KR102175878B1 - Fviii 및 vwf 인자를 포함하는 키메라 단백질 및 그 용도 - Google Patents
Fviii 및 vwf 인자를 포함하는 키메라 단백질 및 그 용도 Download PDFInfo
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Abstract
본원에 따른 키메라 단백질은 FVIII에 결합된 vWF 도메인으로 인해, 투여시 체내 반감기가 월등히 증가하여, 혈우병 A형의 치료제로서 환자의 편리성이 증대되는 것은 물론, 치료비의 절감을 가져올 수 있다.
Description
본원은 혈우병의 치료에 사용되는 재조합 Factor VIII과 관련된 기술이다.
혈우병은 응고인자의 부족으로 지혈이 되지 않아 출혈이 지속적으로 생기는 질환이다. 혈액 내 응고인자는 I~XIII으로 구성되어 있으며, 이중 혈우병과 관련 있는 인자는 VIII, IX 및 XI이다. 혈우병은 상기 각 인자의 유전자 결함으로 발생되며, 부족한 응고인자의 종류에 따라 Factor VIII이 부족한 혈우병 A, Factor IX이 부족한 혈우병 B 및 Factor XI이 부족한 혈우병 C로 구분되며, 혈우병 A가 약 80-85%를 차지한다.
혈우병은 유형에 따라 투여약제가 상이하며, 동일한 유형의 경우에도 투여부위에 따라 투여량과 방법이 다르다. 하지만 혈우병 치료의 목적은 지혈이며 치료는 효소대체요법(ERT, Enzyme Replacement Therapy)을 이용하여 예방적(prophylaxis)과 대증적(on demand)의 두 가지 측면에서 수행되는데, 현재 치료법은 예방적 측면으로 옮겨가는 추세이다.
ERT를 위해 사용되는 효소로 과거에는 전혈과 혈장에서 추출하여 농축한 FVIII (혈우병 A), FIX 인자 (혈우병 B)가 사용되었다. 하지만 전혈과 혈장에서 혈우인자를 추출할 때, AIDS나 HBV와 같은 바이러스에 감염된 사람의 혈액이 일부 사용되었고, 이러한 바이러스가 혈우인자(coagulant factor) 추출과정에서 적절히 제거되지 못함으로 인한 바이러스 감염과 같은 부작용과 합병증의 문제가 발생하였다. 따라서 최근에는 유전자 재조합 기술로 생산된 다양한 인자가 개발되어 사용되고 있으며, 예를 들면 온전한 형태의 FVIII, 또는 B 도메인이 결실된 형태, 특정 잔기에서의 변형을 통해 반감기가 증가된 다양한 FVIII 등이 개발되고 있다.
FVIII을 예방적 요법으로 사용할 때, FVIII의 혈중 반감기가 12시간(human)으로 짧아 2~3일에 한 번 투여해야 하는 불편함이 있다. 또한, I.V 투여이기 때문에 투여 횟수 감소가 환자 편의성 측면에서 봤을 때, 중요하다. 이에 FVIII의 혈중 반감기를 늘려준 long-acting FVIII이 최근 개발되었다.
그 중 한 가지는 FVIII의 C-말단에 항체의 Fc 도메인을 융합한 FVIII-Fc 융합 단백질(FVIII-Fc)이 개시되어 있고, 이 물질은 FcRn recycling을 이용하여 반감기를 증가시키는 전략을 사용하였다. 또한 FVIII에 PEG(polyethylene glycol)를 FVIII의 생체 내 제거(clearance)를 담당하는 것으로 알려진 LRP(Low density lipoprotein receptor related protein)과의 결합부위 근처에 접합시킨 페길화 FVIII이 몇 건 개시되어 있다. 하지만, 이들 long-acting FVIII들의 임상결과 반감기가 기존 12시간에서 평균 18시간 정도로 약 1.5배 밖에 증가하지 못 했으며, 이는 기존 주 3~4회의 투여 횟수가 주 2~3회 투여 횟수로 줄어든 것을 의미한다. 이렇게 FVIII-Fc와 페길화 FVIII의 반감기가 크게 증가하지 못 하는 이유는 FVIII이 혈중에서 vWF와 비공유 결합(non-covalent interaction)을 강하게 하고 있는 상태로 혈중에서 순환(circulation)하고 있기 때문이다. FVIII-Fc와 페길화 FVIII의 경우에도 혈중에서 혈중에 존재하는 vWF와 비공유 결합을 하게 되고, 결국 vWF의 반감기에 영향을 받아 vWF의 반감기(12시간, human)에 수렴하게 되는 한계가 있다 (Tiede A. Journal of Thrombosis and Haemostasis, 13 (Suppl. 1): S176-S179).
현재까지 개발된 치료제는 여전히 기능적 측면에서 투여 주기(정맥 주사)가 짧고, 또한 충분히 길지 못한 반감기로 인한 체내 효력 소실에 따라 잦은 출혈 빈도가 문제점으로 지적되고 있다. 따라서 투여 횟수를 최소 주 1회까지 줄여 주기 위한 반감기가 3배 이상 증가한 long-acting FVIII의 개발이 필요하다.
본원은 체내 체류 시간이 향상되어 주 1회 이하로 투여 가능한 FVIII을 제공하고자 한다.
한 양태에서 본원은 경쇄 및 중쇄를 포함하며 B 도메인의 전장 또는 그 일부가 결실된 인간 혈액응고 8인자(Factor VIII, FVIII); 및 하나 이상의 vWF D’D3 도메인을 포함하는 키메라 단백질을 제공한다. 본원에 따른 단백질은 야생형 FVIII 단백질과 비교하여 최소 2배 반감기가 연장된다.
본원에 따른 키메라 단백질에서, 상기 경쇄 및 중쇄는 한 개 폴리펩타이드로 또는 두 개의 폴리펩타이드로 발현될 수 있다.
본원에 따라 상기 한 개의 폴리펩타이드로 발현되는 경우, 상기 FVIII 및 상기 vWF D’D3 도메인을 연결하며, 그 사이에 위치하는 효소절단부위 갖는 연결체를 포함하며, 상기 vWF D’D3 도메인은 상기 연결체를 기준으로 그 N-말단이 상기 FVIII의 N-말단 또는 C-말단 방향, 또는 상기 vWF D’D3 도메인 및 상기 FVIII는 각각 상기 연결체를 기준으로 그 N-말단 방향 또는 C- 말단 방향에 위치한다. 일 구현예에서 예를 들면 N 부터 C 방향으로 FVIII-연결체-D’D3 또는 D’D3-연결체-FVIII의 순서로 배열될 수 있다.
본원에 따라 두 개의 폴리펩타이드로 발현되는 경우, 상기 vWF D’D3 도메인은 그 N-말단이 상기 중쇄에 포함된 상기 B 도메인의 C-말단에 연결될 수 있다.
일 구현예에서 본원에 따른 키메라 단백질에 포함될 수 있는 일부가 결실된 B 도메인은 서열번호 1의 서열에서, 1648 및 1649 잔기 위치를 기준으로 상기 잔기를 포함하여 N-말단 및/또는 C-말단 방향으로 각각 최소 5개의 아미노산이 결실된 것; 또는 상기 B 도메인이 최소 4개, 또는 4개 내지 6개의 당화부위를 포함하도록 일부가 결실된 것을 포함한다.
다른 구현예에서 본원에 따른 키메라 단백질에 포함될 수 있는 일부가 결실된 B 도메인은 서열번호 01의 서열을 기준으로 아미노산 잔기 741 내지 902 및 1654 내지 1689의 서열, 또는 서열번호 01의 서열을 기준으로 아미노산 잔기 741 내지 936의 서열로 표시된다.
또 다른 구현예에서 본원에 따른 키메라 단백질은 하나 이상의 Fc를 추가로 포함하며, 상기 Fc는 상기 vWF D’D3 도메인 및/또는 상기 FVIII에 연결될 수 있다.
본원에 따른 다른 구현예에서 본원에 따른 키메라 단백질은 두 개 이상의 vWF D’D3 도메인 및/또는 두 개 이상의 Fc를 추가로 포함할 수 있다. 이 경우, 복수 개의 vWF D’D3 도메인간, 그리고 복수 개의 Fc 간에는 서로 효소절단부위를 포함하거나 포함하지 않는 연결체로 연결되거나 및/또는 서로간에 공유결합을 통한 복합체, 예를 들면 두 개를 포함하는 경우 이량체를 형성할 수 있다. 여기에 사용된 효소절단부위 및 연결체는 본 명세서에 기재된 것을 참조 할 수 있다.
또 다른 구현예에서 본원에 따른 키메라 단백질에서 효소절단 부위를 포함하는 연결체가 사용되는 경우, 상기 효소절단부위는 트롬빈, FVIIa, FXa 또는 FXIa에 의해 절단될 수 있는 서열이고, 그 외의 서열은 [G4S]n 의 아미노산 서열로 표시되며, 상기 식에서 n= 0, 또는 1 내지 100 사이 정수, 특히 n은 4, 6, 8, 10, 16, 21, 22 또는 58이다.
또 다른 구현예에서 본원에 따른 키메라 단백질에서 연결체에 포함된 효소절단 부위에서 상기 트롬빈에 의해 절단될 수 있는 아미노산 서열은 DFLAEGGGVR (서열번호 36), TTKIKPR (서열번호 37), 또는 LVPRGS (서열번호 38)이고, 상기 FVIIa에 의해 절단될 수 있는 아미노산 서열은 ASKPQGRIVGG (서열번호 39), 상기 FXa에 의해 절단될 수 있는 아미노산 서열은 IDGR (서열번호 40) 또는 IEGR (서열번호 41)이고, 상기 FXIa에 의해 절단될 수 있는 서열은 SKLTRAETVF (서열번호 42)로 표시될 수 있다.
다른 양태에서 본원에 따른 키메라 단백질은 제 1 및 제 2 폴리펩타이드의 형태로 발현되며, 제 1 폴리펩타이드는 상기 FVIII의 중쇄 및 B 도메인의 전부 또는 일부를 포함하고, 상기 제2 폴리펩타이드는 상기 FVIII의 경쇄 또는 B 도메인의 일부를 포함하고, 상기 vWF D’D3 도메인은 상기 제 1 폴리펩타이드의 N-말단 또는 C-말단 방향, 또는 제 2 폴리펩타이드의 N-말단 또는 C-말단에 위치한다. vWF D’D3 도메인이 제1 폴리펩타이드 N-말단 또는 제 2 폴리펩타이드의 C-말단에 연결되는 경우 scFVIII의 N-말단 또는 C-말단에 연결되는 것과 유사한 구조를 갖으며, 이 경우 FVIII가 단쇄 또는 이쇄인지가 차이나는 부분이다.
일 구현예에서 제 1 및 제 2 폴레펩타이드의 형태로 발현되는 경우, 상기 vWF D’D3 도메인은 제 1 폴리펩타이드의 C-말단 방향에 위치한다.
본원에 따른 키메라 단백질은 하나 이상, 특히 두 개의 vWF D’D3 도메인을 포함할 수 있으며, vWF D’D3 도메인 간에 복합체(이량체)를 형성할 수 있다.
선택적으로 본원에 따른 키메라 단백질은 하나 이상의 Fc, 특히 두개의 Fc를 추가로 포함하며, 상기 Fc는 상기 vWF D’D3 도메인 및/또는 상기 제 2 폴리펩타이드에 연결될 수 있다. Fc 간에 복합체(이량체)를 형성할 수 있다.
일구현예에서 본원에 따른 키메라 단백질에서 Fc 또는 상기 vWF D’D3 도메인은 효소절단부위를 포함하거나 또는 포함하지 않는 연결체로 연결된다.
일 구현예에서 본원에 따른 키메라 단백질은 한 개의 폴리펩타이드로 발현되는 경우 서열번호 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21 또는 25로 표시되는 아미노산 서열 또는 이와 생물학적 기능이 균등하거나 또는 이와 90% 이상 상동성이 있는 아미노산 서열도 본원에 포함될 수 있다.
다른 구현예에서 본원에 따른 키메라 단백질은 두 개의 폴리펩타이드로 발현되는 경우 서열번호 4 및 5 (각각 중쇄 및 경쇄), 서열번호 23 및 24 (각각 중쇄 및 경쇄), 서열번호 26 및 27 (각각 중쇄 및 경쇄), 서열번호 28 및 29 (각각 중쇄 및 경쇄), 서열번호 30 및 31 (각각 중쇄 및 경쇄), 서열번호 32 및 33 (각각 중쇄 및 경쇄), 또는 서열번호 34 및 35 (각각 중쇄 및 경쇄)로 표시되는 아미노산 서열 또는 이와 생물학적 기능이 균등하거나 또는 이와 90% 이상 상동성이 있는 아미노산 서열도 본원에 포함될 수 있다.
본원에 따른 키메라 단백질은 FVIII의 A 도메인 및/또는 B 도메인 단편의 일부 아미노산 잔기에서 친수성 폴리머와 컨쥬게이션될 수 있고, 상기 컨쥬게이션되는 위치는 상기 B 도메인에서 서열번호 01의 서열을 기준으로 아미노산 잔기 754, 781, 782, 788, 789, 825, 및 897로 구성되는 군으로부터 선택되는 하나 이상의 위치이고; 상기 A 도메인에서 서열번호 01의 서열을 기준으로 아미노산 잔기 491, 495, 498 및 1806으로 구성되는 군으로부터 선택되는 하나 이상의 위치이고, 상기 컨쥬게이션되는 위치의 잔기, 특히 컨쥬게이션 되는 모든 위치의 잔기는 친수성 폴리머와의 접합을 위해 시스테인으로 치환된다.
일 구현예에서 친수성 폴리머는 폴리에틸렌글리콜(Polyethylene glycol), 폴리에틸렌 옥사이드, 덱스트란이다.
다른 구현예에서, 친수성 폴리머는 폴리에틸렌글리콜이고, 평균 분자량이 최소 20kDa이상, 특히 40kDa 또는 60kDa의 PEG가 사용된다.
일 구현예에서 본원에 따른 키메라 단백질은 FVIII의 B 도메인 단편, 특히 782번 잔기에서 개질된다.
다른 양태에서 본원은 상술한 키메라 단백질을 코딩하는 핵산, 상기 핵산을 포함하는 벡터, 상기 벡터를 포함하는 세포를 제공한다.
또 다른 양태에서 본원은 키메라 단백질, 이를 코딩하는 핵산, 상기 핵산을 포함하는 벡터, 상기 벡터를 포함하는 세포의 용도를 제공한다.
본원에 따른 발명의 용도는 혈우병 치료용 조성물, 혈우병 치료 방법, 혈액 응고용 조성물, 혈액 응고 방법, 또는 본원에 따른 단백질의 생산 방법을 포함한다.
본원에 따른 FVIII와 vWF 도메인이 결합된 키메라 단백질 또는 PEG로 개질된 형태의 키메라 단백질은 투여 시 체내 반감기가 월등히 증가하여, 기존 주 2~4회의 투여 횟수를 주 1회 이하로 줄여주어, 혈우병 A형의 치료제로서 환자의 편리성이 증대되는 것은 물론, 치료비의 절감을 가져올 수 있다.
도 1은 본원에 따른 반감기가 증가된 재조합 FVIII의 반감기 증가 기전을 도식적으로 나타낸 것이다. FVIII의 생체 내 제거에 가장 큰 영향을 미치는 것으로 알려진 LRP 결합부위를 PEG와 vWF의 일부 도메인인 D’D3로 가려(masking)주는 것이 첫 번째 전략이다. 이와 동시에 본원에 따른 키메라 단백질에 vWF의 일부 도메인인 D’D3를 공유적으로 결합시켜 주어 혈중에 있는 내인성 vWF가 키메라 단백질에 결합하는 것을 막아주는 것이 두 번째 전략이다.
도 2a 및 2b는 각각 본원에 따른 단쇄(단일 폴리펩타이드) 및 이쇄(이중 폴리펩타이드)로 발현되는 키메라 단백질의 다양한 분자구조를 도식적으로 나타낸 것이다.
도 3은 본원의 일 구현예에 따른 단일 폴리펩타이드로 발현되는 키메라 단백질을 코딩하는 벡터를 도식적으로 나타낸 것이다.
도 4는 본원의 일 구현예에 따른 다양한 길이의 연결체를 포함하는 키메라 단백질 발현 및 정제 후 SDS-PAGE로 분석한 결과이다.
도 5는 본원의 일 구현예에 따른 다양한 길이의 연결체를 포함하는 키메라 단백질을 각각 중쇄, 경쇄 및 D’D3에 대한 항체로 분석한 웨스턴블랏 결과이며, 사용된 각 항체는 다음과 같다: 중쇄: Green mountain, GMA-012; 경쇄: Abcam, 41188 및 D’D3: Abcam, 96340.
도 6은 본원의 일 구현예에 따른 scFVIII/D’D3-60, -120 및 -300을 PEG로 개질한 페길화 키메라 단백질을 정제 후 SDS-PAGE로 분석한 결과이다.
도 7은 본원의 일 구현예에 따른 키메라 FVIII (scFVIII/D’D3-120-TH1), 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120-TH1)의 vWF 결합 분석 결과로 비교군으로 Advate®를 사용하였다.
도 8은 본원의 일 구현예에 따른 키메라 FVIII (scFVIII/D’D3-120-TH1), 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120-TH1)의 LRP 결합 분석 결과로 비교군으로 Advate®를 사용하였다.
도 9는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120)의 HA mice에서의 PK 분석 결과로 비교군으로 Advate®를 사용하였다.
도 10은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (40 kDa PEG-scFVIII/D’D3-120, 60 kDa PEG-scFVIII/D’D3-120)의 HA mice에서의 PK 분석 결과로 비교군으로 Advate®를 사용하였다.
도 11은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3/Fc-120, Fc)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 12는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-300, PEG-scFVIII/D’D3/D’D3-120)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 13은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-300)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 14는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 15는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (20 kDa linear PEG-scFVIII/D’D3-120-TH1, 20 kDa branch PEG-scFVIII/D’D3-120-TH1, 40 kDa branch PEG-scFVIII/D’D3-120-TH1)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 16은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120-TH1)의 트롬빈 생성 분석 결과이다.
도 17은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2)의 꼬리클립모델(tail clip model)에서의 급성효능연구결과로 비교군으로 Advate®를 사용하였다.
도 18은 본원의 일 구현예에 따라 CHO 세포에서 발현하여 제조한 페길화 키메라 FVIII (20 kDa linear PEG-scFVIII/D’D3-120-TH1, 40 kDa linear PEG-scFVIII/D’D3-120-TH1)의 HA 마우스에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 2a 및 2b는 각각 본원에 따른 단쇄(단일 폴리펩타이드) 및 이쇄(이중 폴리펩타이드)로 발현되는 키메라 단백질의 다양한 분자구조를 도식적으로 나타낸 것이다.
도 3은 본원의 일 구현예에 따른 단일 폴리펩타이드로 발현되는 키메라 단백질을 코딩하는 벡터를 도식적으로 나타낸 것이다.
도 4는 본원의 일 구현예에 따른 다양한 길이의 연결체를 포함하는 키메라 단백질 발현 및 정제 후 SDS-PAGE로 분석한 결과이다.
도 5는 본원의 일 구현예에 따른 다양한 길이의 연결체를 포함하는 키메라 단백질을 각각 중쇄, 경쇄 및 D’D3에 대한 항체로 분석한 웨스턴블랏 결과이며, 사용된 각 항체는 다음과 같다: 중쇄: Green mountain, GMA-012; 경쇄: Abcam, 41188 및 D’D3: Abcam, 96340.
도 6은 본원의 일 구현예에 따른 scFVIII/D’D3-60, -120 및 -300을 PEG로 개질한 페길화 키메라 단백질을 정제 후 SDS-PAGE로 분석한 결과이다.
도 7은 본원의 일 구현예에 따른 키메라 FVIII (scFVIII/D’D3-120-TH1), 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120-TH1)의 vWF 결합 분석 결과로 비교군으로 Advate®를 사용하였다.
도 8은 본원의 일 구현예에 따른 키메라 FVIII (scFVIII/D’D3-120-TH1), 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120-TH1)의 LRP 결합 분석 결과로 비교군으로 Advate®를 사용하였다.
도 9는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120)의 HA mice에서의 PK 분석 결과로 비교군으로 Advate®를 사용하였다.
도 10은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (40 kDa PEG-scFVIII/D’D3-120, 60 kDa PEG-scFVIII/D’D3-120)의 HA mice에서의 PK 분석 결과로 비교군으로 Advate®를 사용하였다.
도 11은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3/Fc-120, Fc)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 12는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-300, PEG-scFVIII/D’D3/D’D3-120)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 13은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-300)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 14는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 15는 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (20 kDa linear PEG-scFVIII/D’D3-120-TH1, 20 kDa branch PEG-scFVIII/D’D3-120-TH1, 40 kDa branch PEG-scFVIII/D’D3-120-TH1)의 HA mice에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
도 16은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120-TH1)의 트롬빈 생성 분석 결과이다.
도 17은 본원의 일 구현예에 따라 제조한 페길화 키메라 FVIII (PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2)의 꼬리클립모델(tail clip model)에서의 급성효능연구결과로 비교군으로 Advate®를 사용하였다.
도 18은 본원의 일 구현예에 따라 CHO 세포에서 발현하여 제조한 페길화 키메라 FVIII (20 kDa linear PEG-scFVIII/D’D3-120-TH1, 40 kDa linear PEG-scFVIII/D’D3-120-TH1)의 HA 마우스에서의 PK 결과로 비교군으로 Advate®를 사용하였다.
본원은 FVIII이 내인성 vWF 단백질과 복합체를 형성하는 것을 방지 또는 억제하는 것과 동시에 FVIII이 LRP(Low density lipoprotein receptor related protein)에 결합하는 것을 막아주어 FVIII 단백질의 반감기를 연장하는 기술에 관한 것이다. 이로 인해 LRP 및 프로테아제 의존적 FVIII 제거를 지연시키는 것은 물론 vWF 의존적 제거를 동시에 감소시켜 반감기가 증가된다.
따라서 한 양태에서 본원은 인간 혈액응고 8인자(Factor VIII 또는 FVIII) 및 하나 이상의 vWF D’D3 도메인이 융합된 키메라 단백질에 관한 것이다.
본원에 따른 키메라 단백질은 천연형태에서 경쇄와 중쇄로 구성되는 FVIII의 경쇄 및 중쇄를 하나의 폴리펩타이드로 발현되는 단쇄(sc, single chain)로, 또는 경쇄 및 중쇄가 각각 별개의 폴리펩타이드로 발현되는 이쇄(dc, dual chain)의 형태를 취할 수 있다.
일 구현예에서 본원에 따른 키메라 단백질이 단쇄(단일 폴리펩타이드)로 구현되며, 이 경우 vWF D’D3 도메인은 FVIII의 N- 또는 C- 말단 방향에 위치하며, 효소에 의한 절단 부위를 포함하는 아미노산 링커(또는 연결체)로 연결된다.
본원에 따른 키메라 단백질은 또한 이쇄(이중 폴리펩타이드)로 구현되며, 이 경우 vWF D’D3 도메인은 FVIII의 중쇄 또는 경쇄의 N- 또는 C- 말단 방향에 연결될 수 있다. 일 구현예에서는 특히 B 도메인의 C- 말단 방향에 연결된다.
본원에 따른 키메라 단백질에 포함될 수 있는 FVIII 단백질은 vWF와의 결합을 통해 본원에 따른 목적을 달성하는 한, 다양한 길이 또는 유래의 것이 사용될 수 있다.
FVIII는 A1-A2-B-A3-C1-C2 도메인으로 구성되며, 간세포에서 단쇄의 단백질로 합성된다. 합성 후 프로세싱을 거쳐 성숙되어 중쇄와 경쇄로 구성된 280 kDa의 헤테로이량체를 형성한다. 이 중 경쇄는 80kDa이며 A3-C1-C2 도메인으로 구성되고, 중쇄는 A1-A2-B 도메인으로 구성되고 분자량은 90-200kDa으로 B-도메인의 길이에 따라 분자량의 차이가 크다. 이러한 헤테로이량체는 혈액 내에서 vWF와 결합하여 비활성화 상태로 존재하다가, 혈관 손상과 같은 자극에 노출 시, 트롬빈에 의해 알지닌 잔기 뒤, 즉 서열번호 01의 서열을 기준으로 372, 740 및 1689번 잔기 뒤에서 절단된다. 그 결과 vWF로부터 분리되고, 활성화되어 A1, A2 및 A3-C1-C2의 삼량체가 형성된다. 이어 삼량체는 FIXa에 의한 FX의 활성화를 촉매하고, 빠르게 분해된다.
본원에 따른 키메라 단백질에 포함될 수 있는 FVIII 단백질은 응고 경로에서 전장 야생형 FVIII 인자의 기능을 보유하는 전장, 이의 기능적 단편, 변이체, 유사체 또는 유도체를 포함한다. FVIII 단백질은 FVIII 폴리펩타이드 또는 FVIII과 상호 교환적으로 사용된다. FVIII 기능의 예는 혈액 응고를 활성화시키는 능력, IX 인자에 대한 보조인자로서 작용하는 능력, 또는 Ca2+ 및 인지질의 존재 하에 IX 인자와 테나세 복합체(tenase complex)를 형성하는(이후 X 인자를 활성화 형태 Xa로 전환시킴) 능력을 포함하지만, 이들로 제한되지는 않는다.
또한 다양한 유래의 FVIII 단백질이 사용될 수 있으며, 예를 들면 인간, 돼지과, 고양이과, 래트 또는 쥣과 FVIII 단백질일 수 있다. FVIII 아미노산 및 핵산 서열의 비제한적인 예는 예컨대 GenBank NOs: NM_000132, NP_000123, 1012296A AAA52420.1, CAA25619.1, AAA52484.1, 1012298A, EAW72647.1, EAW72646.1, XP_001498954.1, ACK44290.1, ACO95359.1, NP_001138980.1, ABZ10503.1, NP_032003.2로 개시되어 있다.
본원에 따른 일 구현예에서는 인간 유래의 FVIII이 사용되며, 예를 들면 인간 FVIII은 서열번호 01의 서열(신호펩타이드는 포함하지 않음)로 표시될 수 있으며, 야생형 서열에서 중쇄는 서열번호 01의 서열을 기준으로 1-740 잔기, B 도메인은 a3 도메인을 포함하여 741-1689 잔기, 경쇄는 1690-2332 잔기까지이다.
또한 FVIII의 변이체, 유도체, 변형체, 돌연변이체, 복합체 또는 유사체 또한 당해 분야에 공지되어 있으며, 또 본 발명에 포함된다. 예를 들어, 미국 특허번호 제5,668,108호에 개시된 바와 같은 FVIII의 변이체는 위치 1241에 있는 아스파라긴산이 글루탐산에 의해 치환되고 첨부된 핵산 변화를 갖는 FVIII의 변이체를 개시하고; 미국 특허번호 제5,149,637호는 글리코실화되거나 또는 글리코실화되지 않은 C-말단 단편을 포함하는 FVIII 변이체를 기재하며; 또 미국 특허번호 제5,661,008호는 적어도 3개 아미노산 잔기에 의해 아미노산 1649 내지 2332에 연결된 아미노산 1-740을 포함하는 FVIII 변이체를 개시한다.
본원에 따른 키메라 단백질에 포함될 수 있는 FVIII은 B 도메인이 결실된 FVIII을 포함하며, 이러한 FVIII은 예를 들면 미국 특허 제6,316,226호, 제6,346,513호, 제7,041,635호, 제5,789,203호, 제6,060,447호, 제5,595,886호, 제6,228,620호, 제5,972,885호, 제6,048,720호, 제5,543,502호, 제5,610,278호, 제5,171,844호, 제5,112,950호, 제4,868,112호 및 제6,458,563호에 개시된 것을 포함하나 이로 제한하는 것은 아니다.
또한 본원에 따른 키메라 단백질에 포함될 수 있는 FVIII은 B 도메인의 일부가 결실된 FVIII으로 중쇄 및 경쇄를 포함하나 퓨린(furin)과 같은 단백질분해효소에 의한 절단부위를 포함하지 않도록 일부 잔기가 결실된다.
일 구현예에서 일부가 결실된 B 도메인은 서열번호 01의 서열의 1648 및 1649 잔기 위치를 기준으로, 이를 포함하여 N-말단 및 C-말단 방향으로 각각 최소 5개의 아미노산이 결실된 것을 포함하거나 및/또는 4개 내지 6개의 당화부위를 포함하도록 일부가 결실된 B 도메인 단편을 포함한다.
일 구현예에서 본원에 따른 키메라 단백질에 포함될 수 있는 B 도메인의 일부가 결실된 FVIII에서, 상기 B 도메인 단편은 서열번호 01의 서열을 기준으로 아미노산 잔기 741 내지 902 및 1654 내지 1689를 포함하는 것이다.
다른 구현예에서는 이러한 B 도메인 단편은 서열번호 01의 서열을 기준으로 아미노산 잔기 741 내지 902 및 1654 내지 1689의 아미노산 서열을 포함하고 상기 잔기 중 일부, 특히 782번 아이소루이신 잔기가 친수성 폴리머로의 개질을 위해 시스테인으로 치환된 것을 포함하는 것이다.
또 다른 구현예에서 본원에 따른 키메라단백질에 포함될 수 있는, B 도메인의 일부가 결실된 FVIII는 서열번호 03으로 표시되는 아미노산 서열 또는 이와 90% 이상의 상동성이 있는 아미노산 서열을 갖는다. 상기 서열번호 03의 아미노산 서열에 포함된 B 도메인은 서열번호 01을 기준으로 741 - 902번 잔기를 포함한다.
본원에 따른 키메라 단백질에 포함될 수 있는 FVIII은 A 도메인 및/또는 B 도메인 단편의 일부 아미노산 잔기에서 친수성 폴리머와 컨쥬게이션되어 개질될 수 있다.
특히 본원에서는 개질을 통해 본원에 따른 키메라 단백질의 LRP 결합을 방해하기 위한 것으로, 부위 특이적으로 진행되며, 이를 달성할 수 있는 다양한 위치에서 개질될 수 있다.
본원에 따른 일 구현예에서 본원에 따른 키메라 단백질에서 컨쥬게이션되는 위치는 상기 A 도메인에서 서열번호 01의 서열을 기준으로 아미노산 잔기 491, 495, 498 및 1806으로 구성되는 군으로부터 선택되는 하나 이상의 위치; 그리고 상기 B 도메인에서 서열번호 01의 서열을 기준으로 아미노산 잔기 754, 781, 782, 788, 789, 825, 897로 구성되는 군으로부터 선택되는 하나 이상의 위치이다.
본원에 따른 다른 구현예에서 특히 B 도메인, 특히 상술한 위치에서 페길화된다.
본원에 따른 다른 구현예에서는 B 도메인의 782번 아이소루이신 잔기에서 개질되며, 특히 페길화되고, 페길화를 위해, 후술하는 바와 같이 상기 잔기가 시스테인으로 치환되거나 또는 시스테인기가 삽입될 수 있다.
본원에 따른 FVIII의 개질에 사용될 수 있는 친수성 폴리머는 본원에 따른 목적을 달성하는 한 당업계에 공지된 다양한 폴리머가 사용될 수 있으며, 예를 들면 폴리에틸렌글리콜(Polyethylene glycol), 폴리에틸렌 옥사이드, 덱스트란을 포함하나 이로 제한하는 것은 아니다.
본원에 따른 일 구현예에서는 PEG가 사용되며, 특히 20kDa 이상의 PEG가 사용된다. 본원에 따르면, PEG는 LRP 결합영역 근처에 결합되어 PEG hindrance에 의해 FVIII이 LRP에 결합하는 것을 저해 시키는 기능을 하며, 따라서 PEG의 길이가 너무 짧으면 이러한 hindrance를 충분히 주지 못 할 가능성이 있기 때문에 PEG의 길이가 20kDa 이상인 것이 중요하다. 일 구현예에서는 특히 40kDa, 60kDa 또는 그 이상의 PEG가 사용된다.
본원에 따른 개질에서 페길화에 사용되는 PEG는 공지된 다양한 작용기 (functional group)을 이용하여 해당 잔기에 연결될 수 있다.
본원에 따른 일 구현예에서 PEG는 부위 특이적으로 시스테인과 같은 잔기에 존재하는 자유 -SH기를 통해 연결되며, 이 경우 예를 들면 thiol, orthopyridyl disulfide, acryloyl, sulfone 또는 maleimide기, 특히 maleimide기를 통해 연결될 수 있으나, 이로 제한하는 것은 아니다.
본원에 따른 FVIII은 친수성 폴리머로의 개질을 위해 친수성 폴리머에 연결되는 잔기는 다른 잔기로 치환 또는 삽입 될 수 있으며, 치환 또는 삽입되는 잔기는 개질에 사용되는 물질에 따라 상이하며, 당업자라면 적절한 것을 선정할 수 있을 것이다.
본원에 따른 일 구현예에서는 개질로서 페길화가 사용되며, 이 경우 개질되는 잔기는 시스테인으로 치환 또는 시스테인 잔기가 적절한 위치에 삽입될 수 있다.
본원에 따른 일 구현예에서는 B 도메인의 782번 아이소루이신 잔기에서 개질되며, 특히 페길화되고, 페길화를 위해 상기 잔기가 시스테인으로 치환된다.
페길화 반응은 공지된 방법을 이용하여 수행될 수 있으며, 예를 들면 자유 시스테인의 마스킹을 위해 사용된 시스테인이나 글루타치온은 페길화 반응 전에 제거되어 도입한 시스테인의 free thiol이 복원되어야 하며, 이를 위해 TCEP, DTT, beta-mercaptoethanol, 시스테인, 글루타티온(환원형) 등의 환원제를 처리하는 과정과 이 과정에서 FVIII 본연의 이황화 본드가 환원적으로 절단된 것을 복원하기 위한 산화 과정을 포함하여 수행할 수 있다. 개질을 위해 시스테인으로 치환된 단백질의 발현은 당업계에 공지된 방법을 이용하여 수행될 수 있으며, 예를 들면 페길화되는 단백질의 경우, 도입된 자유 시스테인은 발현 도중이나 세포 밖으로 배출 후 배양액의 free thiol을 포함하는 저분자 물질인 시스테인 또는 글루타치온과 이황화 결합을 형성하기 때문에, 자유 시스테인을 마스킹하여 페길화 전 까지 안정화 시켜 페길화 효율을 높이는 것이 필요하다.
본원에 따른 키메라 단백질은 vWF D’D3 도메인을 포함한다.
vWF(또는 F8vWF)는 바이벨펠라드 소체(Weibel-Palade body)에서 내피조직, 거핵세포(혈소판의 α-과립) 및 내피하(subendothelian) 결합조직에서 생성되고 혈장에 존재하는 대형 다합체 당단백질이다. 염기성 vWF 단량체는 2813개(시그널펩타이드 포함하는 경우)의 아미노산으로 구성되고, 모든 단량체는 특이적 기능을 갖는 다수의 특이적 도메인, 즉 (FVIII 인자에 결합하는) D'/D3 도메인, (혈소판 GPIb-수용체, 헤파린 및/또는 가능하게는 콜라겐에 결합하는) A1 도메인, (콜라겐에 결합하는) A3 도메인, C1 도메인(RGD 도메인이 활성화될 때 이것은 혈소판 인테그린 αIIbβ3에 결합) 및 단백질의 C-말단의 "시스테인 매듭" 도메인을 포함하며, 시스테인 매듭은 혈소판 유래 성장 인자(PDGF), 형질전환 성장 인자-β(TGFβ) 및 β-인간 융모성 고나도트로핀(βHCG)에서도 발견된다.
vWF 아미노산 서열 및 vWF 또는 그의 일부를 코딩하는 핵산 서열의 비제한적인 예는 예컨대 GenBank NO: NP_000543, NM_000552, AAE20723, AAB59512, P04275, EAW88815, ACP57027, EAW88816, 및 AAD04919; 미국 특허번호 제5,260,274호; Titani etal., Biochemistry, 25:3171-84 (1986); 및 Sadler etal., PNAS, 82:6391-6398 (1985)로 공지되어 있다.
예를 들면 전장 인간 vWF의 아미노산 서열은 서열번호 02 (시그널펩타이드는 포함하지 않음)로 나타낸다. 서열번호 02를 기준으로 vWF의 "프로펩티드" 부분(D1-D2)은 아미노산 잔기 1 부터 Arg-741까지이고, 또 "성숙" vWF 단백질은 아미노산 잔기 742 내지 2791 까지이다. 개별 도메인은 D': 742 - 842; D3: 843 - 1248; A1: 1249 - 1457; A2: 1458 - 1650; A3: 1651 - 1850; D4: 1851 - 2232; C1: 2233 - 2311; C2: 2312 - 2380; C3: 2407 - 2474; C4: 2475 - 2555; C5: 2556 - 2624; C6: 2625 - 2700 및 CK: 2701 - 2791로 표시될 수 있다 (Lenting. P. J. BLOOD, 26 MARCH 2015. VOLUME 125, NUMBER 13). 또는 대안적인 vWF 도메인 맵핑 및 명명 시스템은 EXPASY 단백질 데이터베이스 명명법 (worldwideweb.uniprot.org/uniprot/P04275)에 의해 D1: 12 - 218; D2: 365 - 576; D': 754 - 805; D3: 844 - 1052; A1: 1255 - 1431; A2: 1476 - 1643; A3: 1669 - 1849; D4: 1927 - 2131; B: 2233 - 2306 (EXPASY에서는 C1으로 명명됨); C1: 2407 - 2473 (EXPASY에서는 C2로 명명됨); C2: 2558 - 2623 (EXPASY에서는 C3으로 명명됨); 및 CK: 2701 - 2790로 표시될 수 있다.
본원에 따른 키메라 단백질에 포함되는 vWF D'D3 도메인은 FVIII와 융합되어 본원에 따른 재조합 FVIII이 내인성 vWF (전장 vWF)에 결합하는 것을 억제한다.
본원에 따른 키메라 단백질에 포함되는 vWF D'D3 도메인은 이러한 활성을 보유하는 한 다양한 길이 또는 유래의 것이 사용되며, 이의 변이체, 유도체, 변형체, 돌연변이체, 복합체 또는 유사체를 포함한다.
일 구현예에서 본원에 따른 키메라 단백질에 포함되는 vWF의 D'D3 도메인은 서열번호 02를 기준으로 742번 내지 1248번 아미노산 또는 이와 적어도 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% 또는 100% 상동성을 나타내고, 본원에 따른 재조합 FVIII에 대한 내인성 vWF의 결합을 방지한다.
앞서 언급한 바와 같이 본원에 따른 키메라 단백질은 FVIII의 경쇄 및 중쇄를 하나의 폴리펩타이드로 발현하는 단일 폴리펩타이드 또는 경쇄 및 중쇄 각각을 별개의 폴리펩타이드로 발현하는 이중 폴리펩타이드로 구현될 수 있다.
도 2a를 참조하면 본원에 따른 키메라 단백질이 단쇄 또는 단일 폴리펩타이드로 구현되는 경우, vWF D’D3 도메인은 FVIII의 N- 또는 C- 말단 방향에 위치할 수 있으며, 체내에서 D’D3의 분리를 위해 효소절단 부위를 포함하는 아미노산 링커(또는 연결체)로 연결될 수 있다.
일 구현예에서 본원에 따른 키메라 단백질에 포함되는 FVIII과 D'D3 도메인은 연결체로서 [G4S]n 의 아미노산 서열로 표시되는(상기 식에서 G는 글라이신, S는 세린이고, n은 1 내지 100 사이 정수) 연결체와 연결될 수 있으며, 상기 연결체는 효소절단 부위를 포함한다. 연결체는 D’D3와 FVIII를 연결하며, D’D3가 FVIII의 vWF 결합 위치에 정확하게 결합할 수 있게 유연성을 부여해 주는 기능을 갖고 있다. 구체적으로 본원에서는 D’D3를 FVIII에 융합시킬 수 있는 위치는 FVIII 중쇄의 N-말단, C-말단, FVIII 경쇄의 N-말단 또는 C-말단 이고, D’D3 도메인에서 N-말단에 해당하는 D’도메인이 FVIII의 A3 도메인 쪽에 결합하고, D3 도메인이 FVIII의 C2 도메인 쪽에 결합하는 것을 고려하면, FVIII 경쇄의 C-말단에 D’D3를 융합시켰을 때가 D’도메인과 FVIII의 A3 도메인의 거리가 가장 멀 때인 것으로 분석되었다. 따라서 D’D3를 FVIII에 융합시킬 때 D’D3가 FVIII에 제대로 잘 결합하게 하기 위해서 유연성 있는 링커를 선택하였다. 그리고 기존에 발표된 FVIII의 결정구조를 근거로 C2 도메인의 C-말단부터 A3 도메인의 N-말단까지 직선 거리를 측정하고, 이 거리를 아미노산 개수로 환산하여 링커의 길이를 선별하였다. 그렇기 때문에 링커의 길이가 상기 직선 거리보다 짧게 되면, D’도메인이 FVIII의 A3 도메인까지 가지 못 하기 때문에 상기 연결체의 길이가 너무 짧으면 D’D3가 FVIII 결합 위치에 정확하게 결합할 수 없다. 그러므로 상술한 조건을 만족하는 충분한 길이의 연결체가 필요하다. D’D3를 융합 하는 위치에 따라 상기 연결체의 최적 길이는 변할 수 있다. 이러한 기능을 갖는 한 다양한 길이의 연결체가 사용될 수 있다. 일 구현예에서는 n은 최소 4이다. 다른 구현예는 상기 식에서 n은 4, 6, 8, 10, 16, 21, 22 또는 58 일 수 있다.
또한, 상기 연결체는 효소절단 부위를 포함하고 있기 때문에 FVIII이 활성화 될 때, vWF D’D3, 또는 vWF D’D3 및 연결체가 FVIII으로부터 잘려 나갈 수 있게 해주는 기능도 갖고 있다.
본원에 따른 연결체에 포함되는 효소절단 부위는 연결체의 N- 또는 C-말단 방향에 연결될 수 있다.
본원에 따른 일 구현예에서 효소절단부위는 트롬빈에 의해 절단될 수 있는 부위로, 이러한 부위는 DFLAEGGGVR (서열번호 36), TTKIKPR (서열번호 37), 또는 LVPRGS (서열번호 38)로 표시될 수 있다. 이 경우, 본원에 따른 키메라 단백질은 투여시 체내에서 트롬빈에 의해 절단되어 FVIIIa로 활성화되고 활성화 IX 인자(FIXa)와 함께 X 인자를 활성화하여 활성화 X 인자(FXa)로 전환시킨다.
또한, 본원에 따른 연결체에 포함될 수 있는 효소절단부위는 트롬빈 외에 coagulant cascade에 있는 FVIIa, FXa, 또는 FXIa에 의해 잘릴 수 있는 부위도 포함될 수 있으며, 이러한 부위는 ASKPQGRIVGG (서열번호 39), IDGR (서열번호 40), IEGR (서열번호 41), SKLTRAETVF (서열번호 42)로 표시될 수 있다.
본원에 따른 일 구현예에서 효소절단 부위는 링커의 N-말단 방향 쪽에 포함되며, 트롬빈과 같은 효소에 의해 이 부위가 잘리면 연결체와 D'D3 도메인이 FVIII으로부터 분리된다.
본원에 따른 다른 구현예에서는 효소절단 부위는 링커의 C-말단 방향에 포함되며, 트롬빈에 의해 이 부위가 잘리어 연결체와 D’D3 도메인이 FVIII으로부터 분리된다.
도 2b를 참조하면, 앞서 언급한 바와 같이 본원에 따른 키메라 단백질은 중쇄 및 경쇄 각각이 별개의 폴리펩타이드로 발현되는 이쇄 또는 제 1 및 제 2의 두 개의 폴리펩타이드의 형태로 구현되며, 이 경우, 경쇄와 중쇄는 금속 이온 매개 비공유 결합에 의해 회합될 수 있다.
본원에 따른 키메라 단백질이 이쇄로 구현되는 경우, 제 1 폴리펩타이드는 FVIII의 중쇄 A1, A2 및 B 도메인의 전부 또는 일부를 포함하고, 제2 폴리펩타이드는 FVIII의 경쇄 A3, C1, C2 및/또는 B 도메인의 일부를 포함한다.
본원에 따른 단백질이 이쇄로 구현되는 경우 제 1 폴리펩타이드에 포함되는 B 도메인은 앞서 언급한 바 및 후술하는 바를 참조할 수 있다.
본원에 따른 단백질이 이쇄로 구현되는 경우 제 2 폴리펩타이드에는 B 도메인이 포함되지 않을 수 있거나 또는 B 도메인의 일부가 사용될 수 있다. 후자의 경우, 예를 들면 B 도메인 C-말단 잔기부터 N-말단 방향으로 86개의 잔기를 포함한다. 제 2 폴리펩타이드로 발현된 B 도메인 일부는 세포외로 분비될 때 떨어져 나간다.
본원에 따른 키메라 단백질이 이쇄로 구현되는 경우, vWF D’D3 도메인은 제 1 폴리펩타이드의 C 말단 방향에 연결된다. 이 경우 B 도메인이 일종의 연결체로서 작용할 수 있다. 또한 본원에 따른 단백질이 이쇄로 구현되는 경우, 단쇄로 발현되는 경우와 비교하여 동일하거나 또는 긴 길이의 B 도메인이 사용될 수 있다.
본원에 따른 일 구현예에서 단쇄의 경우 서열번호 1의 서열을 기준으로 741-902 및 1654-1689 잔기의 B 도메인이 사용되고, 이쇄의 경우 서열번호 01을 기준으로 741-936의 196개 잔기 길이의 B 도메인이 사용된다. 이쇄로 발현되는 경우 B 도메인에 D’도메인이 연결될 수 있고, 본 이론으로 제한되는 것은 아니나, 융합되는 단백질간의 연결이 구조적으로 보다 자연스럽게 이어주기 위함이다. 구체적으로 일 구현예에서 본원에 채용된 196개 B 도메인의 C-말단의 아미노산 잔기 서열은 - SGGPLSLS이다. D’도메인의 N-말단의 아미노산 서열은 SLSCR PPMVKLVCPA- 로 시작한다. 따라서 상기 SLS의 3개의 아미노산 잔기가 겹쳐지도록 하기 위해 196개의 B 도메인이 사용되었다. 196개의 B 도메인은 -SGGPLSLS 중에 SGGPL 잔기까지만을 포함하여 계산한 것이다.
본원에 따른 키메라 단백질은 vWF D’D3를 하나 이상 포함할 수 있다.
D’D3는 vWF의 N-말단에 위치한 2개의 도메인으로, D3 도메인은 실제 다른 분자의 vWF와 D3 도메인을 통해 이량체를 형성한다. 따라서 본원에서는 D’D3를 단량체 또는 이량체 형태로 FVIII에 융합될 수 있다.
복수개의 vWF D’D3를 포함하는 경우, 예를 들면 두 개를 포함하는 경우, 제 1 D’D3는 FVIII에 연결되고, 제 2 D’D3는 제 1 D’D3에 연결되는 방식으로 구현될 수 있다.
복수개의 vWF D’D3를 포함하는 경우, 제 1 및 제 2 D’D3는 연결체를 통해서 연결될 수 있거나, 또는 제 2 D’D3는 별개로 발현되어, 키메라 단백질과 복합체(complex)를 형성하도록 할 수 있다. 예를 들면 scFVIII/D’D3/D’D3-120 키메라 단백질에서와 같이 연결체를 이용하여 연결되는 경우, 상기 연결체는 앞서 언급한 바를 참조할 수 있으나 효소절단 부위는 포함하지 않는다.
또한 D’D3를 dimer 형태로 사용하는 경우 D’D3는 D3 도메인에 있는 S-S bond를 통해 이량체가 형성될 수 있다. 복수개의 D’D3를 예를 들면 이쇄의 경우와 같이 별개의 벡터에서 발현하는 경우, FVIII-D’D3의 DNA와 D’D3 DNA를 하나의 세포에서 동시 발현시켜, 세포내에서 D’D3끼리 이량체가 만들어 지고, 세포 밖으로 분비되게 한 후, 이를 분리 정제하여 사용할 수 있다.
나아가 본원에 따른 키메라 단백질은 하나 이상의 Fc 단백질을 추가로 포함할 수 있다. Fc를 추가로 D’D3와 함께 FVIII에 융합하게 되면, D’D3에 의해 혈중 vWF가 결합하지 못 하는 효과와 함께 Fc에 의해 FcRn 리사이클링을 통한 반감기 증가 효과를 추가로 기대 할 수 있다. 일 구현예에서는 특히 약 50 kDa 정도 되는 Fc가 융합되며, Fc에 의한 입체적 방해(steric hindrance)에 의해 혈중 vWF가 이 물질에 결합하는 것을 더욱 어렵게 할 수 있다.
따라서, 위와 같은 기능을 본원에 따른 목적을 달성하는 한 다양한 길이 및 유래의 Fc가 포함될 수 있다. 본원에 따른 일 구현예에서는 서열번호 22로 표시되는 Fc, 인간 기원의 IgG 유래의 Fc가 사용된다.
본원에 따른 키메라 단백질이 단쇄로 구현되는 경우, Fc는 vWF D’D3에 연결될 수 있다. 이 경우 연결체를 이용하여 연결될 수 있으며, 상기 연결체는 앞서 언급한 바를 참조할 수 있으나 효소절단 부위는 포함하지 않는다.
본원에 따른 키메라 단백질이 이쇄로 구현되는 경우, Fc는 중쇄의 일부로 발현되는 vWF D’D3에 및/또는 경쇄의 C 말단 방향에 연결될 수 있다. 이 경우 연결체를 이용하여 연결될 수 있으며, 상기 연결체는 앞서 언급한 바를 참조할 수 있으며, 특히 경쇄에 연결되는 경우에는 효소절단 부위를 포함하며, D’D3에 연결되는 경우에는 효소절단 부위는 포함하지 않는다.
본원에 따른 키메라 단백질에 두 개 이상의 Fc가 포함될 수 있다.
복수개의 Fc를 포함하는 경우, 예를 들면 두 개를 포함하는 경우, 일 구현예에서 제 1 Fc는 키메라 FVIII에 연결되고, 제 2 Fc는 제 1 Fc에 연결되는 방식으로 구현될 수 있다. 다른 구현예에서 제 1 Fc는 이쇄로 발현되는 키메라 FVIII의 중쇄에 연결되고, 제 2 Fc는 경쇄에(그 역순도 가능) 연결되는 방식으로 구현될 수 있으며, 이 경우 복수의 Fc 간에 이량체와 같은 복합체가 형성될 수 있다.
복수개의 Fc를 포함하는 경우, 제 1 및 제 2 Fc는 연결체를 통해서 연결될 수 있거나, 또는 제 2 Fc는 하나의 세포에서 별개의 벡터로 발현되고, 세포 내에서 Fc 간에 이량체가 만들어지고, 이후 세포 외로 분비된 것을 정제하여 사용할 수 있다.
앞서 언급한 바와 같이 본원에 따른 키메라 단백질은 단쇄 또는 이쇄로 구현될 수 있다.
일 구현예에서 본원에 따른 단쇄의 키메라 단백질은 서열번호 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21 또는 25로 표시될 수 있다. 상기 단백질은 별개의 D’D3 및/또는 Fc와 복합체를 형성할 수 있으며, 일 구현예에서 서열번호 19의 키메라 단백질은 서열번호 20으로 표시되는 하나 이상의 D’D3와 복합체를 형성할 수 있으며, 또는 서열번호 21의 키메라 단백질은 서열번호 22로 표시되는 하나 이상의 Fc와 복합체를 형성할 수 있다.
일 구현예에서 본원에 따른 이쇄의 키메라 단백질은 서열번호 4 및 5 (각각 중쇄 및 경쇄), 서열번호 23 및 24 (각각 중쇄 및 경쇄), 서열번호 26 및 27 (각각 중쇄 및 경쇄), 서열번호 28 및 29 (각각 중쇄 및 경쇄), 서열번호 30 및 31 (각각 중쇄 및 경쇄), 서열번호 32 및 33 (각각 중쇄 및 경쇄), 또는 서열번호 34 및 35 (각각 중쇄 및 경쇄)로 표시될 수 있다. 상기 단백질은 별개의 D’D3 및/또는 Fc와 복합체를 형성할 수 있으며, 일 구현예에서 서열번호 28 및 29 (각각 중쇄 및 경쇄) 또는 서열번호 32 및 33 (각각 중쇄 및 경쇄)의 키메라 단백질은 서열번호 22로 표시되는 하나 이상의 Fc와 복합체를 형성할 수 있다.
본원에는 상술한 서열번호 04 내지 35로 표시되는 아미노산 서열은 물론 이와 90% 이상 상동성 또는 실질적으로 동일한 서열의 단백질도 포함된다.
이러한 실질적 동일성 또는 상동성은 당업계에 공지된 서열비교 방법을 이용하여 결정될 수 있다. 본원에 개시된 서열과 임의의 다른 서열을 최대한 대응되도록 얼라인하고, 당업계에서 통상적으로 이용되는 알고리즘을 이용하여 얼라인된 서열을 분석한 경우에, 최소 80% 상동성, 특히 85% 이상, 더욱 특히 90%, 더더욱 특히 95% 이상의 상동성을 나타내는 서열을 의미한다. 서열비교를 위한 얼라인먼트 방법은 당업계에 공지되어 있다. 예를 들면 Smith and Waterman, Adv. Appl. Math. (1981) 2:482; Needleman and Wunsch, J. Mol. Bio. (1970) 48:443; Pearson and Lipman, Methods in Mol. Biol. (1988) 24: 307-31; Higgins and Sharp, Gene (1988) 73:237-44; Higgins and Sharp, CABIOS (1989) 5:151-3; Corpet et al., Nuc. Acids Res. (1988) 16:10881-90; Huang et al., Comp. Appl. BioSci. (1992) 8:155-65 및 Pearson et al., Meth. Mol. Biol. (1994) 24:307-31에 개시되어 있다. NCBI Basic Local Alignment Search Tool(BLAST)(Altschul et al., J. Mol. Biol. (1990) 215:403-10)은 NCBI 등에서 접근 가능하며, blast, blastp, blasm, blastx, tblastn 및 tblastx와 같은 서열 분석 프로그램과 연동되어 이용할 수 있다. BLSAT는 www.ncbi.nlm.nih.gov/BLAST/에서 접속 가능하며, 이 프로그램을 이용한 서열 상동성 비교 방법은 www.ncbi.nlm.nih.gov/BLAST/blast_help.html에서 확인할 수 있다.
본원에 따른 키메라 단백질은 반감기 측면에서 야생형의 FVIII와 비교하여 최소 1.5배, 최소 2배, 최소 3배 이상의 반감기가 향상된 것이다. 반감기를 측정하는 방법은 개시되어 있으며, 당업자라면 본원의 기재, 당업계의 기술 및 참고문헌의 기재를 고려하여 적절한 방법을 선택하여 반감기 및 활성을 측정하고 이에 기반하여 본원의 범위에 포함되는 키메라단백질을 결정할 수 있을 것이다.
다른 양태에서 본원은 또한 본원에 개시된 키메라 단백질을 코딩하는 핵산분자 또는 폴리뉴클레오타이드, 상기 폴리뉴클레오타이드를 포함하는 벡터 또는 상기 벡터로 형질전환된 세포(주)에 관한 것이다.
본원에 따른 핵산분자는, 본원에 따른 키메라 단백질이 발현되는 세포의 종류에 맞추어 코돈최적화(Codon optimized) 된 것을 포함하는 것이다. 본원에 따른 일 구현예에서는 CHO 코돈에 최적화되며, 이러한 핵산 서열은 서열번호 58, 59, 60, 62, 63, 64, 65, 또는 66의 서열로 표시될 수 있다.
본원에 따른 키메라 단백질을 코딩하는 핵산분자는 또한 상술한 바와 같이 실질적으로 동일한 생물학적 균등물에 해당하는 FVIII을 코딩하는 것을 또한 포함하는 것이다.
또한 본원에 따른 키메라단백질을 코딩하는 핵산분자는 다양한 목적을 위해 다양한 발현 벡터에 클로닝되어 사용될 수 있다. 발현 벡터의 구체적 구성은 본원에 따른 키메라단백질을 발현하고자 하는 숙주 세포에 따라 달라질 수 있으나, 본원에 따른 핵산분자의 mRNA로의 발현 또는 mRNA의 단백질로의 발현을 조절하는 서열, 예를 들면 프로모터 및/또는 인핸서 등과 같은 조절서열을 포함한다. 이러한 또는 다양한 다른 목적으로 사용될 수 있는 다양한 벡터 및 조절서열이 공지되어 있으며, 당업자라면 본원의 구체적 목적 및 효과를 고려하여 적절한 것으로 선택할 수 있을 것이며, 예를 들면 본원의 실시예 및 도면에 기재된 것을 포함할 수 있으나 이로 제한하는 것은 아니다.
본원에 따른 키메라 단백질을 코딩하는 핵산분자를 포함하는 벡터는 당업계에 공지된 방법을 통하여 제조될 수 있으며, 예를 들면 본원에 따른 키메라 단백질을 코딩하는 핵산분자를 벡터의 프로모터 및/또는 인핸서에 작동 가능하게 연결한다. 일 구현예에서는 세포에서 외래 유전자를 발현시킬 수 있는 재조합 발현 벡터에 삽입되며, 이러한 벡터의 예로는 pMSGneo, pcDNA3.1(+) 등 일반 단백질 발현 벡터를 들 수 있으나, 이에 제한되지 않는다. pMSGneo 벡터의 경우 핵 기질(nuclear matrix)에 결합하는 MAR(Matrix attachment region) 인자가 포함된 발현 벡터로서, 상기 MAR 인자는 발현 벡터에 사용되는 경우 위치-독립적 발현(position-independent expression)을 유도하여 유전자 발현을 증대시키는 역할을 한다. 따라서, pMSGneo 벡터를 이용하는 경우 안정적이고 높은 발현량을 얻을 수 있다. 또한, pcDNA3.1(+) 벡터는 강력한 CMV 프로모터가 함유되어 있어 단백질 발현에 널리 사용되고 있다. 본원에 따른 일 구현예에서는 도 3에 기재된 것이 사용된다. 본원에 따른 키메라단백질은 상술한 바와 같이 단일 폴리펩타이드 또는 두 개의 폴리펩타이드로 발현될 수 있다.
또한, 본원에 따른 키메라 단백질을 코딩하는 핵산분자를 포함하는 상기 재조합 발현 벡터는 다양한 목적을 위해 적절한 숙주세포에 형질 전환될 수 있다. 숙주세포는 본원에 따른 벡터의 증폭 및/또는 벡터에 포함된 핵산분자가 발현될 수 있는, 원핵 및 진핵 세포를 모두 포함하는 것이다. 이러한 목적으로 사용될 수 있는 다양한 세포가 공지되어 있으며, 당업자라면 본원의 구체적 목적 및 효과를 고려하여 적절한 것으로 선택할 수 있을 것이며, 예를 들면 본원의 실시예 및 도면에 기재된 것을 포함할 수 있으나 이로 제한하는 것은 아니다. 예를 들면 대장균(E.coli), 포유동물 세포(Mammalian cell), 효모(Yeast), 식물 세포(Plant cell), 곤충 세포(Insect cell)를 포함한다. 본원에 따른 일 구현예에서는 재조합 FVIII의 발현을 위해, 진핵세포, 특히 포유동물 세포주가 사용된다. 당업자라면 본원에 따른 재조합 FVIII의 특징을 고려하여 이러한 특징을 갖는 단백질을 발현할 수 있는 적절한 세포주를 선별할 수 있을 것이며, 예를 들면 이러한 포유동물 세포주로는 당업계에 공지된 CHO, BHK, COS7, HEK 세포주 등을 사용할 수 있으며, 바람직하게는, CHO 세포주, 특히 CHO-S 세포주 또는 CHO-K1 세포주 또는 CHO-DG44 세포주, 또는 HEK 세포주 특히 HEK293 세포주가 사용될 수 있으나 이로 제한하는 것은 아니다.
본원에 따른 벡터는 발현을 위해 상기와 같은 숙주세포로 전달된다. 이러한 숙주세포로의 벡터의 전달 방법은 당업계에 알려져 있으며, 예를 들면, 칼슘포스페이트 침전법, 샷건 방법, 리포좀을 이용한 방법, 나노니들 또는 전기천공법등 당업계의 공지된 방법을 이용하여 수행될 수 있으나, 이로 제한하는 것은 아니다.
다른 양태에서 본원은 또한 상술한 본원에 따른 벡터를 진핵세포에 전달이입하는 단계; 상기 세포를 배양액 중에서 배양하는 단계; 상기 배양액을 수집하여 시스테인(cysteine) 도입된 키메라 단백질을 정제하는 단계; 및 상기 정제된 시스테인(cysteine) 도입된 단백질을 페길화 완충액으로 처리하는 단계; 도입된 시스테인(cysteine)의 자유 티올기를 환원 과정을 통해 복원하는 단계; 환원 과정 중 분리된 키메라 단백질 내부의 이황화 결합을 복원하기 위하여 산화시키는 단계; 환원형의 티올기로 복원된 키메라 단백질 시스테인(cysteine)에 특이적으로 페길화시키는 단계 및 페길화 키메라 단백질을 분리하는 단계를 포함하는, 키메라 단백질의 생산방법에 관한 것이다.
본원에 따른 방법에 사용되는 구체적인 벡터, 세포 및 각 단계의 처리 과정은 본원 실시예에 기재된 것을 참고할 수 있다.
본원에 따른 키메라 단백질은 혈액응고가 필요한 질환, 혈우병, 특히 혈우병 A 환자의 혈액응고 또는 혈우병 A의 치료에 유용하게 사용될 수 있다.
이에 다른 양태에서 본원은 또한 상술한 바와 같은 본원에 따른 키메라 단백질, 이를 코딩하는 핵산 분자, 이를 포함하는 벡터, 또는 상기 핵산분자 또는 벡터를 포함하는 세포, 또는 상기 구성에 및 약학적으로 허용 가능한 담체를 포함하는 약학적 조성물을 혈우병 환자 특히 A형 환자 또는 혈액의 응고가 필요한 환자에게 치료적으로 유효한 양으로 투여하는 단계를 포함하는 혈우병, 특히 A형 혈우병의 치료 방법 또는 혈액응고 방법에 관한 것이다.
본원에 따른 키메라 단백질 또는 이를 코딩하는 핵산 분자 또는 이를 코딩하는 벡터는 약학적으로 허용 가능한 담체를 함께 포함하는 혈우병, 특히 A형 혈우병의 치료용 약학 조성물의 형태로 제공될 수 있다.
또한 본원의 약학 조성물은 단독으로, 또는 기타 약물치료 및 생물학적 반응조절제를 사용하는 방법들과 병용하여 사용할 수 있다.
본원의 조성물은 상기 언급한 유효성분 이외에 추가로 약학적 또는 생리학적으로 허용 가능한 담체를 1종 이상 포함하여 제조할 수 있다.
본원에서 사용된 용어 담체란 사용되는 투여량 및 농도에 노출되는 세포 또는 포유동물에 무독성인 약학적으로 허용 가능한 담체, 부형제, 또는 안정화제를 의미하는 것이다. 이러한 담체의 예로는 식염수, 링거액, 완충 식염수, 포스페이트, 시트레이트 및 다른 유기산과 같은 완충액, 아스코르브산을 비롯한 산화방지제, 저분자량(약 10 잔기 미만) 폴리펩타이드, 단백질, 예를 들어 혈청 알부민, 젤라틴 또는 면역글로불린; 친수성 중합체, 예를 들어 폴리비닐피롤리돈, 아미노산, 예를 들어 글리신, 글루타민, 아스파라긴, 아르기닌 또는 라이신, 단당류, 이당류 및 글루코스, 만노스 또는 덱스트린을 비롯한 기타 탄수화물, 킬레이트화제, 예를 들어 EDTA, 당 알콜, 예를 들어 만니톨또는 소르비톨, 염 형성 카운터 이온, 예를 들어 나트륨, 및(또는) 비 이온계 계면활성제, 예를 들어 트윈, 폴리에틸렌 글리콜(PEG) 및 플루로닉스(PLURONICS)를 들 수 있다.
필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형으로 제제화할 수 있다. 더 나아가 당해 기술분야의 적정한 방법으로 또는 레밍턴의 문헌(Remington's Pharmaceutical Science(최근판), Mack Publishing Company, Easton PA)에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제형화할 수 있다.
본원의 조성물은 목적하는 방법에 따라 특히 비경구투여(예를 들어, 정맥 내, 피하, 복강 내)가 바람직하다. 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.
본원에 따른 조성물은 치료적으로 유효한 양으로 투여된다. 본원에서, "치료적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본원의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.
본원에서 사용되는 용어, "치료"란 본 발명의 조성물의 투여에 의해 질환에 의한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다. 본원에 따른 방법이 사용되는 대상체는 인간을 포함하는 영장류이나, 이로 제한하는 것은 아니다.
본원에 따른 재조합 키메라 단백질은 그 자체 또는 상술한 바와 같은 조성물의 형태로 혈우병 또는 혈액응고가 필요한 대상체에게 치료적으로 유효한 양으로 투여되어, 혈액 응고 방법 또는 혈우병의 치료 방법의 형태로 제공될 수 있으며, 앞서 기재한 바를 참고할 수 있다.
이하, 본 발명의 이해를 돕기 위해서 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐 본 발명이 하기의 실시예에 한정되는 것은 아니다.
실시예
실시예 1. 키메라 FVIII 구축
실시예 1-1. scFVIII (single-chain FVIII) 제작
단쇄 형태로 발현되는 FVIII의 구축을 위해, furin 절단 위치인 1648 잔기(전장 FVIII 아미노산 서열을 나타내는 서열번호 01의 아미노산 서열기준)를 포함하는 B 도메인의 일부가 결실된 FVIII를 구축하였다. 구축된 각각의 scFVIII은 B 도메인의 N-말단(서열번호 01의 741번째 아미노산 잔기)로부터 특정 길이의 B 도메인을 포함하고 포함된 B 도메인은 원래 보유하고 있는 당쇄를 그대로 포함하도록 제작되었으며, 위치 특이적 페길화(site-specific PEGylation)를 위해 B 도메인에 위치한 782번 아이소루이신 잔기를 시스테인으로 개질하였다.
제조 방법은 다음과 같다.
하기 표 1의 B 도메인이 결실된 각 scFVIII은 인간 FVIII 핵산서열 (NM000132)을 기준으로 합성하였다. 먼저 scFVIII 유전자 발현벡터를 제조하기 위하여 인간 FVIII 핵산서열(NM000132)을 기준으로 CSL 형태(Leader-중쇄-B 도메인 (741-764, 1653-1689)-경쇄)를 5’에 NotI-AsisI과 3’에 PacI-XhoI 효소절단 부위를 포함하도록 GeneArt 사를 통해 유전자 전체를 합성하였다. 합성된 유전자를 pcDSW 벡터의 NotI/XhoI site에 클로닝하여 pcDSW-CSL 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 AsisI/PacI 효소를 이용하여 절단 후 절단된 밴드 크기 비교를 통해 확인하였다. 이후 B 도메인이 결실된 각 scFVIII는 FVIII 중쇄의 A2 도메인 내에 존재하는 BamHI site (GGATCC) 부터 경쇄 3’에 PacI 효소절단 부위를 포함하도록 GeneArt 사를 통해 유전자를 합성한 후 기존에 완성된 pcDSW-CSL 발현벡터를 효소 BamHI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 BamHI/PacI site로 자르고 pcDSW-CSL 발현벡터의 BamHI/PacI site에 클로닝하여 pcDSW-scFVIII 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 BamHI/PacI 효소를 이용하여 절단 후 크기 비교를 통해 확인하였다.
[표 1] scFVIII constructed in the present disclosure
Cysteine이 도입된 scFVIII G4 (B3, 서열번호 3)은 cysteine 치환 부위 및 FVIII 중쇄 A2 도메인 내에 존재하는 BamHI site (GGATCC) 부터 경쇄 말단 3’에 PacI 효소절단 부위가 포함되도록 GeneArt 사를 통해 유전자를 합성한 후, 앞에서 제작한 발현벡터 pcDSW-scFVIII G4에 효소 BamHI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후, 합성된 유전자를 BamHI/PacI site에 클로닝하여 cysteine이 치환된 pcDSW-scFVIII G4 (B3) 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 서열분석을 통해 확인하였다.
실시예 1-2. dcFVIII (dual-chain FVIII) 제작
D’D3를 포함하는 이량체 형태로 발현되는 FVIII 구축을 위해 D’D3를 포함하는 중쇄와 경쇄를 각각 다른 벡터를 이용하여 하나의 세포에서 발현시켰다. D’D3를 포함하는 중쇄에는 실시예 1-1에서 사용한 B3에 해당하는 B 도메인이 포함되어 있으며, 경쇄는 대한민국 특허 제251286호에 기재된 경쇄를 발현하는 벡터를 참고하여 만들었다.
실시예 1-3. 키메라 FVIII 제작
키메라 FVIII은 실시예 1-1과 1-2에서 만든 단쇄 FVIII과 이량체 FVIII을 기본으로 하여 D’D3를 1개 또는 2개를 융합 하였으며, 융합한 D’D3가 FVIII에 적절히 결합할 수 있게 링커를 사용하였다. 링커는 FVIII의 B 도메인 일부를 사용하거나 [G4S]n 를 이용하였다. 또한 일부는 상기에 명시한 구성에 Fc까지 포함하였다 (도 2, 도 3, 표 2).
[표 2] 본원에서 제작한 키메라 FVIII
키메라 FVIII의 클로닝은 특허 실시예 1-1에서 기술한 scFVIII G4 (B3)를 이용하여 진행하였다. 서열번호 04의 FVIII 중쇄는 FVIII의 A2 도메인에 존재하는 KpnI 효소절단 부위부터 말단 3’에 PacI 효소절단 부위가 포함하도록 GeneArt 사를 통해 유전자를 합성한 후, 실시예 1-1에서 제작한 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 KpnI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 KpnI/PacI에 클로닝하여 서열번호 04의 중쇄에 해당하는 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다. 서열번호 05 경쇄는 5’에 AsisI-Leader 서열과 FVIII 경쇄 C1 도메인에 존재하는 BspEI 효소절단 부위까지 포함하도록 GeneArt 사를 통해 유전자를 합성한 후 실시예 1-1에 제작한 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 AsiSI/BspEI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 AsiSI/BspEI site에 클로닝하여 서열번호 05의 경쇄에 해당하는 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다. 서열번호 06, 07, 08, 09, 10, 11, 12, 13, 14, 15, 16은 FVIII 경쇄의 C1 도메인에 존재하는 BspEI 효소 부위 서열부터 말단 3’에 PacI 효소절단 부위가 포함되도록 GeneArt 사를 통해 유전자를 합성한 후 실시예 1-1에서 제작한 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 BspEI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 BspEI/PacI site에 클로닝하여 서열번호 06, 07, 08, 09, 10, 11, 12, 13, 14, 15, 16에 해당하는 키메라 FVIII 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석 및 서열분석을 통해 확인하였다. 서열번호 14와 비교하여 서열번호 15로 표시되는 펩타이드는 링커에 있는 트롬빈 절단부위의 위치가 상이하고, 서열번호 16으로 표시되는 펩타이드는 트롬빈 절단부위의 위치 및 링커의 서열자체가 상이하며, 이를 각각 TH1 및 TH2로 표시하였다.
서열번호 17, 18, 21은 FVIII 경쇄 C1 도메인에 존재하는 BspEI 효소 부위 서열부터 말단 3’에 PacI 효소절단 부위가 포함되도록 코스모진텍사를 통해 CHO 코돈 최적화 및 유전자를 합성한 후 실시예 1-1에서 제작한 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 BspEI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 BspEI/PacI site에 클로닝하여 서열번호 17, 18, 21에 해당하는 키메라 FVIII 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다.
서열번호 19는 5’에 NotI-AsiSI 효소절단 부위를 추가하고 FVIII 중쇄 A1 도메인에 존재하는 AflII 효소절단 부위까지 포함되도록 코스모진텍사를 통해 CHO 코돈 최적화 및 유전자를 합성한 후, 실시예 1-1에서 제작한 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 AsiSI/AflII을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 AsiSI/AflII site에 클로닝하여 서열번호 19에 해당하는 키메라 FVIII 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다.
서열번호 20은 코스모진텍에서 합성된 서열번호 64로 표시되는 유전자를 효소 MfeI/XbaI을 이용하여 기존 해당 부위 유전자를 제거하고 합성된 서열번호 63으로 표시되는 유전자로부터 효소 MfeI/XbaI을 처리하고 인서트를 확보하여 MfeI/XbaI site에 일차 클로닝 하였다. 일차 클로닝 된 유전자를 발현 벡터로 옮기기 위하여 기 제작된 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 NotI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 일차 클로닝 된 유전자를 효소 NotI/PacI으로 인서트를 절단하고 발현 벡터의 NotI/PacI site에 클로닝하여 서열번호 20에 해당하는 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다.
서열번호 22는 5’에는 AsisI, 3’에는 PacI 효소절단 부위가 각 각 포함되도록 GeneArt 사를 통해 유전자를 합성한 후, 실시예 1-1에서 제작한 scFVIII G4 (B3) 발현벡터 pcDSW-scFVIII G4 (B3)에 효소 AsisI/PacI을 이용하여 기존 FVIII의 해당 부위를 제거한 후 합성된 유전자를 AsisI/PacI site에 클로닝하여 서열번호 22에 해당하는 발현 벡터를 구축하였고, 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다.
이외에 서열번호 15는 CHO 형질전환 세포주를 제작하기 위하여 추가로 CHO 발현벡터를 제작하였다. 서열번호 15의 5’에는 AatII-AscI, 3’에는 BsiWI-PacI 효소절단 부위가 각각 포함되도록 본 유전자를 합성하거나 Biobasics 사, GeneArt 사 그리고 Genscript 사를 통해 CHO 발현에 최적화되도록 유전자를 합성하였다. CHO 발현벡터 pMSID2의 AscI/PacI site에 합성된 유전자를 클로닝하여 서열번호 15에 해당하는 CHO 발현 벡터를 구축하였다. 발현 벡터 구축의 정확성은 효소 절단을 통한 길이 분석을 통해 확인하였다.
실시예 2. 키메라 FVIII 발현
실시예 1에서 구축한 키메라 FVIII을 세포에서 발현하였으며, 발현된 세포는 세포배양배지로 배양된다. 우선 발현을 위해 실시예 1에서 구축한 발현 벡터를 Expi293F™ 세포에 Expi293F™ Expression System Kit (Thermofisher 사, Catalog Number A14635)을 사용하여 전달 이입하였다. 형질전환 방법 및 배양은 30mL scale 기준으로 아래와 같이 실시하였으며, 단백 필요량에 따라 실제 배양 부피는 조절하였다. 형질전환 수행 24시간 전에 Expi293F™ 세포를 2.0 X 106 cells/mL로 Expi293 배양배지를 이용하여 예상 필요량에 맞추어 계대배양 하였다. 형질전환 수행 당일, 세포 수와 세포 생존율(viability)을 측정하여 세포 생존율이 95% 이상일 경우 형질전환을 진행하였다. 125mL 플라스크에 7.5 X 107 cell 되도록 Expi293 배양배지를 첨가하여 25.5mL로 맞춰주었다. 실시예 1에서 구축한 발현 벡터 30μg를 Opti-MEM을 이용하여 총 볼륨이 1.5mL이 되도록 섞어주었다. 80μL transfection reagent 를 Opti-MEM을 이용하여 총 볼륨이 1.5mL이 되도록 섞어주고 상온에서 5분간 incubation 해주었다. 5분 후 transfection reagent가 들어있는 Opti-MEM을 DNA가 들어있는 Opti-MEM에 넣어주고 부드럽게 섞어주었다. 그리고 20 ~ 30분간 상온에서 반응시켜 주었다. 3mL의 DNA: transfection reagent 복합체를 미리 준비해 놓은 125mL flask Expi293F™ 세포에 (Total volume: 28.5mL) 한 방울씩 떨어뜨려주고 37℃, 5% CO2 shaking incubator에서 125 rpm으로 배양하였다. 16 ~ 20시간 후에 Enhancer 1과 Enhancer 2를 각각 150μL, 1.5mL을 넣어주고 34℃, 5% CO2 shaking incubator에서 125 rpm으로 배양하였다. 배양 2일째 세포를 모두 원심분리하여 기존의 배양배지를 완전히 제거하고 30mL의 새로운 배양배지에 세포를 모두 풀어준 후 34℃, 5% CO2 shaking incubator에서 125 rpm으로 배양하였다. 도입 3일차에 키메라 FVIII을 발현하는 세포를 모두 원심 분리하여 배양액을 회수하였다. 배지 회수 후 남은 세포를 동일한 부피의 새로운 배양배지에 세포를 모두 풀어준 후 1일간 34℃에서 배양을 실시하였다. 본 과정을 배양 5일차까지 반복하여 3일, 4일, 5일 배양액을 회수하여 cysteine 도입된 키메라 FVIII의 활성 분석 및 정제를 진행하였다. 위와 유사한 방법으로 CHO-DG44 세포에서 FVIII 발현을 진행하였다.
이외에 CHO 형질전환 세포주를 제작 및 키메라 FVIII을 발현하였다. 형질전환 수행 24시간 전에 CHO DG44 세포를 CDM4CHO 배지 (Hyclone 사, Catalog Number, SH30557.02)에서 5~6 x 105 cells/mL의 양으로 필요량에 맞추어 계대하였다. 형질전환 당일 125mL 플라스크에 1 x 107 cells/mL로 OptiPro 배지 (Thermo fisher 사, Catalog Number 12309-019)를 이용하여 3mL로 분주하였다. 실시 예 1에서 구축한 30μg의 CHO 발현 벡터를 OptiPro을 이용하여 총 볼륨이 1.5mL이 되도록 섞어주었다. 90μL PEI MAX(1 μg/μL)와 OptiPro을 이용하여 총 볼륨이 1.5mL이 되도록 섞어주었다. 5분 뒤에 PEI MAX가 포함된 1.5mL OptiPro을 DNA가 들어있는 1.5mL OptiPro에 넣어주고 부드럽게 섞어주어 30분간 상온에서 반응시켰다. DNA:PEI 복합체 3mL을 미리 준비한 CHO DG44세포에 한 방울씩 떨어뜨려 주었다. 37℃, 5% CO2 shaking incubator에서 140 rpm으로 4시간 동안 배양하였다. 24mL의 CDM4CHO 배지를 첨가하여 140 rpm으로 배양하고 48시간 후에 세포 선별을 수행하였다.
형질전환된 CHO DG44 세포를 20nM MTX가 포함된 CDM4CHO 배양 배지를 이용하여 T-175 플라스크에 5 x 106 cells/mL로 50mL 되도록 현탁하고 37℃, 5% CO2 incubator에서 7일간 정치 배양하였다. 만약, 세포수가 모자라면 총 볼륨을 줄이고 진행하였다. 선별 수행 7일 후 세포수를 측정하여 125mL 플라스크에 5 x 105 cells/mL로 20nM MTX가 포함된 배양 배지를 이용하여 계대하고 37℃, 5% CO2 shaking incubator에서 140 rpm으로 배양하였다. 3 ~ 4일 간격으로 125mL 플라스크에 계대 배양을 실시하고 세포 농도가 1 x 106 cells/mL, 생존율이 90% 이상일 경우 선별이 완료된 것으로 판단하였다.
키메라 FVIII 발현을 위하여 CHO 형질전환 세포주를 3 x 105 cells/mL로 접종하여 37℃, 5% CO2 shaking incubator에서 140 rpm으로 배양하였다. 배양 4일째 세포를 모두 원심 분리하여 기존의 배양 배지를 완전히 제거하고 동일 부피의 새로운 배양배지에 세포를 모두 풀어준 후 31℃, 5% CO2 shaking incubator에서 140 rpm 으로 1일간 배양하였다. 본 배지교환 및 배양과정을 배양 8일차까지 반복하고 6일, 7일 그리고 8일 배양액을 회수하여 키메라 FVIII의 활성 분석 및 정제를 진행하였다.
실시예 3. 키메라 FVIII 발현 확인
실시예 3-1. 키메라 FVIII 발현량 측정
많이 사용하는 FVIII 활성 측정 시험 방법은 one-stage clotting method와 chromogenic method 두 가지이다. 그 중 키메라 FVIII 발현량 측정은 chromogenic method를 이용하여 측정하였다. Chromogenic method는 FIXa, FX, thrombin, calcium, phospholipids와 FVIII 시료를 섞어 준 후, FVIII 시료에 의해 활성화 된 FXa의 양을 측정하기 위해 FXa에 의해 잘리면서 발색하는 chromogenic substrate를 넣어주고, chromogenic substrate의 발색 정도에 따라 FVIII의 활성을 측정하는 방법이다.
Chromogenic method의 실험 방법은 CHROMOGENIX(社)에서 판매하고 있는 chromogenic assay kit를 endpoint 방법으로 사용하였고, 제조사가 제공한 시험방법을 본 시험에 적합하게 변형하여 시험을 진행하였다. 시료를 1× 희석버퍼를 사용하여 standard range (0.25-1 IU/mL)에 들어 오도록 희석한다. 표준품 (calibration plasma)을 0, 0.25, 0.5, 1 IU/mL 농도가 되게 1× 희석버퍼를 이용하여 준비하였다. 이렇게 준비한 시료와 표준품 10μL를 1× 희석버퍼 790μL로 희석한 후, 96 well plate에 50μL씩 분주하고, 37℃에서 5분간 정치시켰다. Automatic dispenser pipette을 사용하여 factor reagent를 50μL씩 각 well에 분주한 후 2분간 37℃에서 2분간 반응 시켰다. 50μL substrate를 각 well에 분주한 후 2분간 37℃에서 2분간 정치하여 발색 시켰다. 발색된 시료에 50μL 2% citric acid를 넣어주어 반응을 정지 시켰다. 405nm 파장에서 흡광도를 측정하여 선형 calibration curve를 그린 후, 샘플의 흡광도를 calibration curve에 대입하여 시료의 활성을 측정하였다.
[표 3] 키메라 FVIII 발현량
D’D3를 융합 하기 전인 scFVIII-B3의 발현량은 1.6에서 8.0 IU/mL 사이로 측정되었었다. D’D3를 융합한 키메라 FVIII 중 단쇄 형태로 발현되는 경우 expression level이 D’D3를 융합하기 전인 scFVIII-B3와 유사한 것을 확인하였다. 하지만, 이량체로 발현되는 경우 expression level이 0.1 IU/mL로 기존 scFVIII-B3 보다 10배 이상 낮은 것을 확인하였으며, 측정된 결과는 표 3에 기재되었다.
실시예 3-2. 발현 패턴 분석
본원에 구축된 키메라 FVIII을 실시예 4에 따라 정제한 후, 정제 시료를 웨스턴블랏 분석을 통해 발현 패턴을 확인하였다.
배양액과 LDS sample buffer를 각각 3:1의 비율로 혼합하여 4-12% bis-tris (BT) 젤에 혼합한 시료 30μL를 로딩한 후 150 volt로 약 1시간 동안 러닝한다. 러닝이 완료된 젤을 nitrocellulose (NC) membrane에 transfer한 후, blocking 버퍼 (Thermo Scientific)에 한 시간 정치하였다. 제 8인자의 중쇄 (GMA-012, Green Mountain Antibodies), 경쇄 (ab41188, Abcam) 및 D’D3 (96340, Abcam)을 인식하는 primary 항체를 TBST 버퍼에 희석하여 제조한 후, blocking이 완료된 NC membrane과 1시간 반응시켰다. TBST 버퍼를 사용하여 5분 간격으로 3회 washing을 진행한다. Secondary 항체 (Goat anti-mouse IgG-HRP conjugate)는 TBST 버퍼로 희석하여 제조한 후, NC membrane과 10분간 반응시킨다. TBST 버퍼를 사용하여 5분 간격으로 5회 세척하였다. ECL Prime Western Blotting Detection Reagent을 뿌려주어 NC membrane을 발색시킨 후, 암실에서 Hyperfilm ECL (GE healthcare)에 membrane을 노출하여 현상하였다.
그 결과 단쇄로 발현되는 키메라 FVIII은 모두 본원에서 디자인 한 형태로 발현되는 것을 확인하였고, 링커 길이에 따른 발현 패턴 변화는 없는 것으로 나타났다 (도 5).
실시예 4. 키메라 FVIII 생산
키메라 FVIII의 생산 공정은 크게 5단계로 구성되어 있으며, 실시예 2에서 배양한 배양액을 이용하여 정제를 진행하였다.
첫 번째 단계에서는 배양액에서 키메라 FVIII을 분리정제 하는 공정으로 FVIII 정제용으로 GE에서 개발한 VIIISelect resin을 이용하였다. VIIISelect resin을 컬럼에 패킹한 후, 2% citric acid로 컬럼을 세척하였다. 평형 버퍼 (20 mM Histidine, 5 mM CaCl2, 1 M NaCl, 0.02% Tween® 80, pH 7.0)를 흘려주어 평형을 잡는다. 배양액에 최종 1 M NaCl 농도가 되도록 5 M NaCl 버퍼를 넣어주고, pH를 7.0으로 적정한 후, 컬럼에 로딩하였다. 배양액 로딩 후, 평형 버퍼를 UV가 baseline으로 떨어질 때까지 흘려주어 평형을 잡았다. 용출 버퍼 (20 mM Histidine, 5 mM CaCl2, 0.9 M Arginine, 45% propylene glycol, 0.02% Tween® 80, pH 6.5)를 흘려주어 키메라 FVIII을 용출하였다.
두 번째 단계에서는 첫 번째 단계의 용출액에 섞여 있는 propylene glycol을 제거해주고, 일부 섞여 있는 분순물을 제거해주는 공정으로 GE사의 Q fast flow resin을 이용하였다. Q fast flow resin을 컬럼에 패킹한 후, 세척 버퍼 (0.5 M NaOH, 1 M NaCl)를 흘려주어 컬럼을 세척하였다. 평형 버퍼 (20 mM Histidine, 5 mM CaCl2, 0.02% Tween® 80, pH 7.0)를 흘려주어 평형을 잡았다. VIIISelect 공정의 용출액을 평형 버퍼로 10배 희석한 후, pH를 7.0으로 적정하고, 컬럼에 로딩하였다. 로딩 후, 평형 버퍼를 UV가 baseline으로 떨어질 때까지 흘려주어 평형을 잡았다. 용출 버퍼 (20 mM Histidine, 5 mM CaCl2, 500 mM NaCl, 0.02% Tween® 80, pH 7.0)를 흘려주어 키메라 FVIII을 용출하였다.
세 번째 단계에서는 정제한 키메라 FVIII에 PEG를 접합시켜 주는 공정이다. 정제된 키메라 FVIII 용액에 최종농도가 0.1 mM이 되도록 TCEP를 첨가한 후 4℃에서 1시간 정치시켜 삽입한 cysteine을 환원시켰다. 페길화 버퍼 (20 mM Histidine, 5 mM CaCl2, 200 mM NaCl, 0.02% Tween® 80, pH 7.0)로 평형이 잡힌 PD-10 컬럼 (GE healthcare)을 사용하여 잔존 TCEP를 제거하였다. 환원된 키메라 FVIII 자체의 이황화 결합을 산화시켜주기 위하여 4℃에서 2시간 정치 시켰다. PEG 접합은 DMSO에 녹인 PEG 용액(50 mg/mL)을 단백질 당 PEG 비율을 1:20이 되도록 넣어준 후 4℃에서 12-16시간 정치시켰다.
네 번째 단계에서는 PEGylation 과정 중에 생긴 impurity인 PEG가 붙지 않은 키메라 FVIII과 PEG가 2개 이상 붙은 키메라 FVIII을 제거하는 공정으로 GE사의 Superdex 200 resin을 이용하였다. Pre-packing 된 superdex 200 컬럼에 평형 버퍼 (20 mM Histidine, 5 mM CaCl2, 200 mM NaCl, 0.02% Tween® 80, pH 7.0)를 흘려주어 평형을 잡았다. PEGylation 공정 시료를 sample loop를 이용하여 컬럼에 로딩하였다. 로딩 후, 평형 버퍼를 흘려주며 peak을 수집하고, SDS-PAGE로 분석하여 적절한 fraction을 풀링(pooling) 하였다.
다섯 번째 단계에서는 네 번째 단계에서 풀링한 페길화 키메라 FVIII을 농축하는 공정이다. 이를 위해 Millipore사의 amicon 30kDa를 이용하여 25 IU/mL 농도가 되게 농축을 하였다.
정제 정도와 페길화 정도를 분석하기 위하여 공정 중간 시료와 최종 시료에 대한 SDS-PAGE를 수행하였다 (도 4, 도 6). scFVIII/D’D3의 경우 linker 길이에 따른 정제 패턴은 거의 유사한 것을 확인할 수 있다. 첫 번째 공정인 VIIISelect 공정에서 대부분의 HCP가 제거 되는 것을 볼 수 있으며, 두 번째 공정인 Q FF 공정에서 monomer 형태인 scFVIII/D’D3만 정제 되는 것을 확인하였다. 그 후, PEGylation 공정을 진행 한 후, 정제한 결과 PEG-scFVIII/D’D3만 정제 된 것을 확인하였다.
실시예 5. 키메라 FVIII 물질 비역가(specific activity) 확인
키메라 FVIII의 비역가를 확인하기 위해 chromogenic method로 키메라 FVIII의 activity를 측정하였고, BSA를 standard로 사용하여 BCA 방법으로 키메라 FVIII의 단백정량을 측정하였다. 이렇게 측정한 activity 결과 값을 단백정량 값으로 나누어 주어 specific activity를 측정하였으며, 그 결과는 표 4와 같다.
[표 4] 키메라 FVIII 비역가(specific activity)
scFVIII/D’D3의 경우 linker 길이를 바꾸어 주어도 specific activity 값은 큰 차이가 없는 것을 확인하였으며, 본 실시예에서 측정한 specific activity는 다른 recombinant FVIII의 specific activity 값들과 유사한 것을 확인하였다.
실시예 6. 키메라 FVIII의 LRP, vWF binding affinity 측정 실험
키메라 FVIII과 LRP 또는 vWF와의 binding affinity를 ELISA 방법을 이용하여 측정하였다. vWF (HCVWF-0191, Haemtech, 200 μg/mL) 또는 LRP (#04-03, Biomac, 260 μg/mL)를 사용하여 96 well plate에 well 당 100μL씩 coating한 후, 상온에서 2시간 동안 정치한다. 2시간 incubation 후, PBS에 0.1% Tween® 20을 첨가한 washing buffer로 washing을 수행한다. PBS에 1% BSA를 첨가한 blocking buffer를 well 당 200μL씩 넣고, 상온에서 1시간 동안 incubation하여 blocking을 수행한다. 1시간 incubation 후, PBS에 0.1% Tween® 20을 첨가한 washing buffer로 washing을 수행한다. FVIII sample을 blocking buffer로 적절한 농도로 희석한 후, well 당 100μL씩 loading 후, 상온에서 1시간 동안 incubation 한다. PBS에 0.1% Tween® 20을 첨가한 washing buffer로 washing을 수행한다. Anti-Factor VIII biotin (SAF8C-APBIO, Affinity Biologicals, 100 μg/mL)을 blocking buffer로 1/2000 희석하여 well 당 100μL씩 loading 하고, 상온에서 1시간 동안 정치한다. PBS에 0.1% Tween® 20을 첨가한 washing buffer로 washing을 수행한다. Streptavidin-HRP (S2438, Sigma-Aldrich)를 blocking buffer로 1/3000 희석하여 well 당 100μL씩 loading 하고 상온에서 45분 동안 incubation 한다. PBS에 0.1% Tween® 20을 첨가한 washing buffer로 washing을 수행한다. TMB substrate (52-00-03, KPL)를 well 당 100μL씩 처리 후, 10분 동안 incubation 한다. Stop solution (1 N 황산)을 well 당 100μL씩 넣어주어 반응 정지시킨 후, 490 nm 흡광도를 측정한다.
실험 결과, 키메라 FVIII의 LRP, vWF에 대한 결합이 ADVATE® 보다 현저히 감소되는 것을 확인하였다. 또한, 키메라 FVIII이 페길화 되면 LRP, vWF 결합은 더 감소되는 것을 확인하였다 (도 7, 도 8).
실시예 7. 동물 PK 시험: PEG-scFVIII/D’D3-120
페길화 키메라 FVIII의 체내 반감기를 측정하기 위해 혈우병 A형 마우스 모델 (HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화 하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII (본 실시예에서는 PEG-scFVIII/D’D3-120을 이용하였음)을 125 IU/kg 용량을 정맥으로 단 회 투여 하였다. PK 혈액 샘플은 Advate® 투여군에서는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, PEG-scFVIII/D’D3-120 투여군에서는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 PEG-scFVIII/D’D3-120의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다. PK 분석 결과(도 9) Advate®의 반감기는 약 7.86시간으로 측정되었고, PEG-scFVIII/D’D3-120의 반감기는 Advate® 보다 3.3배 긴 약 26.12시간으로 측정되었다. PEG-scFVIII/D’D3-120의 평균 머무름시간(MRT)도 Advate®에 비해 3.6배 증가하였다. 또한 PEG-scFVIII/D’D3-120의 소실속도(CL)는 Advate®보다 약 2배 감소하였고, 농도곡선하면적(AUC)은 약 2배 증가하는 것으로 나타났다.
[표 5] 페길화 키메라 FVIII 약동학적 파라미터
실제 예방적 요법으로 FVIII의 투여 시, 혈중 FVIII의 농도가 1% 또는 3% 이상으로 유지되도록 하기 때문에 반감기가 긴 지속형 FVIII을 사용할 경우 혈중 FVIII의 농도가 1% 또는 3% 이하로 떨어지는 시간을 지연시킬 수 있고 이는 투여 횟수의 감소로 이어질 수 있다. 지금까지 개발된 지속형 FVIII의 경우 HA mice에서 반감기의 증가폭은 대조군 대비 2배 이하 수준이었다. 이는 상기에서 언급한 것처럼 FVIII이 혈중에서 vWF와 강한 비공유 결합을 한 상태로 돌아다니기 때문에 vWF의 반감기보다 더 길어지지 못했던 것으로 판단된다. 본원에서 개발한 키메라 FVIII은 혈중 vWF와의 결합을 저해하는 것과 동시에 키메라 FVIII이 LRP에 의한 소실도 막아주기 때문에 지금까지 개발 된 지속형 FVIII이 넘지 못 했던 대조군 대비 반감기가 2배 이상 증가한 것으로 판단되며, 이는 매우 의미 있는 결과라고 판단한다.
실시예 8. 동물 PK 시험: 다양한 크기의PEG 접합된 scFVIII/D’D3-120
실시예 7과 유사하게 혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화하였다 (n = 3 / time point). 이 후 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량을 정맥으로 단 회 투여 하였다. 본 실시예에서는 scFVIII/D’D3-120에 서로 다른 size의 PEG를 접합시켜 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 7.88시간으로 측정되었고, 40kDa PEG-scFVIII/D’D3-120의 반감기는 약 20.93시간으로 측정되었고, 60kDa PEG-scFVIII/D’D3-120의 반감기는 약 28.09시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 10 및 표 6과 같다.
[표 6] 페길화 키메라 FVIII 약동학적 파라미터
실시예 9. 동물 PK 시험: PEG-scFVIII/D’D3-60과 Fc가 융합된 scFVIII/D’D3-120에 PEG 접합
혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화 하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량을 정맥으로 단 회 투여 하였다. 본 실시예에서는 PEG-scFVIII/D’D3-60과 Fc가 융합된 scFVIII/D’D3-120에 PEG를 접합시킨 물질을 이용하여 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 7.08시간으로 측정되었고, PEG-scFVIII/D’D3-60의 반감기는 약 16.53시간으로 측정되었고, PEG-scFVIII/D’D3/Fc-120, Fc의 반감기는 약 18.89시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 11 및 표 7과 같다.
[표 7] 페길화 키메라 FVIII 약동학적 파라미터
실시예 10. 동물 PK 시험: PEG-scFVIII/D’D3-300과 D’D3를 이량체 형태로 FVIII의 c-말단에 융합한 scFVIII/D’D3-120에 PEG 접합
혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량을 정맥으로 단 회 투여 하였다. 본 실시예에서는 PEG-scFVIII/D’D3-300과 D’D3를 이량체 형태로 FVIII의 c-말단에 융합한 scFVIII/D’D3-120에 PEG를 접합시킨 물질을 이용하여 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 9.02시간으로 측정되었고, PEG-scFVIII/D’D3-300의 반감기는 약 15.76시간으로 측정되었고, PEG-scFVIII/D’D3/D’D3-120의 반감기는 약 18.00시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 12 및 표 8과 같다.
[표 8] 페길화 키메라 FVIII 약동학적 파라미터
실시예 11. 동물 PK 시험: PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-300
혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량을 정맥으로 단 회 투여 하였다. 본 실시예에서는 PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-300을 이용하여 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 6.99시간으로 측정되었고, PEG-scFVIII/D’D3-60, PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-300의 반감기는 각각 약 20.07, 24.48, 17.13시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 13 및 표 9와 같다.
[표 9] 페길화 키메라 FVIII 약동학적 파라미터
실시예 12. 동물 PK 시험: PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2
혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화 하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량을 정맥으로 단 회 투여 하였다. 본 실시예에서는 PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2를 이용하여 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 6.26시간으로 측정되었고, PEG-scFVIII/D’D3-120, PEG-scFVIII/D’D3-120-TH1, PEG-scFVIII/D’D3-120-TH2의 반감기는 각각 약 21.28, 24.19, 25.28시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 14 및 표 10과 같다.
[표 10] 페길화 키메라 FVIII 약동학적 파라미터
실시예 13. 동물 PK 시험: 다양한 크기 및 형태의 PEG 접합된 scFVIII/D’D3-120-TH1
혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화 하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량으로 정맥으로 단 회 투여 하였다. 본 실시예에서는 scFVIII/D’D3-120-TH1에 여러 가지 PEG를 접합시킨 물질을 이용하여 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 7.66시간으로 측정되었고, 20kDa linear PEG-scFVIII/D’D3-120-TH1, 20kDa branch PEG-scFVIII/D’D3-120-TH1, 40kDa branch PEG-scFVIII/D’D3-120-TH1의 반감기는 각각 약 22.98, 16.10, 27.33시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 15 및 표 11과 같다.
[표 11] 페길화 키메라 FVIII 약동학적 파라미터
실시예 14. Thrombin generation assay
페길화 키메라 FVIII의 thrombin generation profile을 확인하기 위해 thrombin generation assay를 진행하였다. Fluoroskan Ascent™ FL microplate fluorometer and luminometer (5210460, Thermofisher) 기기를 미리 37℃로 warming 해 놓고, 37℃ pre-warmed D.W로 기계의 line을 washing 하였다. FVIII deficient plasma (OTXW17, Siemens), Calibrator (TS20.00, Thrombinoscope), PPP-low-reagent (TS31.00, Thrombinoscope)를 상온에서 15분 이상 방치하여 warming 시킨 후, 37℃ pre-warmed D.W 1mL로 reconstitution 하였다. 96 well plate에 Calibrator와 PPP-low-reagent를 20μL씩 넣어주고, 앞에서 준비한 Fluoroskan 기기에 plate를 넣어 37℃로 가온해 놓았다. Sample dilution은 앞에서 준비한 FVIII deficient plasma를 이용하여 적절한 농도로 희석하였다. FluCa-Kit (TS50.00, Thrombinoscope)안의 Fluo-buffer를 37℃로 warming 해 놓았다. 준비한 sample을 Fluoroskan 기기 안에 들어있는 plate에 80μL씩 넣고, 37℃로 warming 된 Fluo-buffer 1 vial에 40μL의 Fluo-substrate를 넣어 준비하였다. 준비된 Flu-Ca를 기계에 넣은 후, 미리 짜놓은 software를 실행하면 각 well 당 20μL의 Flu-Ca가 분주 되고, thrombin generation을 진행하였다.
Thrombin generation assay 결과 FVIII deficient plasma에서는 thrombin generation profile이 현저히 낮은 반면, 키메라 FVIII에서는 thrombin generation profile이 농도 의존적으로 점차 높아지는 것을 확인하였다. 키메라 FVIII 1 IU/mL의 thrombin generation profile은 normal plasma의 thrombin generation profile과 유사한 것을 확인할 수 있었다 (도 16).
실시예 15. 동물 급성 효능(acute efficacy) 시험
키메라 FVIII의 효력을 확인하기 위해 혈우병 A형 마우스(HA mice)의 tail clip model에서 acute efficacy 시험을 진행하였다. Wild type mice (WT)는 19 - 25 g 중량의 수컷 C57BL/6 mouse를 오리엔트 바이오에서 공급받은 개체를 이용하였고, HA mice는 녹십자에서 breeding한 개체로 19 - 25 g 중량의 수컷 mouse를 선별하여 이용하였다. 시험 당일, 15mL conical tube (1개/마리)에 saline을 14mL까지 채운 후, 가열된 water bath에 담가서 saline의 온도가 37℃가 되도록 유지시켰고, 투여 전 mouse의 체중을 측정하였다. Pentobarbital sodium을 IP로 60 mg/kg dose로 투여하여 마취시킨 후, 음성 대조물질은 0.9% 생리 식염수를, 양성대조 물질인 Advate®와 페길화 키메라 FVIII은 100 IU/kg dose로 jugular vein으로 5 mL/kg의 볼륨으로 단 회 투여하였다. 5분 후, HA mice 꼬리의 말단에서 4mm 되는 지점을 절단하고, 꼬리를 saline buffer에 넣어 30분 동안 혈액을 모았다. 30분 동안 모은 혈액 시료를 1,500g 에서 5분간 원심 분리하여 상층액을 제거한 후, 10mL pipet을 이용하여 conical tube에 3차 증류수를 10mL까지 넣고, vortex를 이용하여 혈액을 완전히 용혈 시켰다. Blood loss를 측정하기 위해 용혈 된 샘플을 hemoglobin assay kit (MAK115-1KT, Sigma-Aldrich)의 방법으로 분석하였다.
Wild type mice (WT)의 경우 hemoglobin의 농도가 약 100 nM 인 것을 확인하였고, HA mice (KO)에서 음성대조군인 경우 hemoglobin의 농도가 약 1,000 nM 인 것을 확인하였다. Advate®와 페길화 키메라 FVIII의 경우 hemoglobin의 농도가 약 140 - 450 nM 인 것을 확인하였으며, 이는 음성대조군에 비해 약 55% 감소한 수치이다. Advate®와 페길화 키메라 FVIII 간에는 통계적으로 차이가 없는 것을 확인하였다 (도 17).
실시예 16. 동물 PK 시험: CHO 세포에서 발현한 scFVIII/D’D3-120-TH1에 다양한 크기 및 형태의 PEG 접합
혈우병 A형 마우스 모델(HA mice)을 이용한 PK 시험을 진행하였다. HA mice는 8주령의 쥐(20 ± 2 g)를 이용하였고, 투여 전 개체 별 무게 측정 후, 그룹화 하였다 (n = 3 / time point). 대조 물질인 Advate®와 페길화 키메라 FVIII을 125 IU/kg 용량으로 정맥으로 단 회 투여 하였다. 본 실시예에서는 CHO 세포에서 발현한 scFVIII/D’D3-120-TH1에 여러 가지 PEG를 접합시킨 물질을 이용하여 PK 시험을 진행하였다. 대조 물질인 Advate® 투여군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 9, 16, 22, 30 hr에, 시험군의 혈액 시료는 투여 후 0, 0.083, 2, 6, 16, 30, 40, 48, 64, 72, 96 hr에 200μL sampling 하였다. 채혈 직후 샘플을 항응고제 (sodium citrate 3.2% buffer)와 1:9 부피로 섞은 후 원심분리로 혈장을 분리하였다. Advate®와 페길화 키메라 FVIII의 역가는 Chromogenic method를 사용하여 측정하였고, 약동학적 파라미터는 WinNonLin software(version 6.4)의 non-compartmental (NCA) model을 적용하여 산출하였다.
PK 분석 결과 Advate®의 반감기는 약 8.22시간으로 측정되었고, 20kDa linear PEG-scFVIII/D’D3-120-TH1, 40kDa linear PEG-scFVIII/D’D3-120-TH1의 반감기는 각각 약 25.01, 26.88시간으로 측정되었다. 약동학적 프로파일 및 약동학적 파라미터는 도 18 및 표 12와 같다.
[표 12] 페길화 키메라 FVIII 약동학적 파라미터
이상에서 본원의 예시적인 실시예에 대하여 상세하게 설명하였지만 본원의 권리범위는 이에 한정되는 것은 아니고 다음의 청구범위에서 정의하고 있는 본원의 기본 개념을 이용한 당업자의 여러 변형 및 개량 형태 또한 본원의 권리범위에 속하는 것이다.
본 발명에서 사용되는 모든 기술용어는, 달리 정의되지 않는 이상, 본 발명의 관련 분야에서 통상의 당업자가 일반적으로 이해하는 바와 같은 의미로 사용된다. 본 명세서에 참고문헌으로 기재되는 모든 간행물의 내용은 본 발명에 도입된다.
<110> Mogam Institute for Biomedical Research
<120> Chimeric protein of Factor VIII and Von Willebrand Factor and its
use
<130> PCT201705004P
<150> US 62/354,255
<151> 2016-06-24
<160> 80
<170> KoPatentIn 3.0
<210> 1
<211> 2332
<212> PRT
<213> Homo sapiens
<220>
<221> PEPTIDE
<222> (1)..(2332)
<223> Full length Factor VIII
<400> 1
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Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
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Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
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Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
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Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
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Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
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Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
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Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
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Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
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His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
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Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
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His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
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His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
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Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
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Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
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Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
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Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
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Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
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Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
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Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
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Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
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Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
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Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
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Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
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Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
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Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
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Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
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Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
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Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
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Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
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Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
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Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
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Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
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Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
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Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Xaa Gln Asn
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Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
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His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
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Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
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Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
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Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
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Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
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Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
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Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
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Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
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Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Glu Glu Asn Asn Asp
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Ser Lys Leu Leu Glu Ser Gly Leu Met Asn Ser Gln Glu Ser Ser Trp
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Gly Lys Asn Val Ser Ser Thr Glu Ser Gly Arg Leu Phe Lys Gly Lys
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Arg Ala His Gly Pro Ala Leu Leu Thr Lys Asp Asn Ala Leu Phe Lys
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Val Ser Ile Ser Leu Leu Lys Thr Asn Lys Thr Ser Asn Asn Ser Ala
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Thr Asn Arg Lys Thr His Ile Asp Gly Pro Ser Leu Leu Ile Glu Asn
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Ser Pro Ser Val Trp Gln Asn Ile Leu Glu Ser Asp Thr Glu Phe Lys
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Lys Val Thr Pro Leu Ile His Asp Arg Met Leu Met Asp Lys Asn Ala
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Thr Ala Leu Arg Leu Asn His Met Ser Asn Lys Thr Thr Ser Ser Lys
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Asn Met Glu Met Val Gln Gln Lys Lys Glu Gly Pro Ile Pro Pro Asp
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Ala Gln Asn Pro Asp Met Ser Phe Phe Lys Met Leu Phe Leu Pro Glu
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Ser Ala Arg Trp Ile Gln Arg Thr His Gly Lys Asn Ser Leu Asn Ser
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Gly Gln Gly Pro Ser Pro Lys Gln Leu Val Ser Leu Gly Pro Glu Lys
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Ser Val Glu Gly Gln Asn Phe Leu Ser Glu Lys Asn Lys Val Val Val
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Gly Lys Gly Glu Phe Thr Lys Asp Val Gly Leu Lys Glu Met Val Phe
1155 1160 1165
Pro Ser Ser Arg Asn Leu Phe Leu Thr Asn Leu Asp Asn Leu His Glu
1170 1175 1180
Asn Asn Thr His Asn Gln Glu Lys Lys Ile Gln Glu Glu Ile Glu Lys
1185 1190 1195 1200
Lys Glu Thr Leu Ile Gln Glu Asn Val Val Leu Pro Gln Ile His Thr
1205 1210 1215
Val Thr Gly Thr Lys Asn Phe Met Lys Asn Leu Phe Leu Leu Ser Thr
1220 1225 1230
Arg Gln Asn Val Glu Gly Ser Tyr Asp Gly Ala Tyr Ala Pro Val Leu
1235 1240 1245
Gln Asp Phe Arg Ser Leu Asn Asp Ser Thr Asn Arg Thr Lys Lys His
1250 1255 1260
Thr Ala His Phe Ser Lys Lys Gly Glu Glu Glu Asn Leu Glu Gly Leu
1265 1270 1275 1280
Gly Asn Gln Thr Lys Gln Ile Val Glu Lys Tyr Ala Cys Thr Thr Arg
1285 1290 1295
Ile Ser Pro Asn Thr Ser Gln Gln Asn Phe Val Thr Gln Arg Ser Lys
1300 1305 1310
Arg Ala Leu Lys Gln Phe Arg Leu Pro Leu Glu Glu Thr Glu Leu Glu
1315 1320 1325
Lys Arg Ile Ile Val Asp Asp Thr Ser Thr Gln Trp Ser Lys Asn Met
1330 1335 1340
Lys His Leu Thr Pro Ser Thr Leu Thr Gln Ile Asp Tyr Asn Glu Lys
1345 1350 1355 1360
Glu Lys Gly Ala Ile Thr Gln Ser Pro Leu Ser Asp Cys Leu Thr Arg
1365 1370 1375
Ser His Ser Ile Pro Gln Ala Asn Arg Ser Pro Leu Pro Ile Ala Lys
1380 1385 1390
Val Ser Ser Phe Pro Ser Ile Arg Pro Ile Tyr Leu Thr Arg Val Leu
1395 1400 1405
Phe Gln Asp Asn Ser Ser His Leu Pro Ala Ala Ser Tyr Arg Lys Lys
1410 1415 1420
Asp Ser Gly Val Gln Glu Ser Ser His Phe Leu Gln Gly Ala Lys Lys
1425 1430 1435 1440
Asn Asn Leu Ser Leu Ala Ile Leu Thr Leu Glu Met Thr Gly Asp Gln
1445 1450 1455
Arg Glu Val Gly Ser Leu Gly Thr Ser Ala Thr Asn Ser Val Thr Tyr
1460 1465 1470
Lys Lys Val Glu Asn Thr Val Leu Pro Lys Pro Asp Leu Pro Lys Thr
1475 1480 1485
Ser Gly Lys Val Glu Leu Leu Pro Lys Val His Ile Tyr Gln Lys Asp
1490 1495 1500
Leu Phe Pro Thr Glu Thr Ser Asn Gly Ser Pro Gly His Leu Asp Leu
1505 1510 1515 1520
Val Glu Gly Ser Leu Leu Gln Gly Thr Glu Gly Ala Ile Lys Trp Asn
1525 1530 1535
Glu Ala Asn Arg Pro Gly Lys Val Pro Phe Leu Arg Val Ala Thr Glu
1540 1545 1550
Ser Ser Ala Lys Thr Pro Ser Lys Leu Leu Asp Pro Leu Ala Trp Asp
1555 1560 1565
Asn His Tyr Gly Thr Gln Ile Pro Lys Glu Glu Trp Lys Ser Gln Glu
1570 1575 1580
Lys Ser Pro Glu Lys Thr Ala Phe Lys Lys Lys Asp Thr Ile Leu Ser
1585 1590 1595 1600
Leu Asn Ala Cys Glu Ser Asn His Ala Ile Ala Ala Ile Asn Glu Gly
1605 1610 1615
Gln Asn Lys Pro Glu Ile Glu Val Thr Trp Ala Lys Gln Gly Arg Thr
1620 1625 1630
Glu Arg Leu Cys Ser Gln Asn Pro Pro Val Leu Lys Arg His Gln Arg
1635 1640 1645
Glu Ile Thr Arg Thr Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp Tyr
1650 1655 1660
Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile Tyr
1665 1670 1675 1680
Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr Arg
1685 1690 1695
His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met Ser
1700 1705 1710
Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val Pro
1715 1720 1725
Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe Thr
1730 1735 1740
Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu Gly
1745 1750 1755 1760
Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe Arg
1765 1770 1775
Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser Tyr
1780 1785 1790
Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val Lys
1795 1800 1805
Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met Ala
1810 1815 1820
Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser Asp
1825 1830 1835 1840
Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu Leu
1845 1850 1855
Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val Thr
1860 1865 1870
Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys Ser
1875 1880 1885
Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys Asn
1890 1895 1900
Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His Ala
1905 1910 1915 1920
Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala Gln
1925 1930 1935
Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu Asn
1940 1945 1950
Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys Lys
1955 1960 1965
Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe Glu
1970 1975 1980
Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu Cys
1985 1990 1995 2000
Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu Val
2005 2010 2015
Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His Ile
2020 2025 2030
Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala Pro
2035 2040 2045
Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser Thr
2050 2055 2060
Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met Ile
2065 2070 2075 2080
Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser Leu
2085 2090 2095
Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys Trp
2100 2105 2110
Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe Gly
2115 2120 2125
Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro Ile
2130 2135 2140
Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg Ser
2145 2150 2155 2160
Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser Met
2165 2170 2175
Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr Ala
2180 2185 2190
Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys Ala
2195 2200 2205
Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val Asn
2210 2215 2220
Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys Val
2225 2230 2235 2240
Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met Tyr
2245 2250 2255
Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp Thr
2260 2265 2270
Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln Asp
2275 2280 2285
Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr Arg
2290 2295 2300
Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu Arg
2305 2310 2315 2320
Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr
2325 2330
<210> 2
<211> 2791
<212> PRT
<213> Homo sapiens
<220>
<221> PEPTIDE
<222> (1)..(2791)
<223> Full length vWF
<400> 2
Ala Glu Gly Thr Arg Gly Arg Ser Ser Thr Ala Arg Cys Ser Leu Phe
1 5 10 15
Gly Ser Asp Phe Val Asn Thr Phe Asp Gly Ser Met Tyr Ser Phe Ala
20 25 30
Gly Tyr Cys Ser Tyr Leu Leu Ala Gly Gly Cys Gln Lys Arg Ser Phe
35 40 45
Ser Ile Ile Gly Asp Phe Gln Asn Gly Lys Arg Val Ser Leu Ser Val
50 55 60
Tyr Leu Gly Glu Phe Phe Asp Ile His Leu Phe Val Asn Gly Thr Val
65 70 75 80
Thr Gln Gly Asp Gln Arg Val Ser Met Pro Tyr Ala Ser Lys Gly Leu
85 90 95
Tyr Leu Glu Thr Glu Ala Gly Tyr Tyr Lys Leu Ser Gly Glu Ala Tyr
100 105 110
Gly Phe Val Ala Arg Ile Asp Gly Ser Gly Asn Phe Gln Val Leu Leu
115 120 125
Ser Asp Arg Tyr Phe Asn Lys Thr Cys Gly Leu Cys Gly Asn Phe Asn
130 135 140
Ile Phe Ala Glu Asp Asp Phe Met Thr Gln Glu Gly Thr Leu Thr Ser
145 150 155 160
Asp Pro Tyr Asp Phe Ala Asn Ser Trp Ala Leu Ser Ser Gly Glu Gln
165 170 175
Trp Cys Glu Arg Ala Ser Pro Pro Ser Ser Ser Cys Asn Ile Ser Ser
180 185 190
Gly Glu Met Gln Lys Gly Leu Trp Glu Gln Cys Gln Leu Leu Lys Ser
195 200 205
Thr Ser Val Phe Ala Arg Cys His Pro Leu Val Asp Pro Glu Pro Phe
210 215 220
Val Ala Leu Cys Glu Lys Thr Leu Cys Glu Cys Ala Gly Gly Leu Glu
225 230 235 240
Cys Ala Cys Pro Ala Leu Leu Glu Tyr Ala Arg Thr Cys Ala Gln Glu
245 250 255
Gly Met Val Leu Tyr Gly Trp Thr Asp His Ser Ala Cys Ser Pro Val
260 265 270
Cys Pro Ala Gly Met Glu Tyr Arg Gln Cys Val Ser Pro Cys Ala Arg
275 280 285
Thr Cys Gln Ser Leu His Ile Asn Glu Met Cys Gln Glu Arg Cys Val
290 295 300
Asp Gly Cys Ser Cys Pro Glu Gly Gln Leu Leu Asp Glu Gly Leu Cys
305 310 315 320
Val Glu Ser Thr Glu Cys Pro Cys Val His Ser Gly Lys Arg Tyr Pro
325 330 335
Pro Gly Thr Ser Leu Ser Arg Asp Cys Asn Thr Cys Ile Cys Arg Asn
340 345 350
Ser Gln Trp Ile Cys Ser Asn Glu Glu Cys Pro Gly Glu Cys Leu Val
355 360 365
Thr Gly Gln Ser His Phe Lys Ser Phe Asp Asn Arg Tyr Phe Thr Phe
370 375 380
Ser Gly Ile Cys Gln Tyr Leu Leu Ala Arg Asp Cys Gln Asp His Ser
385 390 395 400
Phe Ser Ile Val Ile Glu Thr Val Gln Cys Ala Asp Asp Arg Asp Ala
405 410 415
Val Cys Thr Arg Ser Val Thr Val Arg Leu Pro Gly Leu His Asn Ser
420 425 430
Leu Val Lys Leu Lys His Gly Ala Gly Val Ala Met Asp Gly Gln Asp
435 440 445
Val Gln Leu Pro Leu Leu Lys Gly Asp Leu Arg Ile Gln His Thr Val
450 455 460
Thr Ala Ser Val Arg Leu Ser Tyr Gly Glu Asp Leu Gln Met Asp Trp
465 470 475 480
Asp Gly Arg Gly Arg Leu Leu Val Lys Leu Ser Pro Val Tyr Ala Gly
485 490 495
Lys Thr Cys Gly Leu Cys Gly Asn Tyr Asn Gly Asn Gln Gly Asp Asp
500 505 510
Phe Leu Thr Pro Ser Gly Leu Ala Glu Pro Arg Val Glu Asp Phe Gly
515 520 525
Asn Ala Trp Lys Leu His Gly Asp Cys Gln Asp Leu Gln Lys Gln His
530 535 540
Ser Asp Pro Cys Ala Leu Asn Pro Arg Met Thr Arg Phe Ser Glu Glu
545 550 555 560
Ala Cys Ala Val Leu Thr Ser Pro Thr Phe Glu Ala Cys His Arg Ala
565 570 575
Val Ser Pro Leu Pro Tyr Leu Arg Asn Cys Arg Tyr Asp Val Cys Ser
580 585 590
Cys Ser Asp Gly Arg Glu Cys Leu Cys Gly Ala Leu Ala Ser Tyr Ala
595 600 605
Ala Ala Cys Ala Gly Arg Gly Val Arg Val Ala Trp Arg Glu Pro Gly
610 615 620
Arg Cys Glu Leu Asn Cys Pro Lys Gly Gln Val Tyr Leu Gln Cys Gly
625 630 635 640
Thr Pro Cys Asn Leu Thr Cys Arg Ser Leu Ser Tyr Pro Asp Glu Glu
645 650 655
Cys Asn Glu Ala Cys Leu Glu Gly Cys Phe Cys Pro Pro Gly Leu Tyr
660 665 670
Met Asp Glu Arg Gly Asp Cys Val Pro Lys Ala Gln Cys Pro Cys Tyr
675 680 685
Tyr Asp Gly Glu Ile Phe Gln Pro Glu Asp Ile Phe Ser Asp His His
690 695 700
Thr Met Cys Tyr Cys Glu Asp Gly Phe Met His Cys Thr Met Ser Gly
705 710 715 720
Val Pro Gly Ser Leu Leu Pro Asp Ala Val Leu Ser Ser Pro Leu Ser
725 730 735
His Arg Ser Lys Arg Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
740 745 750
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
755 760 765
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
770 775 780
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
785 790 795 800
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
805 810 815
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
820 825 830
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
835 840 845
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
850 855 860
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
865 870 875 880
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
885 890 895
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
900 905 910
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
915 920 925
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
930 935 940
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
945 950 955 960
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
965 970 975
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
980 985 990
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
995 1000 1005
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1010 1015 1020
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1025 1030 1035 1040
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
1045 1050 1055
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
1060 1065 1070
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
1075 1080 1085
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
1090 1095 1100
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
1105 1110 1115 1120
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
1125 1130 1135
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
1140 1145 1150
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
1155 1160 1165
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
1170 1175 1180
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
1185 1190 1195 1200
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
1205 1210 1215
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
1220 1225 1230
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
1235 1240 1245
Tyr Cys Ser Arg Leu Leu Asp Leu Val Phe Leu Leu Asp Gly Ser Ser
1250 1255 1260
Arg Leu Ser Glu Ala Glu Phe Glu Val Leu Lys Ala Phe Val Val Asp
1265 1270 1275 1280
Met Met Glu Arg Leu Arg Ile Ser Gln Lys Trp Val Arg Val Ala Val
1285 1290 1295
Val Glu Tyr His Asp Gly Ser His Ala Tyr Ile Gly Leu Lys Asp Arg
1300 1305 1310
Lys Arg Pro Ser Glu Leu Arg Arg Ile Ala Ser Gln Val Lys Tyr Ala
1315 1320 1325
Gly Ser Gln Val Ala Ser Thr Ser Glu Val Leu Lys Tyr Thr Leu Phe
1330 1335 1340
Gln Ile Phe Ser Lys Ile Asp Arg Pro Glu Ala Ser Arg Ile Thr Leu
1345 1350 1355 1360
Leu Leu Met Ala Ser Gln Glu Pro Gln Arg Met Ser Arg Asn Phe Val
1365 1370 1375
Arg Tyr Val Gln Gly Leu Lys Lys Lys Lys Val Ile Val Ile Pro Val
1380 1385 1390
Gly Ile Gly Pro His Ala Asn Leu Lys Gln Ile Arg Leu Ile Glu Lys
1395 1400 1405
Gln Ala Pro Glu Asn Lys Ala Phe Val Leu Ser Ser Val Asp Glu Leu
1410 1415 1420
Glu Gln Gln Arg Asp Glu Ile Val Ser Tyr Leu Cys Asp Leu Ala Pro
1425 1430 1435 1440
Glu Ala Pro Pro Pro Thr Leu Pro Pro Asp Met Ala Gln Val Thr Val
1445 1450 1455
Gly Pro Gly Leu Leu Gly Val Ser Thr Leu Gly Pro Lys Arg Asn Ser
1460 1465 1470
Met Val Leu Asp Val Ala Phe Val Leu Glu Gly Ser Asp Lys Ile Gly
1475 1480 1485
Glu Ala Asp Phe Asn Arg Ser Lys Glu Phe Met Glu Glu Val Ile Gln
1490 1495 1500
Arg Met Asp Val Gly Gln Asp Ser Ile His Val Thr Val Leu Gln Tyr
1505 1510 1515 1520
Ser Tyr Met Val Thr Val Glu Tyr Pro Phe Ser Glu Ala Gln Ser Lys
1525 1530 1535
Gly Asp Ile Leu Gln Arg Val Arg Glu Ile Arg Tyr Gln Gly Gly Asn
1540 1545 1550
Arg Thr Asn Thr Gly Leu Ala Leu Arg Tyr Leu Ser Asp His Ser Phe
1555 1560 1565
Leu Val Ser Gln Gly Asp Arg Glu Gln Ala Pro Asn Leu Val Tyr Met
1570 1575 1580
Val Thr Gly Asn Pro Ala Ser Asp Glu Ile Lys Arg Leu Pro Gly Asp
1585 1590 1595 1600
Ile Gln Val Val Pro Ile Gly Val Gly Pro Asn Ala Asn Val Gln Glu
1605 1610 1615
Leu Glu Arg Ile Gly Trp Pro Asn Ala Pro Ile Leu Ile Gln Asp Phe
1620 1625 1630
Glu Thr Leu Pro Arg Glu Ala Pro Asp Leu Val Leu Gln Arg Cys Cys
1635 1640 1645
Ser Gly Glu Gly Leu Gln Ile Pro Thr Leu Ser Pro Ala Pro Asp Cys
1650 1655 1660
Ser Gln Pro Leu Asp Val Ile Leu Leu Leu Asp Gly Ser Ser Ser Phe
1665 1670 1675 1680
Pro Ala Ser Tyr Phe Asp Glu Met Lys Ser Phe Ala Lys Ala Phe Ile
1685 1690 1695
Ser Lys Ala Asn Ile Gly Pro Arg Leu Thr Gln Val Ser Val Leu Gln
1700 1705 1710
Tyr Gly Ser Ile Thr Thr Ile Asp Val Pro Trp Asn Val Val Pro Glu
1715 1720 1725
Lys Ala His Leu Leu Ser Leu Val Asp Val Met Gln Arg Glu Gly Gly
1730 1735 1740
Pro Ser Gln Ile Gly Asp Ala Leu Gly Phe Ala Val Arg Tyr Leu Thr
1745 1750 1755 1760
Ser Glu Met His Gly Ala Arg Pro Gly Ala Ser Lys Ala Val Val Ile
1765 1770 1775
Leu Val Thr Asp Val Ser Val Asp Ser Val Asp Ala Ala Ala Asp Ala
1780 1785 1790
Ala Arg Ser Asn Arg Val Thr Val Phe Pro Ile Gly Ile Gly Asp Arg
1795 1800 1805
Tyr Asp Ala Ala Gln Leu Arg Ile Leu Ala Gly Pro Ala Gly Asp Ser
1810 1815 1820
Asn Val Val Lys Leu Gln Arg Ile Glu Asp Leu Pro Thr Met Val Thr
1825 1830 1835 1840
Leu Gly Asn Ser Phe Leu His Lys Leu Cys Ser Gly Phe Val Arg Ile
1845 1850 1855
Cys Met Asp Glu Asp Gly Asn Glu Lys Arg Pro Gly Asp Val Trp Thr
1860 1865 1870
Leu Pro Asp Gln Cys His Thr Val Thr Cys Gln Pro Asp Gly Gln Thr
1875 1880 1885
Leu Leu Lys Ser His Arg Val Asn Cys Asp Arg Gly Leu Arg Pro Ser
1890 1895 1900
Cys Pro Asn Ser Gln Ser Pro Val Lys Val Glu Glu Thr Cys Gly Cys
1905 1910 1915 1920
Arg Trp Thr Cys Pro Cys Val Cys Thr Gly Ser Ser Thr Arg His Ile
1925 1930 1935
Val Thr Phe Asp Gly Gln Asn Phe Lys Leu Thr Gly Ser Cys Ser Tyr
1940 1945 1950
Val Leu Phe Gln Asn Lys Glu Gln Asp Leu Glu Val Ile Leu His Asn
1955 1960 1965
Gly Ala Cys Ser Pro Gly Ala Arg Gln Gly Cys Met Lys Ser Ile Glu
1970 1975 1980
Val Lys His Ser Ala Leu Ser Val Glu Leu His Ser Asp Met Glu Val
1985 1990 1995 2000
Thr Val Asn Gly Arg Leu Val Ser Val Pro Tyr Val Gly Gly Asn Met
2005 2010 2015
Glu Val Asn Val Tyr Gly Ala Ile Met His Glu Val Arg Phe Asn His
2020 2025 2030
Leu Gly His Ile Phe Thr Phe Thr Pro Gln Asn Asn Glu Phe Gln Leu
2035 2040 2045
Gln Leu Ser Pro Lys Thr Phe Ala Ser Lys Thr Tyr Gly Leu Cys Gly
2050 2055 2060
Ile Cys Asp Glu Asn Gly Ala Asn Asp Phe Met Leu Arg Asp Gly Thr
2065 2070 2075 2080
Val Thr Thr Asp Trp Lys Thr Leu Val Gln Glu Trp Thr Val Gln Arg
2085 2090 2095
Pro Gly Gln Thr Cys Gln Pro Ile Leu Glu Glu Gln Cys Leu Val Pro
2100 2105 2110
Asp Ser Ser His Cys Gln Val Leu Leu Leu Pro Leu Phe Ala Glu Cys
2115 2120 2125
His Lys Val Leu Ala Pro Ala Thr Phe Tyr Ala Ile Cys Gln Gln Asp
2130 2135 2140
Ser Cys His Gln Glu Gln Val Cys Glu Val Ile Ala Ser Tyr Ala His
2145 2150 2155 2160
Leu Cys Arg Thr Asn Gly Val Cys Val Asp Trp Arg Thr Pro Asp Phe
2165 2170 2175
Cys Ala Met Ser Cys Pro Pro Ser Leu Val Tyr Asn His Cys Glu His
2180 2185 2190
Gly Cys Pro Arg His Cys Asp Gly Asn Val Ser Ser Cys Gly Asp His
2195 2200 2205
Pro Ser Glu Gly Cys Phe Cys Pro Pro Asp Lys Val Met Leu Glu Gly
2210 2215 2220
Ser Cys Val Pro Glu Glu Ala Cys Thr Gln Cys Ile Gly Glu Asp Gly
2225 2230 2235 2240
Val Gln His Gln Phe Leu Glu Ala Trp Val Pro Asp His Gln Pro Cys
2245 2250 2255
Gln Ile Cys Thr Cys Leu Ser Gly Arg Lys Val Asn Cys Thr Thr Gln
2260 2265 2270
Pro Cys Pro Thr Ala Lys Ala Pro Thr Cys Gly Leu Cys Glu Val Ala
2275 2280 2285
Arg Leu Arg Gln Asn Ala Asp Gln Cys Cys Pro Glu Tyr Glu Cys Val
2290 2295 2300
Cys Asp Pro Val Ser Cys Asp Leu Pro Pro Val Pro His Cys Glu Arg
2305 2310 2315 2320
Gly Leu Gln Pro Thr Leu Thr Asn Pro Gly Glu Cys Arg Pro Asn Phe
2325 2330 2335
Thr Cys Ala Cys Arg Lys Glu Glu Cys Lys Arg Val Ser Pro Pro Ser
2340 2345 2350
Cys Pro Pro His Arg Leu Pro Thr Leu Arg Lys Thr Gln Cys Cys Asp
2355 2360 2365
Glu Tyr Glu Cys Ala Cys Asn Cys Val Asn Ser Thr Val Ser Cys Pro
2370 2375 2380
Leu Gly Tyr Leu Ala Ser Thr Ala Thr Asn Asp Cys Gly Cys Thr Thr
2385 2390 2395 2400
Thr Thr Cys Leu Pro Asp Lys Val Cys Val His Arg Ser Thr Ile Tyr
2405 2410 2415
Pro Val Gly Gln Phe Trp Glu Glu Gly Cys Asp Val Cys Thr Cys Thr
2420 2425 2430
Asp Met Glu Asp Ala Val Met Gly Leu Arg Val Ala Gln Cys Ser Gln
2435 2440 2445
Lys Pro Cys Glu Asp Ser Cys Arg Ser Gly Phe Thr Tyr Val Leu His
2450 2455 2460
Glu Gly Glu Cys Cys Gly Arg Cys Leu Pro Ser Ala Cys Glu Val Val
2465 2470 2475 2480
Thr Gly Ser Pro Arg Gly Asp Ser Gln Ser Ser Trp Lys Ser Val Gly
2485 2490 2495
Ser Gln Trp Ala Ser Pro Glu Asn Pro Cys Leu Ile Asn Glu Cys Val
2500 2505 2510
Arg Val Lys Glu Glu Val Phe Ile Gln Gln Arg Asn Val Ser Cys Pro
2515 2520 2525
Gln Leu Glu Val Pro Val Cys Pro Ser Gly Phe Gln Leu Ser Cys Lys
2530 2535 2540
Thr Ser Ala Cys Cys Pro Ser Cys Arg Cys Glu Arg Met Glu Ala Cys
2545 2550 2555 2560
Met Leu Asn Gly Thr Val Ile Gly Pro Gly Lys Thr Val Met Ile Asp
2565 2570 2575
Val Cys Thr Thr Cys Arg Cys Met Val Gln Val Gly Val Ile Ser Gly
2580 2585 2590
Phe Lys Leu Glu Cys Arg Lys Thr Thr Cys Asn Pro Cys Pro Leu Gly
2595 2600 2605
Tyr Lys Glu Glu Asn Asn Thr Gly Glu Cys Cys Gly Arg Cys Leu Pro
2610 2615 2620
Thr Ala Cys Thr Ile Gln Leu Arg Gly Gly Gln Ile Met Thr Leu Lys
2625 2630 2635 2640
Arg Asp Glu Thr Leu Gln Asp Gly Cys Asp Thr His Phe Cys Lys Val
2645 2650 2655
Asn Glu Arg Gly Glu Tyr Phe Trp Glu Lys Arg Val Thr Gly Cys Pro
2660 2665 2670
Pro Phe Asp Glu His Lys Cys Leu Ala Glu Gly Gly Lys Ile Met Lys
2675 2680 2685
Ile Pro Gly Thr Cys Cys Asp Thr Cys Glu Glu Pro Glu Cys Asn Asp
2690 2695 2700
Ile Thr Ala Arg Leu Gln Tyr Val Lys Val Gly Ser Cys Lys Ser Glu
2705 2710 2715 2720
Val Glu Val Asp Ile His Tyr Cys Gln Gly Lys Cys Ala Ser Lys Ala
2725 2730 2735
Met Tyr Ser Ile Asp Ile Asn Asp Val Gln Asp Gln Cys Ser Cys Cys
2740 2745 2750
Ser Pro Thr Arg Thr Glu Pro Met Gln Val Ala Leu His Cys Thr Asn
2755 2760 2765
Gly Ser Val Val Tyr His Glu Val Leu Asn Ala Met Glu Cys Lys Cys
2770 2775 2780
Ser Pro Arg Lys Cys Ser Lys
2785 2790
<210> 3
<211> 1581
<212> PRT
<213> Artificial Sequence
<220>
<223> Single chain FVIII with partial B domain
<220>
<221> MISC_FEATURE
<222> (782)
<223> Xaa = Isoleucine or Cysteine
<400> 3
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Xaa Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr
1570 1575 1580
<210> 4
<211> 1443
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC Heavy chain
<400> 4
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe
<210> 5
<211> 772
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC Light chain
<400> 5
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr
770
<210> 6
<211> 2094
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-6
<400> 6
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu Val Cys
1585 1590 1595 1600
Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys Thr Cys
1605 1610 1615
Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser Gly Cys
1620 1625 1630
Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val Ala Leu
1635 1640 1645
Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro Gly Glu
1650 1655 1660
Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg Lys Trp
1665 1670 1675 1680
Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile Gly Met
1685 1690 1695
Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro Gly Glu
1700 1705 1710
Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro Gly Thr
1715 1720 1725
Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser Val Lys
1730 1735 1740
Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile Glu Leu
1745 1750 1755 1760
Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu Thr His
1765 1770 1775
Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu Gly Lys
1780 1785 1790
Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val Val Leu
1795 1800 1805
Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn Phe Asp
1810 1815 1820
Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val Glu Glu
1825 1830 1835 1840
Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln Cys Ala
1845 1850 1855
Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys His Asn
1860 1865 1870
Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile Leu Thr
1875 1880 1885
Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu Pro Tyr
1890 1895 1900
Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile Gly Asp
1905 1910 1915 1920
Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val Cys Ala
1925 1930 1935
Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys Pro Gln
1940 1945 1950
Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys Glu Trp
1955 1960 1965
Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln His Pro
1970 1975 1980
Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His Ala His
1985 1990 1995 2000
Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys Val Asp
2005 2010 2015
Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe Ala Ser
2020 2025 2030
Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys Gln Ile
2035 2040 2045
Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln Glu Pro
2050 2055 2060
Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro Thr Thr
2065 2070 2075 2080
Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2085 2090
<210> 7
<211> 2098
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-10
<400> 7
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val
1585 1590 1595 1600
Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys
1605 1610 1615
Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys
1620 1625 1630
Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg
1635 1640 1645
Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr
1650 1655 1660
Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg
1665 1670 1675 1680
Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser
1685 1690 1695
Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu
1700 1705 1710
Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser
1715 1720 1725
Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His
1730 1735 1740
Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly
1745 1750 1755 1760
Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys
1765 1770 1775
Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu
1780 1785 1790
Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile
1795 1800 1805
Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys
1810 1815 1820
Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu
1825 1830 1835 1840
Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser
1845 1850 1855
Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala
1860 1865 1870
Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys
1875 1880 1885
Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp
1890 1895 1900
Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu
1905 1910 1915 1920
Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala
1925 1930 1935
His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr
1940 1945 1950
Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr
1955 1960 1965
Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr
1970 1975 1980
Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly
1985 1990 1995 2000
Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln
2005 2010 2015
Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg
2020 2025 2030
Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu
2035 2040 2045
His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala
2050 2055 2060
Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val
2065 2070 2075 2080
Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His
2085 2090 2095
Asp Phe
<210> 8
<211> 2108
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-20
<400> 8
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu Val Cys Pro Ala
1605 1610 1615
Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys Thr Cys Gln Asn
1620 1625 1630
Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser Gly Cys Leu Cys
1635 1640 1645
Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val Ala Leu Glu Arg
1650 1655 1660
Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro Gly Glu Thr Val
1665 1670 1675 1680
Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg Lys Trp Asn Cys
1685 1690 1695
Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile Gly Met Ala His
1700 1705 1710
Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro Gly Glu Cys Gln
1715 1720 1725
Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro Gly Thr Phe Arg
1730 1735 1740
Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser Val Lys Cys Lys
1745 1750 1755 1760
Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile Glu Leu Phe Asp
1765 1770 1775
Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu Thr His Phe Glu
1780 1785 1790
Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu Gly Lys Ala Leu
1795 1800 1805
Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val Val Leu Lys Gln
1810 1815 1820
Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn Phe Asp Gly Ile
1825 1830 1835 1840
Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val Glu Glu Asp Pro
1845 1850 1855
Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln Cys Ala Asp Thr
1860 1865 1870
Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys His Asn Asn Ile
1875 1880 1885
Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile Leu Thr Ser Asp
1890 1895 1900
Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu Pro Tyr Leu Asp
1905 1910 1915 1920
Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile Gly Asp Cys Ala
1925 1930 1935
Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val Cys Ala Gln His
1940 1945 1950
Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys Pro Gln Ser Cys
1955 1960 1965
Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys Glu Trp Arg Tyr
1970 1975 1980
Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln His Pro Glu Pro
1985 1990 1995 2000
Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His Ala His Cys Pro
2005 2010 2015
Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys Val Asp Pro Glu
2020 2025 2030
Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe Ala Ser Gly Lys
2035 2040 2045
Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys Gln Ile Cys His
2050 2055 2060
Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln Glu Pro Gly Gly
2065 2070 2075 2080
Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro Thr Thr Leu Tyr
2085 2090 2095
Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2100 2105
<210> 9
<211> 2118
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-30
<400> 9
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg
1605 1610 1615
Pro Pro Met Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu
1620 1625 1630
Gly Leu Glu Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met
1635 1640 1645
Ser Met Gly Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg
1650 1655 1660
His Glu Asn Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln
1665 1670 1675 1680
Gly Lys Glu Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr
1685 1690 1695
Cys Val Cys Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp
1700 1705 1710
Ala Thr Cys Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly
1715 1720 1725
Leu Lys Tyr Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp
1730 1735 1740
Tyr Cys Gly Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys
1745 1750 1755 1760
Gly Cys Ser His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu
1765 1770 1775
Val Glu Gly Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys
1780 1785 1790
Arg Pro Met Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg
1795 1800 1805
Tyr Ile Ile Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg
1810 1815 1820
His Leu Ser Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val
1825 1830 1835 1840
Cys Gly Leu Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr
1845 1850 1855
Ser Ser Asn Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser
1860 1865 1870
Trp Lys Val Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp
1875 1880 1885
Ser Ser Pro Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val
1890 1895 1900
Asp Ser Ser Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn
1905 1910 1915 1920
Lys Leu Val Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr
1925 1930 1935
Cys Ser Cys Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile
1940 1945 1950
Ala Ala Tyr Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp
1955 1960 1965
Arg Thr Ala Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg
1970 1975 1980
Glu Asn Gly Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala
1985 1990 1995 2000
Cys Gln Val Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln
2005 2010 2015
Cys Val Glu Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp
2020 2025 2030
Glu Leu Leu Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu
2035 2040 2045
Val Ala Gly Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro
2050 2055 2060
Ser Asp Pro Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu
2065 2070 2075 2080
Thr Cys Glu Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr
2085 2090 2095
Asp Ala Pro Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu
2100 2105 2110
Pro Pro Leu His Asp Phe
2115
<210> 10
<211> 2128
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-40
<400> 10
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1620 1625 1630
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1635 1640 1645
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1650 1655 1660
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1665 1670 1675 1680
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1685 1690 1695
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1700 1705 1710
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1715 1720 1725
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1730 1735 1740
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1745 1750 1755 1760
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1765 1770 1775
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1780 1785 1790
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1795 1800 1805
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1810 1815 1820
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1825 1830 1835 1840
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1845 1850 1855
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1860 1865 1870
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1875 1880 1885
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1890 1895 1900
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1905 1910 1915 1920
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
1925 1930 1935
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
1940 1945 1950
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
1955 1960 1965
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
1970 1975 1980
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
1985 1990 1995 2000
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2005 2010 2015
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2020 2025 2030
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2035 2040 2045
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2050 2055 2060
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2065 2070 2075 2080
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2085 2090 2095
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2100 2105 2110
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2115 2120 2125
<210> 11
<211> 2138
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-50
<400> 11
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Leu Ser Cys Arg Pro Pro Met Val Lys Leu Val Cys Pro Ala Asp Asn
1635 1640 1645
Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys Thr Cys Gln Asn Tyr Asp
1650 1655 1660
Leu Glu Cys Met Ser Met Gly Cys Val Ser Gly Cys Leu Cys Pro Pro
1665 1670 1675 1680
Gly Met Val Arg His Glu Asn Arg Cys Val Ala Leu Glu Arg Cys Pro
1685 1690 1695
Cys Phe His Gln Gly Lys Glu Tyr Ala Pro Gly Glu Thr Val Lys Ile
1700 1705 1710
Gly Cys Asn Thr Cys Val Cys Arg Asp Arg Lys Trp Asn Cys Thr Asp
1715 1720 1725
His Val Cys Asp Ala Thr Cys Ser Thr Ile Gly Met Ala His Tyr Leu
1730 1735 1740
Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro Gly Glu Cys Gln Tyr Val
1745 1750 1755 1760
Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro Gly Thr Phe Arg Ile Leu
1765 1770 1775
Val Gly Asn Lys Gly Cys Ser His Pro Ser Val Lys Cys Lys Lys Arg
1780 1785 1790
Val Thr Ile Leu Val Glu Gly Gly Glu Ile Glu Leu Phe Asp Gly Glu
1795 1800 1805
Val Asn Val Lys Arg Pro Met Lys Asp Glu Thr His Phe Glu Val Val
1810 1815 1820
Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu Gly Lys Ala Leu Ser Val
1825 1830 1835 1840
Val Trp Asp Arg His Leu Ser Ile Ser Val Val Leu Lys Gln Thr Tyr
1845 1850 1855
Gln Glu Lys Val Cys Gly Leu Cys Gly Asn Phe Asp Gly Ile Gln Asn
1860 1865 1870
Asn Asp Leu Thr Ser Ser Asn Leu Gln Val Glu Glu Asp Pro Val Asp
1875 1880 1885
Phe Gly Asn Ser Trp Lys Val Ser Ser Gln Cys Ala Asp Thr Arg Lys
1890 1895 1900
Val Pro Leu Asp Ser Ser Pro Ala Thr Cys His Asn Asn Ile Met Lys
1905 1910 1915 1920
Gln Thr Met Val Asp Ser Ser Cys Arg Ile Leu Thr Ser Asp Val Phe
1925 1930 1935
Gln Asp Cys Asn Lys Leu Val Asp Pro Glu Pro Tyr Leu Asp Val Cys
1940 1945 1950
Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile Gly Asp Cys Ala Cys Phe
1955 1960 1965
Cys Asp Thr Ile Ala Ala Tyr Ala His Val Cys Ala Gln His Gly Lys
1970 1975 1980
Val Val Thr Trp Arg Thr Ala Thr Leu Cys Pro Gln Ser Cys Glu Glu
1985 1990 1995 2000
Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys Glu Trp Arg Tyr Asn Ser
2005 2010 2015
Cys Ala Pro Ala Cys Gln Val Thr Cys Gln His Pro Glu Pro Leu Ala
2020 2025 2030
Cys Pro Val Gln Cys Val Glu Gly Cys His Ala His Cys Pro Pro Gly
2035 2040 2045
Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys Val Asp Pro Glu Asp Cys
2050 2055 2060
Pro Val Cys Glu Val Ala Gly Arg Arg Phe Ala Ser Gly Lys Lys Val
2065 2070 2075 2080
Thr Leu Asn Pro Ser Asp Pro Glu His Cys Gln Ile Cys His Cys Asp
2085 2090 2095
Val Val Asn Leu Thr Cys Glu Ala Cys Gln Glu Pro Gly Gly Leu Val
2100 2105 2110
Val Pro Pro Thr Asp Ala Pro Val Ser Pro Thr Thr Leu Tyr Val Glu
2115 2120 2125
Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2130 2135
<210> 12
<211> 2148
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-60
<400> 12
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro
1635 1640 1645
Met Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu
1650 1655 1660
Glu Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met
1665 1670 1675 1680
Gly Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu
1685 1690 1695
Asn Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys
1700 1705 1710
Glu Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val
1715 1720 1725
Cys Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr
1730 1735 1740
Cys Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys
1745 1750 1755 1760
Tyr Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys
1765 1770 1775
Gly Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys
1780 1785 1790
Ser His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu
1795 1800 1805
Gly Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro
1810 1815 1820
Met Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile
1825 1830 1835 1840
Ile Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu
1845 1850 1855
Ser Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly
1860 1865 1870
Leu Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser
1875 1880 1885
Asn Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys
1890 1895 1900
Val Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser
1905 1910 1915 1920
Pro Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser
1925 1930 1935
Ser Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu
1940 1945 1950
Val Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser
1955 1960 1965
Cys Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala
1970 1975 1980
Tyr Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr
1985 1990 1995 2000
Ala Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn
2005 2010 2015
Gly Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln
2020 2025 2030
Val Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val
2035 2040 2045
Glu Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu
2050 2055 2060
Leu Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala
2065 2070 2075 2080
Gly Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp
2085 2090 2095
Pro Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys
2100 2105 2110
Glu Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala
2115 2120 2125
Pro Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro
2130 2135 2140
Leu His Asp Phe
2145
<210> 13
<211> 2178
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-90
<400> 13
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1635 1640 1645
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1650 1655 1660
Gly Gly Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val
1665 1670 1675 1680
Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys
1685 1690 1695
Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys
1700 1705 1710
Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg
1715 1720 1725
Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr
1730 1735 1740
Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg
1745 1750 1755 1760
Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser
1765 1770 1775
Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu
1780 1785 1790
Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser
1795 1800 1805
Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His
1810 1815 1820
Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly
1825 1830 1835 1840
Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys
1845 1850 1855
Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu
1860 1865 1870
Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile
1875 1880 1885
Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys
1890 1895 1900
Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu
1905 1910 1915 1920
Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser
1925 1930 1935
Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala
1940 1945 1950
Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys
1955 1960 1965
Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp
1970 1975 1980
Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu
1985 1990 1995 2000
Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala
2005 2010 2015
His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr
2020 2025 2030
Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr
2035 2040 2045
Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr
2050 2055 2060
Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly
2065 2070 2075 2080
Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln
2085 2090 2095
Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg
2100 2105 2110
Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu
2115 2120 2125
His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala
2130 2135 2140
Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val
2145 2150 2155 2160
Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His
2165 2170 2175
Asp Phe
<210> 14
<211> 2208
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120
<400> 14
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1635 1640 1645
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1650 1655 1660
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1665 1670 1675 1680
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1685 1690 1695
Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1700 1705 1710
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1715 1720 1725
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1730 1735 1740
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1745 1750 1755 1760
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1765 1770 1775
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1780 1785 1790
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1795 1800 1805
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1810 1815 1820
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1825 1830 1835 1840
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1845 1850 1855
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1860 1865 1870
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1875 1880 1885
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1890 1895 1900
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1905 1910 1915 1920
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1925 1930 1935
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1940 1945 1950
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1955 1960 1965
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1970 1975 1980
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1985 1990 1995 2000
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
2005 2010 2015
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
2020 2025 2030
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
2035 2040 2045
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
2050 2055 2060
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
2065 2070 2075 2080
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2085 2090 2095
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2100 2105 2110
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2115 2120 2125
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2130 2135 2140
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2145 2150 2155 2160
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2165 2170 2175
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2180 2185 2190
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2195 2200 2205
<210> 15
<211> 2208
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH1
<400> 15
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Gly Gly Gly
1570 1575 1580
Ser Leu Val Pro Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1585 1590 1595 1600
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1605 1610 1615
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1620 1625 1630
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1635 1640 1645
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1650 1655 1660
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1665 1670 1675 1680
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1685 1690 1695
Gly Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1700 1705 1710
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1715 1720 1725
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1730 1735 1740
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1745 1750 1755 1760
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1765 1770 1775
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1780 1785 1790
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1795 1800 1805
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1810 1815 1820
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1825 1830 1835 1840
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1845 1850 1855
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1860 1865 1870
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1875 1880 1885
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1890 1895 1900
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1905 1910 1915 1920
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1925 1930 1935
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1940 1945 1950
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1955 1960 1965
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1970 1975 1980
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1985 1990 1995 2000
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
2005 2010 2015
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
2020 2025 2030
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
2035 2040 2045
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
2050 2055 2060
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
2065 2070 2075 2080
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2085 2090 2095
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2100 2105 2110
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2115 2120 2125
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2130 2135 2140
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2145 2150 2155 2160
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2165 2170 2175
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2180 2185 2190
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2195 2200 2205
<210> 16
<211> 2208
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH2
<400> 16
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Ser Lys
1570 1575 1580
Asn Asn Ala Ile Glu Pro Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly
1585 1590 1595 1600
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1605 1610 1615
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1620 1625 1630
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1635 1640 1645
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1650 1655 1660
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1665 1670 1675 1680
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1685 1690 1695
Gly Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1700 1705 1710
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1715 1720 1725
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1730 1735 1740
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1745 1750 1755 1760
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1765 1770 1775
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1780 1785 1790
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1795 1800 1805
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1810 1815 1820
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1825 1830 1835 1840
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1845 1850 1855
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1860 1865 1870
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1875 1880 1885
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1890 1895 1900
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1905 1910 1915 1920
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1925 1930 1935
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1940 1945 1950
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1955 1960 1965
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1970 1975 1980
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1985 1990 1995 2000
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
2005 2010 2015
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
2020 2025 2030
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
2035 2040 2045
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
2050 2055 2060
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
2065 2070 2075 2080
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2085 2090 2095
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2100 2105 2110
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2115 2120 2125
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2130 2135 2140
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2145 2150 2155 2160
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2165 2170 2175
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2180 2185 2190
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2195 2200 2205
<210> 17
<211> 2388
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-300
<400> 17
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1635 1640 1645
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1650 1655 1660
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1665 1670 1675 1680
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1685 1690 1695
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1700 1705 1710
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1715 1720 1725
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1730 1735 1740
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1745 1750 1755 1760
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1765 1770 1775
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1780 1785 1790
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1795 1800 1805
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1810 1815 1820
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1825 1830 1835 1840
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1845 1850 1855
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1860 1865 1870
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro
1875 1880 1885
Met Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu
1890 1895 1900
Glu Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met
1905 1910 1915 1920
Gly Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu
1925 1930 1935
Asn Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys
1940 1945 1950
Glu Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val
1955 1960 1965
Cys Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr
1970 1975 1980
Cys Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys
1985 1990 1995 2000
Tyr Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys
2005 2010 2015
Gly Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys
2020 2025 2030
Ser His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu
2035 2040 2045
Gly Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro
2050 2055 2060
Met Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile
2065 2070 2075 2080
Ile Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu
2085 2090 2095
Ser Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly
2100 2105 2110
Leu Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser
2115 2120 2125
Asn Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys
2130 2135 2140
Val Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser
2145 2150 2155 2160
Pro Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser
2165 2170 2175
Ser Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu
2180 2185 2190
Val Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser
2195 2200 2205
Cys Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala
2210 2215 2220
Tyr Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr
2225 2230 2235 2240
Ala Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn
2245 2250 2255
Gly Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln
2260 2265 2270
Val Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val
2275 2280 2285
Glu Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu
2290 2295 2300
Leu Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala
2305 2310 2315 2320
Gly Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp
2325 2330 2335
Pro Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys
2340 2345 2350
Glu Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala
2355 2360 2365
Pro Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro
2370 2375 2380
Leu His Asp Phe
2385
<210> 18
<211> 2745
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3/D'D3-120
<400> 18
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1635 1640 1645
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1650 1655 1660
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1665 1670 1675 1680
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1685 1690 1695
Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1700 1705 1710
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1715 1720 1725
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1730 1735 1740
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1745 1750 1755 1760
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1765 1770 1775
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1780 1785 1790
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1795 1800 1805
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1810 1815 1820
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1825 1830 1835 1840
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1845 1850 1855
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1860 1865 1870
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1875 1880 1885
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1890 1895 1900
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1905 1910 1915 1920
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1925 1930 1935
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1940 1945 1950
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1955 1960 1965
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1970 1975 1980
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1985 1990 1995 2000
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
2005 2010 2015
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
2020 2025 2030
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
2035 2040 2045
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
2050 2055 2060
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
2065 2070 2075 2080
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2085 2090 2095
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2100 2105 2110
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2115 2120 2125
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2130 2135 2140
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2145 2150 2155 2160
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2165 2170 2175
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2180 2185 2190
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2195 2200 2205
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
2210 2215 2220
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gly Ser Leu
2225 2230 2235 2240
Ser Cys Arg Pro Pro Met Val Lys Leu Val Cys Pro Ala Asp Asn Leu
2245 2250 2255
Arg Ala Glu Gly Leu Glu Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu
2260 2265 2270
Glu Cys Met Ser Met Gly Cys Val Ser Gly Cys Leu Cys Pro Pro Gly
2275 2280 2285
Met Val Arg His Glu Asn Arg Cys Val Ala Leu Glu Arg Cys Pro Cys
2290 2295 2300
Phe His Gln Gly Lys Glu Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly
2305 2310 2315 2320
Cys Asn Thr Cys Val Cys Arg Asp Arg Lys Trp Asn Cys Thr Asp His
2325 2330 2335
Val Cys Asp Ala Thr Cys Ser Thr Ile Gly Met Ala His Tyr Leu Thr
2340 2345 2350
Phe Asp Gly Leu Lys Tyr Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu
2355 2360 2365
Val Gln Asp Tyr Cys Gly Ser Asn Pro Gly Thr Phe Arg Ile Leu Val
2370 2375 2380
Gly Asn Lys Gly Cys Ser His Pro Ser Val Lys Cys Lys Lys Arg Val
2385 2390 2395 2400
Thr Ile Leu Val Glu Gly Gly Glu Ile Glu Leu Phe Asp Gly Glu Val
2405 2410 2415
Asn Val Lys Arg Pro Met Lys Asp Glu Thr His Phe Glu Val Val Glu
2420 2425 2430
Ser Gly Arg Tyr Ile Ile Leu Leu Leu Gly Lys Ala Leu Ser Val Val
2435 2440 2445
Trp Asp Arg His Leu Ser Ile Ser Val Val Leu Lys Gln Thr Tyr Gln
2450 2455 2460
Glu Lys Val Cys Gly Leu Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn
2465 2470 2475 2480
Asp Leu Thr Ser Ser Asn Leu Gln Val Glu Glu Asp Pro Val Asp Phe
2485 2490 2495
Gly Asn Ser Trp Lys Val Ser Ser Gln Cys Ala Asp Thr Arg Lys Val
2500 2505 2510
Pro Leu Asp Ser Ser Pro Ala Thr Cys His Asn Asn Ile Met Lys Gln
2515 2520 2525
Thr Met Val Asp Ser Ser Cys Arg Ile Leu Thr Ser Asp Val Phe Gln
2530 2535 2540
Asp Cys Asn Lys Leu Val Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile
2545 2550 2555 2560
Tyr Asp Thr Cys Ser Cys Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys
2565 2570 2575
Asp Thr Ile Ala Ala Tyr Ala His Val Cys Ala Gln His Gly Lys Val
2580 2585 2590
Val Thr Trp Arg Thr Ala Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg
2595 2600 2605
Asn Leu Arg Glu Asn Gly Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys
2610 2615 2620
Ala Pro Ala Cys Gln Val Thr Cys Gln His Pro Glu Pro Leu Ala Cys
2625 2630 2635 2640
Pro Val Gln Cys Val Glu Gly Cys His Ala His Cys Pro Pro Gly Lys
2645 2650 2655
Ile Leu Asp Glu Leu Leu Gln Thr Cys Val Asp Pro Glu Asp Cys Pro
2660 2665 2670
Val Cys Glu Val Ala Gly Arg Arg Phe Ala Ser Gly Lys Lys Val Thr
2675 2680 2685
Leu Asn Pro Ser Asp Pro Glu His Cys Gln Ile Cys His Cys Asp Val
2690 2695 2700
Val Asn Leu Thr Cys Glu Ala Cys Gln Glu Pro Gly Gly Leu Val Val
2705 2710 2715 2720
Pro Pro Thr Asp Ala Pro Val Ser Pro Thr Thr Leu Tyr Val Glu Asp
2725 2730 2735
Ile Ser Glu Pro Pro Leu His Asp Phe
2740 2745
<210> 19
<211> 2949
<212> PRT
<213> Artificial Sequence
<220>
<223> D'D3/scFVIII-120 of the complex of D'D3/scFVIII-120 and D'D3
<400> 19
Ala Glu Gly Thr Arg Gly Arg Ser Ser Thr Ala Arg Cys Ser Leu Phe
1 5 10 15
Gly Ser Asp Phe Val Asn Thr Phe Asp Gly Ser Met Tyr Ser Phe Ala
20 25 30
Gly Tyr Cys Ser Tyr Leu Leu Ala Gly Gly Cys Gln Lys Arg Ser Phe
35 40 45
Ser Ile Ile Gly Asp Phe Gln Asn Gly Lys Arg Val Ser Leu Ser Val
50 55 60
Tyr Leu Gly Glu Phe Phe Asp Ile His Leu Phe Val Asn Gly Thr Val
65 70 75 80
Thr Gln Gly Asp Gln Arg Val Ser Met Pro Tyr Ala Ser Lys Gly Leu
85 90 95
Tyr Leu Glu Thr Glu Ala Gly Tyr Tyr Lys Leu Ser Gly Glu Ala Tyr
100 105 110
Gly Phe Val Ala Arg Ile Asp Gly Ser Gly Asn Phe Gln Val Leu Leu
115 120 125
Ser Asp Arg Tyr Phe Asn Lys Thr Cys Gly Leu Cys Gly Asn Phe Asn
130 135 140
Ile Phe Ala Glu Asp Asp Phe Met Thr Gln Glu Gly Thr Leu Thr Ser
145 150 155 160
Asp Pro Tyr Asp Phe Ala Asn Ser Trp Ala Leu Ser Ser Gly Glu Gln
165 170 175
Trp Cys Glu Arg Ala Ser Pro Pro Ser Ser Ser Cys Asn Ile Ser Ser
180 185 190
Gly Glu Met Gln Lys Gly Leu Trp Glu Gln Cys Gln Leu Leu Lys Ser
195 200 205
Thr Ser Val Phe Ala Arg Cys His Pro Leu Val Asp Pro Glu Pro Phe
210 215 220
Val Ala Leu Cys Glu Lys Thr Leu Cys Glu Cys Ala Gly Gly Leu Glu
225 230 235 240
Cys Ala Cys Pro Ala Leu Leu Glu Tyr Ala Arg Thr Cys Ala Gln Glu
245 250 255
Gly Met Val Leu Tyr Gly Trp Thr Asp His Ser Ala Cys Ser Pro Val
260 265 270
Cys Pro Ala Gly Met Glu Tyr Arg Gln Cys Val Ser Pro Cys Ala Arg
275 280 285
Thr Cys Gln Ser Leu His Ile Asn Glu Met Cys Gln Glu Arg Cys Val
290 295 300
Asp Gly Cys Ser Cys Pro Glu Gly Gln Leu Leu Asp Glu Gly Leu Cys
305 310 315 320
Val Glu Ser Thr Glu Cys Pro Cys Val His Ser Gly Lys Arg Tyr Pro
325 330 335
Pro Gly Thr Ser Leu Ser Arg Asp Cys Asn Thr Cys Ile Cys Arg Asn
340 345 350
Ser Gln Trp Ile Cys Ser Asn Glu Glu Cys Pro Gly Glu Cys Leu Val
355 360 365
Thr Gly Gln Ser His Phe Lys Ser Phe Asp Asn Arg Tyr Phe Thr Phe
370 375 380
Ser Gly Ile Cys Gln Tyr Leu Leu Ala Arg Asp Cys Gln Asp His Ser
385 390 395 400
Phe Ser Ile Val Ile Glu Thr Val Gln Cys Ala Asp Asp Arg Asp Ala
405 410 415
Val Cys Thr Arg Ser Val Thr Val Arg Leu Pro Gly Leu His Asn Ser
420 425 430
Leu Val Lys Leu Lys His Gly Ala Gly Val Ala Met Asp Gly Gln Asp
435 440 445
Val Gln Leu Pro Leu Leu Lys Gly Asp Leu Arg Ile Gln His Thr Val
450 455 460
Thr Ala Ser Val Arg Leu Ser Tyr Gly Glu Asp Leu Gln Met Asp Trp
465 470 475 480
Asp Gly Arg Gly Arg Leu Leu Val Lys Leu Ser Pro Val Tyr Ala Gly
485 490 495
Lys Thr Cys Gly Leu Cys Gly Asn Tyr Asn Gly Asn Gln Gly Asp Asp
500 505 510
Phe Leu Thr Pro Ser Gly Leu Ala Glu Pro Arg Val Glu Asp Phe Gly
515 520 525
Asn Ala Trp Lys Leu His Gly Asp Cys Gln Asp Leu Gln Lys Gln His
530 535 540
Ser Asp Pro Cys Ala Leu Asn Pro Arg Met Thr Arg Phe Ser Glu Glu
545 550 555 560
Ala Cys Ala Val Leu Thr Ser Pro Thr Phe Glu Ala Cys His Arg Ala
565 570 575
Val Ser Pro Leu Pro Tyr Leu Arg Asn Cys Arg Tyr Asp Val Cys Ser
580 585 590
Cys Ser Asp Gly Arg Glu Cys Leu Cys Gly Ala Leu Ala Ser Tyr Ala
595 600 605
Ala Ala Cys Ala Gly Arg Gly Val Arg Val Ala Trp Arg Glu Pro Gly
610 615 620
Arg Cys Glu Leu Asn Cys Pro Lys Gly Gln Val Tyr Leu Gln Cys Gly
625 630 635 640
Thr Pro Cys Asn Leu Thr Cys Arg Ser Leu Ser Tyr Pro Asp Glu Glu
645 650 655
Cys Asn Glu Ala Cys Leu Glu Gly Cys Phe Cys Pro Pro Gly Leu Tyr
660 665 670
Met Asp Glu Arg Gly Asp Cys Val Pro Lys Ala Gln Cys Pro Cys Tyr
675 680 685
Tyr Asp Gly Glu Ile Phe Gln Pro Glu Asp Ile Phe Ser Asp His His
690 695 700
Thr Met Cys Tyr Cys Glu Asp Gly Phe Met His Cys Thr Met Ser Gly
705 710 715 720
Val Pro Gly Ser Leu Leu Pro Asp Ala Val Leu Ser Ser Pro Leu Ser
725 730 735
His Arg Ser Lys Arg Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
740 745 750
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
755 760 765
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
770 775 780
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
785 790 795 800
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
805 810 815
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
820 825 830
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
835 840 845
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
850 855 860
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
865 870 875 880
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
885 890 895
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
900 905 910
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
915 920 925
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
930 935 940
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
945 950 955 960
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
965 970 975
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
980 985 990
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
995 1000 1005
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1010 1015 1020
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1025 1030 1035 1040
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
1045 1050 1055
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
1060 1065 1070
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
1075 1080 1085
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
1090 1095 1100
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
1105 1110 1115 1120
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
1125 1130 1135
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
1140 1145 1150
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
1155 1160 1165
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
1170 1175 1180
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
1185 1190 1195 1200
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
1205 1210 1215
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
1220 1225 1230
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
1235 1240 1245
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1250 1255 1260
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1265 1270 1275 1280
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1285 1290 1295
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1300 1305 1310
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1315 1320 1325
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1330 1335 1340
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1345 1350 1355 1360
Gly Ser Leu Val Pro Arg Gly Ser Ala Thr Arg Arg Tyr Tyr Leu Gly
1365 1370 1375
Ala Val Glu Leu Ser Trp Asp Tyr Met Gln Ser Asp Leu Gly Glu Leu
1380 1385 1390
Pro Val Asp Ala Arg Phe Pro Pro Arg Val Pro Lys Ser Phe Pro Phe
1395 1400 1405
Asn Thr Ser Val Val Tyr Lys Lys Thr Leu Phe Val Glu Phe Thr Asp
1410 1415 1420
His Leu Phe Asn Ile Ala Lys Pro Arg Pro Pro Trp Met Gly Leu Leu
1425 1430 1435 1440
Gly Pro Thr Ile Gln Ala Glu Val Tyr Asp Thr Val Val Ile Thr Leu
1445 1450 1455
Lys Asn Met Ala Ser His Pro Val Ser Leu His Ala Val Gly Val Ser
1460 1465 1470
Tyr Trp Lys Ala Ser Glu Gly Ala Glu Tyr Asp Asp Gln Thr Ser Gln
1475 1480 1485
Arg Glu Lys Glu Asp Asp Lys Val Phe Pro Gly Gly Ser His Thr Tyr
1490 1495 1500
Val Trp Gln Val Leu Lys Glu Asn Gly Pro Met Ala Ser Asp Pro Leu
1505 1510 1515 1520
Cys Leu Thr Tyr Ser Tyr Leu Ser His Val Asp Leu Val Lys Asp Leu
1525 1530 1535
Asn Ser Gly Leu Ile Gly Ala Leu Leu Val Cys Arg Glu Gly Ser Leu
1540 1545 1550
Ala Lys Glu Lys Thr Gln Thr Leu His Lys Phe Ile Leu Leu Phe Ala
1555 1560 1565
Val Phe Asp Glu Gly Lys Ser Trp His Ser Glu Thr Lys Asn Ser Leu
1570 1575 1580
Met Gln Asp Arg Asp Ala Ala Ser Ala Arg Ala Trp Pro Lys Met His
1585 1590 1595 1600
Thr Val Asn Gly Tyr Val Asn Arg Ser Leu Pro Gly Leu Ile Gly Cys
1605 1610 1615
His Arg Lys Ser Val Tyr Trp His Val Ile Gly Met Gly Thr Thr Pro
1620 1625 1630
Glu Val His Ser Ile Phe Leu Glu Gly His Thr Phe Leu Val Arg Asn
1635 1640 1645
His Arg Gln Ala Ser Leu Glu Ile Ser Pro Ile Thr Phe Leu Thr Ala
1650 1655 1660
Gln Thr Leu Leu Met Asp Leu Gly Gln Phe Leu Leu Phe Cys His Ile
1665 1670 1675 1680
Ser Ser His Gln His Asp Gly Met Glu Ala Tyr Val Lys Val Asp Ser
1685 1690 1695
Cys Pro Glu Glu Pro Gln Leu Arg Met Lys Asn Asn Glu Glu Ala Glu
1700 1705 1710
Asp Tyr Asp Asp Asp Leu Thr Asp Ser Glu Met Asp Val Val Arg Phe
1715 1720 1725
Asp Asp Asp Asn Ser Pro Ser Phe Ile Gln Ile Arg Ser Val Ala Lys
1730 1735 1740
Lys His Pro Lys Thr Trp Val His Tyr Ile Ala Ala Glu Glu Glu Asp
1745 1750 1755 1760
Trp Asp Tyr Ala Pro Leu Val Leu Ala Pro Asp Asp Arg Ser Tyr Lys
1765 1770 1775
Ser Gln Tyr Leu Asn Asn Gly Pro Gln Arg Ile Gly Arg Lys Tyr Lys
1780 1785 1790
Lys Val Arg Phe Met Ala Tyr Thr Asp Glu Thr Phe Lys Thr Arg Glu
1795 1800 1805
Ala Ile Gln His Glu Ser Gly Ile Leu Gly Pro Leu Leu Tyr Gly Glu
1810 1815 1820
Val Gly Asp Thr Leu Leu Ile Ile Phe Lys Asn Gln Ala Ser Arg Pro
1825 1830 1835 1840
Tyr Asn Ile Tyr Pro His Gly Ile Thr Asp Val Arg Pro Leu Tyr Ser
1845 1850 1855
Arg Arg Leu Pro Lys Gly Val Lys His Leu Lys Asp Phe Pro Ile Leu
1860 1865 1870
Pro Gly Glu Ile Phe Lys Tyr Lys Trp Thr Val Thr Val Glu Asp Gly
1875 1880 1885
Pro Thr Lys Ser Asp Pro Arg Cys Leu Thr Arg Tyr Tyr Ser Ser Phe
1890 1895 1900
Val Asn Met Glu Arg Asp Leu Ala Ser Gly Leu Ile Gly Pro Leu Leu
1905 1910 1915 1920
Ile Cys Tyr Lys Glu Ser Val Asp Gln Arg Gly Asn Gln Ile Met Ser
1925 1930 1935
Asp Lys Arg Asn Val Ile Leu Phe Ser Val Phe Asp Glu Asn Arg Ser
1940 1945 1950
Trp Tyr Leu Thr Glu Asn Ile Gln Arg Phe Leu Pro Asn Pro Ala Gly
1955 1960 1965
Val Gln Leu Glu Asp Pro Glu Phe Gln Ala Ser Asn Ile Met His Ser
1970 1975 1980
Ile Asn Gly Tyr Val Phe Asp Ser Leu Gln Leu Ser Val Cys Leu His
1985 1990 1995 2000
Glu Val Ala Tyr Trp Tyr Ile Leu Ser Ile Gly Ala Gln Thr Asp Phe
2005 2010 2015
Leu Ser Val Phe Phe Ser Gly Tyr Thr Phe Lys His Lys Met Val Tyr
2020 2025 2030
Glu Asp Thr Leu Thr Leu Phe Pro Phe Ser Gly Glu Thr Val Phe Met
2035 2040 2045
Ser Met Glu Asn Pro Gly Leu Trp Ile Leu Gly Cys His Asn Ser Asp
2050 2055 2060
Phe Arg Asn Arg Gly Met Thr Ala Leu Leu Lys Val Ser Ser Cys Asp
2065 2070 2075 2080
Lys Asn Thr Gly Asp Tyr Tyr Glu Asp Ser Tyr Glu Asp Ile Ser Ala
2085 2090 2095
Tyr Leu Leu Ser Lys Asn Asn Ala Ile Glu Pro Arg Ser Phe Ser Gln
2100 2105 2110
Asn Ser Arg His Pro Ser Thr Arg Gln Lys Gln Phe Asn Ala Thr Thr
2115 2120 2125
Ile Pro Glu Asn Asp Ile Glu Lys Thr Asp Pro Trp Phe Ala His Arg
2130 2135 2140
Thr Pro Met Pro Lys Cys Gln Asn Val Ser Ser Ser Asp Leu Leu Met
2145 2150 2155 2160
Leu Leu Arg Gln Ser Pro Thr Pro His Gly Leu Ser Leu Ser Asp Leu
2165 2170 2175
Gln Glu Ala Lys Tyr Glu Thr Phe Ser Asp Asp Pro Ser Pro Gly Ala
2180 2185 2190
Ile Asp Ser Asn Asn Ser Leu Ser Glu Met Thr His Phe Arg Pro Gln
2195 2200 2205
Leu His His Ser Gly Asp Met Val Phe Thr Pro Glu Ser Gly Leu Gln
2210 2215 2220
Leu Arg Leu Asn Glu Lys Leu Gly Thr Thr Ala Ala Thr Glu Leu Lys
2225 2230 2235 2240
Lys Leu Asp Phe Lys Val Ser Ser Thr Ser Asn Asn Leu Ile Ser Thr
2245 2250 2255
Ile Pro Ser Asp Asn Leu Ala Ala Gly Thr Asp Asn Thr Ser Thr Leu
2260 2265 2270
Gln Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu
2275 2280 2285
Met Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser
2290 2295 2300
Pro Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val
2305 2310 2315 2320
Glu Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg
2325 2330 2335
Asn Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe
2340 2345 2350
Gln Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu
2355 2360 2365
Leu Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val
2370 2375 2380
Glu Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr
2385 2390 2395 2400
Ser Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly
2405 2410 2415
Ala Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr
2420 2425 2430
Phe Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp
2435 2440 2445
Cys Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val
2450 2455 2460
His Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu
2465 2470 2475 2480
Asn Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe
2485 2490 2495
Phe Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met
2500 2505 2510
Glu Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr
2515 2520 2525
Phe Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp
2530 2535 2540
Thr Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr
2545 2550 2555 2560
Leu Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser
2565 2570 2575
Gly His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu
2580 2585 2590
Tyr Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser
2595 2600 2605
Lys Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His
2610 2615 2620
Ala Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr
2625 2630 2635 2640
Pro Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala
2645 2650 2655
Ser Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr
2660 2665 2670
Ser Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile
2675 2680 2685
Lys Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln
2690 2695 2700
Gly Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile
2705 2710 2715 2720
Met Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser
2725 2730 2735
Thr Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile
2740 2745 2750
Lys His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu
2755 2760 2765
His Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met
2770 2775 2780
Gly Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys
2785 2790 2795 2800
Ala Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met
2805 2810 2815
Phe Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg
2820 2825 2830
Ser Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln
2835 2840 2845
Val Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly
2850 2855 2860
Val Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser
2865 2870 2875 2880
Ser Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys
2885 2890 2895
Val Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn
2900 2905 2910
Ser Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln
2915 2920 2925
Ser Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu
2930 2935 2940
Ala Gln Asp Leu Tyr
2945
<210> 20
<211> 1248
<212> PRT
<213> Artificial Sequence
<220>
<223> D'D3 of the complex of D'D3/scFVIII-120 and D'D3
<400> 20
Ala Glu Gly Thr Arg Gly Arg Ser Ser Thr Ala Arg Cys Ser Leu Phe
1 5 10 15
Gly Ser Asp Phe Val Asn Thr Phe Asp Gly Ser Met Tyr Ser Phe Ala
20 25 30
Gly Tyr Cys Ser Tyr Leu Leu Ala Gly Gly Cys Gln Lys Arg Ser Phe
35 40 45
Ser Ile Ile Gly Asp Phe Gln Asn Gly Lys Arg Val Ser Leu Ser Val
50 55 60
Tyr Leu Gly Glu Phe Phe Asp Ile His Leu Phe Val Asn Gly Thr Val
65 70 75 80
Thr Gln Gly Asp Gln Arg Val Ser Met Pro Tyr Ala Ser Lys Gly Leu
85 90 95
Tyr Leu Glu Thr Glu Ala Gly Tyr Tyr Lys Leu Ser Gly Glu Ala Tyr
100 105 110
Gly Phe Val Ala Arg Ile Asp Gly Ser Gly Asn Phe Gln Val Leu Leu
115 120 125
Ser Asp Arg Tyr Phe Asn Lys Thr Cys Gly Leu Cys Gly Asn Phe Asn
130 135 140
Ile Phe Ala Glu Asp Asp Phe Met Thr Gln Glu Gly Thr Leu Thr Ser
145 150 155 160
Asp Pro Tyr Asp Phe Ala Asn Ser Trp Ala Leu Ser Ser Gly Glu Gln
165 170 175
Trp Cys Glu Arg Ala Ser Pro Pro Ser Ser Ser Cys Asn Ile Ser Ser
180 185 190
Gly Glu Met Gln Lys Gly Leu Trp Glu Gln Cys Gln Leu Leu Lys Ser
195 200 205
Thr Ser Val Phe Ala Arg Cys His Pro Leu Val Asp Pro Glu Pro Phe
210 215 220
Val Ala Leu Cys Glu Lys Thr Leu Cys Glu Cys Ala Gly Gly Leu Glu
225 230 235 240
Cys Ala Cys Pro Ala Leu Leu Glu Tyr Ala Arg Thr Cys Ala Gln Glu
245 250 255
Gly Met Val Leu Tyr Gly Trp Thr Asp His Ser Ala Cys Ser Pro Val
260 265 270
Cys Pro Ala Gly Met Glu Tyr Arg Gln Cys Val Ser Pro Cys Ala Arg
275 280 285
Thr Cys Gln Ser Leu His Ile Asn Glu Met Cys Gln Glu Arg Cys Val
290 295 300
Asp Gly Cys Ser Cys Pro Glu Gly Gln Leu Leu Asp Glu Gly Leu Cys
305 310 315 320
Val Glu Ser Thr Glu Cys Pro Cys Val His Ser Gly Lys Arg Tyr Pro
325 330 335
Pro Gly Thr Ser Leu Ser Arg Asp Cys Asn Thr Cys Ile Cys Arg Asn
340 345 350
Ser Gln Trp Ile Cys Ser Asn Glu Glu Cys Pro Gly Glu Cys Leu Val
355 360 365
Thr Gly Gln Ser His Phe Lys Ser Phe Asp Asn Arg Tyr Phe Thr Phe
370 375 380
Ser Gly Ile Cys Gln Tyr Leu Leu Ala Arg Asp Cys Gln Asp His Ser
385 390 395 400
Phe Ser Ile Val Ile Glu Thr Val Gln Cys Ala Asp Asp Arg Asp Ala
405 410 415
Val Cys Thr Arg Ser Val Thr Val Arg Leu Pro Gly Leu His Asn Ser
420 425 430
Leu Val Lys Leu Lys His Gly Ala Gly Val Ala Met Asp Gly Gln Asp
435 440 445
Val Gln Leu Pro Leu Leu Lys Gly Asp Leu Arg Ile Gln His Thr Val
450 455 460
Thr Ala Ser Val Arg Leu Ser Tyr Gly Glu Asp Leu Gln Met Asp Trp
465 470 475 480
Asp Gly Arg Gly Arg Leu Leu Val Lys Leu Ser Pro Val Tyr Ala Gly
485 490 495
Lys Thr Cys Gly Leu Cys Gly Asn Tyr Asn Gly Asn Gln Gly Asp Asp
500 505 510
Phe Leu Thr Pro Ser Gly Leu Ala Glu Pro Arg Val Glu Asp Phe Gly
515 520 525
Asn Ala Trp Lys Leu His Gly Asp Cys Gln Asp Leu Gln Lys Gln His
530 535 540
Ser Asp Pro Cys Ala Leu Asn Pro Arg Met Thr Arg Phe Ser Glu Glu
545 550 555 560
Ala Cys Ala Val Leu Thr Ser Pro Thr Phe Glu Ala Cys His Arg Ala
565 570 575
Val Ser Pro Leu Pro Tyr Leu Arg Asn Cys Arg Tyr Asp Val Cys Ser
580 585 590
Cys Ser Asp Gly Arg Glu Cys Leu Cys Gly Ala Leu Ala Ser Tyr Ala
595 600 605
Ala Ala Cys Ala Gly Arg Gly Val Arg Val Ala Trp Arg Glu Pro Gly
610 615 620
Arg Cys Glu Leu Asn Cys Pro Lys Gly Gln Val Tyr Leu Gln Cys Gly
625 630 635 640
Thr Pro Cys Asn Leu Thr Cys Arg Ser Leu Ser Tyr Pro Asp Glu Glu
645 650 655
Cys Asn Glu Ala Cys Leu Glu Gly Cys Phe Cys Pro Pro Gly Leu Tyr
660 665 670
Met Asp Glu Arg Gly Asp Cys Val Pro Lys Ala Gln Cys Pro Cys Tyr
675 680 685
Tyr Asp Gly Glu Ile Phe Gln Pro Glu Asp Ile Phe Ser Asp His His
690 695 700
Thr Met Cys Tyr Cys Glu Asp Gly Phe Met His Cys Thr Met Ser Gly
705 710 715 720
Val Pro Gly Ser Leu Leu Pro Asp Ala Val Leu Ser Ser Pro Leu Ser
725 730 735
His Arg Ser Lys Arg Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
740 745 750
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
755 760 765
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
770 775 780
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
785 790 795 800
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
805 810 815
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
820 825 830
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
835 840 845
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
850 855 860
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
865 870 875 880
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
885 890 895
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
900 905 910
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
915 920 925
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
930 935 940
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
945 950 955 960
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
965 970 975
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
980 985 990
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
995 1000 1005
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1010 1015 1020
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1025 1030 1035 1040
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
1045 1050 1055
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
1060 1065 1070
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
1075 1080 1085
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
1090 1095 1100
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
1105 1110 1115 1120
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
1125 1130 1135
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
1140 1145 1150
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
1155 1160 1165
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
1170 1175 1180
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
1185 1190 1195 1200
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
1205 1210 1215
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
1220 1225 1230
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
1235 1240 1245
<210> 21
<211> 2465
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3/Fc-120 of the complex of scFVIII/D'D3/Fc-120Fc and
Fc
<400> 21
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1635 1640 1645
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1650 1655 1660
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1665 1670 1675 1680
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1685 1690 1695
Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1700 1705 1710
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1715 1720 1725
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1730 1735 1740
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1745 1750 1755 1760
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1765 1770 1775
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1780 1785 1790
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1795 1800 1805
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1810 1815 1820
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1825 1830 1835 1840
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1845 1850 1855
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1860 1865 1870
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1875 1880 1885
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1890 1895 1900
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1905 1910 1915 1920
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1925 1930 1935
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1940 1945 1950
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1955 1960 1965
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1970 1975 1980
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1985 1990 1995 2000
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
2005 2010 2015
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
2020 2025 2030
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
2035 2040 2045
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
2050 2055 2060
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
2065 2070 2075 2080
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2085 2090 2095
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2100 2105 2110
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2115 2120 2125
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2130 2135 2140
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2145 2150 2155 2160
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2165 2170 2175
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2180 2185 2190
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2195 2200 2205
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
2210 2215 2220
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Lys
2225 2230 2235 2240
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
2245 2250 2255
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
2260 2265 2270
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
2275 2280 2285
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
2290 2295 2300
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
2305 2310 2315 2320
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
2325 2330 2335
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
2340 2345 2350
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
2355 2360 2365
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
2370 2375 2380
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
2385 2390 2395 2400
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
2405 2410 2415
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
2420 2425 2430
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
2435 2440 2445
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
2450 2455 2460
Lys
246
<210> 22
<211> 227
<212> PRT
<213> Artificial Sequence
<220>
<223> Fc of the complex of scFVIII/D'D3/Fc-120 and Fc
<400> 22
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
1 5 10 15
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Lys
225
<210> 23
<211> 1980
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/D'D3-HC Heavy chain
<400> 23
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1445 1450 1455
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1460 1465 1470
Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu Val Cys Pro Ala
1475 1480 1485
Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys Thr Cys Gln Asn
1490 1495 1500
Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser Gly Cys Leu Cys
1505 1510 1515 1520
Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val Ala Leu Glu Arg
1525 1530 1535
Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro Gly Glu Thr Val
1540 1545 1550
Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg Lys Trp Asn Cys
1555 1560 1565
Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile Gly Met Ala His
1570 1575 1580
Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro Gly Glu Cys Gln
1585 1590 1595 1600
Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro Gly Thr Phe Arg
1605 1610 1615
Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser Val Lys Cys Lys
1620 1625 1630
Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile Glu Leu Phe Asp
1635 1640 1645
Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu Thr His Phe Glu
1650 1655 1660
Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu Gly Lys Ala Leu
1665 1670 1675 1680
Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val Val Leu Lys Gln
1685 1690 1695
Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn Phe Asp Gly Ile
1700 1705 1710
Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val Glu Glu Asp Pro
1715 1720 1725
Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln Cys Ala Asp Thr
1730 1735 1740
Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys His Asn Asn Ile
1745 1750 1755 1760
Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile Leu Thr Ser Asp
1765 1770 1775
Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu Pro Tyr Leu Asp
1780 1785 1790
Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile Gly Asp Cys Ala
1795 1800 1805
Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val Cys Ala Gln His
1810 1815 1820
Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys Pro Gln Ser Cys
1825 1830 1835 1840
Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys Glu Trp Arg Tyr
1845 1850 1855
Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln His Pro Glu Pro
1860 1865 1870
Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His Ala His Cys Pro
1875 1880 1885
Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys Val Asp Pro Glu
1890 1895 1900
Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe Ala Ser Gly Lys
1905 1910 1915 1920
Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys Gln Ile Cys His
1925 1930 1935
Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln Glu Pro Gly Gly
1940 1945 1950
Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro Thr Thr Leu Tyr
1955 1960 1965
Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
1970 1975 1980
<210> 24
<211> 772
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/D'D3-HC Light chain
<400> 24
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr
770
<210> 25
<211> 2722
<212> PRT
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3/Fc/Fc-120
<400> 25
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Thr Leu Gln Ser Asp Gln Glu Glu Ile Asp
900 905 910
Tyr Asp Asp Thr Ile Ser Val Glu Met Lys Lys Glu Asp Phe Asp Ile
915 920 925
Tyr Asp Glu Asp Glu Asn Gln Ser Pro Arg Ser Phe Gln Lys Lys Thr
930 935 940
Arg His Tyr Phe Ile Ala Ala Val Glu Arg Leu Trp Asp Tyr Gly Met
945 950 955 960
Ser Ser Ser Pro His Val Leu Arg Asn Arg Ala Gln Ser Gly Ser Val
965 970 975
Pro Gln Phe Lys Lys Val Val Phe Gln Glu Phe Thr Asp Gly Ser Phe
980 985 990
Thr Gln Pro Leu Tyr Arg Gly Glu Leu Asn Glu His Leu Gly Leu Leu
995 1000 1005
Gly Pro Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe
1010 1015 1020
Arg Asn Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser
1025 1030 1035 1040
Tyr Glu Glu Asp Gln Arg Gln Gly Ala Glu Pro Arg Lys Asn Phe Val
1045 1050 1055
Lys Pro Asn Glu Thr Lys Thr Tyr Phe Trp Lys Val Gln His His Met
1060 1065 1070
Ala Pro Thr Lys Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser
1075 1080 1085
Asp Val Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu
1090 1095 1100
Leu Val Cys His Thr Asn Thr Leu Asn Pro Ala His Gly Arg Gln Val
1105 1110 1115 1120
Thr Val Gln Glu Phe Ala Leu Phe Phe Thr Ile Phe Asp Glu Thr Lys
1125 1130 1135
Ser Trp Tyr Phe Thr Glu Asn Met Glu Arg Asn Cys Arg Ala Pro Cys
1140 1145 1150
Asn Ile Gln Met Glu Asp Pro Thr Phe Lys Glu Asn Tyr Arg Phe His
1155 1160 1165
Ala Ile Asn Gly Tyr Ile Met Asp Thr Leu Pro Gly Leu Val Met Ala
1170 1175 1180
Gln Asp Gln Arg Ile Arg Trp Tyr Leu Leu Ser Met Gly Ser Asn Glu
1185 1190 1195 1200
Asn Ile His Ser Ile His Phe Ser Gly His Val Phe Thr Val Arg Lys
1205 1210 1215
Lys Glu Glu Tyr Lys Met Ala Leu Tyr Asn Leu Tyr Pro Gly Val Phe
1220 1225 1230
Glu Thr Val Glu Met Leu Pro Ser Lys Ala Gly Ile Trp Arg Val Glu
1235 1240 1245
Cys Leu Ile Gly Glu His Leu His Ala Gly Met Ser Thr Leu Phe Leu
1250 1255 1260
Val Tyr Ser Asn Lys Cys Gln Thr Pro Leu Gly Met Ala Ser Gly His
1265 1270 1275 1280
Ile Arg Asp Phe Gln Ile Thr Ala Ser Gly Gln Tyr Gly Gln Trp Ala
1285 1290 1295
Pro Lys Leu Ala Arg Leu His Tyr Ser Gly Ser Ile Asn Ala Trp Ser
1300 1305 1310
Thr Lys Glu Pro Phe Ser Trp Ile Lys Val Asp Leu Leu Ala Pro Met
1315 1320 1325
Ile Ile His Gly Ile Lys Thr Gln Gly Ala Arg Gln Lys Phe Ser Ser
1330 1335 1340
Leu Tyr Ile Ser Gln Phe Ile Ile Met Tyr Ser Leu Asp Gly Lys Lys
1345 1350 1355 1360
Trp Gln Thr Tyr Arg Gly Asn Ser Thr Gly Thr Leu Met Val Phe Phe
1365 1370 1375
Gly Asn Val Asp Ser Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro
1380 1385 1390
Ile Ile Ala Arg Tyr Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg
1395 1400 1405
Ser Thr Leu Arg Met Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser
1410 1415 1420
Met Pro Leu Gly Met Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr
1425 1430 1435 1440
Ala Ser Ser Tyr Phe Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys
1445 1450 1455
Ala Arg Leu His Leu Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val
1460 1465 1470
Asn Asn Pro Lys Glu Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys
1475 1480 1485
Val Thr Gly Val Thr Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met
1490 1495 1500
Tyr Val Lys Glu Phe Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp
1505 1510 1515 1520
Thr Leu Phe Phe Gln Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln
1525 1530 1535
Asp Ser Phe Thr Pro Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr
1540 1545 1550
Arg Tyr Leu Arg Ile His Pro Gln Ser Trp Val His Gln Ile Ala Leu
1555 1560 1565
Arg Met Glu Val Leu Gly Cys Glu Ala Gln Asp Leu Tyr Leu Val Pro
1570 1575 1580
Arg Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1585 1590 1595 1600
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1605 1610 1615
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1620 1625 1630
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1635 1640 1645
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1650 1655 1660
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1665 1670 1675 1680
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1685 1690 1695
Ser Gly Gly Gly Gly Ser Leu Ser Cys Arg Pro Pro Met Val Lys Leu
1700 1705 1710
Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu Cys Thr Lys
1715 1720 1725
Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly Cys Val Ser
1730 1735 1740
Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn Arg Cys Val
1745 1750 1755 1760
Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu Tyr Ala Pro
1765 1770 1775
Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys Arg Asp Arg
1780 1785 1790
Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys Ser Thr Ile
1795 1800 1805
Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr Leu Phe Pro
1810 1815 1820
Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly Ser Asn Pro
1825 1830 1835 1840
Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser His Pro Ser
1845 1850 1855
Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly Gly Glu Ile
1860 1865 1870
Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met Lys Asp Glu
1875 1880 1885
Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile Leu Leu Leu
1890 1895 1900
Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser Ile Ser Val
1905 1910 1915 1920
Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu Cys Gly Asn
1925 1930 1935
Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn Leu Gln Val
1940 1945 1950
Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val Ser Ser Gln
1955 1960 1965
Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro Ala Thr Cys
1970 1975 1980
His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser Cys Arg Ile
1985 1990 1995 2000
Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val Asp Pro Glu
2005 2010 2015
Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys Glu Ser Ile
2020 2025 2030
Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr Ala His Val
2035 2040 2045
Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala Thr Leu Cys
2050 2055 2060
Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly Tyr Glu Cys
2065 2070 2075 2080
Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val Thr Cys Gln
2085 2090 2095
His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu Gly Cys His
2100 2105 2110
Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu Gln Thr Cys
2115 2120 2125
Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly Arg Arg Phe
2130 2135 2140
Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro Glu His Cys
2145 2150 2155 2160
Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu Ala Cys Gln
2165 2170 2175
Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro Val Ser Pro
2180 2185 2190
Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu His Asp Phe
2195 2200 2205
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
2210 2215 2220
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Lys
2225 2230 2235 2240
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
2245 2250 2255
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
2260 2265 2270
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
2275 2280 2285
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
2290 2295 2300
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
2305 2310 2315 2320
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
2325 2330 2335
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
2340 2345 2350
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
2355 2360 2365
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
2370 2375 2380
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
2385 2390 2395 2400
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
2405 2410 2415
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
2420 2425 2430
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
2435 2440 2445
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
2450 2455 2460
Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
2465 2470 2475 2480
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp
2485 2490 2495
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
2500 2505 2510
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
2515 2520 2525
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
2530 2535 2540
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
2545 2550 2555 2560
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
2565 2570 2575
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
2580 2585 2590
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
2595 2600 2605
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
2610 2615 2620
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
2625 2630 2635 2640
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
2645 2650 2655
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
2660 2665 2670
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
2675 2680 2685
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
2690 2695 2700
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
2705 2710 2715 2720
Gly Lys
<210> 26
<211> 1700
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC/Fc-LC Heavy chain
<400> 26
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1445 1450 1455
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1460 1465 1470
Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
1475 1480 1485
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
1490 1495 1500
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
1505 1510 1515 1520
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
1525 1530 1535
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
1540 1545 1550
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
1555 1560 1565
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
1570 1575 1580
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
1585 1590 1595 1600
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
1605 1610 1615
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
1620 1625 1630
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
1635 1640 1645
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
1650 1655 1660
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
1665 1670 1675 1680
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
1685 1690 1695
Ser Pro Gly Lys
1700
<210> 27
<211> 1029
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC/Fc-LC Light chain
<400> 27
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr Leu Val Pro Arg Gly Ser Gly Gly Gly Gly Ser Gly
770 775 780
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
785 790 795 800
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
805 810 815
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
820 825 830
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
835 840 845
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
850 855 860
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
865 870 875 880
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
885 890 895
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
900 905 910
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
915 920 925
Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
930 935 940
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
945 950 955 960
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
965 970 975
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
980 985 990
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
995 1000 1005
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
1010 1015 1020
Leu Ser Pro Gly Lys
1025
<210> 28
<211> 1700
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC, Fc Heavy chain
<400> 28
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1445 1450 1455
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1460 1465 1470
Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
1475 1480 1485
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
1490 1495 1500
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
1505 1510 1515 1520
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
1525 1530 1535
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
1540 1545 1550
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
1555 1560 1565
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
1570 1575 1580
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
1585 1590 1595 1600
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
1605 1610 1615
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
1620 1625 1630
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
1635 1640 1645
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
1650 1655 1660
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
1665 1670 1675 1680
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
1685 1690 1695
Ser Pro Gly Lys
1700
<210> 29
<211> 772
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC, Fc Light chain
<400> 29
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr
770
<210> 30
<211> 1957
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc/Fc-HC Heavy chain
<400> 30
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1445 1450 1455
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
1460 1465 1470
Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
1475 1480 1485
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
1490 1495 1500
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
1505 1510 1515 1520
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
1525 1530 1535
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
1540 1545 1550
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
1555 1560 1565
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
1570 1575 1580
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
1585 1590 1595 1600
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
1605 1610 1615
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
1620 1625 1630
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
1635 1640 1645
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
1650 1655 1660
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
1665 1670 1675 1680
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
1685 1690 1695
Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1700 1705 1710
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
1715 1720 1725
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
1730 1735 1740
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
1745 1750 1755 1760
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
1765 1770 1775
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
1780 1785 1790
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
1795 1800 1805
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
1810 1815 1820
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
1825 1830 1835 1840
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
1845 1850 1855
Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
1860 1865 1870
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
1875 1880 1885
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
1890 1895 1900
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
1905 1910 1915 1920
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
1925 1930 1935
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
1940 1945 1950
Leu Ser Pro Gly Lys
1955
<210> 31
<211> 772
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc/Fc-HC Light chain
<400> 31
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr
770
<210> 32
<211> 1443
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc-LC, Fc Heavy chain
<400> 32
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe
<210> 33
<211> 1029
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc-LC, Fc Light chain
<400> 33
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr Leu Val Pro Arg Gly Ser Gly Gly Gly Gly Ser Gly
770 775 780
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
785 790 795 800
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
805 810 815
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
820 825 830
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
835 840 845
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
850 855 860
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
865 870 875 880
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
885 890 895
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
900 905 910
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
915 920 925
Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
930 935 940
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
945 950 955 960
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
965 970 975
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
980 985 990
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
995 1000 1005
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
1010 1015 1020
Leu Ser Pro Gly Lys
1025
<210> 34
<211> 1443
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc/Fc-LC Heavy chain
<400> 34
Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser Trp Asp Tyr
1 5 10 15
Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg Phe Pro Pro
20 25 30
Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val Tyr Lys Lys
35 40 45
Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile Ala Lys Pro
50 55 60
Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln Ala Glu Val
65 70 75 80
Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser His Pro Val
85 90 95
Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser Glu Gly Ala
100 105 110
Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp Asp Lys Val
115 120 125
Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu Lys Glu Asn
130 135 140
Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser Tyr Leu Ser
145 150 155 160
His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile Gly Ala Leu
165 170 175
Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr Gln Thr Leu
180 185 190
His Lys Phe Ile Leu Leu Phe Ala Val Phe Asp Glu Gly Lys Ser Trp
195 200 205
His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp Ala Ala Ser
210 215 220
Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr Val Asn Arg
225 230 235 240
Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val Tyr Trp His
245 250 255
Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile Phe Leu Glu
260 265 270
Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser Leu Glu Ile
275 280 285
Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met Asp Leu Gly
290 295 300
Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His Asp Gly Met
305 310 315 320
Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro Gln Leu Arg
325 330 335
Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp Leu Thr Asp
340 345 350
Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser Pro Ser Phe
355 360 365
Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr Trp Val His
370 375 380
Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro Leu Val Leu
385 390 395 400
Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn Asn Gly Pro
405 410 415
Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met Ala Tyr Thr
420 425 430
Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu Ser Gly Ile
435 440 445
Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu Leu Ile Ile
450 455 460
Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro His Gly Ile
465 470 475 480
Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys Gly Val Lys
485 490 495
His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe Lys Tyr Lys
500 505 510
Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp Pro Arg Cys
515 520 525
Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg Asp Leu Ala
530 535 540
Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu Ser Val Asp
545 550 555 560
Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val Ile Leu Phe
565 570 575
Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu Asn Ile Gln
580 585 590
Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp Pro Glu Phe
595 600 605
Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val Phe Asp Ser
610 615 620
Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp Tyr Ile Leu
625 630 635 640
Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe Ser Gly Tyr
645 650 655
Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr Leu Phe Pro
660 665 670
Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro Gly Leu Trp
675 680 685
Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly Met Thr Ala
690 695 700
Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp Tyr Tyr Glu
705 710 715 720
Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys Asn Asn Ala
725 730 735
Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro Ser Thr Arg
740 745 750
Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp Ile Glu Lys
755 760 765
Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys Cys Gln Asn
770 775 780
Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser Pro Thr Pro
785 790 795 800
His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr Glu Thr Phe
805 810 815
Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn Ser Leu Ser
820 825 830
Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly Asp Met Val
835 840 845
Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu Lys Leu Gly
850 855 860
Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys Val Ser Ser
865 870 875 880
Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn Leu Ala Ala
885 890 895
Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met Pro Val His
900 905 910
Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys Ser Ser Pro
915 920 925
Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Cys Arg Pro Pro Met
930 935 940
Val Lys Leu Val Cys Pro Ala Asp Asn Leu Arg Ala Glu Gly Leu Glu
945 950 955 960
Cys Thr Lys Thr Cys Gln Asn Tyr Asp Leu Glu Cys Met Ser Met Gly
965 970 975
Cys Val Ser Gly Cys Leu Cys Pro Pro Gly Met Val Arg His Glu Asn
980 985 990
Arg Cys Val Ala Leu Glu Arg Cys Pro Cys Phe His Gln Gly Lys Glu
995 1000 1005
Tyr Ala Pro Gly Glu Thr Val Lys Ile Gly Cys Asn Thr Cys Val Cys
1010 1015 1020
Arg Asp Arg Lys Trp Asn Cys Thr Asp His Val Cys Asp Ala Thr Cys
1025 1030 1035 1040
Ser Thr Ile Gly Met Ala His Tyr Leu Thr Phe Asp Gly Leu Lys Tyr
1045 1050 1055
Leu Phe Pro Gly Glu Cys Gln Tyr Val Leu Val Gln Asp Tyr Cys Gly
1060 1065 1070
Ser Asn Pro Gly Thr Phe Arg Ile Leu Val Gly Asn Lys Gly Cys Ser
1075 1080 1085
His Pro Ser Val Lys Cys Lys Lys Arg Val Thr Ile Leu Val Glu Gly
1090 1095 1100
Gly Glu Ile Glu Leu Phe Asp Gly Glu Val Asn Val Lys Arg Pro Met
1105 1110 1115 1120
Lys Asp Glu Thr His Phe Glu Val Val Glu Ser Gly Arg Tyr Ile Ile
1125 1130 1135
Leu Leu Leu Gly Lys Ala Leu Ser Val Val Trp Asp Arg His Leu Ser
1140 1145 1150
Ile Ser Val Val Leu Lys Gln Thr Tyr Gln Glu Lys Val Cys Gly Leu
1155 1160 1165
Cys Gly Asn Phe Asp Gly Ile Gln Asn Asn Asp Leu Thr Ser Ser Asn
1170 1175 1180
Leu Gln Val Glu Glu Asp Pro Val Asp Phe Gly Asn Ser Trp Lys Val
1185 1190 1195 1200
Ser Ser Gln Cys Ala Asp Thr Arg Lys Val Pro Leu Asp Ser Ser Pro
1205 1210 1215
Ala Thr Cys His Asn Asn Ile Met Lys Gln Thr Met Val Asp Ser Ser
1220 1225 1230
Cys Arg Ile Leu Thr Ser Asp Val Phe Gln Asp Cys Asn Lys Leu Val
1235 1240 1245
Asp Pro Glu Pro Tyr Leu Asp Val Cys Ile Tyr Asp Thr Cys Ser Cys
1250 1255 1260
Glu Ser Ile Gly Asp Cys Ala Cys Phe Cys Asp Thr Ile Ala Ala Tyr
1265 1270 1275 1280
Ala His Val Cys Ala Gln His Gly Lys Val Val Thr Trp Arg Thr Ala
1285 1290 1295
Thr Leu Cys Pro Gln Ser Cys Glu Glu Arg Asn Leu Arg Glu Asn Gly
1300 1305 1310
Tyr Glu Cys Glu Trp Arg Tyr Asn Ser Cys Ala Pro Ala Cys Gln Val
1315 1320 1325
Thr Cys Gln His Pro Glu Pro Leu Ala Cys Pro Val Gln Cys Val Glu
1330 1335 1340
Gly Cys His Ala His Cys Pro Pro Gly Lys Ile Leu Asp Glu Leu Leu
1345 1350 1355 1360
Gln Thr Cys Val Asp Pro Glu Asp Cys Pro Val Cys Glu Val Ala Gly
1365 1370 1375
Arg Arg Phe Ala Ser Gly Lys Lys Val Thr Leu Asn Pro Ser Asp Pro
1380 1385 1390
Glu His Cys Gln Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu
1395 1400 1405
Ala Cys Gln Glu Pro Gly Gly Leu Val Val Pro Pro Thr Asp Ala Pro
1410 1415 1420
Val Ser Pro Thr Thr Leu Tyr Val Glu Asp Ile Ser Glu Pro Pro Leu
1425 1430 1435 1440
His Asp Phe
<210> 35
<211> 1286
<212> PRT
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc/Fc-LC Light chain
<400> 35
Ala Thr Asp Pro Leu Ala Trp Asp Asn His Tyr Gly Thr Gln Ile Pro
1 5 10 15
Lys Glu Glu Trp Lys Ser Gln Glu Lys Ser Pro Glu Lys Thr Ala Phe
20 25 30
Lys Lys Lys Asp Thr Ile Leu Ser Leu Asn Ala Cys Glu Ser Asn His
35 40 45
Ala Ile Ala Ala Ile Asn Glu Gly Gln Asn Lys Pro Glu Ile Glu Val
50 55 60
Thr Trp Ala Lys Gln Gly Arg Thr Glu Arg Leu Cys Ser Gln Asn Pro
65 70 75 80
Pro Val Leu Lys Arg His Gln Arg Glu Ile Thr Arg Thr Thr Leu Gln
85 90 95
Ser Asp Gln Glu Glu Ile Asp Tyr Asp Asp Thr Ile Ser Val Glu Met
100 105 110
Lys Lys Glu Asp Phe Asp Ile Tyr Asp Glu Asp Glu Asn Gln Ser Pro
115 120 125
Arg Ser Phe Gln Lys Lys Thr Arg His Tyr Phe Ile Ala Ala Val Glu
130 135 140
Arg Leu Trp Asp Tyr Gly Met Ser Ser Ser Pro His Val Leu Arg Asn
145 150 155 160
Arg Ala Gln Ser Gly Ser Val Pro Gln Phe Lys Lys Val Val Phe Gln
165 170 175
Glu Phe Thr Asp Gly Ser Phe Thr Gln Pro Leu Tyr Arg Gly Glu Leu
180 185 190
Asn Glu His Leu Gly Leu Leu Gly Pro Tyr Ile Arg Ala Glu Val Glu
195 200 205
Asp Asn Ile Met Val Thr Phe Arg Asn Gln Ala Ser Arg Pro Tyr Ser
210 215 220
Phe Tyr Ser Ser Leu Ile Ser Tyr Glu Glu Asp Gln Arg Gln Gly Ala
225 230 235 240
Glu Pro Arg Lys Asn Phe Val Lys Pro Asn Glu Thr Lys Thr Tyr Phe
245 250 255
Trp Lys Val Gln His His Met Ala Pro Thr Lys Asp Glu Phe Asp Cys
260 265 270
Lys Ala Trp Ala Tyr Phe Ser Asp Val Asp Leu Glu Lys Asp Val His
275 280 285
Ser Gly Leu Ile Gly Pro Leu Leu Val Cys His Thr Asn Thr Leu Asn
290 295 300
Pro Ala His Gly Arg Gln Val Thr Val Gln Glu Phe Ala Leu Phe Phe
305 310 315 320
Thr Ile Phe Asp Glu Thr Lys Ser Trp Tyr Phe Thr Glu Asn Met Glu
325 330 335
Arg Asn Cys Arg Ala Pro Cys Asn Ile Gln Met Glu Asp Pro Thr Phe
340 345 350
Lys Glu Asn Tyr Arg Phe His Ala Ile Asn Gly Tyr Ile Met Asp Thr
355 360 365
Leu Pro Gly Leu Val Met Ala Gln Asp Gln Arg Ile Arg Trp Tyr Leu
370 375 380
Leu Ser Met Gly Ser Asn Glu Asn Ile His Ser Ile His Phe Ser Gly
385 390 395 400
His Val Phe Thr Val Arg Lys Lys Glu Glu Tyr Lys Met Ala Leu Tyr
405 410 415
Asn Leu Tyr Pro Gly Val Phe Glu Thr Val Glu Met Leu Pro Ser Lys
420 425 430
Ala Gly Ile Trp Arg Val Glu Cys Leu Ile Gly Glu His Leu His Ala
435 440 445
Gly Met Ser Thr Leu Phe Leu Val Tyr Ser Asn Lys Cys Gln Thr Pro
450 455 460
Leu Gly Met Ala Ser Gly His Ile Arg Asp Phe Gln Ile Thr Ala Ser
465 470 475 480
Gly Gln Tyr Gly Gln Trp Ala Pro Lys Leu Ala Arg Leu His Tyr Ser
485 490 495
Gly Ser Ile Asn Ala Trp Ser Thr Lys Glu Pro Phe Ser Trp Ile Lys
500 505 510
Val Asp Leu Leu Ala Pro Met Ile Ile His Gly Ile Lys Thr Gln Gly
515 520 525
Ala Arg Gln Lys Phe Ser Ser Leu Tyr Ile Ser Gln Phe Ile Ile Met
530 535 540
Tyr Ser Leu Asp Gly Lys Lys Trp Gln Thr Tyr Arg Gly Asn Ser Thr
545 550 555 560
Gly Thr Leu Met Val Phe Phe Gly Asn Val Asp Ser Ser Gly Ile Lys
565 570 575
His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr Ile Arg Leu His
580 585 590
Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met Glu Leu Met Gly
595 600 605
Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met Glu Ser Lys Ala
610 615 620
Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe Thr Asn Met Phe
625 630 635 640
Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu Gln Gly Arg Ser
645 650 655
Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu Trp Leu Gln Val
660 665 670
Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr Thr Gln Gly Val
675 680 685
Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe Leu Ile Ser Ser
690 695 700
Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln Asn Gly Lys Val
705 710 715 720
Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro Val Val Asn Ser
725 730 735
Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile His Pro Gln Ser
740 745 750
Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu Gly Cys Glu Ala
755 760 765
Gln Asp Leu Tyr Leu Val Pro Arg Gly Ser Gly Gly Gly Gly Ser Gly
770 775 780
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
785 790 795 800
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
805 810 815
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
820 825 830
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
835 840 845
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
850 855 860
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
865 870 875 880
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
885 890 895
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
900 905 910
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
915 920 925
Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
930 935 940
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
945 950 955 960
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
965 970 975
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
980 985 990
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
995 1000 1005
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
1010 1015 1020
Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1025 1030 1035 1040
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
1045 1050 1055
Gly Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
1060 1065 1070
Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
1075 1080 1085
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
1090 1095 1100
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
1105 1110 1115 1120
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
1125 1130 1135
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
1140 1145 1150
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
1155 1160 1165
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
1170 1175 1180
Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn
1185 1190 1195 1200
Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
1205 1210 1215
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
1220 1225 1230
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
1235 1240 1245
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
1250 1255 1260
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
1265 1270 1275 1280
Ser Leu Ser Pro Gly Lys
1285
<210> 36
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Thrombin site
<400> 36
Asp Phe Leu Ala Glu Gly Gly Gly Val Arg
1 5 10
<210> 37
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Thrombin site
<400> 37
Thr Thr Lys Ile Lys Pro Arg
1 5
<210> 38
<211> 6
<212> PRT
<213> Artificial Sequence
<220>
<223> Thrombin site
<400> 38
Leu Val Pro Arg Gly Ser
1 5
<210> 39
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> FVIIa site
<400> 39
Ala Ser Lys Pro Gln Gly Arg Ile Val Gly Gly
1 5 10
<210> 40
<211> 4
<212> PRT
<213> Artificial Sequence
<220>
<223> FXa site
<400> 40
Ile Asp Gly Arg
1
<210> 41
<211> 4
<212> PRT
<213> Artificial Sequence
<220>
<223> FXa site
<400> 41
Ile Glu Gly Arg
1
<210> 42
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> FXa site
<400> 42
Ser Lys Leu Thr Arg Ala Glu Thr Val Phe
1 5 10
<210> 43
<211> 6999
<212> DNA
<213> Homo sapiens
<220>
<221> gene
<222> (1)..(6999)
<223> mature FVIII
<400> 43
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaaatacaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cttgagtgaa 2820
gaaaataatg attcaaagtt gttagaatca ggtttaatga atagccaaga aagttcatgg 2880
ggaaaaaatg tatcgtcaac agagagtggt aggttattta aagggaaaag agctcatgga 2940
cctgctttgt tgactaaaga taatgcctta ttcaaagtta gcatctcttt gttaaagaca 3000
aacaaaactt ccaataattc agcaactaat agaaagactc acattgatgg cccatcatta 3060
ttaattgaga atagtccatc agtctggcaa aatatattag aaagtgacac tgagtttaaa 3120
aaagtgacac ctttgattca tgacagaatg cttatggaca aaaatgctac agctttgagg 3180
ctaaatcata tgtcaaataa aactacttca tcaaaaaaca tggaaatggt ccaacagaaa 3240
aaagagggcc ccattccacc agatgcacaa aatccagata tgtcgttctt taagatgcta 3300
ttcttgccag aatcagcaag gtggatacaa aggactcatg gaaagaactc tctgaactct 3360
gggcaaggcc ccagtccaaa gcaattagta tccttaggac cagaaaaatc tgtggaaggt 3420
cagaatttct tgtctgagaa aaacaaagtg gtagtaggaa agggtgaatt tacaaaggac 3480
gtaggactca aagagatggt ttttccaagc agcagaaacc tatttcttac taacttggat 3540
aatttacatg aaaataatac acacaatcaa gaaaaaaaaa ttcaggaaga aatagaaaag 3600
aaggaaacat taatccaaga gaatgtagtt ttgcctcaga tacatacagt gactggcact 3660
aagaatttca tgaagaacct tttcttactg agcactaggc aaaatgtaga aggttcatat 3720
gacggggcat atgctccagt acttcaagat tttaggtcat taaatgattc aacaaataga 3780
acaaagaaac acacagctca tttctcaaaa aaaggggagg aagaaaactt ggaaggcttg 3840
ggaaatcaaa ccaagcaaat tgtagagaaa tatgcatgca ccacaaggat atctcctaat 3900
acaagccagc agaattttgt cacgcaacgt agtaagagag ctttgaaaca attcagactc 3960
ccactagaag aaacagaact tgaaaaaagg ataattgtgg atgacacctc aacccagtgg 4020
tccaaaaaca tgaaacattt gaccccgagc accctcacac agatagacta caatgagaag 4080
gagaaagggg ccattactca gtctccctta tcagattgcc ttacgaggag tcatagcatc 4140
cctcaagcaa atagatctcc attacccatt gcaaaggtat catcatttcc atctattaga 4200
cctatatatc tgaccagggt cctattccaa gacaactctt ctcatcttcc agcagcatct 4260
tatagaaaga aagattctgg ggtccaagaa agcagtcatt tcttacaagg agccaaaaaa 4320
aataaccttt ctttagccat tctaaccttg gagatgactg gtgatcaaag agaggttggc 4380
tccctgggga caagtgccac aaattcagtc acatacaaga aagttgagaa cactgttctc 4440
ccgaaaccag acttgcccaa aacatctggc aaagttgaat tgcttccaaa agttcacatt 4500
tatcagaagg acctattccc tacggaaact agcaatgggt ctcctggcca tctggatctc 4560
gtggaaggga gccttcttca gggaacagag ggagcgatta agtggaatga agcaaacaga 4620
cctggaaaag ttccctttct gagagtagca acagaaagct ctgcaaagac tccctccaag 4680
ctattggatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 4740
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 4800
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 4860
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 4920
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 4980
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 5040
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 5100
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 5160
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 5220
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 5280
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 5340
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 5400
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 5460
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 5520
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 5580
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 5640
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 5700
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 5760
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 5820
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 5880
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 5940
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 6000
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 6060
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 6120
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 6180
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 6240
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 6300
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 6360
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 6420
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 6480
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 6540
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 6600
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 6660
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 6720
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 6780
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 6840
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 6900
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 6960
atggaggttc tgggctgcga ggcacaggac ctctactga 6999
<210> 44
<211> 8376
<212> DNA
<213> Homo sapiens
<220>
<221> gene
<222> (1)..(8376)
<223> full length vWF
<400> 44
gcagaaggaa ctcgcggcag gtcatccacg gcccgatgca gccttttcgg aagtgacttc 60
gtcaacacct ttgatgggag catgtacagc tttgcgggat actgcagtta cctcctggca 120
gggggctgcc agaaacgctc cttctcgatt attggggact tccagaatgg caagagagtg 180
agcctctccg tgtatcttgg ggaatttttt gacatccatt tgtttgtcaa tggtaccgtg 240
acacaggggg accaaagagt ctccatgccc tatgcctcca aagggctgta tctagaaact 300
gaggctgggt actacaagct gtccggtgag gcctatggct ttgtggccag gatcgatggc 360
agcggcaact ttcaagtcct gctgtcagac agatacttca acaagacctg cgggctgtgt 420
ggcaacttta acatctttgc tgaagatgac tttatgaccc aagaagggac cttgacctcg 480
gacccttatg actttgccaa ctcatgggct ctgagcagtg gagaacagtg gtgtgaacgg 540
gcatctcctc ccagcagctc atgcaacatc tcctctgggg aaatgcagaa gggcctgtgg 600
gagcagtgcc agcttctgaa gagcacctcg gtgtttgccc gctgccaccc tctggtggac 660
cccgagcctt ttgtggccct gtgtgagaag actttgtgtg agtgtgctgg ggggctggag 720
tgcgcctgcc ctgccctcct ggagtacgcc cggacctgtg cccaggaggg aatggtgctg 780
tacggctgga ccgaccacag cgcgtgcagc ccagtgtgcc ctgctggtat ggagtatagg 840
cagtgtgtgt ccccttgcgc caggacctgc cagagcctgc acatcaatga aatgtgtcag 900
gagcgatgcg tggatggctg cagctgccct gagggacagc tcctggatga aggcctctgc 960
gtggagagca ccgagtgtcc ctgcgtgcat tccggaaagc gctaccctcc cggcacctcc 1020
ctctctcgag actgcaacac ctgcatttgc cgaaacagcc agtggatctg cagcaatgaa 1080
gaatgtccag gggagtgcct tgtcacaggt caatcacact tcaagagctt tgacaacaga 1140
tacttcacct tcagtgggat ctgccagtac ctgctggccc gggattgcca ggaccactcc 1200
ttctccattg tcattgagac tgtccagtgt gctgatgacc gcgacgctgt gtgcacccgc 1260
tccgtcaccg tccggctgcc tggcctgcac aacagccttg tgaaactgaa gcatggggca 1320
ggagttgcca tggatggcca ggacgtccag ctccccctcc tgaaaggtga cctccgcatc 1380
cagcatacag tgacggcctc cgtgcgcctc agctacgggg aggacctgca gatggactgg 1440
gatggccgcg ggaggctgct ggtgaagctg tcccccgtct atgccgggaa gacctgcggc 1500
ctgtgtggga attacaatgg caaccagggc gacgacttcc ttaccccctc tgggctggcg 1560
gagccccggg tggaggactt cgggaacgcc tggaagctgc acggggactg ccaggacctg 1620
cagaagcagc acagcgatcc ctgcgccctc aacccgcgca tgaccaggtt ctccgaggag 1680
gcgtgcgcgg tcctgacgtc ccccacattc gaggcctgcc atcgtgccgt cagcccgctg 1740
ccctacctgc ggaactgccg ctacgacgtg tgctcctgct cggacggccg cgagtgcctg 1800
tgcggcgccc tggccagcta tgccgcggcc tgcgcgggga gaggcgtgcg cgtcgcgtgg 1860
cgcgagccag gccgctgtga gctgaactgc ccgaaaggcc aggtgtacct gcagtgcggg 1920
accccctgca acctgacctg ccgctctctc tcttacccgg atgaggaatg caatgaggcc 1980
tgcctggagg gctgcttctg ccccccaggg ctctacatgg atgagagggg ggactgcgtg 2040
cccaaggccc agtgcccctg ttactatgac ggtgagatct tccagccaga agacatcttc 2100
tcagaccatc acaccatgtg ctactgtgag gatggcttca tgcactgtac catgagtgga 2160
gtccccggaa gcttgctgcc tgacgctgtc ctcagcagtc ccctgtctca tcgcagcaaa 2220
aggagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 2280
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 2340
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 2400
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 2460
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 2520
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 2580
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 2640
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 2700
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 2760
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 2820
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 2880
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 2940
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 3000
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 3060
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 3120
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 3180
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 3240
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 3300
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 3360
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 3420
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 3480
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 3540
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 3600
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 3660
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 3720
tcggaaccgc cgttgcacga tttctactgc agcaggctac tggacctggt cttcctgctg 3780
gatggctcct ccaggctgtc cgaggctgag tttgaagtgc tgaaggcctt tgtggtggac 3840
atgatggagc ggctgcgcat ctcccagaag tgggtccgcg tggccgtggt ggagtaccac 3900
gacggctccc acgcctacat cgggctcaag gaccggaagc gaccgtcaga gctgcggcgc 3960
attgccagcc aggtgaagta tgcgggcagc caggtggcct ccaccagcga ggtcttgaaa 4020
tacacactgt tccaaatctt cagcaagatc gaccgccctg aagcctcccg catcaccctg 4080
ctcctgatgg ccagccagga gccccaacgg atgtcccgga actttgtccg ctacgtccag 4140
ggcctgaaga agaagaaggt cattgtgatc ccggtgggca ttgggcccca tgccaacctc 4200
aagcagatcc gcctcatcga gaagcaggcc cctgagaaca aggccttcgt gctgagcagt 4260
gtggatgagc tggagcagca aagggacgag atcgttagct acctctgtga ccttgcccct 4320
gaagcccctc ctcctactct gccccccgac atggcacaag tcactgtggg cccggggctc 4380
ttgggggttt cgaccctggg gcccaagagg aactccatgg ttctggatgt ggcgttcgtc 4440
ctggaaggat cggacaaaat tggtgaagcc gacttcaaca ggagcaagga gttcatggag 4500
gaggtgattc agcggatgga tgtgggccag gacagcatcc acgtcacggt gctgcagtac 4560
tcctacatgg tgactgtgga gtaccccttc agcgaggcac agtccaaagg ggacatcctg 4620
cagcgggtgc gagagatccg ctaccagggc ggcaacagga ccaacactgg gctggccctg 4680
cggtacctct ctgaccacag cttcttggtc agccagggtg accgggagca ggcgcccaac 4740
ctggtctaca tggtcaccgg aaatcctgcc tctgatgaga tcaagaggct gcctggagac 4800
atccaggtgg tgcccattgg agtgggccct aatgccaacg tgcaggagct ggagaggatt 4860
ggctggccca atgcccctat cctcatccag gactttgaga cgctcccccg agaggctcct 4920
gacctggtgc tgcagaggtg ctgctccgga gaggggctgc agatccccac cctctcccct 4980
gcacctgact gcagccagcc cctggacgtg atccttctcc tggatggctc ctccagtttc 5040
ccagcttctt attttgatga aatgaagagt ttcgccaagg ctttcatttc aaaagccaat 5100
atagggcctc gtctcactca ggtgtcagtg ctgcagtatg gaagcatcac caccattgac 5160
gtgccatgga acgtggtccc ggagaaagcc catttgctga gccttgtgga cgtcatgcag 5220
cgggagggag gccccagcca aatcggggat gccttgggct ttgctgtgcg atacttgact 5280
tcagaaatgc atggtgccag gccgggagcc tcaaaggcgg tggtcatcct ggtcacggac 5340
gtctctgtgg attcagtgga tgcagcagct gatgccgcca ggtccaacag agtgacagtg 5400
ttccctattg gaattggaga tcgctacgat gcagcccagc tacggatctt ggcaggccca 5460
gcaggcgact ccaacgtggt gaagctccag cgaatcgaag acctccctac catggtcacc 5520
ttgggcaatt ccttcctcca caaactgtgc tctggatttg ttaggatttg catggatgag 5580
gatgggaatg agaagaggcc cggggacgtc tggaccttgc cagaccagtg ccacaccgtg 5640
acttgccagc cagatggcca gaccttgctg aagagtcatc gggtcaactg tgaccggggg 5700
ctgaggcctt cgtgccctaa cagccagtcc cctgttaaag tggaagagac ctgtggctgc 5760
cgctggacct gcccctgcgt gtgcacaggc agctccactc ggcacatcgt gacctttgat 5820
gggcagaatt tcaagctgac tggcagctgt tcttatgtcc tatttcaaaa caaggagcag 5880
gacctggagg tgattctcca taatggtgcc tgcagccctg gagcaaggca gggctgcatg 5940
aaatccatcg aggtgaagca cagtgccctc tccgtcgagc tgcacagtga catggaggtg 6000
acggtgaatg ggagactggt ctctgttcct tacgtgggtg ggaacatgga agtcaacgtt 6060
tatggtgcca tcatgcatga ggtcagattc aatcaccttg gtcacatctt cacattcact 6120
ccacaaaaca atgagttcca actgcagctc agccccaaga cttttgcttc aaagacgtat 6180
ggtctgtgtg ggatctgtga tgagaacgga gccaatgact tcatgctgag ggatggcaca 6240
gtcaccacag actggaaaac acttgttcag gaatggactg tgcagcggcc agggcagacg 6300
tgccagccca tcctggagga gcagtgtctt gtccccgaca gctcccactg ccaggtcctc 6360
ctcttaccac tgtttgctga atgccacaag gtcctggctc cagccacatt ctatgccatc 6420
tgccagcagg acagttgcca ccaggagcaa gtgtgtgagg tgatcgcctc ttatgcccac 6480
ctctgtcgga ccaacggggt ctgcgttgac tggaggacac ctgatttctg tgctatgtca 6540
tgcccaccat ctctggtcta caaccactgt gagcatggct gtccccggca ctgtgatggc 6600
aacgtgagct cctgtgggga ccatccctcc gaaggctgtt tctgccctcc agataaagtc 6660
atgttggaag gcagctgtgt ccctgaagag gcctgcactc agtgcattgg tgaggatgga 6720
gtccagcacc agttcctgga agcctgggtc ccggaccacc agccctgtca gatctgcaca 6780
tgcctcagcg ggcggaaggt caactgcaca acgcagccct gccccacggc caaagctccc 6840
acgtgtggcc tgtgtgaagt agcccgcctc cgccagaatg cagaccagtg ctgccccgag 6900
tatgagtgtg tgtgtgaccc agtgagctgt gacctgcccc cagtgcctca ctgtgaacgt 6960
ggcctccagc ccacactgac caaccctggc gagtgcagac ccaacttcac ctgcgcctgc 7020
aggaaggagg agtgcaaaag agtgtcccca ccctcctgcc ccccgcaccg tttgcccacc 7080
cttcggaaga cccagtgctg tgatgagtat gagtgtgcct gcaactgtgt caactccaca 7140
gtgagctgtc cccttgggta cttggcctca actgccacca atgactgtgg ctgtaccaca 7200
accacctgcc ttcccgacaa ggtgtgtgtc caccgaagca ccatctaccc tgtgggccag 7260
ttctgggagg agggctgcga tgtgtgcacc tgcaccgaca tggaggatgc cgtgatgggc 7320
ctccgcgtgg cccagtgctc ccagaagccc tgtgaggaca gctgtcggtc gggcttcact 7380
tacgttctgc atgaaggcga gtgctgtgga aggtgcctgc catctgcctg tgaggtggtg 7440
actggctcac cgcgggggga ctcccagtct tcctggaaga gtgtcggctc ccagtgggcc 7500
tccccggaga acccctgcct catcaatgag tgtgtccgag tgaaggagga ggtctttata 7560
caacaaagga acgtctcctg cccccagctg gaggtccctg tctgcccctc gggctttcag 7620
ctgagctgta agacctcagc gtgctgccca agctgtcgct gtgagcgcat ggaggcctgc 7680
atgctcaatg gcactgtcat tgggcccggg aagactgtga tgatcgatgt gtgcacgacc 7740
tgccgctgca tggtgcaggt gggggtcatc tctggattca agctggagtg caggaagacc 7800
acctgcaacc cctgccccct gggttacaag gaagaaaata acacaggtga atgttgtggg 7860
agatgtttgc ctacggcttg caccattcag ctaagaggag gacagatcat gacactgaag 7920
cgtgatgaga cgctccagga tggctgtgat actcacttct gcaaggtcaa tgagagagga 7980
gagtacttct gggagaagag ggtcacaggc tgcccaccct ttgatgaaca caagtgtctg 8040
gctgagggag gtaaaattat gaaaattcca ggcacctgct gtgacacatg tgaggagcct 8100
gagtgcaacg acatcactgc caggctgcag tatgtcaagg tgggaagctg taagtctgaa 8160
gtagaggtgg atatccacta ctgccagggc aaatgtgcca gcaaagccat gtactccatt 8220
gacatcaacg atgtgcagga ccagtgctcc tgctgctctc cgacacggac ggagcccatg 8280
caggtggccc tgcactgcac caatggctct gttgtgtacc atgaggttct caatgccatg 8340
gagtgcaaat gctcccccag gaagtgcagc aagtga 8376
<210> 45
<211> 4749
<212> DNA
<213> Artificial Sequence
<220>
<223> Single chain FVIII with partial B domain
<220>
<221> misc_feature
<222> (2344)
<223> W= A or T
<220>
<221> misc_feature
<222> (2345)
<223> K= T or G
<220>
<221> misc_feature
<222> (2346)
<223> W= A or T
<220>
<221> misc_feature
<222> (2344)..(2346)
<223> encoding Isoleucine or Cysteine
<400> 45
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaawkwcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tactgataa 4749
<210> 46
<211> 4335
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC Heavy chain
<400> 46
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttct gataa 4335
<210> 47
<211> 2322
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC light chain
<400> 47
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctactgat aa 2322
<210> 48
<211> 6288
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-6
<400> 48
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cagcctatcc tgtcggcccc ccatggtcaa gctggtgtgt 4800
cccgctgaca acctgcgggc tgaagggctc gagtgtacca aaacgtgcca gaactatgac 4860
ctggagtgca tgagcatggg ctgtgtctct ggctgcctct gccccccggg catggtccgg 4920
catgagaaca gatgtgtggc cctggaaagg tgtccctgct tccatcaggg caaggagtat 4980
gcccctggag aaacagtgaa gattggctgc aacacttgtg tctgtcggga ccggaagtgg 5040
aactgcacag accatgtgtg tgatgccacg tgctccacga tcggcatggc ccactacctc 5100
accttcgacg ggctcaaata cctgttcccc ggggagtgcc agtacgttct ggtgcaggat 5160
tactgcggca gtaaccctgg gacctttcgg atcctagtgg ggaataaggg atgcagccac 5220
ccctcagtga aatgcaagaa acgggtcacc atcctggtgg agggaggaga gattgagctg 5280
tttgacgggg aggtgaatgt gaagaggccc atgaaggatg agactcactt tgaggtggtg 5340
gagtctggcc ggtacatcat tctgctgctg ggcaaagccc tctccgtggt ctgggaccgc 5400
cacctgagca tctccgtggt cctgaagcag acataccagg agaaagtgtg tggcctgtgt 5460
gggaattttg atggcatcca gaacaatgac ctcaccagca gcaacctcca agtggaggaa 5520
gaccctgtgg actttgggaa ctcctggaaa gtgagctcgc agtgtgctga caccagaaaa 5580
gtgcctctgg actcatcccc tgccacctgc cataacaaca tcatgaagca gacgatggtg 5640
gattcctcct gtagaatcct taccagtgac gtcttccagg actgcaacaa gctggtggac 5700
cccgagccat atctggatgt ctgcatttac gacacctgct cctgtgagtc cattggggac 5760
tgcgcctgct tctgcgacac cattgctgcc tatgcccacg tgtgtgccca gcatggcaag 5820
gtggtgacct ggaggacggc cacattgtgc ccccagagct gcgaggagag gaatctccgg 5880
gagaacgggt atgagtgtga gtggcgctat aacagctgtg cacctgcctg tcaagtcacg 5940
tgtcagcacc ctgagccact ggcctgccct gtgcagtgtg tggagggctg ccatgcccac 6000
tgccctccag ggaaaatcct ggatgagctt ttgcagacct gcgttgaccc tgaagactgt 6060
ccagtgtgtg aggtggctgg ccggcgtttt gcctcaggaa agaaagtcac cttgaatccc 6120
agtgaccctg agcactgcca gatttgccac tgtgatgttg tcaacctcac ctgtgaagcc 6180
tgccaggagc cgggaggcct ggtggtgcct cccacagatg ccccggtgag ccccaccact 6240
ctgtatgtgg aggacatctc ggaaccgccg ttgcacgatt tctgataa 6288
<210> 49
<211> 6300
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-10
<400> 49
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggtagcctat cctgtcggcc ccccatggtc 4800
aagctggtgt gtcccgctga caacctgcgg gctgaagggc tcgagtgtac caaaacgtgc 4860
cagaactatg acctggagtg catgagcatg ggctgtgtct ctggctgcct ctgccccccg 4920
ggcatggtcc ggcatgagaa cagatgtgtg gccctggaaa ggtgtccctg cttccatcag 4980
ggcaaggagt atgcccctgg agaaacagtg aagattggct gcaacacttg tgtctgtcgg 5040
gaccggaagt ggaactgcac agaccatgtg tgtgatgcca cgtgctccac gatcggcatg 5100
gcccactacc tcaccttcga cgggctcaaa tacctgttcc ccggggagtg ccagtacgtt 5160
ctggtgcagg attactgcgg cagtaaccct gggacctttc ggatcctagt ggggaataag 5220
ggatgcagcc acccctcagt gaaatgcaag aaacgggtca ccatcctggt ggagggagga 5280
gagattgagc tgtttgacgg ggaggtgaat gtgaagaggc ccatgaagga tgagactcac 5340
tttgaggtgg tggagtctgg ccggtacatc attctgctgc tgggcaaagc cctctccgtg 5400
gtctgggacc gccacctgag catctccgtg gtcctgaagc agacatacca ggagaaagtg 5460
tgtggcctgt gtgggaattt tgatggcatc cagaacaatg acctcaccag cagcaacctc 5520
caagtggagg aagaccctgt ggactttggg aactcctgga aagtgagctc gcagtgtgct 5580
gacaccagaa aagtgcctct ggactcatcc cctgccacct gccataacaa catcatgaag 5640
cagacgatgg tggattcctc ctgtagaatc cttaccagtg acgtcttcca ggactgcaac 5700
aagctggtgg accccgagcc atatctggat gtctgcattt acgacacctg ctcctgtgag 5760
tccattgggg actgcgcctg cttctgcgac accattgctg cctatgccca cgtgtgtgcc 5820
cagcatggca aggtggtgac ctggaggacg gccacattgt gcccccagag ctgcgaggag 5880
aggaatctcc gggagaacgg gtatgagtgt gagtggcgct ataacagctg tgcacctgcc 5940
tgtcaagtca cgtgtcagca ccctgagcca ctggcctgcc ctgtgcagtg tgtggagggc 6000
tgccatgccc actgccctcc agggaaaatc ctggatgagc ttttgcagac ctgcgttgac 6060
cctgaagact gtccagtgtg tgaggtggct ggccggcgtt ttgcctcagg aaagaaagtc 6120
accttgaatc ccagtgaccc tgagcactgc cagatttgcc actgtgatgt tgtcaacctc 6180
acctgtgaag cctgccagga gccgggaggc ctggtggtgc ctcccacaga tgccccggtg 6240
agccccacca ctctgtatgt ggaggacatc tcggaaccgc cgttgcacga tttctgataa 6300
6300
<210> 50
<211> 6330
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-20
<400> 50
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggtagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 4860
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 4920
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 4980
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 5040
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 5100
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 5160
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 5220
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 5280
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 5340
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 5400
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 5460
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 5520
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 5580
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 5640
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 5700
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 5760
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 5820
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 5880
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 5940
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 6000
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 6060
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 6120
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 6180
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 6240
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 6300
tcggaaccgc cgttgcacga tttctgataa 6330
<210> 51
<211> 6360
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-30
<400> 51
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggttcaggag gtggaggttc aggaggtgga ggtagcctat cctgtcggcc ccccatggtc 4860
aagctggtgt gtcccgctga caacctgcgg gctgaagggc tcgagtgtac caaaacgtgc 4920
cagaactatg acctggagtg catgagcatg ggctgtgtct ctggctgcct ctgccccccg 4980
ggcatggtcc ggcatgagaa cagatgtgtg gccctggaaa ggtgtccctg cttccatcag 5040
ggcaaggagt atgcccctgg agaaacagtg aagattggct gcaacacttg tgtctgtcgg 5100
gaccggaagt ggaactgcac agaccatgtg tgtgatgcca cgtgctccac gatcggcatg 5160
gcccactacc tcaccttcga cgggctcaaa tacctgttcc ccggggagtg ccagtacgtt 5220
ctggtgcagg attactgcgg cagtaaccct gggacctttc ggatcctagt ggggaataag 5280
ggatgcagcc acccctcagt gaaatgcaag aaacgggtca ccatcctggt ggagggagga 5340
gagattgagc tgtttgacgg ggaggtgaat gtgaagaggc ccatgaagga tgagactcac 5400
tttgaggtgg tggagtctgg ccggtacatc attctgctgc tgggcaaagc cctctccgtg 5460
gtctgggacc gccacctgag catctccgtg gtcctgaagc agacatacca ggagaaagtg 5520
tgtggcctgt gtgggaattt tgatggcatc cagaacaatg acctcaccag cagcaacctc 5580
caagtggagg aagaccctgt ggactttggg aactcctgga aagtgagctc gcagtgtgct 5640
gacaccagaa aagtgcctct ggactcatcc cctgccacct gccataacaa catcatgaag 5700
cagacgatgg tggattcctc ctgtagaatc cttaccagtg acgtcttcca ggactgcaac 5760
aagctggtgg accccgagcc atatctggat gtctgcattt acgacacctg ctcctgtgag 5820
tccattgggg actgcgcctg cttctgcgac accattgctg cctatgccca cgtgtgtgcc 5880
cagcatggca aggtggtgac ctggaggacg gccacattgt gcccccagag ctgcgaggag 5940
aggaatctcc gggagaacgg gtatgagtgt gagtggcgct ataacagctg tgcacctgcc 6000
tgtcaagtca cgtgtcagca ccctgagcca ctggcctgcc ctgtgcagtg tgtggagggc 6060
tgccatgccc actgccctcc agggaaaatc ctggatgagc ttttgcagac ctgcgttgac 6120
cctgaagact gtccagtgtg tgaggtggct ggccggcgtt ttgcctcagg aaagaaagtc 6180
accttgaatc ccagtgaccc tgagcactgc cagatttgcc actgtgatgt tgtcaacctc 6240
acctgtgaag cctgccagga gccgggaggc ctggtggtgc ctcccacaga tgccccggtg 6300
agccccacca ctctgtatgt ggaggacatc tcggaaccgc cgttgcacga tttctgataa 6360
6360
<210> 52
<211> 6390
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-40
<400> 52
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4860
ggtagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 4920
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 4980
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 5040
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 5100
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 5160
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 5220
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 5280
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 5340
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 5400
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 5460
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 5520
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 5580
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 5640
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 5700
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 5760
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 5820
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 5880
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 5940
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 6000
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 6060
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 6120
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 6180
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 6240
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 6300
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 6360
tcggaaccgc cgttgcacga tttctgataa 6390
<210> 53
<211> 6420
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-50
<400> 53
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4860
ggttcaggag gtggaggttc aggaggtgga ggtagcctat cctgtcggcc ccccatggtc 4920
aagctggtgt gtcccgctga caacctgcgg gctgaagggc tcgagtgtac caaaacgtgc 4980
cagaactatg acctggagtg catgagcatg ggctgtgtct ctggctgcct ctgccccccg 5040
ggcatggtcc ggcatgagaa cagatgtgtg gccctggaaa ggtgtccctg cttccatcag 5100
ggcaaggagt atgcccctgg agaaacagtg aagattggct gcaacacttg tgtctgtcgg 5160
gaccggaagt ggaactgcac agaccatgtg tgtgatgcca cgtgctccac gatcggcatg 5220
gcccactacc tcaccttcga cgggctcaaa tacctgttcc ccggggagtg ccagtacgtt 5280
ctggtgcagg attactgcgg cagtaaccct gggacctttc ggatcctagt ggggaataag 5340
ggatgcagcc acccctcagt gaaatgcaag aaacgggtca ccatcctggt ggagggagga 5400
gagattgagc tgtttgacgg ggaggtgaat gtgaagaggc ccatgaagga tgagactcac 5460
tttgaggtgg tggagtctgg ccggtacatc attctgctgc tgggcaaagc cctctccgtg 5520
gtctgggacc gccacctgag catctccgtg gtcctgaagc agacatacca ggagaaagtg 5580
tgtggcctgt gtgggaattt tgatggcatc cagaacaatg acctcaccag cagcaacctc 5640
caagtggagg aagaccctgt ggactttggg aactcctgga aagtgagctc gcagtgtgct 5700
gacaccagaa aagtgcctct ggactcatcc cctgccacct gccataacaa catcatgaag 5760
cagacgatgg tggattcctc ctgtagaatc cttaccagtg acgtcttcca ggactgcaac 5820
aagctggtgg accccgagcc atatctggat gtctgcattt acgacacctg ctcctgtgag 5880
tccattgggg actgcgcctg cttctgcgac accattgctg cctatgccca cgtgtgtgcc 5940
cagcatggca aggtggtgac ctggaggacg gccacattgt gcccccagag ctgcgaggag 6000
aggaatctcc gggagaacgg gtatgagtgt gagtggcgct ataacagctg tgcacctgcc 6060
tgtcaagtca cgtgtcagca ccctgagcca ctggcctgcc ctgtgcagtg tgtggagggc 6120
tgccatgccc actgccctcc agggaaaatc ctggatgagc ttttgcagac ctgcgttgac 6180
cctgaagact gtccagtgtg tgaggtggct ggccggcgtt ttgcctcagg aaagaaagtc 6240
accttgaatc ccagtgaccc tgagcactgc cagatttgcc actgtgatgt tgtcaacctc 6300
acctgtgaag cctgccagga gccgggaggc ctggtggtgc ctcccacaga tgccccggtg 6360
agccccacca ctctgtatgt ggaggacatc tcggaaccgc cgttgcacga tttctgataa 6420
6420
<210> 54
<211> 6450
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-60
<400> 54
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4860
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4920
ggtagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 4980
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 5040
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 5100
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 5160
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 5220
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 5280
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 5340
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 5400
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 5460
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 5520
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 5580
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 5640
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 5700
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 5760
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 5820
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 5880
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 5940
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 6000
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 6060
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 6120
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 6180
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 6240
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 6300
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 6360
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 6420
tcggaaccgc cgttgcacga tttctgataa 6450
<210> 55
<211> 6540
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-90
<400> 55
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4860
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4920
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4980
ggttcaggag gtggaggttc aggaggtgga ggtagcctat cctgtcggcc ccccatggtc 5040
aagctggtgt gtcccgctga caacctgcgg gctgaagggc tcgagtgtac caaaacgtgc 5100
cagaactatg acctggagtg catgagcatg ggctgtgtct ctggctgcct ctgccccccg 5160
ggcatggtcc ggcatgagaa cagatgtgtg gccctggaaa ggtgtccctg cttccatcag 5220
ggcaaggagt atgcccctgg agaaacagtg aagattggct gcaacacttg tgtctgtcgg 5280
gaccggaagt ggaactgcac agaccatgtg tgtgatgcca cgtgctccac gatcggcatg 5340
gcccactacc tcaccttcga cgggctcaaa tacctgttcc ccggggagtg ccagtacgtt 5400
ctggtgcagg attactgcgg cagtaaccct gggacctttc ggatcctagt ggggaataag 5460
ggatgcagcc acccctcagt gaaatgcaag aaacgggtca ccatcctggt ggagggagga 5520
gagattgagc tgtttgacgg ggaggtgaat gtgaagaggc ccatgaagga tgagactcac 5580
tttgaggtgg tggagtctgg ccggtacatc attctgctgc tgggcaaagc cctctccgtg 5640
gtctgggacc gccacctgag catctccgtg gtcctgaagc agacatacca ggagaaagtg 5700
tgtggcctgt gtgggaattt tgatggcatc cagaacaatg acctcaccag cagcaacctc 5760
caagtggagg aagaccctgt ggactttggg aactcctgga aagtgagctc gcagtgtgct 5820
gacaccagaa aagtgcctct ggactcatcc cctgccacct gccataacaa catcatgaag 5880
cagacgatgg tggattcctc ctgtagaatc cttaccagtg acgtcttcca ggactgcaac 5940
aagctggtgg accccgagcc atatctggat gtctgcattt acgacacctg ctcctgtgag 6000
tccattgggg actgcgcctg cttctgcgac accattgctg cctatgccca cgtgtgtgcc 6060
cagcatggca aggtggtgac ctggaggacg gccacattgt gcccccagag ctgcgaggag 6120
aggaatctcc gggagaacgg gtatgagtgt gagtggcgct ataacagctg tgcacctgcc 6180
tgtcaagtca cgtgtcagca ccctgagcca ctggcctgcc ctgtgcagtg tgtggagggc 6240
tgccatgccc actgccctcc agggaaaatc ctggatgagc ttttgcagac ctgcgttgac 6300
cctgaagact gtccagtgtg tgaggtggct ggccggcgtt ttgcctcagg aaagaaagtc 6360
accttgaatc ccagtgaccc tgagcactgc cagatttgcc actgtgatgt tgtcaacctc 6420
acctgtgaag cctgccagga gccgggaggc ctggtggtgc ctcccacaga tgccccggtg 6480
agccccacca ctctgtatgt ggaggacatc tcggaaccgc cgttgcacga tttctgataa 6540
6540
<210> 56
<211> 6630
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120
<400> 56
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctggttc cgcgtggatc cgggggtgga ggttcaggag gtggaggttc aggaggtgga 4800
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4860
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4920
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 4980
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 5040
ggttcaggag gtggaggttc aggaggtgga ggttcaggag gtggaggttc aggaggtgga 5100
ggtagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 5160
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 5220
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 5280
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 5340
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 5400
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 5460
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 5520
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 5580
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 5640
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 5700
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 5760
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 5820
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 5880
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 5940
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 6000
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 6060
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 6120
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 6180
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 6240
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 6300
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 6360
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 6420
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 6480
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 6540
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 6600
tcggaaccgc cgttgcacga tttctgataa 6630
<210> 57
<211> 6630
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH1
<400> 57
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacggtggag gttcactggt tccgcgtgga tccgggggtg gaggttcagg aggtggaggt 4800
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 4860
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 4920
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 4980
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 5040
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 5100
ggtagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 5160
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 5220
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 5280
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 5340
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 5400
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 5460
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 5520
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 5580
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 5640
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 5700
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 5760
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 5820
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 5880
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 5940
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 6000
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 6060
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 6120
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 6180
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 6240
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 6300
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 6360
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 6420
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 6480
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 6540
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 6600
tcggaaccgc cgttgcacga tttctgataa 6630
<210> 58
<211> 6630
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH1
<400> 58
gccactagaa ggtactacct gggggctgtg gagctgtcct gggattacat gcagtccgac 60
ctgggagagc tgcccgttga tgctagattc ccacctaggg tgcccaaatc tttccccttt 120
aatacatctg tggtgtacaa aaagacactg tttgtggagt ttaccgatca cctcttcaat 180
attgccaagc ctcgaccacc ctggatgggt ctccttggcc ctacaattca ggccgaggtc 240
tacgacaccg tggtaatcac cctgaagaac atggcatcac accccgtgag cctgcacgct 300
gtgggcgtgt cttactggaa ggccagtgaa ggcgccgaat atgatgacca gacaagccag 360
agagaaaagg aggacgataa ggtgttccct ggcggatctc acacatatgt gtggcaggtg 420
ctgaaggaga atggacccat ggcatctgac cccctttgcc taacttatag ttatctttcc 480
catgtggacc ttgtcaagga cctgaattct gggcttatcg gagcactgct cgtgtgtaga 540
gagggttcat tggctaagga gaagactcag acactgcaca agttcatcct gctgttcgct 600
gtcttcgatg aggggaagtc ttggcacagc gaaaccaaga attccctgat gcaagatcgc 660
gatgccgcat ccgctcgcgc atggcctaag atgcatacag tgaatgggta cgtcaataga 720
tccttgcctg gcctgattgg ctgtcacaga aaatctgtgt actggcacgt gattggtatg 780
gggaccaccc cagaagtgca tagtatcttt ttggaagggc atacctttct ggtgcggaac 840
cacagacagg cctccctgga aattagtcca atcacattcc tgacagccca gacactgctg 900
atggatctgg ggcagtttct cctgttctgt catatctcta gccaccagca tgacgggatg 960
gaggcctatg tgaaggtgga tagttgtcca gaagagcctc agttgagaat gaagaacaac 1020
gaggaggctg aagactatga tgatgatttg accgactctg aaatggatgt tgtcagattt 1080
gacgacgata atagcccaag ctttatccag atcagatcag tggcaaagaa gcaccctaag 1140
acctgggtgc attatatcgc tgccgaagag gaggactggg actacgcccc tcttgtgctg 1200
gcccctgacg ataggagcta taagtcacag tacctcaaca acggacctca gcgaattggc 1260
cggaagtata aaaaggttcg gtttatggct tacaccgatg aaaccttcaa gactagggag 1320
gccattcaac acgagtccgg tattctcggc cccctcctgt atggcgaagt gggggacaca 1380
ctgctgatca tctttaagaa ccaggcttcc cgcccctaca acatctaccc tcacggtatt 1440
accgatgtgc ggccactgta ttctaggaga ctccctaaag gagtgaagca cctcaaagat 1500
ttccctattc tgcctggtga gatctttaaa tacaaatgga ccgtcaccgt ggaggatggg 1560
cccacaaaat ccgaccccag gtgtctgacc aggtattata gtagctttgt taatatggag 1620
agggatctcg ccagcggact gatcggaccc ctgctgattt gctacaagga atctgttgat 1680
cagagaggta accagatcat gagtgataag agaaacgtga tactgttttc agtgttcgat 1740
gaaaacagat catggtactt gactgaaaat attcagcggt ttctgccaaa ccccgctgga 1800
gttcagcttg aggaccctga atttcaagct agtaacatca tgcacagcat taatggatat 1860
gtttttgatt cattgcagct gtctgtttgc ctgcacgagg tggcctattg gtatattctt 1920
agcattggtg ctcagactga cttcctgtcc gtgttcttta gcgggtacac cttcaagcac 1980
aagatggtct atgaggacac attgactctc tttccattct ccggtgagac tgtctttatg 2040
tctatggaga acccagggtt gtggattctg ggctgccata actctgattt tagaaacaga 2100
ggcatgacag ctctgctaaa ggtgagttct tgcgataaga acacaggaga ctattatgag 2160
gacagttacg aagacatctc agcgtacctt ctaagtaaga ataatgccat tgaaccaaga 2220
agtttctctc agaattcccg ccatccctcc acaaggcaga aacagttcaa cgccacaact 2280
atccctgaga atgatattga aaagacagac ccttggttcg ctcacagaac tccaatgcct 2340
aaatgccaga atgtctcctc aagcgacctg ctgatgctgc tgagacagag ccccacccct 2400
cacgggttgt ccctctctga cctgcaggaa gcaaagtatg agacttttag cgatgatccc 2460
tccccaggag caattgactc taacaattcc ctgagtgaga tgacccactt cagaccacag 2520
ttgcaccata gcggcgacat ggttttcaca cctgaaagcg gcctgcagct tcgtctgaat 2580
gagaagctcg gcacaacagc tgcaactgaa cttaagaaac tcgattttaa agtgagctcc 2640
accagtaata atctgatctc caccatccct agcgataact tggccgccgg gacagataat 2700
acttctactc tgcagagtga ccaggaggaa atcgactatg acgatactat tagcgtggag 2760
atgaagaagg aagattttga tatctatgac gaagatgaga atcagagccc caggtctttt 2820
cagaaaaaga ccagacacta tttcatcgca gctgtcgagc ggttgtggga ctacggcatg 2880
tctagttctc cccatgtgct caggaacagg gcccagtctg gcagcgtccc tcagttcaag 2940
aaggtcgtgt tccaggaatt cactgacggc tcttttacac agccactgta tcgtggagag 3000
ctcaatgaac atctgggtct gctgggtccc tacattcgcg cagaagtgga ggataacatc 3060
atggtgactt tcagaaatca ggcaagtaga ccttacagct tctactcaag cttaattagt 3120
tacgaggagg accagagaca aggcgctgag cctaggaaga actttgttaa acctaatgag 3180
acaaagacct acttctggaa ggtgcagcat catatggctc ctacaaagga tgagttcgac 3240
tgtaaagcat gggcttactt ttcagatgtg gatctggaaa aggacgttca ttcgggtctg 3300
atcggacctc tactggtgtg ccacaccaac accctaaacc cagctcatgg acgacaggtg 3360
acagtgcagg aattcgctct gttctttacc atcttcgatg agactaagtc atggtacttc 3420
actgaaaata tggagcgtaa ttgtcgagct ccatgtaaca ttcagatgga ggatcctact 3480
ttcaaagaga attacagatt tcatgccatt aatggctata ttatggatac tctgccaggc 3540
ctggtcatgg cccaggatca gaggattcga tggtacctgc tcagtatggg tagcaatgaa 3600
aacattcatt caatccattt ttctggtcat gtgttcactg tgagaaagaa agaggagtat 3660
aaaatggctc tgtataatct gtatcctgga gtgtttgaga cagttgaaat gctgccctca 3720
aaagcaggta tctggagagt ggaatgcttg atcggtgagc atctgcatgc cggtatgagc 3780
accctgttct tggtctactc caataagtgt cagacgcctc tggggatggc cagcgggcac 3840
atccgagatt tccagatcac cgccagcgga cagtacggac agtgggcacc aaagctggcc 3900
aggctccact atagcggctc tatcaacgcg tggtccacta aagaaccctt tagctggatc 3960
aaagtcgatc tgttggcccc aatgatcatc catgggatta agactcaggg tgctaggcag 4020
aaattcagca gcctgtacat atcccagttc atcattatgt atagcttgga cggaaagaaa 4080
tggcagacat atagagggaa ctctaccggc accttaatgg tgttcttcgg aaacgtcgac 4140
agcagcggca ttaagcataa tatctttaat cccccaatca ttgctcgtta tattaggctc 4200
caccctaccc attattctat ccggagtacc ctgcgaatgg aactgatggg ctgtgacctt 4260
aatagctgct ctatgcctct cggcatggaa tctaaagcaa tttccgacgc ccagatcaca 4320
gcctcttcct actttacaaa catgttcgca acctggtctc catctaaggc aagactgcac 4380
ttgcagggcc ggagtaatgc ttggaggcct caggtgaaca atcccaaaga gtggctgcag 4440
gttgattttc agaagacaat gaaggtgact ggggtgacca cacaaggtgt taagagcctg 4500
ttaacatcta tgtacgtgaa ggagtttctg atcagctctt ctcaggatgg ccatcaatgg 4560
accctatttt tccagaacgg aaaagttaag gtgttccagg gaaaccagga cagctttacc 4620
cccgtggtga atagtttgga ccctccactc ctgacacgtt atcttaggat ccacccacaa 4680
agctgggttc atcagattgc cctgagaatg gaagtcctgg ggtgcgaggc acaggacctg 4740
tacgggggag gttctctcgt gccaagaggt agtggagggg gcggatccgg aggaggtggc 4800
agcggcggag gaggatctgg agggggcggg tcaggggggg gcggctccgg cggtggtgga 4860
agcggaggcg ggggaagcgg cggcggaggg agcggaggtg gcggatccgg cgggggcggc 4920
agcggcggag gtggtagcgg ggggggtggc tctggtggag ggggatctgg cggtggcggc 4980
agtggagggg gaggttctgg aggcggcgga tctggaggag gaggatcagg cggcggaggt 5040
agtggaggag ggggaagcgg cggcggaggc agcggtggcg gtggctcagg cggcggagga 5100
ggttcactca gttgcaggcc tcccatggtg aaactggtgt gtcccgctga taacctgagg 5160
gccgagggac tggagtgcac taagacatgc caaaactacg atctggagtg tatgagcatg 5220
ggatgtgtga gtggctgtct atgtcctcct ggcatggtgc ggcacgagaa tcggtgcgtc 5280
gctctagagc gttgtccatg cttccaccag ggcaaggagt acgctcctgg cgagacagta 5340
aaaatcggtt gtaacacttg tgtttgcaga gataggaagt ggaactgtac agatcacgtc 5400
tgcgacgcca catgttccac tattggcatg gctcactacc tgaccttcga cggtctgaag 5460
tatctgttcc ctggcgaatg tcagtatgtg ctggtacagg attactgcgg gagtaatccc 5520
ggtactttta ggatcctggt cggaaataag ggttgtagtc acccttccgt aaagtgtaag 5580
aaacgggtga ccatccttgt cgaggggggg gaaatcgagc tgtttgacgg cgaggtgaat 5640
gtgaagaggc ctatgaagga tgagactcat tttgaggtgg tggagtctgg ccgctatatt 5700
attctgctgc tcggcaaagc actctctgtt gtgtgggatc ggcacctgtc aatcagcgtg 5760
gtgcttaagc agacatacca ggagaaagtg tgcggcctct gtggaaactt cgacgggatc 5820
cagaacaatg atctgaccag ctctaatctc caggtggagg aggatccagt ggactttgga 5880
aattcctgga aagtgagttc ccagtgtgca gacaccagaa aggtccctct ggactctagc 5940
cccgctacct gtcacaataa catcatgaag cagacaatgg ttgacagctc ttgtcgcatt 6000
ttgactagtg atgtgttcca ggattgtaat aagctggttg acccagaacc ctatttggat 6060
gtgtgtatct atgacacatg ctcttgtgag tctatcggtg actgtgcctg tttctgcgat 6120
acaatcgccg cttacgcgca tgtttgcgca cagcacggta aagtggtaac ttggcgaacg 6180
gcaacacttt gtccccagtc ctgtgaggaa cgtaatcttc gtgaaaacgg ctatgagtgt 6240
gagtggcgat ataatagctg cgcgccagca tgtcaagtga catgccagca tcctgagcct 6300
ctagcttgtc cagtgcagtg cgttgagggt tgccatgctc actgtccacc cggcaagatt 6360
ctggatgaac ttctgcagac atgcgtggat ccagaggatt gtccagtttg tgaagtggcc 6420
ggccgtaggt ttgctagtgg taaaaaggtg accctgaatc ccagtgaccc agagcattgc 6480
cagatctgcc actgcgacgt ggtgaacctg acatgtgaag cttgccagga acctggaggg 6540
ctggttgtgc ctcctaccga cgcaccagtg tctcccacta cactgtatgt ggaggatatc 6600
agtgagccac ctctgcatga tttctgataa 6630
<210> 59
<211> 6630
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH1
<400> 59
gccaccagaa gatattacct gggcgccgtg gaactgtcct gggactacat gcagtctgat 60
ctgggcgagc tgcccgtgga tgccagattt cctccaagag tgcccaagag cttccccttc 120
aacacctccg tggtgtacaa gaaaaccctg ttcgtggaat tcaccgacca cctgttcaat 180
atcgccaagc ctagacctcc ttggatgggc ctgctgggac ctacaattca ggccgaggtg 240
tacgacaccg tggtcatcac cctgaagaac atggcttctc accccgtgtc tctgcacgcc 300
gtgggagtgt cttattggaa ggcttctgag ggcgccgagt acgacgatca gaccagccag 360
agagagaaag aggacgacaa ggttttccct ggcggctccc acacctatgt gtggcaggtc 420
ctgaaagaaa acggccccat ggcctctgat cctctgtgcc tgacctactc ctacctgtct 480
cacgtggacc tggtcaagga cctgaattct ggactgatcg gcgctctgct cgtgtgcaga 540
gagggaagcc tggccaaaga aaagacccag acactgcaca agttcatcct gctgttcgcc 600
gtgttcgacg agggcaagtc ctggcactcc gagacaaaga actctctgat gcaggacagg 660
gacgccgcct ctgctagagc ttggcctaag atgcacaccg tgaacggcta cgtgaacaga 720
tccctgcctg gcctgatcgg atgccacaga aagtctgtgt actggcacgt gatcggcatg 780
ggcacaacac ctgaggtgca ctccatcttt ctggaaggcc acaccttcct cgtgcggaac 840
catagacagg ccagcctgga aatctcccct atcacctttc tgaccgctca gaccctgctg 900
atggatctgg gccagtttct gctgttctgt cacatctcca gccaccagca cgacggcatg 960
gaagcttacg tgaaggtgga cagctgcccc gaggaacccc agctgagaat gaagaacaac 1020
gaggaagccg aggactacga cgacgacctg accgactctg agatggatgt cgtcagattc 1080
gacgacgata actcccctag cttcatccag attcggagcg tggccaagaa gcaccctaag 1140
acttgggtgc actatatcgc cgccgaggaa gaggattggg attacgctcc tctggtgctg 1200
gcccctgacg acagatccta caagtcccag tacctgaaca acggccctca gcggatcggc 1260
cggaagtata agaaagtgcg gttcatggcc tacaccgacg agacattcaa gaccagagag 1320
gctatccagc acgagtccgg aattctggga cctctgctgt atggcgaagt gggcgacaca 1380
ctgctgatca tcttcaagaa ccaggcctct cggccctaca acatctaccc tcatggcatc 1440
acagacgtgc ggcctctgta ctctagaagg ctgcctaagg gcgtgaagca cctgaaggac 1500
ttccctattc tgcccggcga gatcttcaag tacaagtgga ccgtgaccgt ggaagatggc 1560
cccaccaagt ctgaccctag atgtctgacc cggtactact cctccttcgt gaacatggaa 1620
agagatctgg cctccggcct gattggccct ctgctgatct gctacaaaga atccgtggac 1680
cagcggggca accagatcat gtccgacaag cggaacgtga tcctgttctc tgtgttcgat 1740
gagaaccggt cctggtatct gaccgagaac atccagcggt ttctgcccaa tcctgctggc 1800
gtgcagctgg aagatcctga gttccaggcc tccaacatca tgcacagcat caacggctat 1860
gtgttcgact ccctgcagct gtctgtgtgt ctgcacgagg tggcctactg gtacatcctg 1920
tctatcggag cccagaccga cttcctgtcc gtgttcttct ccggctacac cttcaagcac 1980
aagatggtgt acgaggacac cctgacactg ttccctttct ccggcgaaac cgtgttcatg 2040
tccatggaaa accccggcct gtggatcctg ggctgccaca actctgactt ccggaacaga 2100
ggcatgaccg ctctgctgaa ggtgtcctcc tgtgacaaga acaccggcga ctactacgag 2160
gactcttacg aggatatctc cgcctacctg ctgtccaaga acaatgccat cgagccccgg 2220
tccttcagcc agaactctag acacccttcc accagacaga agcagttcaa cgccaccacc 2280
attccagaga acgacatcga aaagaccgat ccttggtttg cccacagaac ccctatgcct 2340
aagtgccaga acgtgtcctc cagcgacctg ctcatgctgc tgagacagtc tccaacacct 2400
cacggcctgt ctctgtctga cctgcaagag gctaagtacg agactttcag cgacgaccct 2460
tctcctggcg ccatcgactc taacaactcc ctgtccgaga tgacccactt cagacctcag 2520
ctgcaccact ccggcgatat ggtgtttacc cctgagtctg gcctgcagct cagactgaat 2580
gagaagctgg gcaccaccgc tgccaccgag ctgaagaaac tggacttcaa ggtgtccagc 2640
acctccaaca acctgatcag cacaatccct tccgacaacc tggccgctgg caccgataat 2700
acctctacac tgcagtccga ccaagaggaa atcgattacg acgacaccat cagcgtcgag 2760
atgaagaaag aagatttcga catctacgac gaggacgaga accagtctcc tcggagcttc 2820
cagaagaaaa ccagacacta ctttatcgcc gctgtggaac ggctgtggga ctacggaatg 2880
tcctcttctc ctcacgtgct gcggaataga gcccagtctg gctccgtgcc tcagttcaag 2940
aaagtggtgt tccaagagtt caccgacggc agcttcaccc agcctctgta tagaggcgag 3000
ctgaacgagc atctgggact gctgggccct tatatcagag ccgaggtcga ggataacatc 3060
atggtcacct tcagaaacca ggccagcaga ccctactcct tctactccag cctgatctcc 3120
tacgaagagg accagagaca aggcgccgag cctcggaaga acttcgtgaa gcctaacgag 3180
actaagacct acttttggaa ggtgcagcac cacatggctc ctaccaagga cgagttcgat 3240
tgcaaggcct gggcctactt ctccgatgtg gatctggaaa aggacgtgca ctccgggctg 3300
atcggaccac tgcttgtgtg tcacaccaac acactgaacc ctgctcacgg caggcaagtg 3360
acagtgcaag agttcgccct gttcttcacc atcttcgacg aaacaaagtc ctggtacttc 3420
accgaaaaca tggaacggaa ctgcagagcc ccttgcaaca tccagatgga agatcccacc 3480
ttcaaagaga actaccggtt ccacgccatc aatggctaca tcatggatac cctgccaggc 3540
ctcgtgatgg ctcaggatca gcgcatcaga tggtatctgc tgagcatggg ctccaacgag 3600
aatatccact ccatccactt cagcggccac gtgttcaccg tgcggaaaaa agaagagtac 3660
aagatggctc tgtacaatct gtaccccggc gtgttcgaga cagtggaaat gctgccttct 3720
aaggccggca tctggcgcgt ggaatgtctg atcggagagc atctgcatgc cggaatgtct 3780
accctgttcc tggtgtactc caacaagtgt cagacccctc tcggcatggc ttccggccat 3840
atcagagact tccagatcac cgcttccgga cagtatggcc agtgggctcc taagctggcc 3900
agactgcact actctggctc catcaacgcc tggtctacca aagagccctt cagctggatc 3960
aaggtggacc tgctggctcc catgatcatc cacggcatca agacacaggg cgctagacag 4020
aagttcagct ccctgtacat ctcccagttc atcatcatgt actccctgga cggcaagaag 4080
tggcagacct acagaggcaa ctccaccggc acactcatgg tgttcttcgg caacgtggac 4140
tcctccggca ttaagcacaa catcttcaac cctccaatca ttgcccggta catccggctg 4200
caccccacac actactccat cagatctacc ctgagaatgg aactgatggg ctgcgacctg 4260
aactcctgtt ctatgcccct cggaatggaa tccaaggcca tcagcgacgc ccagattacc 4320
gcctcctcct acttcaccaa catgttcgcc acttggagcc cctctaaggc caggctgcat 4380
ctgcagggca gatctaatgc ttggcggcct caagtgaaca accccaaaga gtggctgcag 4440
gtcgactttc agaaaaccat gaaggtcaca ggcgtgacaa cccagggcgt caagtccctg 4500
ctgacctcta tgtacgtgaa agagttcctg atctctagca gccaggacgg ccaccagtgg 4560
accctgtttt tccagaacgg caaagtgaag gtgttccagg gcaatcagga ttctttcacc 4620
cctgtggtca acagcctgga tcctccactg ctgaccagat acctgcggat ccatcctcag 4680
tcctgggtgc accagattgc tctgcgaatg gaagtgctgg gatgtgaagc ccaagatctg 4740
tacggcggag gctctctggt tcctagagga tctggcggtg gtggttctgg cggcggagga 4800
agcggaggcg gaggtagtgg tggcggtgga agtggcggtg gcggatccgg tggcggcgga 4860
tctggcggag gcggcagcgg cggtggtggt agcggcggag gcggctcagg tggcggaggc 4920
agtggcggcg gaggttcagg cggtggtgga agtggcggcg gtggctctgg cggtggcggc 4980
tcaggcggtg gtggctccgg cggaggcgga agcggcggag gcggcagcgg aggcggaggc 5040
tctggtggcg gaggttccgg tggcggaggc agcggcggag gcggctctgg cggaggtggt 5100
ggatctttgt cttgtcggcc tcctatggtc aagctcgtgt gtcccgccga taatctgaga 5160
gccgaaggac tggaatgcac caagacctgc cagaactacg acctcgagtg catgtctatg 5220
ggctgtgtgt ccggatgtct gtgccctcct ggcatggtcc gacacgagaa tagatgcgtg 5280
gccctggaaa gatgcccctg cttccaccaa ggcaaagagt acgctcctgg cgagactgtg 5340
aagatcggct gcaacacctg tgtgtgtcgg gacagaaagt ggaactgcac agaccacgtg 5400
tgcgacgcca catgctctac catcggaatg gcccactacc tgaccttcga cggcctgaag 5460
tacctgtttc caggcgagtg ccagtatgtg ctggtgcagg actactgtgg cagcaacccc 5520
ggcaccttta gaatcctcgt gggcaacaag ggctgctctc acccctccgt gaaatgcaag 5580
aagcgggtca caatcctggt cgaaggcgga gagatcgagc tgttcgacgg cgaagtgaac 5640
gtgaagcggc ccatgaagga cgagacacac ttcgaggttg tggaatccgg ccggtacatt 5700
atcctgctgc tgggcaaagc tctgtccgtc gtgtgggata gacacctgtc catttccgtg 5760
gtgctgaagc agacctatca agagaaagtg tgcggcctgt gcggcaactt tgacggcatc 5820
cagaacaatg acctgacctc cagcaatctc caggtcgaag aggatcccgt ggatttcggc 5880
aactcctgga aagtgtccag ccagtgcgcc gacacaagaa aggtgccact ggattctagc 5940
cctgccacct gtcacaacaa tatcatgaag caaaccatgg tggactccag ctgccggatc 6000
ctgacatccg atgtgtttca ggactgcaac aagctggtgg accccgagcc ttacctggac 6060
gtgtgtatct acgatacctg ctcctgcgag tctatcggcg actgcgcctg cttttgcgac 6120
acaatcgctg cttacgccca tgtgtgcgct cagcatggaa aggtggtcac ttggagaacc 6180
gccacactgt gtccccagtc ctgcgaggaa agaaacctgc gcgagaatgg ctatgagtgc 6240
gagtggcggt acaactcttg tgcccctgcc tgtcaagtga cctgtcagca tcctgaacca 6300
ctggcttgtc ccgtgcagtg cgtggaaggc tgtcatgctc attgcccacc aggcaagatc 6360
ctggatgagc tgctgcagac ctgtgtggat ccagaggatt gtcctgtgtg cgaagtggcc 6420
ggcagaagat tcgcctccgg aaagaaagtg accctgaatc ctagcgaccc tgagcactgc 6480
cagatctgcc actgtgatgt cgtgaacctg acctgcgagg cttgtcaaga acctggcgga 6540
ctggtggtgc ctcctacaga tgctcctgtg tctcccacca cactgtacgt cgaggacatc 6600
tctgagccac ctctgcacga tttctgatag 6630
<210> 60
<211> 6630
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH1
<400> 60
gctactcgtc gttactacct gggagctgtc gagctgagct gggattacat gcagtctgac 60
ctgggagagc tgcctgtgga cgctaggttt ccacctcggg tgcctaagtc cttccccttc 120
aacaccagcg tggtgtataa gaagaccctg ttcgtggagt ttacagatca cctgttcaac 180
atcgctaagc ctagaccacc ctggatgggc ctgctgggcc caacaatcca ggccgaggtg 240
tacgacaccg tggtcatcac cctgaagaac atggcttccc accctgtgag cctgcatgct 300
gtgggcgtgt cctactggaa ggccagcgag ggcgctgagt atgacgatca gacctctcag 360
cgcgagaagg aggacgataa ggtgtttcca ggcggctccc atacctacgt gtggcaggtg 420
ctgaaggaga atggcccaat ggcttctgat cccctgtgcc tgacctactc ttatctgtcc 480
cacgtggacc tggtgaagga tctgaacagc ggcctgatcg gcgctctgct ggtgtgcagg 540
gagggctctc tggccaagga gaagacccag acactgcata agttcatcct gctgttcgcc 600
gtgtttgacg agggcaagag ctggcactct gagacaaaga attctctgat gcaggacaga 660
gatgctgctt ccgccagggc ttggccaaag atgcatacag tgaacggcta cgtgaataga 720
tccctgcctg gcctgatcgg atgccacagg aagagcgtgt attggcatgt gatcggcatg 780
ggcaccacac cagaggtgca ctctatcttc ctggagggcc atacctttct ggtgaggaat 840
caccggcagg cttctctgga gatctccccc atcaccttcc tgacagccca gaccctgctg 900
atggatctgg gccagttcct gctgttttgc cacatctcca gccaccagca tgatggcatg 960
gaggcttacg tgaaggtgga ctcctgtcct gaggagccac agctgaggat gaagaacaat 1020
gaggaggccg aggactatga cgatgacctg accgactctg agatggatgt ggtgaggttc 1080
gatgacgata actcccccag ctttatccag atccggtccg tggccaagaa gcaccctaag 1140
acatgggtgc attacatcgc cgctgaggag gaggactggg attatgctcc tctggtgctg 1200
gccccagacg atcggtccta caagagccag tatctgaaca atggccccca gagaatcggc 1260
cgcaagtaca agaaggtgag attcatggct tatacagatg agacattcaa gacacgcgag 1320
gccatccagc acgagagcgg aatcctggga cctctgctgt acggagaagt gggcgacacc 1380
ctgctgatca tctttaagaa ccaggccagc agaccctaca atatctatcc tcatggcatc 1440
acagatgtgc gcccactgta ctctaggcgg ctgcccaagg gcgtgaagca cctgaaggac 1500
ttccccatcc tgcctggcga gatctttaag tataagtgga ccgtgacagt ggaggatggc 1560
ccaaccaaga gcgaccccag atgcctgaca cgctactatt cttccttcgt gaatatggag 1620
cgggatctgg cttctggcct gatcggacca ctgctgatct gttacaagga gtctgtggat 1680
cagaggggca accagatcat gtccgacaag cggaatgtga tcctgttcag cgtgtttgac 1740
gagaacaggt cttggtatct gactgagaac atccagcggt tcctgcctaa tccagctggc 1800
gtgcagctgg aggatcctga gtttcaggcc tctaacatca tgcattccat caatggctac 1860
gtgttcgaca gcctgcagct gtccgtgtgc ctgcacgagg tggcttactg gtatatcctg 1920
tccatcggcg cccagaccga tttcctgtcc gtgttcttta gcggctacac cttcaagcat 1980
aagatggtgt atgaggacac cctgacactg ttcccatttt ccggcgagac agtgttcatg 2040
agcatggaga atcccggcct gtggatcctg ggctgccaca acagcgattt caggaatcgg 2100
ggcatgaccg ccctgctgaa ggtgagctct tgtgacaaga acacaggcga ctactatgag 2160
gatagctacg aggacatctc tgcttatctg ctgtccaaga acaatgccat cgagcccaga 2220
tctttttccc agaattctag gcatccttcc acccggcaga agcagttcaa cgctaccaca 2280
atcccagaga atgatatcga gaagaccgac ccctggtttg cccataggac acccatgcct 2340
aagtgtcaga acgtgtccag ctctgatctg ctgatgctgc tgaggcagtc tccaacccca 2400
cacggcctga gcctgtctga cctgcaggag gctaagtacg agacattcag cgacgatcct 2460
tctccaggcg ccatcgattc caacaattcc ctgagcgaga tgacccactt caggcctcag 2520
ctgcaccata gcggcgacat ggtgtttaca ccagagtctg gcctgcagct gcggctgaac 2580
gagaagctgg gcaccacagc cgctaccgag ctgaagaagc tggactttaa ggtgtccagc 2640
acctccaaca atctgatctc cacaatccct agcgataacc tggccgctgg caccgacaat 2700
acctctacac tgcagtccga tcaggaggag atcgactacg acgatacaat cagcgtggag 2760
atgaagaagg aggacttcga tatctatgac gaggatgaga accagtctcc aagatccttc 2820
cagaagaaga cccgccatta ctttatcgcc gctgtggaga ggctgtggga ttatggcatg 2880
tcttccagcc ctcacgtgct gagaaatcgc gcccagtctg gctccgtgcc acagttcaag 2940
aaggtggtgt tccaggagtt taccgacggc agctttacac agccactgta cagaggcgag 3000
ctgaacgagc acctgggcct gctgggacca tatatcaggg ctgaggtgga ggataacatc 3060
atggtgacct tcagaaatca ggccagccgc ccatactctt tttattcttc cctgatctct 3120
tacgaggagg accagaggca gggagctgag cctcggaaga acttcgtgaa gccaaatgag 3180
acaaagacat acttttggaa ggtgcagcac catatggccc ccaccaagga cgagttcgat 3240
tgcaaggcct gggcttactt ctccgacgtg gatctggaga aggacgtgca ctccggcctg 3300
attggaccac tgctggtgtg ccataccaac acactgaatc ctgctcacgg caggcaggtg 3360
accgtgcagg agttcgccct gttctttacc atctttgatg agacaaagtc ctggtacttc 3420
acagagaaca tggagaggaa ttgtcgggct ccctgcaata tccagatgga ggaccctacc 3480
ttcaaggaga actaccggtt tcatgctatc aatggctata tcatggatac actgccaggc 3540
ctggtcatgg cccaggacca gaggatccgg tggtacctgc tgagcatggg ctctaacgag 3600
aatatccact ccatccattt cagcggccac gtgtttaccg tgagaaagaa ggaggagtat 3660
aagatggccc tgtacaacct gtatccaggc gtgttcgaga cagtggagat gctgcccagc 3720
aaggctggca tctggagggt ggagtgcctg atcggagagc acctgcatgc cggcatgtct 3780
accctgtttc tggtgtactc caataagtgt cagacacctc tgggcatggc ttctggccat 3840
atcagagatt tccagatcac cgcttccgga cagtacggac agtgggctcc aaagctggct 3900
cgcctgcact atagcggctc tatcaacgcc tggtctacca aggagccatt ttcctggatc 3960
aaggtggacc tgctggctcc catgatcatc catggcatca agacacaggg cgccagacag 4020
aagttcagct ctctgtacat cagccagttt atcatcatgt attctctgga tggcaagaag 4080
tggcagacct accgcggcaa tagcaccggc acactgatgg tgttctttgg caacgtggac 4140
tccagcggca tcaagcacaa catcttcaat cctccaatca tcgctagata catccgcctg 4200
caccccaccc attattccat caggagcaca ctgcggatgg agctgatggg ctgcgatctg 4260
aactcctgta gcatgcctct gggcatggag tctaaggcca tctccgacgc tcagatcacc 4320
gcctcttcct acttcaccaa tatgtttgct acatggtccc cctccaaggc ccggctgcat 4380
ctgcagggca ggtctaacgc ctggcggcct caggtgaaca atccaaagga gtggctgcag 4440
gtggattttc agaagaccat gaaggtgaca ggcgtgacca cacagggcgt gaagagcctg 4500
ctgacctcta tgtatgtgaa ggagttcctg atcagctctt cccaggacgg ccaccagtgg 4560
acactgttct ttcagaacgg caaggtgaag gtgttccagg gcaatcagga ttcctttacc 4620
ccagtggtga acagcctgga cccccctctg ctgacaaggt acctgcggat ccacccccag 4680
tcctgggtgc atcagatcgc tctgagaatg gaggtgctgg gatgcgaggc tcaggacctg 4740
tatggaggcg gcagcctggt gccccgcggc agtggcggcg gcggcagcgg cggcggcggc 4800
tccggcggcg gtggttctgg tggtggtggt tccggtggtg gcggctccgg cggcggaggt 4860
tctggtggtg gtggatccgg tggaggcggc tccggcggcg ggggttctgg tggtggtggg 4920
tccggtgggg gcggctccgg cggcggcggt tctggtggtg gtggctccgg tggcggtggc 4980
tccggcggcg gtggatctgg tggtggtggt agcggtggcg gaggctccgg cggcggtggg 5040
tctggtggtg gtggttcagg cggcggcggc tccggaggag gaggaagcgg cggaggagga 5100
ggcagcctgt cttgcaggcc acccatggtg aagctggtgt gccctgctga taatctgcgg 5160
gccgagggcc tggagtgtac caagacatgc cagaactacg acctggagtg catgtctatg 5220
ggatgcgtgt ccggatgcct gtgccctcca ggcatggtga gacatgagaa ccgctgcgtg 5280
gctctggaga gatgcccttg ttttcaccag ggcaaggagt atgccccagg cgagacagtg 5340
aagatcggct gcaatacatg cgtgtgcagg gacaggaagt ggaactgtac cgatcacgtg 5400
tgcgacgcta cctgcagcac aatcggcatg gcccactacc tgacattcga tggcctgaag 5460
tatctgtttc caggcgagtg ccagtacgtg ctggtgcagg actattgtgg ctctaatcct 5520
ggcaccttca gaatcctggt gggcaacaag ggctgctctc acccatccgt gaagtgtaag 5580
aagcgcgtga caatcctggt ggagggcggc gagatcgagc tgtttgatgg cgaagtgaat 5640
gtgaagaggc ccatgaagga cgagacacat ttcgaggtgg tggagtccgg ccggtacatc 5700
atcctgctgc tgggcaaggc cctgagcgtg gtgtgggaca ggcacctgtc catcagcgtg 5760
gtgctgaagc agacctatca ggagaaggtg tgcggcctgt gcggcaactt tgatggcatc 5820
cagaacaatg acctgacaag ctctaatctg caggtggagg aggaccctgt ggatttcggc 5880
aactcttgga aggtgtccag ccagtgcgct gataccagga aggtgcccct ggactcttcc 5940
cctgccacct gtcacaacaa tatcatgaag cagacaatgg tggatagctc ttgccggatc 6000
ctgacatccg acgtgttcca ggactgtaac aagctggtgg accccgagcc ttacctggac 6060
gtgtgcatct atgacacctg cagctgtgag tctatcggcg attgcgcttg tttctgcgac 6120
acaatcgccg cttacgctca cgtgtgcgcc cagcatggca aggtggtgac ctggaggacc 6180
gccaccctgt gcccacagtc ctgcgaggag agaaacctgc gcgagaatgg ctacgagtgc 6240
gagtggcggt ataacagctg tgcccccgct tgccaggtga cctgtcagca cccagagccc 6300
ctggcttgcc ctgtgcagtg cgtggaggga tgccacgctc attgtccacc tggcaagatc 6360
ctggatgagc tgctgcagac ctgcgtggac cccgaggatt gtcccgtgtg cgaggtggct 6420
ggcagacgct tcgcttccgg caagaaggtg acactgaatc caagcgatcc cgagcactgt 6480
cagatctgtc attgcgacgt ggtgaacctg acctgtgagg cttgccagga gcctggaggc 6540
ctggtggtgc cacccacaga cgcccctgtc agcccaacta ccctgtatgt ggaagatatt 6600
tcagagcccc ccctgcacga tttctgataa 6630
<210> 61
<211> 6630
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-120-TH2
<400> 61
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc aatcaatgcc tggagcacca aggagccctt ttcttggatc 3960
aaggtggatc tgttggcacc aatgattatt cacggcatca agacccaggg tgcccgtcag 4020
aagttctcca gcctctacat ctctcagttt atcatcatgt atagtcttga tgggaagaag 4080
tggcagactt atcgaggaaa ttccactgga accttaatgg tcttctttgg caatgtggat 4140
tcatctggga taaaacacaa tatttttaac cctccaatta ttgctcgata catccgtttg 4200
cacccaactc attatagcat tcgcagcact cttcgcatgg agttgatggg ctgtgattta 4260
aatagttgca gcatgccatt gggaatggag agtaaagcaa tatcagatgc acagattact 4320
gcttcatcct actttaccaa tatgtttgcc acctggtctc cttcaaaagc tcgacttcac 4380
ctccaaggga ggagtaatgc ctggagacct caggtgaata atccaaaaga gtggctgcaa 4440
gtggacttcc agaagacaat gaaagtcaca ggagtaacta ctcagggagt aaaatctctg 4500
cttaccagca tgtatgtgaa ggagttcctc atctccagca gtcaagatgg ccatcagtgg 4560
actctctttt ttcagaatgg caaagtaaag gtttttcagg gaaatcaaga ctccttcaca 4620
cctgtggtga actctctaga cccaccgtta ctgactcgct accttcgaat tcacccccag 4680
agttgggtgc accagattgc cctgaggatg gaggttctgg gctgcgaggc acaggacctc 4740
tacctgagta aaaacaatgc cattgaacca agagggggtg gaggttcagg aggtggaggt 4800
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 4860
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 4920
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 4980
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 5040
tcaggaggtg gaggttcagg aggtggaggt tcaggaggtg gaggttcagg aggtggaggt 5100
ggtagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 5160
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 5220
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 5280
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 5340
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 5400
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 5460
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 5520
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 5580
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 5640
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 5700
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 5760
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 5820
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 5880
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 5940
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 6000
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 6060
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 6120
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 6180
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 6240
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 6300
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 6360
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 6420
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 6480
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 6540
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 6600
tcggaaccgc cgttgcacga tttctgataa 6630
<210> 62
<211> 7170
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3-300
<400> 62
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc catcaacgca tggtccacca aggagccatt cagctggatc 3960
aaggtggacc tcctcgcacc tatgatcatc cacggcatca agacccaggg cgcaaggcag 4020
aagttctcca gcctctacat ctcccagttc atcatcatgt actccctgga cggcaagaag 4080
tggcagacct accgcggcaa ctccaccggc accctcatgg tcttcttcgg caacgtcgac 4140
tcctccggca tcaagcacaa catcttcaac cctcctatca tcgccaggta catccgtctc 4200
caccccaccc actactccat ccgaagcacc ctgcgaatgg agctcatggg ctgcgacctc 4260
aactcctgct ccatgcccct gggcatggag agcaaggcaa tctccgacgc tcagatcacc 4320
gcatcctcct acttcaccaa catgttcgct acctggagcc ctagcaaggc ccggctccac 4380
cttcagggca ggtccaacgc atggcgtcca caggtcaaca accctaagga gtggctgcag 4440
gtcgacttcc agaagactat gaaggtcacc ggcgtgacca cccagggagt caagtccctc 4500
ctcacctcca tgtacgtcaa ggagttcctc atctcctcca gccaggacgg ccaccagtgg 4560
accctcttct tccagaacgg aaaggtgaag gtgttccagg gaaaccagga ctccttcacc 4620
cccgtcgtca actccctcga ccccccactc ctgacccgtt acctgcgtat ccatccacag 4680
agctgggtcc atcagatagc cctgagaatg gaggtgctgg gatgcgaggc tcaggacctg 4740
tacctggtcc caagaggatc tggaggtgga ggcagcggcg gagggggatc aggcggaggc 4800
ggatcaggcg gcggtggctc tggaggtggc ggatctggag gaggtggttc aggaggaggt 4860
ggaagcggtg gtggaggttc aggtggagga ggttcaggag gtggtgggtc aggtgggggg 4920
ggatcaggag gtgggggaag tggtggtgga ggatctgggg gtggtggaag tggtggaggt 4980
ggtagtggtg gaggaggctc aggtggtggg ggttctggag gaggaggttc tggtggggga 5040
ggttcaggcg gaggtggaag tgggggtgga ggtagtggag gcggtggtag tggcggtggt 5100
ggcagcggag gaggggggtc tggaggaggc ggttcaggag gcggagggtc tggaggcgga 5160
ggaagtgggg ggggtggatc aggaggggga ggaagtggcg gtggagggag tggaggaggg 5220
ggaagtggag gcggcggttc tggagggggc ggatctgggg gaggaggctc tggaggcggg 5280
ggttcaggag gggggggaag cggcggcgga ggcagtggtg ggggcggcag tggcgggggc 5340
gggagtggtg gcggggggtc agggggcggg gggagtgggg gagggggctc aggcggcggg 5400
gggagcgggg gaggtggttc tgggggcggg ggctcaggtg ggggcgggag cggaggcggt 5460
gggtcaggcg gaggagggtc aggcgggggg ggcagtggcg gtggggggtc tggtggcggt 5520
gggagtgggg gcggtggtag cggcggtggt ggatcagggg ggggcggctc tggcggcggg 5580
ggctctggcg gaggtggttc cgggggcggc gggagcggcg gcggtgggtc cggcgggggc 5640
ggcagccttt cttgtcgacc acctatggta aaacttgttt gtcctgctga taatcttaga 5700
gcagaaggct tggaatgtac aaaaacttgt caaaattatg atttggaatg tatgagtatg 5760
ggctgtgtaa gtgggtgttt gtgtcctcca ggcatggttc gacatgaaaa tcgatgtgta 5820
gctcttgaaa gatgtccatg ttttcatcaa ggcaaagaat atgctcctgg cgaaactgta 5880
aaaattggct gtaatacatg tgtatgtaga gatagaaaat ggaattgtac agatcatgta 5940
tgtgatgcaa cttgttctac aattgggatg gcacattatt tgacatttga tggcttgaaa 6000
tatttgtttc caggcgaatg tcaatatgta ttggttcaag attattgtgg ctctaatcca 6060
ggcactttta ggatacttgt tggcaataaa ggctgttctc atccaagtgt taaatgtaaa 6120
aaaagggtta ctattcttgt tgaaggcggc gaaattgaac tttttgatgg cgaagttaac 6180
gttaaaaggc caatgaaaga tgaaacacat tttgaggttg ttgaaagtgg ccgctatatt 6240
attcttttgt tggggaaagc tctttctgtg gtgtgggatc ggcatctttc tatatctgtg 6300
gtgttgaaac aaacatatca agaaaaggtg tgtggcctgt gtggcaactt tgacgggatt 6360
cagaacaacg acctgaccag cagcaacctg caagtggagg aggaccctgt ggactttggc 6420
aacagctgga aggtgagctc ccagtgcgcc gacacccgca aggtgccttt ggactcttct 6480
ccagctacat gccacaataa tattatgaag cagacaatgg tggacagcag ctgccgcatt 6540
ctgacaagcg acgtgtttca ggactgcaat aagctggtgg accccgagcc ttacctggac 6600
gtctgcatat acgacacttg ctcctgcgag agcattggcg attgcgcctg cttttgcgat 6660
acaattgccg cctacgccca cgtctgcgcc cagcacggca aggtggtgac atggcgcaca 6720
gccactctgt gccctcagag ctgcgaggag cggaatctgc gggagaatgg ctacgagtgc 6780
gagtggcgct ataattcctg cgctcccgcc tgccaggtca cttgccagca ccccgagcct 6840
ctggcttgcc cagtgcagtg cgtggaaggc tgtcatgctc attgccctcc aggcaagatt 6900
ctggatgagc tgctgcagac atgcgtcgat cccgaggatt gccctgtatg cgaggtggct 6960
gggcggcggt ttgcctctgg gaaaaaggtg actctgaatc cctctgatcc cgagcactgt 7020
caaatatgcc actgcgatgt ggtgaatctg acttgcgagg cctgtcaaga gcccggcggc 7080
ctggtggtgc cccccacaga tgcccccgtg tctcccacta ctctgtatgt ggaagatatt 7140
agcgagcccc ccctgcacga tttttgataa 7170
<210> 63
<211> 8241
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3/D'D3-120
<400> 63
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggaag catcaacgca tggtccacca aggagccttt ctcctggatc 3960
aaggtggacc tgcttgcacc tatgatcatc cacggaatca agacccaggg cgctcgtcaa 4020
aagttctcct ccctgtacat ctcccagttc atcatcatgt actccctgga cggaaagaag 4080
tggcagacct acaggggaaa ctccaccggc accctgatgg tcttcttcgg aaacgtggac 4140
tccagcggaa tcaagcacaa catcttcaac cctcccatca tcgcccgtta catccgtctc 4200
caccctaccc actactccat ccgttccacc cttcgtatgg agctgatggg atgcgatctc 4260
aacagctgct ccatgcccct cggaatggaa agcaaggcaa tctccgacgc tcaaatcacc 4320
gcttcctcct acttcaccaa catgttcgcc acctggtccc catcaaaggc acgacttcac 4380
ctgcaaggac gttccaacgc atggcgacct caagtaaaca accctaagga gtggctgcaa 4440
gtggacttcc agaagaccat gaaggtgacc ggagtgacca cccaaggagt aaagtccctc 4500
ctcacctcca tgtacgtcaa ggagttcctg atctcctcat cacaagacgg acatcagtgg 4560
acccttttct tccagaacgg caaggtgaag gtcttccagg gcaaccagga ctccttcacc 4620
cctgtagtca actccctcga ccctccattg ctcacccgat acttgcgaat ccatccacag 4680
tcctgggttc atcagatcgc actccgaatg gaagtcctgg gctgtgaggc acaagacctt 4740
tacctggtgc ctagaggaag cggaggcgga ggaagtggag gcggtggatc tggtggcggg 4800
ggatctggtg gtggaggtag tggtggagga ggttcaggtg gaggtggttc tggaggtggt 4860
ggttctggtg gcggaggttc aggaggtgga ggttcagggg gtggtggatc aggaggtggc 4920
ggttctggtg ggggaggatc tggaggagga ggcagtggtg gtggcggtag tggaggaggt 4980
ggtagtggtg gtgggggttc aggagggggt ggttcaggag gaggcggatc tggaggtggg 5040
gggagtggcg gaggaggtag tggcggaggg ggatcaggag gagggggatc tggcggtggt 5100
gggtcacttt cttgtcgacc tccaatggta aaacttgtat gtccagctga taatcttaga 5160
gcagaaggcc ttgaatgtac aaaaacatgt caaaattatg atcttgaatg tatgtctatg 5220
ggttgtgtat caggttgtct ttgtccacct ggaatggtac gacatgaaaa tagatgtgta 5280
gcacttgaaa gatgtccttg ttttcatcaa gggaaagaat atgctccagg ggaaactgta 5340
aaaatagggt gtaatacttg tgtatgcagg gatagaaagt ggaactgcac cgaccacgtg 5400
tgtgacgcta cctgttccac catcggcatg gctcattacc tgaccttcga cgggcttaaa 5460
tacctgttcc ccggggagtg tcaatacgtg cttgtgcaag actactgcgg ctcaaaccca 5520
gggacattca gaatcctcgt cggcaacaag gggtgttccc atccttccgt taagtgcaag 5580
aagagggtga ccatcctggt ggaaggcggg gaaatcgaac ttttcgacgg ggaagttaac 5640
gtgaagcgcc caatgaagga cgagacccat ttcgaggtgg tcgaaagcgg gagatacatc 5700
atcctgctgc tcgggaaggc tttgtccgtt gtttgggaca gacacctctc catctcagtg 5760
gtcctcaagc agacctacca ggaaaaggtg tgcggcttgt gtgggaactt cgacggcata 5820
cagaacaacg acctcacctc atcaaatctc caggtcgagg aagaccctgt cgactttggg 5880
aacagctgga aggtgagcag ccagtgcgca gataccagaa aggttccact cgacagcagc 5940
ccagcaactt gtcataacaa catcatgaag cagaccatgg tggacagcag ctgcaggatt 6000
ctgaccagcg acgtctttca ggactgcaac aagctggtcg acccagagcc atacctcgac 6060
gtttgcattt acgacacatg cagctgcgag agcattgggg actgcgcatg cttttgcgac 6120
accattgctg cttacgccca tgtgtgcgct cagcatggga aggttgttac atggaggaca 6180
gctacactgt gcccacagag ctgcgaagaa agaaacctca gggagaacgg gtacgaatgc 6240
gagtggaggt acaacagctg cgctccagca tgtcaggtaa cttgccagca cccagaacct 6300
cttgcttgcc cagttcaatg cgttgaaggg tgccatgctc attgcccacc aggcaaaatt 6360
ctggacgaac tgctccagac ttgcgtcgat ccagaagact gcccagtatg cgaggttgct 6420
ggcaggaggt ttgcaagtgg gaaaaaggtt acactcaacc caagcgaccc agaacactgc 6480
cagatatgcc actgcgacgt tgttaacctg acttgcgagg catgccagga acctggcggc 6540
cttgttgttc ctccaacaga tgctccagtt agcccaacaa cattgtatgt ggaggacatt 6600
agcgagcccc ctttgcacga ctttggcggg ggaggcagtg ggggcggggg cagtggcggg 6660
ggcggctctg gggggggcgg ctcagggggc ggtggtagtg gcggcggggg cgggagtttg 6720
tcttgtaggc ctcctatggt aaaattggtt tgtcctgctg ataatttgag ggctgaaggg 6780
ttggagtgta caaaaacttg tcagaattat gatttggaat gcatgtctat gggctgtgtt 6840
agtggctgtt tgtgtcctcc tggcatggtc cggcatgaaa atcggtgtgt ggctttggaa 6900
cggtgtccct gttttcatca gggcaaagag tatgcccctg gcgagacagt caaaattggc 6960
tgtaatacat gtgtgtgtcg ggatcggaaa tggaattgta cagatcatgt gtgtgatgcc 7020
acttgttcta ctattggcat ggcccactat ttgacttttg acggcctgaa gtatctgttt 7080
cccggcgagt gccagtatgt gctggtgcag gactattgcg gcagcaaccc tggcacattt 7140
cgcatactgg tggggaataa gggctgcagt caccctagtg tgaagtgcaa gaaacgcgtg 7200
actattctgg tggagggggg cgaaattgag ttgtttgacg gcgaggtgaa tgtgaagcgg 7260
cctatgaagg atgagactca ctttgaggtg gtggagagcg gccggtatat tatactcctg 7320
ctgggcaaag ccctgagcgt cgtgtgggat cgccacttgt ctattagcgt ggtgttgaaa 7380
cagacttatc aggagaaagt ctgcgggctg tgcggcaatt ttgatgggat tcagaataat 7440
gatctgacta gcagcaatct gcaggtggaa gaagatcccg tcgattttgg caatagttgg 7500
aaagtgtctt ctcagtgcgc cgatacacgg aaagtgcccc tggatagtag tcccgccaca 7560
tgccacaata atattatgaa acagacaatg gtggatagtt cttgccgcat tctgacttct 7620
gatgtgtttc aggattgcaa taaactggtc gatcccgagc cctatctgga tgtgtgcata 7680
tatgatactt gctcttgcga gtctattggc gattgcgcct gcttttgcga tacaattgcc 7740
gcctatgccc acgtgtgcgc ccagcacggc aaagtcgtca cttggcggac tgccactctg 7800
tgcccccagt cttgcgagga gcgcaatctc cgcgagaatg gctatgagtg cgagtggcgc 7860
tataattctt gcgcccccgc ctgccaggtg acttgtcagc accccgagcc cctggcctgt 7920
cccgtgcagt gtgtggaggg ctgtcacgcc cactgtcccc ccggcaaaat tctcgatgag 7980
ctgctgcaga catgtgtgga ccccgaggat tgtcccgtct gtgaggtggc cggccggcgc 8040
tttgcctctg gcaaaaaagt cacactgaat ccctctgatc ccgagcactg tcagatatgt 8100
cattgtgatg tcgtcaatct cacttgtgag gcctgtcagg agcccggcgg cctggtggtg 8160
cctcccacag atgcccccgt gtctcccaca actttgtatg tcgaggatat ttctgagccc 8220
cccctgcatg atttttgata a 8241
<210> 64
<211> 8853
<212> DNA
<213> Artificial Sequence
<220>
<223> D'D3/scFVIII-120
<400> 64
gcagaaggaa cccgtgggag atcctccaca gcacgttgtt cccttttcgg ctccgacttc 60
gtcaacacct tcgacggctc catgtactcc ttcgccggat actgctcata cctgctcgca 120
ggagggtgcc agaagcgttc tttctccatc atcggagact tccagaacgg aaagcgtgtc 180
tccctgtccg tctacctggg ggaattcttc gacatccacc tcttcgtcaa cggaaccgtc 240
acccagggag accagcgtgt cagtatgcct tacgcatcca agggacttta cctggagacc 300
gaggcaggat actacaagct cagcggagag gcatacggct tcgtcgcaag aatcgacggc 360
tccggcaact tccaggtgct gctctccgac cgttacttca acaagacctg cggtctttgc 420
ggtaacttca acatcttcgc cgaggacgac ttcatgaccc aggagggcac ccttacctca 480
gacccatacg acttcgcaaa cagctgggct ctttcctccg gagagcagtg gtgtgagcga 540
gcatctcctc ctagttcttc ctgtaacatc tccagcggtg agatgcagaa gggtctgtgg 600
gagcagtgtc agctcttgaa gtcaacctcc gtcttcgcac gatgccaccc attggtcgat 660
cctgagccat tcgtagctct ttgcgagaag accctctgcg aatgcgcagg aggtcttgaa 720
tgcgcatgtc cagcattgct cgagtacgca cgaacttgcg cacaagaggg tatggtactc 780
tacggttgga ccgaccattc agcatgctcc ccagtatgcc cagcaggaat ggaataccga 840
cagtgtgtat ccccatgtgc tcgaacctgt cagtccctcc acatcaacga gatgtgccag 900
gagcgatgtg tcgacggttg ttcatgccct gagggacaac ttctcgacga gggtttgtgt 960
gtcgagagta ccgagtgtcc atgcgtacac tcaggaaagc gatacccacc aggtacctcc 1020
ctttcaagag actgcaacac ctgcatctgc aggaacagcc agtggatctg ctccaacgag 1080
gagtgccctg gtgaatgtct tgtcaccggt cagtcacact tcaaaagctt cgacaaccgc 1140
tacttcacct tctccggcat ctgccagtac ctgctcgctc gagactgcca ggatcactca 1200
ttctccatcg tgatcgagac cgtgcagtgc gctgatgaca gagacgctgt ctgtacccga 1260
tcagtaaccg tcagactgcc aggtctccat aactcccttg tcaagctcaa gcacggtgct 1320
ggtgtagcta tggacggtca ggacgtacag ctccctttgc tcaagggaga ccttagaatc 1380
cagcacaccg taaccgcttc agtgagactc agctacggtg aggaccttca gatggactgg 1440
gacggtagag gtagactgct cgtcaagctc tcaccagtat acgccggtaa gacctgcgga 1500
ctttgcggta attacaacgg gaaccagggt gacgacttcc tgaccccatc aggtcttgct 1560
gagcctagag tcgaagactt cggtaacgct tggaagctgc acggagactg ccaagacctc 1620
cagaagcagc actcagaccc atgtgctttg aaccctagaa tgaccaggtt tagcgaggag 1680
gcctgcgctg tcttgacctc cccaactttt gaggcttgcc acagagctgt aagcccattg 1740
ccatacctga gaaactgcag gtacgacgtc tgcagctgta gcgacggtag ggagtgtctt 1800
tgcggagctt tggcttccta cgctgcagca tgtgctggaa ggggagttag ggtagcttgg 1860
agggaaccag gaaggtgtga attgaactgc ccaaagggac aggtctactt gcagtgcgga 1920
actccttgca acttgacttg caggtccctg agctacccag acgaggagtg taacgaggct 1980
tgcttggagg gatgtttttg cccaccagga ttgtacatgg acgagcgggg agactgcgtt 2040
ccaaaggctc agtgtccatg ctattatgac ggcgagatct ttcagcccga ggacatcttt 2100
agcgaccacc acactatgtg ctattgcgag gacggcttta tgcactgcac aatgagcggg 2160
gtgcctggca gcctgttgcc agacgccgtt ttgagcagcc ctttgagcca taggagcaag 2220
aggagcttga gctgcaggcc tcctatggtt aagttggtgt gccctgccga taacttgagg 2280
gccgaaggat tggagtgtac aaagacttgc cagaactatg atctggagtg catgagcatg 2340
ggctgcgtga gcgggtgcct gtgccctccc ggaatggtta ggcatgaaaa ccgctgtgtt 2400
gccctggaac ggtgtccttg ttttcaccaa ggaaaggagt atgcccctgg agagacagtt 2460
aagatcgggt gcaatacttg cgtgtgccgc gatcggaagt ggaattgcac agatcacgtg 2520
tgcgatgcca catgcagcac aatcgggatg gcccactatc tgacatttga tggcctgaag 2580
tatctgtttc ccggcgagtg ccagtatgtg ctggtgcagg attattgcgg cagcaatccc 2640
gggacttttc ggatactggt gggcaataag ggctgcagtc accccagcgt gaagtgcaag 2700
aagcgcgtga caattctggt cgagggcgga gagattgagc tgtttgatgg cgaggtgaat 2760
gtcaaacggc ccatgaaaga tgagactcac tttgaggtgg tggagagtgg gcggtatata 2820
atactgcttc ttgggaaagc cctgagtgtg gtgtgggatc gccatctgtc tatatctgtg 2880
gtgctgaaac agacttatca ggagaaagtg tgcgggctgt gcgggaattt tgatggcatt 2940
cagaataatg atctgactag tagtaatctg caggtggaag aagatcccgt ggattttggc 3000
aattcttgga aagtgtcttc tcagtgtgcc gacacacgga aagtgcccct ggattcttct 3060
cccgccactt gtcataataa tataatgaaa cagactatgg tggattctag ttgtcgcatt 3120
ctgactagtg atgtgtttca ggattgtaat aaactggttg atcccgagcc ctatctggat 3180
gtgtgtattt atgatacatg tagttgtgaa agtattggcg attgtgcctg tttttgtgat 3240
acaattgccg cctatgccca tgtgtgtgcc cagcatggca aagtggtgac atggcgcaca 3300
gccacactct gtccccaaag ctgtgaagaa cgcaatctgc gggaaaatgg ctatgaatgt 3360
gaatggcgct ataattcttg tgcccccgcc tgtcaagtga catgtcaaca tcccgaaccc 3420
ctggcctgtc ctgtgcaatg tgtggaaggc tgtcatgctc attgtccccc tgggaaaatt 3480
ctcgacgaac tcctgcaaac atgtgttgat cccgaagatt gtcccgtgtg tgaagtggcc 3540
gggcgccgct ttgccagtgg caaaaaagtg actctgaatc ccagtgatcc cgaacattgt 3600
caaatttgtc attgtgatgt ggtgaatctg acatgtgaag cttgtcaaga accaggcggg 3660
ctcgtggtgc ctcccacaga tgcccccgtg tctcccacaa ctctgtatgt ggaagatatt 3720
tctgaacccc ctctgcatga ttttggaggc ggcggaagtg gcggcggagg cagcggaggc 3780
ggggggtctg ggggtggtgg atctggcggg ggcgggagtg gcggaggagg gtctggcggc 3840
gggggcagcg gcggtggtgg gagtggaggc gggggctcag ggggcggggg atcaggcggg 3900
ggcggctcag gcggaggtgg ttcagggggt ggtggcagtg gggggggcgg gtctgggggc 3960
ggaggaagtg ggggcggcgg gtctggcgga gggggatctg gaggcggtgg ctctggtgga 4020
gggggctctg gcggtggtgg ctctgggggc ggggggagtg gcggcggtgg ctcaggcggt 4080
ggtagtctcg ttcctcgggg ctctgccaca cggcggtatt atctgggcgc cgttgaactc 4140
tcttgggatt atatgcaatc tgatctcggc gaactgcctg ttgatgcccg ctttcctcct 4200
cgggttccta aatcttttcc ctttaataca tctgttgttt ataaaaaaac tctgtttgtt 4260
gaatttactg atcatctgtt taatattgcc aaaccccggc ccccctggat gggcctgctg 4320
ggccccacaa ttcaagccga agtttatgat actgttgtga ttactcttaa gaacatggct 4380
tcccatcctg tcagtcttca tgctgttggt gtatcctact ggaaagcttc tgagggagct 4440
gaatatgatg atcagaccag tcaaagggag aaagaagatg ataaagtctt ccctggtgga 4500
agccatacat atgtctggca ggtcctgaaa gagaatggtc caatggcctc tgacccactg 4560
tgccttacct actcatatct ttctcatgtg gacctggtaa aagacttgaa ttcaggcctc 4620
attggagccc tactagtatg tagagaaggg agtctggcca aggaaaagac acagaccttg 4680
cacaaattta tactactttt tgctgtattt gatgaaggga aaagttggca ctcagaaaca 4740
aagaactcct tgatgcagga tagggatgct gcatctgctc gggcctggcc taaaatgcac 4800
acagtcaatg gttatgtaaa caggtctctg ccaggtctga ttggatgcca caggaaatca 4860
gtctattggc atgtgattgg aatgggcacc actcctgaag tgcactcaat attcctcgaa 4920
ggtcacacat ttcttgtgag gaaccatcgc caggcgtcct tggaaatctc gccaataact 4980
ttccttactg ctcaaacact cttgatggac cttggacagt ttctactgtt ttgtcatatc 5040
tcttcccacc aacatgatgg catggaagct tatgtcaaag tagacagctg tccagaggaa 5100
ccccaactac gaatgaaaaa taatgaagaa gcggaagact atgatgatga tcttactgat 5160
tctgaaatgg atgtggtcag gtttgatgat gacaactctc cttcctttat ccaaattcgc 5220
tcagttgcca agaagcatcc taaaacttgg gtacattaca ttgctgctga agaggaggac 5280
tgggactatg ctcccttagt cctcgccccc gatgacagaa gttataaaag tcaatatttg 5340
aacaatggcc ctcagcggat tggtaggaag tacaaaaaag tccgatttat ggcatacaca 5400
gatgaaacct ttaagactcg tgaagctatt cagcatgaat caggaatctt gggaccttta 5460
ctttatgggg aagttggaga cacactgttg attatattta agaatcaagc aagcagacca 5520
tataacatct accctcacgg aatcactgat gtccgtcctt tgtattcaag gagattacca 5580
aaaggtgtaa aacatttgaa ggattttcca attctgccag gagaaatatt caaatataaa 5640
tggacagtga ctgtagaaga tgggccaact aaatcagatc ctcggtgcct gacccgctat 5700
tactctagtt tcgttaatat ggagagagat ctagcttcag gactcattgg ccctctcctc 5760
atctgctaca aagaatctgt agatcaaaga ggaaaccaga taatgtcaga caagaggaat 5820
gtcatcctgt tttctgtatt tgatgagaac cgaagctggt acctcacaga gaatatacaa 5880
cgctttctcc ccaatccagc tggagtgcag cttgaggatc cagagttcca agcctccaac 5940
atcatgcaca gcatcaatgg ctatgttttt gatagtttgc agttgtcagt ttgtttgcat 6000
gaggtggcat actggtacat tctaagcatt ggagcacaga ctgacttcct ttctgtcttc 6060
ttctctggat ataccttcaa acacaaaatg gtctatgaag acacactcac cctattccca 6120
ttctcaggag aaactgtctt catgtcgatg gaaaacccag gtctatggat tctggggtgc 6180
cacaactcag actttcggaa cagaggcatg accgccttac tgaaggtttc tagttgtgac 6240
aagaacactg gtgattatta cgaggacagt tatgaagata tttcagcata cttgctgagt 6300
aaaaacaatg ccattgaacc aagaagcttc tcccagaatt caagacaccc tagcactagg 6360
caaaagcaat ttaatgccac cacaattcca gaaaatgaca tagagaagac tgacccttgg 6420
tttgcacaca gaacacctat gcctaaatgt caaaatgtct cctctagtga tttgttgatg 6480
ctcttgcgac agagtcctac tccacatggg ctatccttat ctgatctcca agaagccaaa 6540
tatgagactt tttctgatga tccatcacct ggagcaatag acagtaataa cagcctgtct 6600
gaaatgacac acttcaggcc acagctccat cacagtgggg acatggtatt tacccctgag 6660
tcaggcctcc aattaagatt aaatgagaaa ctggggacaa ctgcagcaac agagttgaag 6720
aaacttgatt tcaaagtttc tagtacatca aataatctga tttcaacaat tccatcagac 6780
aatttggcag caggtactga taatacaagt actcttcagt cagatcaaga ggaaattgac 6840
tatgatgata ccatatcagt tgaaatgaag aaggaagatt ttgacattta tgatgaggat 6900
gaaaatcaga gcccccgcag ctttcaaaag aaaacacgac actattttat tgctgcagtg 6960
gagaggctct gggattatgg gatgagtagc tccccacatg ttctaagaaa cagggctcag 7020
agtggcagtg tccctcagtt caagaaagtt gttttccagg aatttactga tggctccttt 7080
actcagccct tataccgtgg agaactaaat gaacatttgg gactcctggg gccatatata 7140
agagcagaag ttgaagataa tatcatggta actttcagaa atcaggcctc tcgtccctat 7200
tccttctatt ctagccttat ttcttatgag gaagatcaga ggcaaggagc agaacctaga 7260
aaaaactttg tcaagcctaa tgaaaccaaa acttactttt ggaaagtgca acatcatatg 7320
gcacccacta aagatgagtt tgactgcaaa gcctgggctt atttctctga tgttgacctg 7380
gaaaaagatg tgcactcagg cctgattgga ccccttctgg tctgccacac taacacactg 7440
aaccctgctc atgggagaca agtgacagta caggaatttg ctctgttttt caccatcttt 7500
gatgagacca aaagctggta cttcactgaa aatatggaaa gaaactgcag ggctccctgc 7560
aatatccaga tggaagatcc cacttttaaa gagaattatc gcttccatgc aatcaatggc 7620
tacataatgg atacactacc tggcttagta atggctcagg atcaaaggat tcgatggtat 7680
ctgctcagca tgggcagcaa tgaaaacatc cattctattc atttcagtgg acatgtgttc 7740
actgtacgaa aaaaagagga gtataaaatg gcactgtaca atctctatcc aggtgttttt 7800
gagacagtgg aaatgttacc atccaaagct ggaatttggc gggtggaatg ccttattggc 7860
gagcatctac atgctgggat gagcacactt tttctggtgt acagcaataa gtgtcagact 7920
cccctgggaa tggcttctgg acacattaga gattttcaga ttacagcttc aggacaatat 7980
ggacagtggg ccccaaagct ggccagactt cattattccg gatcaatcaa tgcctggagc 8040
accaaggagc ccttttcttg gatcaaggtg gatctgttgg caccaatgat tattcacggc 8100
atcaagaccc agggtgcccg tcagaagttc tccagcctct acatctctca gtttatcatc 8160
atgtatagtc ttgatgggaa gaagtggcag acttatcgag gaaattccac tggaacctta 8220
atggtcttct ttggcaatgt ggattcatct gggataaaac acaatatttt taaccctcca 8280
attattgctc gatacatccg tttgcaccca actcattata gcattcgcag cactcttcgc 8340
atggagttga tgggctgtga tttaaatagt tgcagcatgc cattgggaat ggagagtaaa 8400
gcaatatcag atgcacagat tactgcttca tcctacttta ccaatatgtt tgccacctgg 8460
tctccttcaa aagctcgact tcacctccaa gggaggagta atgcctggag acctcaggtg 8520
aataatccaa aagagtggct gcaagtggac ttccagaaga caatgaaagt cacaggagta 8580
actactcagg gagtaaaatc tctgcttacc agcatgtatg tgaaggagtt cctcatctcc 8640
agcagtcaag atggccatca gtggactctc ttttttcaga atggcaaagt aaaggttttt 8700
cagggaaatc aagactcctt cacacctgtg gtgaactctc tagacccacc gttactgact 8760
cgctaccttc gaattcaccc ccagagttgg gtgcaccaga ttgccctgag gatggaggtt 8820
ctgggctgcg aggcacagga cctctactga taa 8853
<210> 65
<211> 3750
<212> DNA
<213> Artificial Sequence
<220>
<223> D'D3
<400> 65
gcagaaggaa ctcgcggcag gtcatccacg gcccgatgca gccttttcgg aagtgacttc 60
gtcaacacct ttgatgggag catgtacagc tttgcgggat actgcagtta cctcctggca 120
gggggctgcc agaaacgctc cttctcgatt attggggact tccagaatgg caagagagtg 180
agcctctccg tgtatcttgg ggaatttttt gacatccatt tgtttgtcaa tggtaccgtg 240
acacaggggg accaaagagt ctccatgccc tatgcctcca aagggctgta tctagaaact 300
gaggctgggt actacaagct gtccggtgag gcctatggct ttgtggccag gatcgatggc 360
agcggcaact ttcaagtcct gctgtcagac agatacttca acaagacctg cgggctgtgt 420
ggcaacttta acatctttgc tgaagatgac tttatgaccc aagaagggac cttgacctcg 480
gacccttatg actttgccaa ctcatgggct ctgagcagtg gagaacagtg gtgtgaacgg 540
gcatctcctc ccagcagctc atgcaacatc tcctctgggg aaatgcagaa gggcctgtgg 600
gagcagtgcc agcttctgaa gagcacctcg gtgtttgccc gctgccaccc tctggtggac 660
cccgagcctt ttgtggccct gtgtgagaag actttgtgtg agtgtgctgg ggggctggag 720
tgcgcctgcc ctgccctcct ggagtacgcc cggacctgtg cccaggaggg aatggtgctg 780
tacggctgga ccgaccacag cgcgtgcagc ccagtgtgcc ctgctggtat ggagtatagg 840
cagtgtgtgt ccccttgcgc caggacctgc cagagcctgc acatcaatga aatgtgtcag 900
gagcgatgcg tggatggctg cagctgccct gagggacagc tcctggatga aggcctctgc 960
gtggagagca ccgagtgtcc ctgcgtgcat tccggaaagc gctaccctcc cggcacctcc 1020
ctctctcgag actgcaacac ctgcatttgc cgaaacagcc agtggatctg cagcaatgaa 1080
gaatgtccag gggagtgcct tgtcacaggt caatcacact tcaagagctt tgacaacaga 1140
tacttcacct tcagtgggat ctgccagtac ctgctggccc gggattgcca ggaccactcc 1200
ttctccattg tcattgagac tgtccagtgt gctgatgacc gcgacgctgt gtgcacccgc 1260
tccgtcaccg tccggctgcc tggcctgcac aacagccttg tgaaactgaa gcatggggca 1320
ggagttgcca tggatggcca ggacgtccag ctccccctcc tgaaaggtga cctccgcatc 1380
cagcatacag tgacggcctc cgtgcgcctc agctacgggg aggacctgca gatggactgg 1440
gatggccgcg ggaggctgct ggtgaagctg tcccccgtct atgccgggaa gacctgcggc 1500
ctgtgtggga attacaatgg caaccagggc gacgacttcc ttaccccctc tgggctggcg 1560
gagccccggg tggaggactt cgggaacgcc tggaagctgc acggggactg ccaggacctg 1620
cagaagcagc acagcgatcc ctgcgccctc aacccgcgca tgaccaggtt ctccgaggag 1680
gcgtgcgcgg tcctgacgtc ccccacattc gaggcctgcc atcgtgccgt cagcccgctg 1740
ccctacctgc ggaactgccg ctacgacgtg tgctcctgct cggacggccg cgagtgcctg 1800
tgcggcgccc tggccagcta tgccgcggcc tgcgcgggga gaggcgtgcg cgtcgcgtgg 1860
cgcgagccag gccgctgtga gctgaactgc ccgaaaggcc aggtgtacct gcagtgcggg 1920
accccctgca acctgacctg ccgctctctc tcttacccgg atgaggaatg caatgaggcc 1980
tgcctggagg gctgcttctg ccccccaggg ctctacatgg atgagagggg ggactgcgtg 2040
cccaaggccc agtgcccctg ttactatgac ggtgagatct tccagccaga agacatcttc 2100
tcagaccatc acaccatgtg ctactgtgag gatggcttca tgcactgtac catgagtgga 2160
gtccccggaa gcttgctgcc tgacgctgtc ctcagcagtc ccctgtctca tcgcagcaaa 2220
aggagcctat cctgtcggcc ccccatggtc aagctggtgt gtcccgctga caacctgcgg 2280
gctgaagggc tcgagtgtac caaaacgtgc cagaactatg acctggagtg catgagcatg 2340
ggctgtgtct ctggctgcct ctgccccccg ggcatggtcc ggcatgagaa cagatgtgtg 2400
gccctggaaa ggtgtccctg cttccatcag ggcaaggagt atgcccctgg agaaacagtg 2460
aagattggct gcaacacttg tgtctgtcgg gaccggaagt ggaactgcac agaccatgtg 2520
tgtgatgcca cgtgctccac gatcggcatg gcccactacc tcaccttcga cgggctcaaa 2580
tacctgttcc ccggggagtg ccagtacgtt ctggtgcagg attactgcgg cagtaaccct 2640
gggacctttc ggatcctagt ggggaataag ggatgcagcc acccctcagt gaaatgcaag 2700
aaacgggtca ccatcctggt ggagggagga gagattgagc tgtttgacgg ggaggtgaat 2760
gtgaagaggc ccatgaagga tgagactcac tttgaggtgg tggagtctgg ccggtacatc 2820
attctgctgc tgggcaaagc cctctccgtg gtctgggacc gccacctgag catctccgtg 2880
gtcctgaagc agacatacca ggagaaagtg tgtggcctgt gtgggaattt tgatggcatc 2940
cagaacaatg acctcaccag cagcaacctc caagtggagg aagaccctgt ggactttggg 3000
aactcctgga aagtgagctc gcagtgtgct gacaccagaa aagtgcctct ggactcatcc 3060
cctgccacct gccataacaa catcatgaag cagacgatgg tggattcctc ctgtagaatc 3120
cttaccagtg acgtcttcca ggactgcaac aagctggtgg accccgagcc atatctggat 3180
gtctgcattt acgacacctg ctcctgtgag tccattgggg actgcgcctg cttctgcgac 3240
accattgctg cctatgccca cgtgtgtgcc cagcatggca aggtggtgac ctggaggacg 3300
gccacattgt gcccccagag ctgcgaggag aggaatctcc gggagaacgg gtatgagtgt 3360
gagtggcgct ataacagctg tgcacctgcc tgtcaagtca cgtgtcagca ccctgagcca 3420
ctggcctgcc ctgtgcagtg tgtggagggc tgccatgccc actgccctcc agggaaaatc 3480
ctggatgagc ttttgcagac ctgcgttgac cctgaagact gtccagtgtg tgaggtggct 3540
ggccggcgtt ttgcctcagg aaagaaagtc accttgaatc ccagtgaccc tgagcactgc 3600
cagatttgcc actgtgatgt tgtcaacctc acctgtgaag cctgccagga gccgggaggc 3660
ctggtggtgc ctcccacaga tgccccggtg agccccacca ctctgtatgt ggaggacatc 3720
tcggaaccgc cgttgcacga tttctgataa 3750
<210> 66
<211> 7401
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3/Fc-120
<400> 66
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc tatcaacgca tggagcacca aggagccatt ctcctggatc 3960
aaggtggacc tcctcgctcc tatgatcatc cacggcatca agacccaggg cgctagacag 4020
aagttcagca gcctctacat ctcccagttc atcatcatgt actccctgga cggcaagaag 4080
tggcagacct accgcggaaa ctccaccgga accctcatgg tcttcttcgg aaacgtggac 4140
agctccggta tcaagcacaa catcttcaac ccccctatca tcgcacgtta catccgtctg 4200
caccctaccc actactccat caggagcacc ctcagaatgg agctgatggg ttgcgacctc 4260
aactcctgct ccatgccact cggaatggag tcaaaggcaa tctccgacgc acaaatcacc 4320
gcatcctcct acttcaccaa catgttcgcc acctggtcac caagtaaggc aaggctccac 4380
ctccaaggac gttcaaacgc atggcgtcca caagtcaaca acccaaagga gtggctccaa 4440
gtggacttcc agaagacaat gaaggtgacc ggcgtgacca cccaaggagt caagtcactg 4500
ctcacctcca tgtacgtcaa ggagttcctg atctccagct cccaagacgg ccaccaatgg 4560
acccttttct tccagaacgg caaggtgaag gtcttccagg gtaaccagga ctccttcacc 4620
ccagtcgtca actccctcga tccaccactt cttaccagat acctccgaat ccaccctcaa 4680
agctgggtcc accaaatcgc acttcgaatg gaggtcctgg gatgcgaagc acaagatctg 4740
tacctggtcc cacgtggaag tggtggtggg ggaagtggtg gaggtggtag tggaggtgga 4800
ggatctggtg gtggaggttc aggtggaggg ggaagtggag gtggtgggtc tggtggagga 4860
ggtagtggtg gcggaggttc tggaggtggg ggatcaggtg gtggaggatc aggaggaggt 4920
ggatcaggtg ggggaggatc aggcggaggt ggttctggag ggggtgggag tggaggaggc 4980
ggatcaggag gcggtggctc aggaggaggc ggctcaggag gcggaggatc tgggggaggg 5040
ggctcaggag gtgggggttc agggggcggt ggttctgggg gcggggggtc tggcggcgga 5100
ggctctcttt catgtcgacc acctatggtt aaacttgtat gtcctgcaga taatcttaga 5160
gctgaagggc ttgaatgtac aaaaacttgt caaaattatg atcttgaatg tatgtcaatg 5220
gggtgtgtta gtgggtgtct ttgtcctcct gggatggtta gacatgaaaa tcgatgtgta 5280
gctttggaaa gatgtccatg ttttcatcaa ggcaaagaat atgcaccagg cgaaacagta 5340
aaaattgggt gtaatacatg tgtatgtcga gatcgaaaat ggaattgtac tgatcatgtt 5400
tgtgatgcaa cctgtagtac catcggcatg gctcactacc ttaccttcga cgggttgaag 5460
tacctgttcc ctggggaatg ccagtacgta ctcgtccagg attactgcgg gtctaaccct 5520
gggacattta gaattctggt cggcaacaag ggctgctccc acccaagtgt caagtgcaag 5580
aagagggtca ccattctggt cgagggcggc gaaatagaac tgttcgacgg ggaagtaaac 5640
gtgaagaggc caatgaagga cgagacccac ttcgaggtgg tggaatccgg gagatacatt 5700
attctgctcc tcgggaaggc cctgagtgtc gtatgggaca ggcacctttc cattagcgtc 5760
gtcctcaagc agacctacca ggaaaaggtc tgcggccttt gtgggaactt tgacggcata 5820
cagaacaacg acctgacctc cagcaacctg caggtagagg aagacccagt agactttggg 5880
aacagctgga aggtctcctc ccagtgtgct gacaccagga aggtccctct tgactctagt 5940
ccagctacct gtcacaacaa cattatgaag cagaccatgg tggacagcag ctgccggatt 6000
ctcacttccg acgtgtttca ggactgcaac aagctggttg accctgagcc ctacctggac 6060
gtgtgtattt acgacacatg ctcctgcgag agcattggcg actgcgcttg cttttgcgac 6120
acaatagccg cctatgccca cgtgtgtgct cagcacggca aagtggtgac ttggaggaca 6180
gctacactgt gtcctcagag ttgtgaggag aggaacctgc gggaaaatgg ctatgagtgc 6240
gaatggcggt ataatagctg cgccccagct tgtcaggtga cttgtcagca cccagaacca 6300
ctggcttgtc ctgtgcagtg tgttgaaggc tgccatgctc attgcccacc aggcaaaatt 6360
ctggacgaac tgctgcagac atgcgttgat cctgaggatt gccctgtatg cgaagtggct 6420
ggccggcggt ttgcttctgg caaaaaagtt actctgaatc cctccgatcc tgagcactgc 6480
cagatatgcc attgcgatgt ggtgaatctg acttgcgagg cctgccagga acctgggggc 6540
ttggttgttc ctcctacaga tgcccctgtt agtcctacta ctctgtatgt ggaggatatt 6600
agcgagcccc ctctgcatga ttttggcggt ggggggtcag gcggaggcgg cagtggcggc 6660
gggggctctg gcggaggtgg gtccggcggt ggcggttctg gcgggggcgg ctctgacaaa 6720
actcacacat gcccaccgtg cccagcacct gaactccttg ggggaccgtc agtcttcctc 6780
ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 6840
gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 6900
gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg 6960
gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 7020
gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag 7080
ccccgagaac cacaggtgta caccctgccc ccatcccggg atgagctgac caagaaccag 7140
gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 7200
agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc 7260
tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc 7320
ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc 7380
ctgtctccgg gtaaatgata a 7401
<210> 67
<211> 687
<212> DNA
<213> Artificial Sequence
<220>
<223> Fc
<400> 67
gacaaaactc acacatgccc accgtgccca gcacctgaac tccttggggg accgtcagtc 60
ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 120
tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 180
ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 240
cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 300
tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 360
gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 420
aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 480
tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 540
gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 600
aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 660
ctctccctgt ctccgggtaa atagtaa 687
<210> 68
<211> 5946
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/D'D3-HC Heavy chain
<400> 68
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttcg gcggtggggg gtcaggcgga ggcggcagtg gcggcggggg ctctggcgga 4380
ggtgggtccg gcggtggcgg ttctggcggg ggcggcggca gcctatcctg tcggcccccc 4440
atggtcaagc tggtgtgtcc cgctgacaac ctgcgggctg aagggctcga gtgtaccaaa 4500
acgtgccaga actatgacct ggagtgcatg agcatgggct gtgtctctgg ctgcctctgc 4560
cccccgggca tggtccggca tgagaacaga tgtgtggccc tggaaaggtg tccctgcttc 4620
catcagggca aggagtatgc ccctggagaa acagtgaaga ttggctgcaa cacttgtgtc 4680
tgtcgggacc ggaagtggaa ctgcacagac catgtgtgtg atgccacgtg ctccacgatc 4740
ggcatggccc actacctcac cttcgacggg ctcaaatacc tgttccccgg ggagtgccag 4800
tacgttctgg tgcaggatta ctgcggcagt aaccctggga cctttcggat cctagtgggg 4860
aataagggat gcagccaccc ctcagtgaaa tgcaagaaac gggtcaccat cctggtggag 4920
ggaggagaga ttgagctgtt tgacggggag gtgaatgtga agaggcccat gaaggatgag 4980
actcactttg aggtggtgga gtctggccgg tacatcattc tgctgctggg caaagccctc 5040
tccgtggtct gggaccgcca cctgagcatc tccgtggtcc tgaagcagac ataccaggag 5100
aaagtgtgtg gcctgtgtgg gaattttgat ggcatccaga acaatgacct caccagcagc 5160
aacctccaag tggaggaaga ccctgtggac tttgggaact cctggaaagt gagctcgcag 5220
tgtgctgaca ccagaaaagt gcctctggac tcatcccctg ccacctgcca taacaacatc 5280
atgaagcaga cgatggtgga ttcctcctgt agaatcctta ccagtgacgt cttccaggac 5340
tgcaacaagc tggtggaccc cgagccatat ctggatgtct gcatttacga cacctgctcc 5400
tgtgagtcca ttggggactg cgcctgcttc tgcgacacca ttgctgccta tgcccacgtg 5460
tgtgcccagc atggcaaggt ggtgacctgg aggacggcca cattgtgccc ccagagctgc 5520
gaggagagga atctccggga gaacgggtat gagtgtgagt ggcgctataa cagctgtgca 5580
cctgcctgtc aagtcacgtg tcagcaccct gagccactgg cctgccctgt gcagtgtgtg 5640
gagggctgcc atgcccactg ccctccaggg aaaatcctgg atgagctttt gcagacctgc 5700
gttgaccctg aagactgtcc agtgtgtgag gtggctggcc ggcgttttgc ctcaggaaag 5760
aaagtcacct tgaatcccag tgaccctgag cactgccaga tttgccactg tgatgttgtc 5820
aacctcacct gtgaagcctg ccaggagccg ggaggcctgg tggtgcctcc cacagatgcc 5880
ccggtgagcc ccaccactct gtatgtggag gacatctcgg aaccgccgtt gcacgatttc 5940
tgataa 5946
<210> 69
<211> 2322
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/D'D3-HC Light chain
<400> 69
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctactgat aa 2322
<210> 70
<211> 8172
<212> DNA
<213> Artificial Sequence
<220>
<223> scFVIII/D'D3/Fc/Fc-120
<400> 70
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagtactc ttcagtcaga tcaagaggaa attgactatg atgataccat atcagttgaa 2760
atgaagaagg aagattttga catttatgat gaggatgaaa atcagagccc ccgcagcttt 2820
caaaagaaaa cacgacacta ttttattgct gcagtggaga ggctctggga ttatgggatg 2880
agtagctccc cacatgttct aagaaacagg gctcagagtg gcagtgtccc tcagttcaag 2940
aaagttgttt tccaggaatt tactgatggc tcctttactc agcccttata ccgtggagaa 3000
ctaaatgaac atttgggact cctggggcca tatataagag cagaagttga agataatatc 3060
atggtaactt tcagaaatca ggcctctcgt ccctattcct tctattctag ccttatttct 3120
tatgaggaag atcagaggca aggagcagaa cctagaaaaa actttgtcaa gcctaatgaa 3180
accaaaactt acttttggaa agtgcaacat catatggcac ccactaaaga tgagtttgac 3240
tgcaaagcct gggcttattt ctctgatgtt gacctggaaa aagatgtgca ctcaggcctg 3300
attggacccc ttctggtctg ccacactaac acactgaacc ctgctcatgg gagacaagtg 3360
acagtacagg aatttgctct gtttttcacc atctttgatg agaccaaaag ctggtacttc 3420
actgaaaata tggaaagaaa ctgcagggct ccctgcaata tccagatgga agatcccact 3480
tttaaagaga attatcgctt ccatgcaatc aatggctaca taatggatac actacctggc 3540
ttagtaatgg ctcaggatca aaggattcga tggtatctgc tcagcatggg cagcaatgaa 3600
aacatccatt ctattcattt cagtggacat gtgttcactg tacgaaaaaa agaggagtat 3660
aaaatggcac tgtacaatct ctatccaggt gtttttgaga cagtggaaat gttaccatcc 3720
aaagctggaa tttggcgggt ggaatgcctt attggcgagc atctacatgc tgggatgagc 3780
acactttttc tggtgtacag caataagtgt cagactcccc tgggaatggc ttctggacac 3840
attagagatt ttcagattac agcttcagga caatatggac agtgggcccc aaagctggcc 3900
agacttcatt attccggatc tatcaacgca tggagcacca aggagccatt ctcctggatc 3960
aaggtggacc tcctcgctcc tatgatcatc cacggcatca agacccaggg cgctagacag 4020
aagttcagca gcctctacat ctcccagttc atcatcatgt actccctgga cggcaagaag 4080
tggcagacct accgcggaaa ctccaccgga accctcatgg tcttcttcgg aaacgtggac 4140
agctccggta tcaagcacaa catcttcaac ccccctatca tcgcacgtta catccgtctg 4200
caccctaccc actactccat caggagcacc ctcagaatgg agctgatggg ttgcgacctc 4260
aactcctgct ccatgccact cggaatggag tcaaaggcaa tctccgacgc acaaatcacc 4320
gcatcctcct acttcaccaa catgttcgcc acctggtcac caagtaaggc aaggctccac 4380
ctccaaggac gttcaaacgc atggcgtcca caagtcaaca acccaaagga gtggctccaa 4440
gtggacttcc agaagacaat gaaggtgacc ggcgtgacca cccaaggagt caagtcactg 4500
ctcacctcca tgtacgtcaa ggagttcctg atctccagct cccaagacgg ccaccaatgg 4560
acccttttct tccagaacgg caaggtgaag gtcttccagg gtaaccagga ctccttcacc 4620
ccagtcgtca actccctcga tccaccactt cttaccagat acctccgaat ccaccctcaa 4680
agctgggtcc accaaatcgc acttcgaatg gaggtcctgg gatgcgaagc acaagatctg 4740
tacctggtcc cacgtggaag tggtggtggg ggaagtggtg gaggtggtag tggaggtgga 4800
ggatctggtg gtggaggttc aggtggaggg ggaagtggag gtggtgggtc tggtggagga 4860
ggtagtggtg gcggaggttc tggaggtggg ggatcaggtg gtggaggatc aggaggaggt 4920
ggatcaggtg ggggaggatc aggcggaggt ggttctggag ggggtgggag tggaggaggc 4980
ggatcaggag gcggtggctc aggaggaggc ggctcaggag gcggaggatc tgggggaggg 5040
ggctcaggag gtgggggttc agggggcggt ggttctgggg gcggggggtc tggcggcgga 5100
ggctctcttt catgtcgacc acctatggtt aaacttgtat gtcctgcaga taatcttaga 5160
gctgaagggc ttgaatgtac aaaaacttgt caaaattatg atcttgaatg tatgtcaatg 5220
gggtgtgtta gtgggtgtct ttgtcctcct gggatggtta gacatgaaaa tcgatgtgta 5280
gctttggaaa gatgtccatg ttttcatcaa ggcaaagaat atgcaccagg cgaaacagta 5340
aaaattgggt gtaatacatg tgtatgtcga gatcgaaaat ggaattgtac tgatcatgtt 5400
tgtgatgcaa cctgtagtac catcggcatg gctcactacc ttaccttcga cgggttgaag 5460
tacctgttcc ctggggaatg ccagtacgta ctcgtccagg attactgcgg gtctaaccct 5520
gggacattta gaattctggt cggcaacaag ggctgctccc acccaagtgt caagtgcaag 5580
aagagggtca ccattctggt cgagggcggc gaaatagaac tgttcgacgg ggaagtaaac 5640
gtgaagaggc caatgaagga cgagacccac ttcgaggtgg tggaatccgg gagatacatt 5700
attctgctcc tcgggaaggc cctgagtgtc gtatgggaca ggcacctttc cattagcgtc 5760
gtcctcaagc agacctacca ggaaaaggtc tgcggccttt gtgggaactt tgacggcata 5820
cagaacaacg acctgacctc cagcaacctg caggtagagg aagacccagt agactttggg 5880
aacagctgga aggtctcctc ccagtgtgct gacaccagga aggtccctct tgactctagt 5940
ccagctacct gtcacaacaa cattatgaag cagaccatgg tggacagcag ctgccggatt 6000
ctcacttccg acgtgtttca ggactgcaac aagctggttg accctgagcc ctacctggac 6060
gtgtgtattt acgacacatg ctcctgcgag agcattggcg actgcgcttg cttttgcgac 6120
acaatagccg cctatgccca cgtgtgtgct cagcacggca aagtggtgac ttggaggaca 6180
gctacactgt gtcctcagag ttgtgaggag aggaacctgc gggaaaatgg ctatgagtgc 6240
gaatggcggt ataatagctg cgccccagct tgtcaggtga cttgtcagca cccagaacca 6300
ctggcttgtc ctgtgcagtg tgttgaaggc tgccatgctc attgcccacc aggcaaaatt 6360
ctggacgaac tgctgcagac atgcgttgat cctgaggatt gccctgtatg cgaagtggct 6420
ggccggcggt ttgcttctgg caaaaaagtt actctgaatc cctccgatcc tgagcactgc 6480
cagatatgcc attgcgatgt ggtgaatctg acttgcgagg cctgccagga acctgggggc 6540
ttggttgttc ctcctacaga tgcccctgtt agtcctacta ctctgtatgt ggaggatatt 6600
agcgagcccc ctctgcatga ttttggcggt ggggggtcag gcggaggcgg cagtggcggc 6660
gggggctctg gcggaggtgg gtccggcggt ggcggttctg gcgggggcgg ctctgacaaa 6720
actcacacat gcccaccgtg cccagcacct gaactccttg ggggaccgtc agtcttcctc 6780
ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 6840
gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 6900
gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg 6960
gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 7020
gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag 7080
ccccgagaac cacaggtgta caccctgccc ccatcccggg atgagctgac caagaaccag 7140
gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 7200
agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc 7260
tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc 7320
ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc 7380
ctgtctccgg gtaaaggcgg tggggggtca ggcggaggcg gcagtggcgg cgggggctct 7440
ggcggaggtg ggtccggcgg tggcggttct ggcgggggcg gctctgacaa aactcacaca 7500
tgcccaccgt gcccagcacc tgaactcctt gggggaccgt cagtcttcct cttcccccca 7560
aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac 7620
gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 7680
aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 7740
ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac 7800
aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa 7860
ccacaggtgt acaccctgcc cccatcccgg gatgagctga ccaagaacca ggtcagcctg 7920
acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg 7980
cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc 8040
ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc 8100
tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg 8160
ggtaaatgat aa 8172
<210> 71
<211> 5106
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC/Fc-LC Heavy chain
<400> 71
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttcg gcggtggggg gtcaggcgga ggcggcagtg gcggcggggg ctctggcgga 4380
ggtgggtccg gcggtggcgg ttctggcggg ggcggctctg acaaaactca cacatgccca 4440
ccgtgcccag cacctgaact ccttggggga ccgtcagtct tcctcttccc cccaaaaccc 4500
aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc 4560
cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc 4620
aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc 4680
gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc 4740
ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag 4800
gtgtacaccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag cctgacctgc 4860
ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg 4920
gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac 4980
agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg 5040
atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa 5100
tgataa 5106
<210> 72
<211> 3093
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC/Fc-LC light chain
<400> 72
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctacctgg tcccacgtgg aagtggcggt 2340
ggggggtcag gcggaggcgg cagtggcggc gggggctctg gcggaggtgg cagtggcggg 2400
ggctctgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct tgggggaccg 2460
tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 2520
gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 2580
gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 2640
acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 2700
tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 2760
gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagctg 2820
accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc 2880
gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 2940
gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 3000
caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 3060
aagagcctct ccctgtctcc gggtaaatga taa 3093
<210> 73
<211> 5106
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC, Fc Heavy chain
<400> 73
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttcg gcggtggggg gtcaggcgga ggcggcagtg gcggcggggg ctctggcgga 4380
ggtgggtccg gcggtggcgg ttctggcggg ggcggctctg acaaaactca cacatgccca 4440
ccgtgcccag cacctgaact ccttggggga ccgtcagtct tcctcttccc cccaaaaccc 4500
aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc 4560
cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc 4620
aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc 4680
gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc 4740
ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag 4800
gtgtacaccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag cctgacctgc 4860
ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg 4920
gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac 4980
agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg 5040
atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa 5100
tgataa 5106
<210> 74
<211> 2322
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc-HC, Fc Light chain
<400> 74
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctactgat aa 2322
<210> 75
<211> 5877
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc/Fc-HC Heavy chain
<400> 75
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttcg gcggtggggg gtcaggcgga ggcggcagtg gcggcggggg ctctggcgga 4380
ggtgggtccg gcggtggcgg ttctggcggg ggcggctctg acaaaactca cacatgccca 4440
ccgtgcccag cacctgaact ccttggggga ccgtcagtct tcctcttccc cccaaaaccc 4500
aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc 4560
cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc 4620
aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc 4680
gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc 4740
ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag 4800
gtgtacaccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag cctgacctgc 4860
ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg 4920
gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt cttcctctac 4980
agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg 5040
atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa 5100
ggcggtgggg ggtcaggcgg aggcggcagt ggcggcgggg gctctggcgg aggtgggtcc 5160
ggcggtggcg gttctggcgg gggcggctct gacaaaactc acacatgccc accgtgccca 5220
gcacctgaac tccttggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc 5280
ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac 5340
cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 5400
ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac 5460
caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc 5520
cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc 5580
ctgcccccat cccgggatga gctgaccaag aaccaggtca gcctgacctg cctggtcaaa 5640
ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac 5700
tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaagctc 5760
accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 5820
gctctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa atgataa 5877
<210> 76
<211> 2322
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3/Fc/Fc-HC Light chain
<400> 76
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctactgat aa 2322
<210> 77
<211> 4335
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc-LC Heavy chain
<400> 77
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttct gataa 4335
<210> 78
<211> 3093
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc-LC Light chain
<400> 78
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctacctgg tcccacgtgg aagtggcggt 2340
ggggggtcag gcggaggcgg cagtggcggc gggggctctg gcggaggtgg cagtggcggg 2400
ggctctgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct tgggggaccg 2460
tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 2520
gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 2580
gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 2640
acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 2700
tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 2760
gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagctg 2820
accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc 2880
gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 2940
gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 3000
caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 3060
aagagcctct ccctgtctcc gggtaaatga taa 3093
<210> 79
<211> 4335
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc/Fc-LC Heavy chain
<400> 79
gccaccagaa gatactacct gggtgcagtg gaactgtcat gggactatat gcaaagtgat 60
ctcggtgagc tgcctgtgga cgcaagattt cctcctagag tgccaaaatc ttttccattc 120
aacacctcag tcgtgtacaa aaagactctg tttgtagaat tcacggatca ccttttcaac 180
atcgctaagc caaggccacc ctggatgggt ctgctaggtc ctaccatcca ggctgaggtt 240
tatgatacag tggtcattac acttaagaac atggcttccc atcctgtcag tcttcatgct 300
gttggtgtat cctactggaa agcttctgag ggagctgaat atgatgatca gaccagtcaa 360
agggagaaag aagatgataa agtcttccct ggtggaagcc atacatatgt ctggcaggtc 420
ctgaaagaga atggtccaat ggcctctgac ccactgtgcc ttacctactc atatctttct 480
catgtggacc tggtaaaaga cttgaattca ggcctcattg gagccctact agtatgtaga 540
gaagggagtc tggccaagga aaagacacag accttgcaca aatttatact actttttgct 600
gtatttgatg aagggaaaag ttggcactca gaaacaaaga actccttgat gcaggatagg 660
gatgctgcat ctgctcgggc ctggcctaaa atgcacacag tcaatggtta tgtaaacagg 720
tctctgccag gtctgattgg atgccacagg aaatcagtct attggcatgt gattggaatg 780
ggcaccactc ctgaagtgca ctcaatattc ctcgaaggtc acacatttct tgtgaggaac 840
catcgccagg cgtccttgga aatctcgcca ataactttcc ttactgctca aacactcttg 900
atggaccttg gacagtttct actgttttgt catatctctt cccaccaaca tgatggcatg 960
gaagcttatg tcaaagtaga cagctgtcca gaggaacccc aactacgaat gaaaaataat 1020
gaagaagcgg aagactatga tgatgatctt actgattctg aaatggatgt ggtcaggttt 1080
gatgatgaca actctccttc ctttatccaa attcgctcag ttgccaagaa gcatcctaaa 1140
acttgggtac attacattgc tgctgaagag gaggactggg actatgctcc cttagtcctc 1200
gcccccgatg acagaagtta taaaagtcaa tatttgaaca atggccctca gcggattggt 1260
aggaagtaca aaaaagtccg atttatggca tacacagatg aaacctttaa gactcgtgaa 1320
gctattcagc atgaatcagg aatcttggga cctttacttt atggggaagt tggagacaca 1380
ctgttgatta tatttaagaa tcaagcaagc agaccatata acatctaccc tcacggaatc 1440
actgatgtcc gtcctttgta ttcaaggaga ttaccaaaag gtgtaaaaca tttgaaggat 1500
tttccaattc tgccaggaga aatattcaaa tataaatgga cagtgactgt agaagatggg 1560
ccaactaaat cagatcctcg gtgcctgacc cgctattact ctagtttcgt taatatggag 1620
agagatctag cttcaggact cattggccct ctcctcatct gctacaaaga atctgtagat 1680
caaagaggaa accagataat gtcagacaag aggaatgtca tcctgttttc tgtatttgat 1740
gagaaccgaa gctggtacct cacagagaat atacaacgct ttctccccaa tccagctgga 1800
gtgcagcttg aggatccaga gttccaagcc tccaacatca tgcacagcat caatggctat 1860
gtttttgata gtttgcagtt gtcagtttgt ttgcatgagg tggcatactg gtacattcta 1920
agcattggag cacagactga cttcctttct gtcttcttct ctggatatac cttcaaacac 1980
aaaatggtct atgaagacac actcacccta ttcccattct caggagaaac tgtcttcatg 2040
tcgatggaaa acccaggtct atggattctg gggtgccaca actcagactt tcggaacaga 2100
ggcatgaccg ccttactgaa ggtttctagt tgtgacaaga acactggtga ttattacgag 2160
gacagttatg aagatatttc agcatacttg ctgagtaaaa acaatgccat tgaaccaaga 2220
agcttctccc agaattcaag acaccctagc actaggcaaa agcaatttaa tgccaccaca 2280
attccagaaa atgacataga gaagactgac ccttggtttg cacacagaac acctatgcct 2340
aaatgtcaaa atgtctcctc tagtgatttg ttgatgctct tgcgacagag tcctactcca 2400
catgggctat ccttatctga tctccaagaa gccaaatatg agactttttc tgatgatcca 2460
tcacctggag caatagacag taataacagc ctgtctgaaa tgacacactt caggccacag 2520
ctccatcaca gtggggacat ggtatttacc cctgagtcag gcctccaatt aagattaaat 2580
gagaaactgg ggacaactgc agcaacagag ttgaagaaac ttgatttcaa agtttctagt 2640
acatcaaata atctgatttc aacaattcca tcagacaatt tggcagcagg tactgataat 2700
acaagttcct taggaccccc aagtatgcca gttcattatg atagtcaatt agataccact 2760
ctatttggca aaaagtcatc tccccttact gagtctggtg gacctctgag cctatcctgt 2820
cggcccccca tggtcaagct ggtgtgtccc gctgacaacc tgcgggctga agggctcgag 2880
tgtaccaaaa cgtgccagaa ctatgacctg gagtgcatga gcatgggctg tgtctctggc 2940
tgcctctgcc ccccgggcat ggtccggcat gagaacagat gtgtggccct ggaaaggtgt 3000
ccctgcttcc atcagggcaa ggagtatgcc cctggagaaa cagtgaagat tggctgcaac 3060
acttgtgtct gtcgggaccg gaagtggaac tgcacagacc atgtgtgtga tgccacgtgc 3120
tccacgatcg gcatggccca ctacctcacc ttcgacgggc tcaaatacct gttccccggg 3180
gagtgccagt acgttctggt gcaggattac tgcggcagta accctgggac ctttcggatc 3240
ctagtgggga ataagggatg cagccacccc tcagtgaaat gcaagaaacg ggtcaccatc 3300
ctggtggagg gaggagagat tgagctgttt gacggggagg tgaatgtgaa gaggcccatg 3360
aaggatgaga ctcactttga ggtggtggag tctggccggt acatcattct gctgctgggc 3420
aaagccctct ccgtggtctg ggaccgccac ctgagcatct ccgtggtcct gaagcagaca 3480
taccaggaga aagtgtgtgg cctgtgtggg aattttgatg gcatccagaa caatgacctc 3540
accagcagca acctccaagt ggaggaagac cctgtggact ttgggaactc ctggaaagtg 3600
agctcgcagt gtgctgacac cagaaaagtg cctctggact catcccctgc cacctgccat 3660
aacaacatca tgaagcagac gatggtggat tcctcctgta gaatccttac cagtgacgtc 3720
ttccaggact gcaacaagct ggtggacccc gagccatatc tggatgtctg catttacgac 3780
acctgctcct gtgagtccat tggggactgc gcctgcttct gcgacaccat tgctgcctat 3840
gcccacgtgt gtgcccagca tggcaaggtg gtgacctgga ggacggccac attgtgcccc 3900
cagagctgcg aggagaggaa tctccgggag aacgggtatg agtgtgagtg gcgctataac 3960
agctgtgcac ctgcctgtca agtcacgtgt cagcaccctg agccactggc ctgccctgtg 4020
cagtgtgtgg agggctgcca tgcccactgc cctccaggga aaatcctgga tgagcttttg 4080
cagacctgcg ttgaccctga agactgtcca gtgtgtgagg tggctggccg gcgttttgcc 4140
tcaggaaaga aagtcacctt gaatcccagt gaccctgagc actgccagat ttgccactgt 4200
gatgttgtca acctcacctg tgaagcctgc caggagccgg gaggcctggt ggtgcctccc 4260
acagatgccc cggtgagccc caccactctg tatgtggagg acatctcgga accgccgttg 4320
cacgatttct gataa 4335
<210> 80
<211> 3864
<212> DNA
<213> Artificial Sequence
<220>
<223> dcFVIII/D'D3-HC/Fc/Fc-LC Light chain
<400> 80
gccaccgatc ctcttgcttg ggataaccac tatggtactc agataccaaa agaagagtgg 60
aaatcccaag agaagtcacc agaaaaaaca gcttttaaga aaaaggatac cattttgtcc 120
ctgaacgctt gtgaaagcaa tcatgcaata gcagcaataa atgagggaca aaataagccc 180
gaaatagaag tcacctgggc aaagcaaggt aggactgaaa ggctgtgctc tcaaaaccca 240
ccagtcttga aacgccatca acgggaaata actcgtacta ctcttcagtc agatcaagag 300
gaaattgact atgatgatac catatcagtt gaaatgaaga aggaagattt tgacatttat 360
gatgaggatg aaaatcagag cccccgcagc tttcaaaaga aaacacgaca ctattttatt 420
gctgcagtgg agaggctctg ggattatggg atgagtagct ccccacatgt tctaagaaac 480
agggctcaga gtggcagtgt ccctcagttc aagaaagttg ttttccagga atttactgat 540
ggctccttta ctcagccctt ataccgtgga gaactaaatg aacatttggg actcctgggg 600
ccatatataa gagcagaagt tgaagataat atcatggtaa ctttcagaaa tcaggcctct 660
cgtccctatt ccttctattc tagccttatt tcttatgagg aagatcagag gcaaggagca 720
gaacctagaa aaaactttgt caagcctaat gaaaccaaaa cttacttttg gaaagtgcaa 780
catcatatgg cacccactaa agatgagttt gactgcaaag cctgggctta tttctctgat 840
gttgacctgg aaaaagatgt gcactcaggc ctgattggac cccttctggt ctgccacact 900
aacacactga accctgctca tgggagacaa gtgacagtac aggaatttgc tctgtttttc 960
accatctttg atgagaccaa aagctggtac ttcactgaaa atatggaaag aaactgcagg 1020
gctccctgca atatccagat ggaagatccc acttttaaag agaattatcg cttccatgca 1080
atcaatggct acataatgga tacactacct ggcttagtaa tggctcagga tcaaaggatt 1140
cgatggtatc tgctcagcat gggcagcaat gaaaacatcc attctattca tttcagtgga 1200
catgtgttca ctgtacgaaa aaaagaggag tataaaatgg cactgtacaa tctctatcca 1260
ggtgtttttg agacagtgga aatgttacca tccaaagctg gaatttggcg ggtggaatgc 1320
cttattggcg agcatctaca tgctgggatg agcacacttt ttctggtgta cagcaataag 1380
tgtcagactc ccctgggaat ggcttctgga cacattagag attttcagat tacagcttca 1440
ggacaatatg gacagtgggc cccaaagctg gccagacttc attattccgg atcaatcaat 1500
gcctggagca ccaaggagcc cttttcttgg atcaaggtgg atctgttggc accaatgatt 1560
attcacggca tcaagaccca gggtgcccgt cagaagttct ccagcctcta catctctcag 1620
tttatcatca tgtatagtct tgatgggaag aagtggcaga cttatcgagg aaattccact 1680
ggaaccttaa tggtcttctt tggcaatgtg gattcatctg ggataaaaca caatattttt 1740
aaccctccaa ttattgctcg atacatccgt ttgcacccaa ctcattatag cattcgcagc 1800
actcttcgca tggagttgat gggctgtgat ttaaatagtt gcagcatgcc attgggaatg 1860
gagagtaaag caatatcaga tgcacagatt actgcttcat cctactttac caatatgttt 1920
gccacctggt ctccttcaaa agctcgactt cacctccaag ggaggagtaa tgcctggaga 1980
cctcaggtga ataatccaaa agagtggctg caagtggact tccagaagac aatgaaagtc 2040
acaggagtaa ctactcaggg agtaaaatct ctgcttacca gcatgtatgt gaaggagttc 2100
ctcatctcca gcagtcaaga tggccatcag tggactctct tttttcagaa tggcaaagta 2160
aaggtttttc agggaaatca agactccttc acacctgtgg tgaactctct agacccaccg 2220
ttactgactc gctaccttcg aattcacccc cagagttggg tgcaccagat tgccctgagg 2280
atggaggttc tgggctgcga ggcacaggac ctctacctgg tcccacgtgg aagtggcggt 2340
ggggggtcag gcggaggcgg cagtggcggc gggggctctg gcggaggtgg cagtggcggg 2400
ggctctgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct tgggggaccg 2460
tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 2520
gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 2580
gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 2640
acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 2700
tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 2760
gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagctg 2820
accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc 2880
gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 2940
gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 3000
caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 3060
aagagcctct ccctgtctcc gggtaaaggc ggtggggggt caggcggagg cggcagtggc 3120
ggcgggggct ctggcggagg tgggtccggc ggtggcggtt ctggcggggg cggctctgac 3180
aaaactcaca catgcccacc gtgcccagca cctgaactcc ttgggggacc gtcagtcttc 3240
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 3300
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 3360
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 3420
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 3480
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 3540
cagccccgag aaccacaggt gtacaccctg cccccatccc gggatgagct gaccaagaac 3600
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 3660
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 3720
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 3780
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 3840
tccctgtctc cgggtaaatg ataa 3864
Claims (32)
- 인간 혈액응고 8인자(Factor VIII, FVIII),
하나 이상의 vWF D’D3 도메인, 및
상기 FVIII 및 상기 vWF D’D3 도메인을 연결하는 링커를 포함하는 키메라 단백질로,
상기 FVIII는 서열번호 1의 아미노산 서열에서 B 도메인의 일부가 결실되고, 782번 위치의 아미노산 잔기가 시스테인으로 치환된 것이며,
상기 링커는 효소절단부위를 포함하고,
상기 키메라 단백질은 vWF를 구성하는 도메인 중 D1, D2, A1, A2, A3, D4, C1, C2, C3, C4, C5, C6 및 CK 중 어느 도메인도 포함하지 않는 것인, 키메라 단백질. - 제1항에 있어서,
상기 FVIII는 서열번호 1의 아미노산 서열에서 1648번 및 1649번 위치의 아미노산 잔기를 기준으로 상기 잔기를 포함하여 N-말단, C-말단, 또는 N-말단 및 C-말단 방향으로 각각 최소 5개의 아미노산 잔기가 결실된 것인, 키메라 단백질. - 제2항에 있어서,
상기 FVIII는 서열번호 1의 아미노산 서열에서 903번 내지 1653번 위치의 아미노산 잔기가 결실된 것인, 키메라 단백질. - 제1항에 있어서,
상기 FVIII는 추가적으로 서열번호 1의 아미노산 서열에서 491, 495, 498, 754, 781, 788, 789, 825, 897 및 1806 위치의 아미노산 잔기로 구성된 군으로부터 선택되는 하나 이상의 아미노산 잔기가 시스테인으로 치환된 것인, 키메라 단백질. - 제1항에 있어서,
상기 시스테인에 친수성 폴리머가 연결된 것인, 키메라 단백질. - 제5항에 있어서,
상기 친수성 폴리머는 폴리에틸렌글리콜(Polyethylene glycol), 폴리에틸렌 옥사이드 또는 덱스트란인 것인, 키메라 단백질. - 제6항에 있어서,
상기 친수성 폴리머는 폴리에텔렌글리콜이고,
상기 폴리에틸렌글리콜의 평균 분자량은 20kDa 이상인 것인, 키메라 단백질. - 제7항에 있어서,
상기 폴리에틸렌글리콜의 평균 분자량은 40kDa 또는 60kDa인 것인, 키메라 단백질. - 제1항에 있어서,
상기 효소절단부위는 트롬빈, FVIIa, FXa 또는 FXIa에 의해 절단될 수 있는 아미노산 서열이고,
상기 링커에서 효소절단부위를 제외한 아미노산 서열은 [G4S]n의 아미노산 서열로 표시되고,
상기 식에서 n = 0 내지 100 사이 정수인, 키메라 단백질. - 제9항에 있어서,
상기 트롬빈에 의해 절단될 수 있는 아미노산 서열은 DFLAEGGGVR (서열번호 36), TTKIKPR (서열번호 37), 또는 LVPRGS (서열번호 38)이고;
상기 FVIIa에 의해 절단될 수 있는 아미노산 서열은 ASKPQGRIVGG (서열번호 39)이며;
상기 FXa에 의해 절단될 수 있는 서열은 IDGR (서열번호 40) 또는 IEGR (서열번호 41)이고; 또는
상기 FXIa에 의해 절단될 수 있는 서열은 SKLTRAETVF (서열번호 42)인, 키메라 단백질. - 제1항에 있어서,
상기 키메라 단백질은 단쇄(single chain) 또는 이쇄(double chain)인 것인, 키메라 단백질. - 제11항에 있어서,
상기 이쇄인 키메라 단백질은 제1 폴리펩타이드 및 제2 폴리펩타이드로 구성된 FVIII를 포함하는 것으로서;
상기 제1 폴리펩타이드는 A1 도메인, A2 도메인 및 B 도메인 단편을 포함하며,
상기 제2 폴리펩타이드는 A3 도메인, C1 도메인, C2 도메인 및 B 도메인 단편을 포함하는 것인, 키메라 단백질. - 제11항에 있어서,
상기 단쇄인 키메라 단백질은 서열번호 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21 또는 25의 아미노산 서열 또는 이와 90% 이상 상동성이 있는 아미노산 서열로 구성되는 것인, 키메라 단백질. - 제12항에 있어서,
상기 이쇄인 키메라 단백질은 서열번호 4 및 5, 서열번호 23 및 24, 서열번호 26 및 27, 서열번호 28 및 29, 서열번호 30 및 31, 서열번호 32 및 33, 또는 서열번호 34 및 35의 아미노산 서열 또는 이와 90% 이상 상동성이 있는 아미노산 서열로 구성되는 것인, 키메라 단백질. - 제13항에 기재된 키메라 단백질을 코딩하는 핵산분자.
- 제14항에 기재된 키메라 단백질을 코딩하는 핵산분자.
- 제15항에 있어서,
상기 핵산분자는 서열번호 48 내지 64, 66, 또는 70의 염기 서열로 구성되되는 것인, 핵산분자. - 제16항에 있어서,
상기 핵산분자는 서열번호 46, 47, 68, 69, 또는 71 내지 80의 염기 서열로 구성되는 것인, 핵산분자. - 제15항 내지 제18항 중 어느 한 항에 기재된 핵산분자를 포함하는 벡터.
- 제19항에 기재된 벡터를 포함하는 세포.
- 제1항 내지 제14항 중 어느 한 항에 기재된 키메라 단백질, 제15항 내지 제 18항 중 어느 한 항에 기재된 핵산분자, 제19항에 기재된 벡터, 또는 제20항에 기재된 세포 및 약학적으로 허용 가능한 담체를 포함하는 A형 혈우병 치료용 약학적 조성물.
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PCT/KR2017/006655 WO2017222337A1 (ko) | 2016-06-24 | 2017-06-23 | Fviii 및 vwf 인자를 포함하는 키메라 단백질 및 그 용도 |
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AU2018366110A1 (en) | 2017-11-07 | 2020-06-04 | Rani Therapeutics, Llc | Clotting factor preparations for delivery into tissue of the intestinal tract using a swallowable drug delivery device |
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AU2019366942A1 (en) * | 2018-10-23 | 2021-06-10 | The Children's Hospital Of Philadelphia | Compositions and methods for modulating factor VIII function |
AU2020270420A1 (en) * | 2019-05-03 | 2021-11-18 | Rani Therapeutics, Llc | Clotting factor preparations for delivery into tissue of the intestinal tract using a swallowable drug delivery device |
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US20190330311A1 (en) | 2019-10-31 |
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