTCHEON, Krista Maureen
JIANG, Ying-Ping
ZHANG, Fen
CHEN, Bo
SPERINDE, Gizette
PARK, Minha
FOORD, Orit
<120> HIGH AFFINITY HUMAN ANTIBODIES TO HUMAN
CYTOMEGALOVIRUS (CMV) GB PROTEIN
<130> 388512012740
<140> Not Yet Assigned
<141> Concurrently Herewith
<150> US 61/335,499
<151> 2010-06-16
<160> 78
<170> FastSEQ for Windows Version 4.0
<210> 1
<211> 420
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed 4A2 heavy chain VH3-30
nucleic acid sequence
<220>
<221> CDS
<222> (1)...(420)
<400> 1
atg gaa ttg ggg ctg agc tgg gtt ttc gtc gtt gct ctt tta aga ggt 48
Met Glu Leu Gly Leu Ser Trp Val Phe Val Val Ala Leu Leu Arg Gly
1 5 10 15
gtc cag tgt caa gtg ttg ttg gag gag tct ggg gga ggc gtg gtc cag 96
Val Gln Cys Gln Val Leu Leu Glu Glu Ser Gly Gly Gly Val Val Gln
20 25 30
cct ggg agg tct ctg aga ctc tcc tgt gca ggc tct gga ttc acc ttc 144
Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe
35 40 45
aat agg cat gga att cac tgg gtc cgc cag gct cca ggc aag ggg ctg 192
Asn Arg His Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
50 55 60
gag tgg gtg act gtt ata tca tct gat gga gca aat caa cag tat gca 240
Glu Trp Val Thr Val Ile Ser Ser Asp Gly Ala Asn Gln Gln Tyr Ala
65 70 75 80
gag tcc gtg aag ggc cga ttc atc atc tcc aga gac aat tcc aag aac 288
Glu Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asn
85 90 95
acg gta tat cta gaa atg aat agc ctg agg aat gac gac acg ggt gtg 336
Thr Val Tyr Leu Glu Met Asn Ser Leu Arg Asn Asp Asp Thr Gly Val
100 105 110
tat ttc tgc gcg aga gac ggt cgt tgt gaa ggc gag agg tgc tac tcc 384
Tyr Phe Cys Ala Arg Asp Gly Arg Cys Glu Gly Glu Arg Cys Tyr Ser
115 120 125
ggt gtc acg gac ttc tgg ggc cag gga aca ctg gtc 420
Gly Val Thr Asp Phe Trp Gly Gln Gly Thr Leu Val
130 135 140
<210> 2
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed 4A2 heavy chain VH3 30
amino acid sequence
<220>
<221> misc_feature
<222> (1)...(19)
<223> secretion signal sequence
<400> 2
Met Glu Leu Gly Leu Ser Trp Val Phe Val Val Ala Leu Leu Arg Gly
1 5 10 15
Val Gln Cys Gln Val Leu Leu Glu Glu Ser Gly Gly Gly Val Val Gln
20 25 30
Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe
35 40 45
Asn Arg His Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
50 55 60
Glu Trp Val Thr Val Ile Ser Ser Asp Gly Ala Asn Gln Gln Tyr Ala
65 70 75 80
Glu Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asn
85 90 95
Thr Val Tyr Leu Glu Met Asn Ser Leu Arg Asn Asp Asp Thr Gly Val
100 105 110
Tyr Phe Cys Ala Arg Asp Gly Arg Cys Glu Gly Glu Arg Cys Tyr Ser
115 120 125
Gly Val Thr Asp Phe Trp Gly Gln Gly Thr Leu Val
130 135 140
<210> 3
<211> 360
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed 4A2 light chain L6
IgKV3-11 nucleic acid sequence
<220>
<221> CDS
<222> (1)...(360)
<400> 3
atg gaa gcc cca gcg cag ctt ctc ttc ctc ctg cta ctc tgg ctc cca 48
Met Glu Ala Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
gat acc acc gga gaa att gta ttg aca cag tct cca gcc acc ctg tct 96
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
ttg tct cca ggg gag aga gcc acc ctc tcc tgc agg gcc agt cag aat 144
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn
35 40 45
att ggc ggc tac ttg gcc tgg ttc caa caa aaa gct ggc cag gct ccc 192
Ile Gly Gly Tyr Leu Ala Trp Phe Gln Gln Lys Ala Gly Gln Ala Pro
50 55 60
agg ctc ctc atc tat gat gca tcc atc agg gcc act ggc atc cca gcc 240
Arg Leu Leu Ile Tyr Asp Ala Ser Ile Arg Ala Thr Gly Ile Pro Ala
65 70 75 80
agg ttc agt ggc agt ggg tct ggg aca gac ttc act ctc acc atc agc 288
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
agc cta gag cct gaa gat ttt gca gtt tat tac tgt cag cag cgt aac 336
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Asn
100 105 110
agt tgg cct cca ctc act ttc ggc 360
Ser Trp Pro Pro Leu Thr Phe Gly
115 120
<210> 4
<211> 120
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed 4A2 light chain L6
IgKV3-11 amino acid sequence
<400> 4
Met Glu Ala Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn
35 40 45
Ile Gly Gly Tyr Leu Ala Trp Phe Gln Gln Lys Ala Gly Gln Ala Pro
50 55 60
Arg Leu Leu Ile Tyr Asp Ala Ser Ile Arg Ala Thr Gly Ile Pro Ala
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Asn
100 105 110
Ser Trp Pro Pro Leu Thr Phe Gly
115 120
<210> 5
<211> 486
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed 19B10 heavy chain VH4-31
D2 J6 nucleic acid sequence
<220>
<221> CDS
<222> (1)...(486)
<400> 5
atg aaa cat ctg tgg ttc ttc ctc ctg ctg gtg gca gct ccc aga tgg 48
Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp
1 5 10 15
gtc ctg tcc cag gtg cag ctg cag cag tcg ggc cca gga ctg gtg aag 96
Val Leu Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys
20 25 30
cct tca cag acc ctg tcc ctc acc tgc act gtc tct ggt ggc tcc atc 144
Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile
35 40 45
agt agc ggt gat ttt tgc tgg aat tgg atc cgc cag ccc cca ggg aag 192
Ser Ser Gly Asp Phe Cys Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys
50 55 60
ggc ctg gag tgg att ggg tac atc tgt tac acc ggg gac acc tac tac 240
Gly Leu Glu Trp Ile Gly Tyr Ile Cys Tyr Thr Gly Asp Thr Tyr Tyr
65 70 75 80
aac ccg ccc ctt aac agt cga gtt acc ata tca gtc gac agg tcc agg 288
Asn Pro Pro Leu Asn Ser Arg Val Thr Ile Ser Val Asp Arg Ser Arg
85 90 95
aac caa atc tcc ctg agg ctg agt tct gtg act gcc gca gac acg gcc 336
Asn Gln Ile Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala
100 105 110
gtg tat tat tgt gcc aga gag gat agg aga caa cta cac tct cgc ccc 384
Val Tyr Tyr Cys Ala Arg Glu Asp Arg Arg Gln Leu His Ser Arg Pro
115 120 125
tac ttc tac tac ggt ttg gac gtc tgg ggc cga ggg acc aag gtc acc 432
Tyr Phe Tyr Tyr Gly Leu Asp Val Trp Gly Arg Gly Thr Lys Val Thr
130 135 140
gtc tcc tca gct tcc acc aag ggc cca tcg gtc ttc ccc ctg gta ccc 480
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Val Pro
145 150 155 160
tct agc 486
Ser Ser
<210> 6
<211> 162
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed 19B10 heavy chain VH4-31
D2 J6 amino acid sequence
<220>
<221> misc_feature
<222> (1)...(19)
<223> secretion signal sequence
<400> 6
Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp
1 5 10 15
Val Leu Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys
20 25 30
Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile
35 40 45
Ser Ser Gly Asp Phe Cys Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys
50 55 60
Gly Leu Glu Trp Ile Gly Tyr Ile Cys Tyr Thr Gly Asp Thr Tyr Tyr
65 70 75 80
Asn Pro Pro Leu Asn Ser Arg Val Thr Ile Ser Val Asp Arg Ser Arg
85 90 95
Asn Gln Ile Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Arg Glu Asp Arg Arg Gln Leu His Ser Arg Pro
115 120 125
Tyr Phe Tyr Tyr Gly Leu Asp Val Trp Gly Arg Gly Thr Lys Val Thr
130 135 140
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Val Pro
145 150 155 160
Ser Ser
<210> 7
<211> 420
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed 19B10 light chain A3
IgKV2 nucleic acid sequence
<220>
<221> CDS
<222> (1)...(420)
<400> 7
atg agg ctc cct gct cag ctt ctg ggg ctg cta atg ctc tgg gtc tct 48
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Ser
1 5 10 15
gga tcc agt ggg gag att gtg atg act cag tct ccg ctc tcc ctg ccc 96
Gly Ser Ser Gly Glu Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro
20 25 30
gtc acc cct gga gag acg gcc tcc atc tcc tgc agg tct agt cag agc 144
Val Thr Pro Gly Glu Thr Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
ctc ctg cat agt aat gga cac aac tat ttg gat tgg tat ctg cag aag 192
Leu Leu His Ser Asn Gly His Asn Tyr Leu Asp Trp Tyr Leu Gln Lys
50 55 60
cca ggg cag tct cca cac ctc ctg atc tat ttg ggt tct att cgg gcc 240
Pro Gly Gln Ser Pro His Leu Leu Ile Tyr Leu Gly Ser Ile Arg Ala
65 70 75 80
tcc ggg gtc cct gac agg ttc agt ggc agt gga aca ggc aca gat ttt 288
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Thr Gly Thr Asp Phe
85 90 95
aca ctg aaa atc agc aga gtg gag gct gag gat gtt ggg gtt tat tac 336
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
tgc atg caa gct cta caa act cct aac act ttt ggc cag ggg acc aag 384
Cys Met Gln Ala Leu Gln Thr Pro Asn Thr Phe Gly Gln Gly Thr Lys
115 120 125
ctg gag atc aga cga act gtg gct gca cca tct gtc 420
Leu Glu Ile Arg Arg Thr Val Ala Ala Pro Ser Val
130 135 140
<210> 8
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed 19B10 light chain A3
IgKV2 amino acid sequence
<400> 8
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Ser
1 5 10 15
Gly Ser Ser Gly Glu Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro
20 25 30
Val Thr Pro Gly Glu Thr Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Leu Leu His Ser Asn Gly His Asn Tyr Leu Asp Trp Tyr Leu Gln Lys
50 55 60
Pro Gly Gln Ser Pro His Leu Leu Ile Tyr Leu Gly Ser Ile Arg Ala
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Thr Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
Cys Met Gln Ala Leu Gln Thr Pro Asn Thr Phe Gly Gln Gly Thr Lys
115 120 125
Leu Glu Ile Arg Arg Thr Val Ala Ala Pro Ser Val
130 135 140
<210> 9
<211> 330
<212> PRT
<213> Homo sapiens
<220>
<221> CHAIN
<222> (1)...(330)
<223> IgG1 heavy chain amino acid sequence of constant
region
<400> 9
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Val Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 10
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB297 heavy chain
variable domain amino acid sequence
<400> 10
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Asn Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Val
35 40 45
Ala Val Ile Ser Lys Asp Gly Asn Glu Lys His Tyr Ala Glu Ser Ala
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Met Glu Met His Ser Leu Thr Pro Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Thr Arg Asp Gly Arg Thr Asp Gly Thr Gly Tyr Ser Gly Ile Leu Asp
100 105 110
Ile Trp Gly Gln Gly Thr Lys Val Ile Val Ser
115 120
<210> 11
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB309 and MAB318 heavy
chain variable domain amino acid sequence
<400> 11
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Thr
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Met Phe Asn Thr Tyr
20 25 30
Asn Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Asn Asp Gly Thr Tyr Lys His Phe Ala Asp Ser Leu
50 55 60
Lys Gly Arg Phe Ser Ile Ser Arg Asp Asp Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu His Met Asn Ser Leu Arg Pro Asp Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ser Val Gly Gly Phe Ser Gly Ile Leu Asp Pro
100 105 110
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 12
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB310 heavy chain
variable domain amino acid sequence
<400> 12
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Thr
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Met Phe Asn Thr Tyr
20 25 30
Asn Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Asn Asp Gly Thr Tyr Lys Tyr Ser Ala Asp Ser Leu
50 55 60
Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu His Met Asn Ser Leu Arg Pro Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ser Val Gly Gly Phe Ser Gly Ile Leu Asp Pro
100 105 110
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 13
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB313 heavy chain
variable domain amino acid sequence
<400> 13
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Ile Gln Pro Gly Arg
1 5 10 15
Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr
20 25 30
Ser Leu His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gln Trp Val
35 40 45
Ala Val Ile Ser Phe Asp Gly Asn Phe Lys His Phe Ala Asp Ser Leu
50 55 60
Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Arg Phe Tyr
65 70 75 80
Leu Gln Met Asn Gly Leu Arg Gly Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ala Val Asp Gly Phe Ser Gly Ile Leu Asp Phe
100 105 110
Trp Gly Gln Gly Thr Leu Val Ser Val Ser Ser
115 120
<210> 14
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB314 heavy chain
variable domain amino acid sequence
<400> 14
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Val Ser Gly Phe Ser Phe Gly Gly Ser
20 25 30
Ala Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Ile
35 40 45
Gly His Ile Arg Ser Gly Ala Asn Asn Phe Glu Thr Ala Tyr Ala Pro
50 55 60
Ser Leu Asp Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu His Met Asn Ser Leu Lys Thr Asp Asp Thr Ala Met Tyr
85 90 95
Phe Cys Thr Thr Gly Leu Ile Ala Ser Gly Asp Ala Asn Phe Asp Tyr
100 105 110
Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
<210> 15
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB316 heavy chain
variable domain amino acid sequence
<400> 15
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Thr Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Phe
20 25 30
Ser Leu His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Gln Trp Val
35 40 45
Ala Val Ile Ser Phe Asp Gly Asn His Lys His Phe Ala Asp Ser Leu
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Ile Asn Asp Leu Arg Gly Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ala Val Asp Gly Phe Ser Gly Ile Leu Asp Phe
100 105 110
Trp Gly Gln Gly Thr Leu Val Ser Val Ser Ser
115 120
<210> 16
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB319 heavy chain
variable domain amino acid sequence
<400> 16
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Asn Leu His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Ile Asp Gly Ser Asp Lys His His Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Val Ser
65 70 75 80
Leu Gln Met Asp Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ser Val Gly Gly Tyr Ser Gly Ile Leu Asp Pro
100 105 110
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 17
<211> 128
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB321 heavy chain
variable domain amino acid sequence
<400> 17
Glu Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Glu
1 5 10 15
Ser Leu Lys Ile Ser Cys Gln Gly Ser Gly Tyr Arg Phe Thr Asn Tyr
20 25 30
Trp Ile Ala Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met
35 40 45
Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr His Pro Ser Phe
50 55 60
Gln Gly Gln Val Thr Ile Ser Ser Asp Lys Ser Leu Asn Thr Ala Tyr
65 70 75 80
Leu Gln Trp Ser Ser Leu Lys Pro Ser Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg His His Cys Leu Ser Thr Asn Cys Gln Thr Ala Val Ala Gly
100 105 110
Tyr Asn Asp Tyr Trp Gly Gln Gly Asn Pro Gly Arg Arg Leu Leu Ser
115 120 125
<210> 18
<211> 124
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB322 heavy chain
variable domain amino acid sequence
<400> 18
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Thr Phe Thr Asn Tyr
20 25 30
Asn Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Thr Ser Lys Asp Gly Asn Glu Lys His Phe Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Glu Met Asn Thr Leu Thr Ala Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Thr Arg Asp Gly Arg Thr Asp Gly Thr Gly Tyr Ser Gly Ile Leu Asp
100 105 110
Ile Trp Gly Gln Gly Thr Lys Val Thr Val Ser Ser
115 120
<210> 19
<211> 124
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB323 heavy chain
variable domain amino acid sequence
<400> 19
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Phe
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Asn Ala Gly Arg Glu Thr His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Met Leu Ser
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Gly Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Thr Asp Gly Ser Gly Tyr Ser Gly Val Leu Asp
100 105 110
Ile Trp Ala Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 20
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB338 heavy chain
variable domain amino acid sequence
<400> 20
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Gly Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Asn Leu His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Ile Asp Gly Thr Asn Lys His His Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Asn
65 70 75 80
Leu Glu Met Ser Arg Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Asp Gly Arg Ser Ile Gly Gly Tyr Ser Gly Ile Phe Asp Pro
100 105 110
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 21
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB343 heavy chain
variable domain amino acid sequence
<400> 21
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr
20 25 30
Asn Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Asn Asp Gly Thr Tyr Lys Tyr Ser Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ser Ile Ser Arg Gly Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Pro Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ser Val Gly Gly Phe Ser Gly Ile Leu Asp Pro
100 105 110
Trp Gly Gln Gly Thr Leu Ala Thr Val Ser Ser
115 120
<210> 22
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB345 heavy chain
variable domain amino acid sequence
<400> 22
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Asn Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Ile Asp Gly Thr Tyr Lys Tyr Ser Ala Asp Ser Val
50 55 60
Ala Gly Arg Phe Ser Leu Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Pro Asp Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Arg Ser Val Gly Gly Phe Ser Gly Ile Leu Asp Pro
100 105 110
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 23
<211> 106
<212> PRT
<213> Homo sapiens
<220>
<221> CHAIN
<222> (1)...(106)
<223> light chain amino acid sequence of constant kappa
region
<400> 23
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
1 5 10 15
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
20 25 30
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
35 40 45
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
50 55 60
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
65 70 75 80
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
85 90 95
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105
<210> 24
<211> 109
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB297 and MAB322 light
chain variable domain amino acid sequence
<400> 24
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Gly Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Val Ser Asn Arg Ala Ala Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Asn Thr Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105
<210> 25
<211> 109
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB309 light chain
variable domain amino acid sequence
<400> 25
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Asp Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Thr Val Gly Arg Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asp Arg Ala Thr Gly Ile Ser Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105
<210> 26
<211> 109
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB310 light chain
variable domain amino acid sequence
<400> 26
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Asp Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Thr Val Gly Arg Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asp Arg Ala Thr Gly Ile Ser Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105
<210> 27
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB313 light chain
variable domain amino acid sequence
<400> 27
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Arg Tyr
20 25 30
Leu Thr Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Glu Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ala Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 28
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB314 light chain
variable domain amino acid sequence
<400> 28
Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Phe Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Thr Val Arg Asn Gly
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ile Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Glu Thr Asp Phe Thr Leu Ser Ile Thr Arg Val Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Arg Leu Ser
85 90 95
Ser Thr Phe Gly Gln Gly Thr Lys Leu Asp Leu Lys
100 105
<210> 29
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB316 light chain
variable domain amino acid sequence
<400> 29
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Arg Tyr
20 25 30
Leu Thr Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Glu Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 30
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB318 light chain
variable domain amino acid sequence
<400> 30
Glu Val Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Asp Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Thr Val Gly Arg Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asp Arg Ala Thr Gly Ile Ser Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Gly Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 31
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB319 light chain
variable domain amino acid sequence
<400> 31
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Glu Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Arg Asn Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 32
<211> 113
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB321 light chain
variable domain amino acid sequence
<400> 32
Glu Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Ile Leu Phe Ser
20 25 30
Ser Lys Asn Gln Asn His Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln
85 90 95
Tyr Tyr Asn Ile Pro His Thr Phe Gly Gly Gly Thr Lys Val Glu Ile
100 105 110
Lys
<210> 33
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB323 light chain
variable domain amino acid sequence
<400> 33
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Asn Arg Tyr
20 25 30
Leu Ala Trp Phe Gln His Arg Pro Gly Gln Pro Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Lys Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 34
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB338 light chain
variable domain amino acid sequence
<400> 34
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Asp Arg Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Gln Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Ala Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Ile Glu Ile Lys
100 105
<210> 35
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB343 light chain
variable domain amino acid sequence
<400> 35
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Asp Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asp Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 36
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB345 light chain
variable domain amino acid sequence
<400> 36
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Asp Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Met
35 40 45
Tyr Asp Ser Ser Val Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Asn Asn Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 37
<211> 1599
<212> DNA
<213> Homo sapiens
<220>
<221> misc_feature
<222> (1)...(1599)
<223> IgG1 heavy chain nucleotide sequence of constant
region
<220>
<221> intron
<222> (289)...(685)
<220>
<221> intron
<222> (731)...(848)
<220>
<221> intron
<222> (1208)...(1277)
<400> 37
gcctccacca agggcccatc agtcttcccc ctggcaccct ctaccaagag cacctctggg 60
ggcacaacgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 120
tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 180
ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 240
tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagag agttggtgag 300
aggccagcac agggagggag ggtgtctgct ggaagccagg ctcagcgctc ctgcctggac 360
gcatcccggc tatgcagtcc cagtccaggg cagcaaggca ggccccgtct gcctcttcac 420
ccggaggcct ctgcccgccc cactcatgct cagggagagg gtcttctggc tttttcccca 480
ggctctgggc aggcacaggc taggtgcccc taacccaggc cctgcacaca aaggggcagg 540
tgctgggctc agacctgcca agagccatat ccgggaggac cctgcccctg acctaagccc 600
accccaaagg ccaaactctc cactccctca gctcggacac cttctctcct cccagattcc 660
agtaactccc aatcttctct ctgcagagcc caaatcttgt gacaaaactc acacatgccc 720
accgtgccca ggtaagccag cccaggcctc gccctccagc tcaaggcggg acaggtgccc 780
tagagtagcc tgcatccagg gacaggcccc agccgggtgc tgacacgtcc acctccatct 840
cttcctcagc acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca 900
aggacaccct catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc 960
acgaagaccc tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca 1020
agacaaagcc gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg 1080
tcctgcacca ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc 1140
tcccagcccc catcgagaaa accatctcca aagccaaagg tgggacccgt ggggtgcgag 1200
ggccacatgg acagaggccg gctcggccca ccctctgccc tgagagtgac cgctgtacca 1260
acctctgtcc ctacagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 1320
gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 1380
gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 1440
cccgtgctgg actccgacgg ctccttcttc ctctatagca agctcaccgt ggacaagagc 1500
aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1560
tacacgcaga agagcctctc cctgtccccg ggtaaatga 1599
<210> 38
<211> 369
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB297 heavy chain
variable domain nucleotide sequence
<400> 38
caggtgcaac tggtgcagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgttcag cctctggatt caccttcagc aactataata tgcactgggt ccgccaggct 120
ccaggcaagg ggccggagtg ggtggcagtt atatcaaaag atggaaacga aaaacactat 180
gcagagtctg cgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240
atggaaatgc acagcctgac acctgaggac acggctatgt attactgtac gagagatggg 300
cgaaccgatg gtactgggta ctccggtatt cttgatatct ggggccaagg gacaaaggtc 360
atcgtctct 369
<210> 39
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB309 and MAB318 heavy
chain variable domain nucleotide sequence
<400> 39
caggtgcagc tggtgcagtc tgggggaggc gtggtccagc ctgggacgtc cctgagactc 60
tcctgtgcag cctctggatt catgttcaat acctataata tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atatcaaatg atggaaccta taagcatttc 180
gctgactccc tgaagggccg attcagcatc tccagagacg attccaagaa cacgctgtat 240
ctgcacatga acagcctgag acctgacgac acggctatat attactgtgc gagagatggc 300
cgtagtgttg gcgggtttag tgggatcctc gacccctggg gccagggaac cctggtcacc 360
gtctcctcag 370
<210> 40
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB310 heavy chain
variable domain nucleotide sequence
<400> 40
caggtgcagc tggtgcagtc tgggggaggc gtggtccagc ctgggacgtc cctgagactc 60
tcctgtgcag cctctggatt catgttcaat acctacaata tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atatcaaatg atggaaccta taagtactcc 180
gctgactccc tgaagggccg attcagcatc tccagagaca attccaagaa cacgttgtat 240
ctgcacatga acagcctgag acctgacgac acggctgtat attactgtgc gagagatggc 300
cgtagtgttg gcgggtttag tgggatcctc gacccctggg gccagggaac cctggtcacc 360
gtctcctcag 370
<210> 41
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB313 heavy chain
variable domain nucleotide sequence
<400> 41
caggtgcagc tggtgcagtc tgggggaggc gtgatccagc ctgggaggtc cctgacactc 60
tcctgtgcag cctctggatt caccttcagt gcctattctc tacactgggt ccgccaggct 120
ccaggcaaag ggctacagtg ggtggcggtt atctcatttg atgggaattt taaacacttc 180
gcagactccc tgaggggccg attcaccatc tccagagaca attccaagaa cagattctat 240
ttgcaaatga atggcctgag aggtgaggac acggctgtat attactgtgc gagagatgga 300
cgtgctgttg acgggtttag tgggatcctc gacttctggg gccagggaac cctagtcagc 360
gtctcctcag 370
<210> 42
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB314 heavy chain
variable domain nucleotide sequence
<400> 42
caggtgcagc tgcaggagtc ggggggaggc ttggtccagc cgggggggtc cctgaaactc 60
tcctgtgcag tctctggatt ctccttcggt ggctctgcaa tgcactgggt ccgccaggct 120
tccgggaaag ggctggagtg gattggccat attagaagcg gagctaataa tttcgagaca 180
gcatatgctc cgtcgctgga tggcaggttc accatctcca gagacgattc aaagaacacg 240
gcgtatctgc acatgaacag cctgaaaacc gatgacacgg ccatgtattt ctgcactacc 300
ggacttatag cgtcaggtga tgcaaatttt gactactggg gccagggaac ccaggtcacc 360
gtctcctcgg 370
<210> 43
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB316 heavy chain
variable domain nucleotide sequence
<400> 43
caggtgcagc tggtgcagtc tgggggaggc gtggtccagc ctgggaggtc cctgacactc 60
tcctgtgcag cctctggatt caccttcagt ggcttttctc tacactgggt ccgccaggct 120
ccaggcaagg ggctacagtg ggtggcggtt atctcatttg atgggaacca taaacacttc 180
gcagactccc tgaagggccg attcaccatc tccagagaca attccaagaa cacattgtat 240
ttgcaaatta atgacctgag aggtgaggac acggctgtat attactgtgc gagagatgga 300
cgtgctgttg acgggtttag tgggattctc gacttctggg gccagggaac cctggtcagc 360
gtctcctcag 370
<210> 44
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB319 heavy chain
variable domain nucleotide sequence
<400> 44
caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgttcag cctcaggatt caccttcagt gactataatc tacactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtc atctcaattg atggaagcga taaacaccac 180
gcagactccg tgaagggccg attcaccgtc tccagagaca attccaagaa cacagtgagt 240
ctacaaatgg acagcctgag acctgaagac acggctgtat attactgtgc gagagatggc 300
cgtagtgtgg gcggctacag tgggatcctc gacccctggg gccagggaac cctggtcacc 360
gtctcctcag 370
<210> 45
<211> 383
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB321 heavy chain
variable domain nucleotide sequence
<400> 45
gaggtgcagc tggtggagtc cggagcagag gtgaaaaagc ccggggagtc tctgaagatc 60
tcctgtcagg gttctggata caggtttacc aattactgga tcgcctgggt gcgccagatg 120
cccgggaaag gcctggagtg gatggggatc atctatcctg gtgactctga taccagatat 180
cacccgtcct tccaaggcca ggtcaccatc tcatccgaca aatccctcaa caccgcctac 240
ctgcagtgga gcagcctgaa gccctcggac accgccgtgt attactgtgc gagacaccac 300
tgccttagta ccaactgcca aaccgcagtg gctggatata atgactactg gggccaggga 360
aaccctggtc gccgtctcct cag 383
<210> 46
<211> 372
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB322 heavy chain
variable domain nucleotide sequence
<400> 46
caggtgcagc tggtggagtc cgggggaggc gtggtccagc ctgggaggtc cctgagactt 60
tcctgttcag cctctggatt caccttcacc aactataaca tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt acgtcaaaag atggaaacga aaaacacttt 180
gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240
ctggaaatga acaccctgac agctgaggac acggcgatat attactgtac gagagatggg 300
cgaaccgatg gtactgggta ctccggtatt cttgatatct ggggccaagg gacaaaggtc 360
accgtctcct ca 372
<210> 47
<211> 372
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB323 heavy chain
variable domain nucleotide sequence
<400> 47
caggtgcagc tggtgcagtc tgggggaggg gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt caccttcagt aactttgcta tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atatcaaatg ctggaaggga aacacactac 180
gcagactccg tgaagggccg attcaccgtc tccagagaca attccaagaa tatgttgtct 240
ctgcaaatga acagcctgag aggtgaggac acggctgtgt attactgtgc gagagatggg 300
cgaaccgatg gtagtggcta ttccggtgtt cttgatatct gggcccaagg gacactggtc 360
actgtctcct ca 372
<210> 48
<211> 369
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB338 heavy chain
variable domain nucleotide sequence
<400> 48
caggtgcagc tggtggagtc cgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgttcag gctctggatt caccttcagt gactataatc tacactgggt ccgccaggct 120
ccaggcaagg ggctggaatg ggtggcagtc atttcaattg atggaactaa taaacaccac 180
gcagactccg tgaagggccg attcaccatc tccagagaca actccaagaa tacagtgaat 240
ctggaaatga gtcggctgaa agcagaagac acggctgtat attactgtgt gagagatggg 300
cgaagtattg gcggctacag tggaatcttc gacccctggg gccagggaac cctggtcacc 360
gtctcctca 369
<210> 49
<211> 369
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB343 heavy chain
variable domain nucleotide sequence
<400> 49
caggtgcagc tgcaggagtc agggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt caccttcaat acctacaata tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atatcaaatg atggaaccta taaatactcc 180
gctgactccg tgaagggccg attcagcatc tccagaggca attccaagaa cacgttgtat 240
ctgcagatga acagcctgag acctgacgac acggctgtat attactgtgc gagagatggg 300
cgtagtgttg gcgggtttag tgggatcctc gacccctggg gccagggaac cctggccacc 360
gtctcctca 369
<210> 50
<211> 370
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB345 heavy chain
variable domain nucleotide sequence
<400> 50
caggtgcagc tggtggagtc cgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt caccttcagt gactacaata tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atttcaattg atggaacgta taaatactcc 180
gctgactccg tggcgggccg attcagtctc tccagagaca attccaagaa cacgttgtat 240
ttgcagatga atagtctgag acctgacgac acggctatat attattgcgc gagagatggg 300
cgtagtgttg gcgggtttag tgggatcctc gacccctggg gccagggaac cctggtcacc 360
gtctcctcag 370
<210> 51
<211> 321
<212> DNA
<213> Homo sapiens
<220>
<221> misc_feature
<222> (1)...(321)
<223> light chain nucleotide sequence of constant kappa
region
<400> 51
actgtggctg caccatctgt cttcatcttc ccgccatctg atgagcagtt gaaatctgga 60
actgctagcg ttgtgtgcct gctgaataac ttctatccca gagaggccaa agtacagtgg 120
aaggtggata acgccctcca atcgggtaac tcccaggaga gtgtcacaga gcaggacagc 180
aaggacagca cctacagcct cagcagcacc ctgacgctga gcaaagcaga ctacgagaaa 240
cacaaagtct acgcctgcga agtcacccat cagggcctga gctcgcccgt cacaaagagc 300
ttcaacaggg gagagtgtta g 321
<210> 52
<211> 327
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB297 and MAB322 light
chain variable domain nucleotide sequence
<400> 52
gaaattgtaa tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttggc ggctacttag cctggtacca acagaaacct 120
gaccaggctc ccaggctcct catctatgat gtttccaata gggccgctgg catcccagcc 180
aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctggagcct 240
gaagattttg cagtttatta ctgtcagcag cggaacacct ggcctccgct cactttcggc 300
ggagggacca aggtggagat caaacga 327
<210> 53
<211> 327
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB309 light chain
variable domain nucleotide sequence
<400> 53
gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga tagagccacc 60
ctctcctgca gggccagtca gactgttggc aggtacttag cctggtacca acaaaaacct 120
ggccaggctc ccaggctcct catctatgat gcttccgaca gggccactgg catctcagcc 180
aggttcagtg gcagtgggtc tgggacagat ttcactctca ccatcagcag cctggagcct 240
gaagattttg cagtctatta ctgtcagcag cggagcagct ggccgccgct cactttcggc 300
ggagggacca aggtggagat caaacga 327
<210> 54
<211> 327
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB310 light chain
variable domain nucleotide sequence
<400> 54
gaaattgtgt tgactcagtc tccagccacc ctgtctttgt ctccagggga tagagccacc 60
ctctcctgca gggccagtca gactgttggc aggtacttag cctggtacca acagaaacct 120
ggccaggctc ccaggctcct catctatgat gcttccgaca gggccactgg catctcagcc 180
aggttcagtg gcagtgggtc tgggacagat ttcactctca ccatcagcag cctagagcct 240
gaagattttg cagtctatta ctgtcagcag cggagcaact ggcctccgct cactttcggc 300
ggagggacca aggtggagat caaacga 327
<210> 55
<211> 327
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB313 light chain
variable domain nucleotide sequence
<400> 55
gaaattgtga tgactcagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttggc agatacttaa cttggttcca gcagaaacct 120
ggccaggctc ccaggctcct catctatgat gcttccgaga gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240
gaagattttg cagtttatta ctgtcaacag cgtgctaact ggcctccgct cactttcggc 300
ggagggacca aggtggagat caaacga 327
<210> 56
<211> 327
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB314 light chain
variable domain nucleotide sequence
<400> 56
gaaattgtga tgacccagtc tccaggcacc ctgtccttgt ttccagggga aagagccacc 60
ctctcctgca gggccagtca gactgttagg aacggctact tagcctggta ccagcagaaa 120
cctggccagg ctcccaggct cctcatctat ggtgcttcca tcagggccac tggcatccca 180
gacaggttca gtggcagtgg gtctgagaca gacttcaccc tcagcatcac cagagtggag 240
cctgaagatt ttgcagttta ttactgtcaa cagtatggaa ggttatcgtc cacttttggc 300
caggggacca agctggacct caaacga 327
<210> 57
<211> 325
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB316 light chain
variable domain nucleotide sequence
<400> 57
gaaattgtga tgacccagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttggc agatacttaa cttggttcca gcagaaacct 120
ggccaggctc ccaggctcct catctatgat gcttccgaga gggccactgg cgtcccagcc 180
aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240
gaagattttg cagtttatta ctgtcaacag cgtagtaact ggcctccgct cactttcggc 300
ggagggacca aggtggagat caaac 325
<210> 58
<211> 325
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB318 light chain
variable domain nucleotide sequence
<400> 58
gaagttgtgc tgacgcagtc tccagccacc ctgtctttgt ctccagggga tagagccacc 60
ctctcctgca gggccagtca gactgttggc aggtacttag cctggtacca acaaaaacct 120
ggccaggctc ccaggctcct catctatgat gcttccgaca gggccactgg catctcagcc 180
aggttcagtg gcagtgggtc tgggacagat ttcactctca ccatcggcag cctggagcct 240
gaagattttg cagtctatta ctgtcagcag cggagcagct ggccgccgct cactttcggc 300
ggagggacca aggtggagat caaac 325
<210> 59
<211> 325
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB319 light chain
variable domain nucleotide sequence
<400> 59
gaaattgtgt tgacgcagtc tccagccacc ctgtctttgt ctccagggga aagggccacc 60
ctctcctgca gggccagtca gagtgttggc agctacttag cctggtatca acagaaacct 120
ggccaggctc ccaggctcct catctatgat gcatccgaga gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240
gaagatgttg cagtttatta ctgtcagcag cgtaacaact ggcctccgct caccttcggc 300
ggagggacca aggtggagat caaac 325
<210> 60
<211> 339
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB321 light chain
variable domain nucleotide sequence
<400> 60
gaaattgtga tgacccagtc tccagactcc cttgctgtgt ctctgggcga gagggccacc 60
atcaactgca agtccagtca gagtatttta ttcagctcca agaatcagaa ccacttagct 120
tggtaccagc agaaaccagg acagcctcct aagctgctga tttactgggc atctacccgg 180
gaatccgggg tccccgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 240
atcagcagcc tccaggctga agatgtggca gtttattact gtcagcaata ttataatatt 300
cctcacactt tcggcggagg gaccaaggtg gagatcaaa 339
<210> 61
<211> 321
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB323 light chain
variable domain nucleotide sequence
<400> 61
gaaattgtgt tgactcagtc tccagccacc ttgtctttgt ctccagggga aagagccacc 60
ctctcctgcc gggccagtca gagtgttaac cgctacttag cctggttcca acacagacct 120
ggccagcctc ccaggctcct catctatgat gcgtccaaga gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 240
gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgctcac tttcggcgga 300
gggaccaagg tggagatcaa g 321
<210> 62
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB338 light chain
variable domain nucleotide sequence
<400> 62
gaaattgtgt tgacccagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60
ctctcctgca gggccagtca gagtgttgac aggtacttag cctggtacca acagaaacct 120
ggccaggctc ccagactcct catctatgat gcatcccaga gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc cgggacagac ttcactctcg ccatcagcag cctggagcct 240
gaagatgttg cagtttatta ctgtcagcag cgtagtaact ggcctccgct caccttcggc 300
ggagggacca aaatagagat caaa 324
<210> 63
<211> 325
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB343 light chain
variable domain nucleotide sequence
<400> 63
gaaatcgtga tgacccagtc tccagccacc ctgtctttgt ctccagggga tagagccacc 60
ctctcctgca gggccagtca gagtgttggc agctacttag cctggtacca acagaaacct 120
ggccaggctc ccaggctcct catctatgat gcttccgaca gggccactgg catcccagcc 180
aggttcagtg gcagtgggtc tgggacagat ttcactctca ccatcagcag cctagagcct 240
gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccgct cactttcggc 300
ggagggacca aggtggagat caaac 325
<210> 64
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> synthetically constructed MAB345 light chain
variable domain nucleotide sequence
<400> 64
gaaattgtga tgacccagtc tccagccacc ctgtctttgt ctccagggga tagagccacc 60
ctctcctgca gggccagtca gagtgttggc agctacttag cctggtacca acagaaacct 120
ggccaggctc ccaggctcct catgtatgat tcttccgtca gggccactgg catcccagcc 180
aggttcagtg gcagcgggtc tgggacagat ttcactctca ccatcagcag cctagagcct 240
gaagattttg cagtttatta ctgtcagcag cgtaacaact ggcctccgct cactttcggc 300
ggagggacca aggtggagat caaa 324
<210> 65
<211> 321
<212> DNA
<213> Homo sapiens
<220>
<221> misc_feature
<222> (1)...(321)
<223> light chain nucleotide sequence of constant kappa
region
<400> 65
actgtggctg caccatctgt cttcatcttc ccgccatctg atgagcagtt gaaatctgga 60
actgctagcg ttgtgtgcct gctgaataac ttctatccca gagaggccaa agtacagtgg 120
aaggtggata acgccctcca atcgggtaac tcccaggaga gtgtcacaga gcaggacagc 180
aaggacagca cctacagcct cagcagcacc ctgacgctga gcaaagcaga ctacgagaaa 240
cacaaagtct acgcctgcga agtcacccat cagggcctga gctcgcccgt cacaaagagc 300
ttcaacaggg gagagtgtta g 321
<210> 66
<211> 318
<212> DNA
<213> Homo sapiens
<220>
<221> misc_feature
<222> (1)...(318)
<223> light chain nucleotide sequence of constant lambda
region
<400> 66
ggtcagccca aggctgcccc ctctgtcact ctgttcccgc cctctagcga ggagcttcaa 60
gccaacaagg ccacactggt gtgtctcata agtgacttct acccgggagc cgtgacagtg 120
gcctggaagg cagatagcag ccccgtcaag gcgggagtgg agaccaccac accctccaaa 180
caaagcaaca acaagtacgc ggccagcagc tatctgagcc tgacgcctga gcagtggaag 240
tcccacagaa gctacagctg ccaggtcacg catgaaggga gcaccgtgga gaagacagtg 300
gtccctgcag aatgctct 318
<210> 67
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR1 region of 4A2 heavy
chain VH3-30
<400> 67
Gly Phe Thr Phe Asn Arg His Gly
1 5
<210> 68
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR1 region of 19B10
heavy chain VH4-31 D2 J6
<400> 68
Gly Ser Ile Ser Ser Glu Asp Phe Cys
1 5
<210> 69
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR2 region of 4A2 heavy
chain VH- 30
<400> 69
Ser Ser Asp Gly Ala Asn Gln
1 5
<210> 70
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR2 region of 19B10
heavy chain VH4-31 D2 J6
<400> 70
Ile Cys Tyr Thr Gly Asp
1 5
<210> 71
<211> 18
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR3 region of 4A2 heavy
chain VH3-30
<400> 71
Ala Arg Asp Gly Arg Cys Glu Gly Glu Arg Cys Tyr Ser Gly Val Thr
1 5 10 15
Asp Phe
<210> 72
<211> 20
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR3 region of 19B10
heavy chain VH4-31 D2 J6
<400> 72
Ala Arg Glu Asp Arg Arg Gln Leu His Ser Arg Pro Tyr Phe Tyr Tyr
1 5 10 15
Gly Leu Asp Val
20
<210> 73
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR1 region of 4A2 light
chain L6 IgKV3-11
<400> 73
Gln Asn Ile Gly Gly Tyr
1 5
<210> 74
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR1 region of 19B10
light chain A3 IgKV2
<400> 74
Gln Ser Leu Leu His Ser Asn Gly His Asn Tyr
1 5 10
<210> 75
<211> 3
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR2 region of 4A2 light
chain L6 IgKV3-11
<400> 75
Asp Ala Ser
1
<210> 76
<211> 2
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR2 region of 19B10
light chain A3 IgKV2
<400> 76
Leu Gly
1
<210> 77
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR3 region of 4A2 light
chain L6 IgKV3-11
<400> 77
Gln Gln Arg Asn Ser Trp Pro Pro Leu Thr
1 5 10
<210> 78
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> synthetically constructed CDR3 region of 19B10
light chain A3 IgKV2
<400> 78
Gln Ala Leu Gln Thr Pro Asn Thr
1 5
Claims (16)
- (a) 사이토메갈로바이러스(CMV) 스트레인 AD169의 gB 단백질에 적어도 10 nM의 친화도로 결합하고,
(b) 인간 항체 또는 단편이며,
(c) 섬유 모세포 배양 중에서 CMV를 중화하는 것인,
분리된 단일클론 항체(mAb) 또는 그의 면역반응성 단편. - 제1항에 있어서, 완전 항체의 형태인 것인 mAb.
- 제1항에 있어서, 이중 특이적 항체인 것인 mAb.
- 제1항에 있어서, CMV의 AD169 스트레인의 gB에 대하여 적어도 1 nM의 친화도를 가지는 것인 mAb 또는 단편.
- 제1항에 있어서, 중쇄는 GFTFNRHG (SEQ ID NO:67) 또는 GSISSEDFC (SEQ ID NO:68)의 CDR1을 가지거나; 및/또는
중쇄는 SSDGANQ (SEQ ID NO:69) 또는 ICYTGD (SEQ ID NO:70)의 CDR2 영역을 가지거나; 및/또는
중쇄는 ARDGRCEGERCYSGVTDF (SEQ ID NO:71) 또는 AREDRRQLHSRPYFYYGLDV (SEQ ID NO:72)의 CDR3 영역을 가지는 것인 항체. - 제1항에 있어서, 경쇄는 QNIGGY (SEQ ID NO:73) 또는 QSLLHSNGHNY (SEQ ID NO:74)의 CDR1 영역을 가지거나; 및/또는
경쇄는 DAS (SEQ ID NO:75) 또는 LG (SEQ ID NO:76)의 CDR2 영역을 가지거나; 및/또는
경쇄는 QQRNSWPPLT (SEQ ID NO:77) 또는 QALQTPNT (SEQ ID NO:78)의 CDR3 영역을 가지는 것인 항체. - 제6항에 있어서, 4A2, 19B10, 313, 338 또는 345인 것인 항체 또는 단편.
- 제1항 내지 제7항 중 어느 하나의 항의 mAb 또는 단편을 인코딩하는 제1뉴클레오타이드 서열을 포함하는 것인 하나 이상의 핵산 분자, 또는 전체 길이에 걸쳐서 상기 제1뉴클레오타이드 서열과 상보적인 제2뉴클레오타이드 서열을 포함하는 것인 하나 이상의 핵산 분자.
- 제1항 내지 제7항 중 어느 하나의 항의 mAb 또는 단편을 생산하는 발현 시스템을 포함하는 재조합 숙주 세포로서, 상기 재조합 숙주 세포는 포유류 세포, 미생물 세포, 곤충 세포 또는 식물 세포인 것인 재조합 숙주 세포.
- mAb 또는 그의 면역반응성 단편을 생산하는 방법으로서, 상기 방법은 제9항의 세포를 배양하는 단계 및 상기 mAb 또는 단편을 수득하는 단계를 포함하는 것인 방법.
- 제1항 내지 제7항 중 어느 하나의 항의 분리된 단일클론 항체 또는 단편을, 약학적으로 허용가능한 부형제와 함께 포함하는 약학적 조성물.
- 제11항에 있어서, 추가의 약학적 제제를 약학적으로 허용가능한 부형제와 함께 더 포함하는 것인 약학적 조성물.
- CMV로 감염된 인간 대상체 중의 CMV를 치료하는 방법으로서, 그러한 치료가 필요한 대상체에게 제1항 내지 제7항 중 어느 하나의 항의 항체 또는 단편의 유효량을 투여하는 것을 포함하는 것인 방법.
- 인간 대상체에 있어서, CMV에 의한 감염에 대한 저항을 증진시키는 방법으로서, 그러한 증진이 필요한 대상체에게 제1항 내지 제7항 중 어느 하나의 항의 항체 또는 단편의 유효량을 투여하는 것을 포함하는 것인 방법.
- 제13항 또는 제14항에 있어서, 상기 대상체는 면역력이 약화된 것인 방법.
- 제13항 또는 제14항에 있어서, 상기 대상체는 임신한 여성인 것인 방법.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US35549910P | 2010-06-16 | 2010-06-16 | |
US61/355,499 | 2010-06-16 | ||
PCT/US2011/040761 WO2011159938A2 (en) | 2010-06-16 | 2011-06-16 | High affinity human antibodies to human cytomegalovirus (cmv) gb protein |
Publications (2)
Publication Number | Publication Date |
---|---|
KR20130086036A true KR20130086036A (ko) | 2013-07-30 |
KR101900435B1 KR101900435B1 (ko) | 2018-09-20 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020137001220A KR101900435B1 (ko) | 2010-06-16 | 2011-06-16 | 인간 사이토메갈로바이러스 gB 단백질에 대한 고친화도 인간 항체 |
Country Status (11)
Country | Link |
---|---|
US (3) | US9017668B2 (ko) |
EP (1) | EP2582389B1 (ko) |
JP (2) | JP6055763B2 (ko) |
KR (1) | KR101900435B1 (ko) |
CN (1) | CN103221063B (ko) |
AU (1) | AU2011268277B2 (ko) |
BR (1) | BR112012032182A2 (ko) |
CA (1) | CA2839420C (ko) |
IL (1) | IL223665B (ko) |
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Families Citing this family (8)
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WO2009114560A2 (en) * | 2008-03-10 | 2009-09-17 | Spaltudaq Corporation | Compositions and methods for the therapy and diagnosis of cytomegalovirus |
EP2516463B1 (en) | 2009-12-23 | 2016-06-15 | 4-Antibody AG | Binding members for human cytomegalovirus |
KR101900435B1 (ko) * | 2010-06-16 | 2018-09-20 | 트렐리스 바이오싸이언스 인코포레이티드 | 인간 사이토메갈로바이러스 gB 단백질에 대한 고친화도 인간 항체 |
US10611800B2 (en) | 2016-03-11 | 2020-04-07 | Pfizer Inc. | Human cytomegalovirus gB polypeptide |
US11440951B2 (en) | 2017-03-13 | 2022-09-13 | The Government Of The United States, As Represented By The Secretary Of The Army | Therapeutic antibodies to Marburg virus |
US11629172B2 (en) | 2018-12-21 | 2023-04-18 | Pfizer Inc. | Human cytomegalovirus gB polypeptide |
CN112898414B (zh) * | 2019-12-04 | 2024-05-10 | 珠海泰诺麦博制药股份有限公司 | 抗人巨细胞病毒抗体及其用途 |
US11857622B2 (en) | 2020-06-21 | 2024-01-02 | Pfizer Inc. | Human cytomegalovirus GB polypeptide |
Family Cites Families (20)
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US6642062B2 (en) | 1998-09-03 | 2003-11-04 | Trellis Bioinformatics, Inc. | Multihued labels |
US20090042291A1 (en) * | 2002-03-01 | 2009-02-12 | Xencor, Inc. | Optimized Fc variants |
US8420353B2 (en) | 2002-03-22 | 2013-04-16 | Aprogen, Inc. | Humanized antibody and process for preparing same |
US7427469B2 (en) * | 2002-11-05 | 2008-09-23 | Institut Pasteur | Method of treating cytomegalovirus with DC-SIGN blockers |
RU2239453C2 (ru) * | 2002-12-03 | 2004-11-10 | Федеральное государственное унитарное предприятие " Научно-производственное объединение по медицинским иммунобиологическим препаратам " Микроген" | Препарат иммуноглобулина человека против цитомегаловируса и способ его получения |
US7595379B2 (en) * | 2003-05-30 | 2009-09-29 | Agensys, Inc. | Antibodies and related molecules that bind to PSCA proteins |
US7413868B2 (en) | 2003-11-05 | 2008-08-19 | Trellis Bioscience, Inc. | Use of particulate labels in bioanalyte detection methods |
AU2005250499B2 (en) | 2004-06-03 | 2011-12-08 | Novimmune S.A. | Anti-CD3 antibodies and methods of use thereof |
WO2007094423A1 (ja) | 2006-02-15 | 2007-08-23 | Evec Incorporated | ヒトサイトメガロウイルスに結合するヒトのモノクローナル抗体並びにその抗原結合部分 |
JP2009531273A (ja) * | 2006-02-28 | 2009-09-03 | オンコセラピー・サイエンス株式会社 | 抗EphA4抗体のエフェクター機能を用いて細胞を障害する方法 |
SI2041177T1 (sl) * | 2006-06-02 | 2012-03-30 | Regeneron Pharma | Visoko afinitetna protitelesa za humani IL receptor |
JP2009544014A (ja) | 2006-07-12 | 2009-12-10 | トレリス バイオサイエンス、インク. | セルスポット・アプリケーション |
GB0700133D0 (en) | 2007-01-04 | 2007-02-14 | Humabs Llc | Human cytomegalovirus neutralising antibodies and use thereof |
CN100575361C (zh) * | 2007-04-28 | 2009-12-30 | 中国人民解放军第三军医大学野战外科研究所 | 抗人巨细胞病毒包膜糖蛋白b的人源化单链抗体 |
EP2190876A1 (en) | 2007-08-22 | 2010-06-02 | Ribovax Biotechnologies SA | Antibodies against human cytomegalovirus (hcmv) |
RU2364627C2 (ru) * | 2007-11-06 | 2009-08-20 | Власик Татьяна Николаевна | Экспрессионный вектор для синтеза белков в клетках млекопитающих |
CA2717803A1 (en) | 2008-03-03 | 2009-09-11 | Dyax Corp. | Metalloproteinase 9 and metalloproteinase 2 binding proteins |
WO2009114560A2 (en) * | 2008-03-10 | 2009-09-17 | Spaltudaq Corporation | Compositions and methods for the therapy and diagnosis of cytomegalovirus |
PE20141398A1 (es) | 2008-07-16 | 2014-10-18 | Inst Research In Biomedicine | Anticuerpos neutralizantes de citomegalovirus humano |
KR101900435B1 (ko) * | 2010-06-16 | 2018-09-20 | 트렐리스 바이오싸이언스 인코포레이티드 | 인간 사이토메갈로바이러스 gB 단백질에 대한 고친화도 인간 항체 |
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JP2017038612A (ja) | 2017-02-23 |
EP2582389A4 (en) | 2014-01-08 |
US20150252099A1 (en) | 2015-09-10 |
CA2839420A1 (en) | 2011-12-22 |
US10030069B2 (en) | 2018-07-24 |
CN103221063B (zh) | 2016-08-24 |
KR101900435B1 (ko) | 2018-09-20 |
CA2839420C (en) | 2023-06-13 |
US9017668B2 (en) | 2015-04-28 |
AU2011268277A1 (en) | 2013-01-31 |
AU2011268277A2 (en) | 2013-07-25 |
RU2613421C2 (ru) | 2017-03-16 |
US9688744B2 (en) | 2017-06-27 |
RU2013101769A (ru) | 2014-07-27 |
IL223665B (en) | 2020-01-30 |
AU2011268277B2 (en) | 2016-10-27 |
JP2013531991A (ja) | 2013-08-15 |
WO2011159938A3 (en) | 2012-04-19 |
BR112012032182A2 (pt) | 2016-10-25 |
US20170204165A1 (en) | 2017-07-20 |
CN103221063A (zh) | 2013-07-24 |
EP2582389B1 (en) | 2018-02-21 |
WO2011159938A2 (en) | 2011-12-22 |
JP6055763B2 (ja) | 2016-12-27 |
US20120020980A1 (en) | 2012-01-26 |
EP2582389A2 (en) | 2013-04-24 |
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