KR20210034055A - Htlv-1 관련 척수증(ham) 치료 또는 예방제, 및 ham의 치료 방법 - Google Patents
Htlv-1 관련 척수증(ham) 치료 또는 예방제, 및 ham의 치료 방법 Download PDFInfo
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- KR20210034055A KR20210034055A KR1020217005032A KR20217005032A KR20210034055A KR 20210034055 A KR20210034055 A KR 20210034055A KR 1020217005032 A KR1020217005032 A KR 1020217005032A KR 20217005032 A KR20217005032 A KR 20217005032A KR 20210034055 A KR20210034055 A KR 20210034055A
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- Prior art keywords
- ser
- leu
- thr
- val
- gly
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Abstract
본 발명은, RGMa 저해 물질을 포함하는, HTLV-1 관련 척수증(HAM)의 치료 또는 예방제, 및 그것을 필요로 하는 HTLV-1 관련 척수증(HAM)환자에 약리학적으로 유효량의 RGMa 저해 물질을 투여하는 것을 포함하는, HAM의 치료 방법을 제공한다.
Description
본 발명은, HTLV-1 관련 척수증(HAM) 치료 또는 예방제에 관한 것이다. 또한, 본 발명은, HAM의 치료 방법에도 관한 것이다.
HTLV-1(Human T-cell Lukemia Virus Type 1; 인간 T세포 백혈병 바이러스)은, 혈액 중의 백혈구의 하나인 T세포 등(주로 CD4 양성 T세포)에 감염하는 바이러스이다.
HTLV-1에 감염된 T세포는, 척수에 만성 염증을 형성하고, 그 결과, 척수 신경 세포의 장애 및 변성을 일으키고, 경성(痙性) 척수마비를 야기한다. HTLV-1에 감염된 세포가 원인이 되어 야기되는 경성 척수마비는, HTLV-1 관련 척수증(HAM으로도 약기함)으로 불리운다.
HAM의 증상으로서는, 신경의 조직 장애에 의한, 양발의 마비, 통증, 배뇨 장애, 끈덕진 변비 등의 증상 등을 예로 들 수 있다. 이러한 증상이 진행되면, 휠체어 생활, 와병 생활을 하게 된다. HAM은 일본의 지정 난병(불치병)에 지정된 질환의 하나이다. 현재, 유효한 HAM의 치료 방법은 확립되어 있지 않고, 다만 대증요법이 행해지고 있다.
치료 방법의 하나로서, 항CCR4항체를 사용하는 치료 방법이, HTLV-1 감염 세포를 감소시켜, HAM의 척수염증을 경감하고, 증상 개선 효과를 얻을 수 있는 것이 증명되어 있다(비특허문헌 1, 특허문헌 5).
RGMa 단백질은, 망막이나 해마의 신경세포의 축색 유도, 신경관의 폐쇄 등에 관여하는 단백질 RGM(Repulsive guidance molecule) 패밀리의 일종이다. RGM의 활동은 이들로 한정되지 않으며, 다양한 기능을 가지는 것으로 알려져 있다.
예를 들면, 특허문헌 1에는, 골수 유래 수상 세포(BMDCs)에 RGM이 발현되고 있는 것, CD4+ T세포 및 CD11b+ 마크로파지에 RGM 수용체가 발현되고 있고, 상기 RGM수 용체에 RGM이 결합함으로써, CD4+ T세포 및 CD11b+ 마크로파지의 세포 접착 활성이 증강하는 것이 개시되어 있다. 또한, 특허문헌 1에는, 항RGM중화항체에 의해, 다발성 경화증 모델 마우스의 임상 증상 및 조직 병변의 양쪽을 경감할 수 있고, 상기 마우스로부터 취득한 비장세포에서는 항원특이적 및 비특이적인 T세포 활성화가 감약한 것이 개시되어 있다.
특허문헌 2 및 3에는, RGMa의 수용체 네오게닌 및 골형성 단백질 2 및 4(BMP-2, BMP-4)로의 RGMa의 결합을 선택적으로 저해하는, RGMa에 대한 중화 모노클로날 항체가 개시되어 있다. 상기 중화 모노클로날 항체에 의하면, 손상을 입고, 염증을 일으킨 인간 중추 신경계, 구체적으로는, 다발성 경화증, 급성 척수손상, 뇌외상 후, 헌팅턴무도병, 파킨슨병, 알츠하이머병 등의 신경변성 질환에 있어서, 파괴된 뉴런 결합의 신경 재생 및 재성장을 촉진할 수 있는 것으로 여겨지고 있다.
특허문헌 4에는, RGMa는 외상성 뇌손상 또는 속이 허헐성 뇌졸중을 앓고 있는 인간의 중추 신경계 미에린, 신선 병소 및 성숙 반흔 조직에 국재화(局在化)하고 있고, 이 신경 변성질환을 진단하는 것을 목적으로, RGMa 단편을 검출하고, 정량화(定量化)하는 방법이 개시되어 있다. 또한, 특허문헌 4에는, RGMa 단편을 검출하는 대상의 신경 변성질환 및 장애로서, 다발성 경화증, 파킨슨병, 알츠하이머병, 테이-삭스병, 니만-피크병, 고쉐병, 헬러증후군, 헌팅톤병, 근위축성 측색경화증, 특발성 염증성 탈수질환, 비타민 B12 결핍증, 교중심수초파괴증, 척수매독, 횡단성 척수염, 데빅병, 진행성 다병소성 백질뇌증, 시신경염, 척수손상, 외상성 뇌손상, 졸중, 녹내장, 당뇨병성 망막증, 노인성 황반변성, 백질 디스트로피를 예로 들 수 있다.
N Engl J Med, 2018, 378, 529-538.
비특허문헌 1, 특허문헌 5에 개시되어 있는 항CCR4항체를 사용하는 방법에 의하면 HAM의 증상을 개선할 수 있지만, 신경파괴가 진행된 HAM 환자에게는 항CCR4항체의 효과가 한정적이다. 이 때문에, HAM을 치료할 수 있는 보다 우수한 방법이 요구되고 있다.
본 발명은, 상기한 문제점을 해결하기 위해 이루어진 것이며, HAM을 치료할 수 있는 치료제 및 치료 방법을 제공하는 것을 목적으로 한다.
본 발명자들은, 상기 문제점을 해결하기 위해 예의(銳意) 검토를 거듭한 결과, RGMa를 저해하는 물질은, HAM의 치료에 유효한 것을 발견하고, 본 발명을 완성하기에 이르렀다.
즉, 본 발명은 하기와 같다.
[1]
RGMa 저해 물질을 포함하는, HTLV-1 관련 척수증(HAM)의 치료 또는 예방제.
[2]
상기 RGMa 저해 물질이, RGMa를 인식하는 항체인, [1]에 기재된 HAM의 치료 또는 예방제.
[3]
그것을 필요로 하는 HTLV-1 관련 척수증(HAM) 환자에게 약리학적으로 유효량의 RGMa 저해 물질을 투여하는 공정을 포함하는, HAM의 치료 방법.
[4]
상기 RGMa 저해 물질이, RGMa를 인식하는 항체인, [3]에 기재된 HAM의 치료 방법.
본 발명에 의하면, 난치성 질환인 HAM을 치료할 수 있다.
도 1은 CD4양성 T세포에 있어서, HTLV-1 감염 세포의 지표인 CADM1과, CD7에 의해 분류된 군 P, D, N을 나타낸 도면이다.
도 2는 정상 T세포(Normal.CD4), HAM 환자 유래 CD4양성 T세포(HAM.CD4), 건강한 사람 유래 CD4양성/CADM1음성/CD7양성 T세포(Normal.P), HTLV-1 감염자 CD4양성/CADM1음성/CD7양성 T세포(P군), HTLV-1 감염자 CD4양성/CADM1양성/CD7양성 T세포(D군), HTLV-1 감염자 CD4양성/CADM1양성/CD7음성 T세포(N군), 급성형 ATL 환자 CD4양성/CADM1양성/CD7음성 T세포(Acute.N), 건강한 사람 CD4양성 T세포(Normal.CD4.1), 무증후성 ATL환자 유래 PBMC(Smoldering), 만성형 ATL환자 유래PBMC(Chronic), 급성형 ATL환자 유래 PBMC(Acute)에서의, RGMa 유전자의 발현 레벨을 나타낸 도면이다.
도 3은 HAM환자 CD4양성 T세포 및 건강한 사람 CD4양성 T세포에서의 RGMa의 발현량을 나타낸 도면이다.
도 4는 HAM환자 PBMC에서의 세포종 사이에서의 RGMa의 발현량을 분석한 결과를 나타낸 도면이다.
도 5는 HAM환자 PBMC의 배양에서의 HTLV-1 바이러스의 발현에 따른 RGMa의 발현 변화를 나타낸 도면이다.
도 6은 RGMa유전자의 전사개시점으로부터 -2,916bp 상류 부근의 H3K27me3 레벨을 나타낸 도면이다.
도 7은 인간 CD4양성 T세포 백혈병세포주 Jurkat에 대하여, HTLV-1 Tax를 코딩하는 cDNA를 삽입하는 렌티바이러스 벡터를 도입했을 때의, RGMa유전자 mRNA의 레벨을 나타낸 도면이다.
도 8은 HTLV-1-tax 발현유도 세포주 JPX-9에서의 Tax 및 RGMa의 단백질 발현의 분석 결과를 나타낸 도면이다.
도 9는 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, 자발적 증식 활성에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 10은 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, HTLV-1 프로바이러스량 변화에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 11은 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, CXCL10 생산에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 12는 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, HAM환자 PBMC의 사이토카인 생산에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 13은 HAM-PBMC이 신경세포주의 아포토시스 유도를 일으킨 것을 나타낸 도면이다.
도 14는 HTLV-1-tax 유도세포주에 의한 신경세포주의 아포토시스 유도에 대한다 RGMa항체의 억제 작용의 결과를 나타낸 도면으로서,
(a)는, NB-1세포와 JPX-9세포(미자극)를 공배양했을 때의, FACS 플롯이다.
(b)는, NB-1세포와 CdCl2 자극 JPX-9세포(HTLV-1-tax 발현세포)를 공배양했을 때의, FACS 플롯이다.
(c)는, (b)의 조건에, 컨트롤 항체를 첨가했을 때의, FACS 플롯이다.
(d)는, (b)의 조건에, 항RGMa항체를 첨가했을 때의, FACS 플롯이다.
도 2는 정상 T세포(Normal.CD4), HAM 환자 유래 CD4양성 T세포(HAM.CD4), 건강한 사람 유래 CD4양성/CADM1음성/CD7양성 T세포(Normal.P), HTLV-1 감염자 CD4양성/CADM1음성/CD7양성 T세포(P군), HTLV-1 감염자 CD4양성/CADM1양성/CD7양성 T세포(D군), HTLV-1 감염자 CD4양성/CADM1양성/CD7음성 T세포(N군), 급성형 ATL 환자 CD4양성/CADM1양성/CD7음성 T세포(Acute.N), 건강한 사람 CD4양성 T세포(Normal.CD4.1), 무증후성 ATL환자 유래 PBMC(Smoldering), 만성형 ATL환자 유래PBMC(Chronic), 급성형 ATL환자 유래 PBMC(Acute)에서의, RGMa 유전자의 발현 레벨을 나타낸 도면이다.
도 3은 HAM환자 CD4양성 T세포 및 건강한 사람 CD4양성 T세포에서의 RGMa의 발현량을 나타낸 도면이다.
도 4는 HAM환자 PBMC에서의 세포종 사이에서의 RGMa의 발현량을 분석한 결과를 나타낸 도면이다.
도 5는 HAM환자 PBMC의 배양에서의 HTLV-1 바이러스의 발현에 따른 RGMa의 발현 변화를 나타낸 도면이다.
도 6은 RGMa유전자의 전사개시점으로부터 -2,916bp 상류 부근의 H3K27me3 레벨을 나타낸 도면이다.
도 7은 인간 CD4양성 T세포 백혈병세포주 Jurkat에 대하여, HTLV-1 Tax를 코딩하는 cDNA를 삽입하는 렌티바이러스 벡터를 도입했을 때의, RGMa유전자 mRNA의 레벨을 나타낸 도면이다.
도 8은 HTLV-1-tax 발현유도 세포주 JPX-9에서의 Tax 및 RGMa의 단백질 발현의 분석 결과를 나타낸 도면이다.
도 9는 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, 자발적 증식 활성에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 10은 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, HTLV-1 프로바이러스량 변화에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 11은 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, CXCL10 생산에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 12는 HAM환자 PBMC에 대한 RGMa항체의 작용에 관하여, HAM환자 PBMC의 사이토카인 생산에 대한 RGMa항체의 작용 결과를 나타낸 도면이다.
도 13은 HAM-PBMC이 신경세포주의 아포토시스 유도를 일으킨 것을 나타낸 도면이다.
도 14는 HTLV-1-tax 유도세포주에 의한 신경세포주의 아포토시스 유도에 대한다 RGMa항체의 억제 작용의 결과를 나타낸 도면으로서,
(a)는, NB-1세포와 JPX-9세포(미자극)를 공배양했을 때의, FACS 플롯이다.
(b)는, NB-1세포와 CdCl2 자극 JPX-9세포(HTLV-1-tax 발현세포)를 공배양했을 때의, FACS 플롯이다.
(c)는, (b)의 조건에, 컨트롤 항체를 첨가했을 때의, FACS 플롯이다.
(d)는, (b)의 조건에, 항RGMa항체를 첨가했을 때의, FACS 플롯이다.
이하에서, 본 발명의 실시형태에 대하여 상세하게 설명한다. 그리고, 본 발명은, 이하의 본 실시형태로 제한되지 않고, 그 요지의 범위 내에서 다양하게 변형되어 실시할 수 있다.
본 발명은, RGMa 저해 물질을 포함하는, HTLV-1 관련 척수증(HAM)의 치료 또는 예방제이다. RGMa 저해 물질은, RGMa 그 자체에 작용하여 RGMa의 활성을 저해하는 물질이라도 되고, RGMa의 발현을 억제하는 물질이라도 된다. RGMa 저해 물질로서는, 예를 들면, RGMa를 저해하는 활성을 가지는 화합물, 및 RGMa를 인식하는 항체 등이 있다.
또한, RGMa 저해 물질로서는, 예를 들면, RGMa를 발현하는 유전자의 siRNA(short interfering RNA), shRNA(short hairpin RNA), 및 안티센스올리고뉴클레오티드 등의 RGMa의 발현을 억제하는 물질이 있다.
RGMa유전자로서는, 예를 들면, 서열번호 1에 표시되는 염기서열로 이루어지는 인간 RGMa유전자, 서열번호 2에 표시되는 염기서열로 이루어지는 RGMa유전자 등이 있지만, 이것으로 한정되는 것은 아니다. 다양한 생물 유래의 RGMa유전자의 염기서열 정보는, GenBank 등의 데이터베이스로부터 취득할 수 있다.
siRNA는, 표적인 RGMa유전자의 발현을 억제할 수 있는 이중가닥 RNA이다. siRNA에서의 염기서열의 길이(염기길이)는 특별히 한정되지 않지만, 바람직하게는 약 30염기 미만, 보다 바람직하게는 약 19∼27 염기, 더욱 바람직하게는 약 21∼25 염기이다.
shRNA는, 단일가닥 RNA 중에 부분적으로 회문형의 염기서열을 포함함으로써, 분자 내에 이중가닥 구조를 가지고, 3'말단에 돌출부를 가지는 짧은 헤어핀 구조로부터로 이루어지는 약 20염기쌍 이상의 분자를 나타낸다. shRNA는, 세포 내에 도입된 후, 세포 내에서 약 20염기의 길이로 분해되어, siRNA와 마찬가지로 표적인 RGMa유전자의 발현을 억제할 수 있다.
siRNA 및 shRNA는, 인공적으로 화학 합성할 수 있다. 또한, siRNA 및 shRNA는, 예를 들면, T7RNA폴리머라아제 및 T7프로모터를 사용하여, 주형DNA로부터 안티 센스 및 센스의 RNA를 인비트로에서 합성할 수 있다.
안티센스올리고뉴클레오티드는, RGMa유전자의 DNA서열 중의 연속하는 약 30염기 미만의 염기서열에 대하여 상보적(相補的)인, 또는 하이브리다이제이션하는 뉴클레오티드이면 되고, DNA 또는 RNA 중 어느 것이라도 된다. 또한, 기능에 지장이 없는 한 수식된 것이라도 된다. 안티센스올리고뉴클레오티드는 통상적인 방법에 의해 합성할 수 있으며, 예를 들면, 시판중인 DNA합성장치에 의해 용이하게 합성할 수 있다.
본 발명의 HAM의 치료 또는 예방제에 포함되는 RGMa 저해 물질로서는, RGMa를 인식하는 항체가 바람직하다. 이하에서, RGMa를 인식하는 항체를 RGMa항체라고도 한다. 본 발명에서의 RGMa항체는, RGMa에 결합하여 그 활성을 저해하는 항체이면 되며, 예를 들면, RGMa에 결합함으로써 RGMa가 RGMa수용체에 결합할 수 없도록 하는 항체 등이 있다.
본 발명에서의 RGMa항체는, 모노클로날 항체라도 되고 폴리클로날 항체라도 된다. 또한, 본 발명에서의 항체는, IgG, IgM, IgA, IgD 및 IgE 중의 어느 아이소타입이라도 된다.
본 발명에서의 RGMa항체는, 예를 들면, 마우스 항체, 인간형 CDR 이식 항체, 인간형 키메라 항체, 인간화 항체, 또는 완전 인간 항체라도 되고, 저분자 항체라도 된다. 이들 항체는, 1종 단독으로 사용해도 되고, 2종 이상을 조합하여 사용할 수도 있다.
인간형 CDR 이식 항체는, 인간 이외의 동물의 항체 CDR을, 인간 항체의 CDR로 치환한 항체이다. 인간형 키메라 항체는, 인간 이외의 동물의 항체에 유래하는 가변 영역과, 인간 항체에 유래하는 정상 영역으로 이루어지는 항체이다. 또한, 인간화 항체란, 인간 이외의 동물의 항체에 있어서, 안정성이 높은 일부 영역을 남기고, 인간의 항체에 유래하는 부분을 조합한 것을 일컬으며, 인간형 키메라 항체, 및 인간형 CDR 이식 항체를 포함하는 개념이다.
본 명세서에 있어서 「저분자 항체」란, 항체의 단편 또는 항체의 단편에 임의의 분자를 결합시킨 것으로서, 원래의 항체와 동일한 에피토프를 인식하는 것을 의미한다. 구체적으로는, VL, VH, CL 및 CH1 영역으로 이루어지는 Fab; 2개의 Fab가 힌지 영역에서 디술피드 결합에 의해 연결되어 있는 F(ab')2; VL 및 VH로 이루어지는 Fv; VL 및 VH를 인공의 폴리펩티드 링커로 연결한 단일가닥 항체인 scFv 외에, sdFv, Diabody, sc(Fv)2를 예로 들 수 있지만, 이들로 한정되지 않는다.
본 발명에 있어서 사용되는 RGMa항체는, RGMa 또는 그의 플래그먼트를 면역원으로서 사용하여, 공지의 방법으로 제작할 수 있다.
얻어진 항체가 RGMa항체인 것은, RGMa 활성을 지표로 하여 확인할 수 있다.
RGMa로서는, 예를 들면, 서열번호 3에 표시되는 아미노산 서열을 포함하는 인간 RGMa, 서열번호 4에 표시되는 아미노산 서열을 포함하는 RGMa 등이 있다. 다양한 생물 유래의 RGMa를 면역원으로서 사용할 수 있다. RGMa의 아미노산 서열은 공지의 데이터베이스인 Protein Data Bank 등으로부터 취득할 수 있다.
본 발명에 있어서 사용되는 RGMa항체가 폴리클로날 항체일 때, 예를 들면, 하기와 같이 하여 제작할 수 있다. 먼저, 항원으로서 RGMa 또는 이들의 플래그먼트를 인산완충 생리식염수(PBS로도 기재함)에 용해하고, 필요에 따라 통상의 아쥬반트, 예를 들면, 프로인드 완전아쥬반트를 적량 혼합한 것을 면역원으로 하여, 마우스, 래트(rat), 토끼, 염소, 및 말 등의 포유동물을 면역한다. 면역 방법은 특별히 한정되지 않지만, 예를 들면, 1회 또는 적당한 간격으로 2회 이상, 피하 주사 또는 복강내 주사하는 방법이 있다. 다음으로, 통상적인 방법에 따리, 면역한 동물로부터 혈액을 채취하여 혈청을 분리하고, 폴리클로날 항체 획분을 정제함으로써 취득할 수 있다.
본 발명에 있어서 사용되는 RGMa항체가 모노클로날 항체일 때, 상기 모노클로날 항체는, 상기 면역된 포유동물로부터 얻은 면역세포, 예를 들면, 비장세포와 미엘로마 세포를 융합시켜서 하이브리도마를 얻고, 상기 하이브리도마의 배양물로부터 항체를 채취함으로써 얻을 수 있다. 또한, 상기 모노클로날 항체는, 항체유전자를 하이브리도마로부터 클로닝하고, 적절한 벡터에 도입하고, 이것을 숙주세포에 도입하고, 유전자재조합 기술을 사용하여 재조합형의 모노클로날 항체를 생산시킬 수도 있다. 또한, 상기 모노클로날 항체는, 파지디스플레이법을 사용하여 제작할 수도 있다.
본 발명에 있어서 사용되는 RGMa항체로서는, 예를 들면, Yamashita, T., Mueller, B.K.& Hata, K. Neogenin and repulsive guidance molecule signaling in the central nervous system. Curr. Opin. Neurobiol.17, 29-34(2007); 일본공개특허 제2014-138599호 공보; 일본공개특허 제2016-175897호 공보; 일본특표 2017-526930호 공보; 국제공개 2016/175236호; 일본특표 2015-508061호 공보 등에 개시되어 있는 항체를 사용할 수 있다.
또한, 본 발명에 있어서 사용되는 RGMa항체로서는, 예를 들면, 가부시키가이샤면역생물연구소(IBL)제, R&D Systems사제 등의 시판품으로서 입수할 수도 있다.
RGMa항체는, 항원 결합 도메인으로서,
GTTPDY(서열번호 7);
FQATHDPLT(서열번호 10);
ARRNEYYGSSFFDY(서열번호 13);
LQGYIPPRT(서열번호 16); 및
상기 서열의 1개와 적어도 50%의 서열 동일성을 가지는 수식 CDR 아미노산 서열;
로 이루어지는 군으로부터 선택되는 아미노산 서열을 포함하는 적어도 1개의 CDR을 포함하는 것이 바람직하다. 상기한 서열 동일성은, 바람직하게는 80% 이상이며, 보다 바람직하게는 90% 이상이다. 그리고, 본 명세서에 있어서 아미노산은 관용적인 1문자 표기 또는 3문자 표기로 표시되는 경우가 있다.
상보성 결정 영역(CDR)이란 면역 글로불린 분자의 가변 영역 중, 항원 결합 부위를 형성하는 영역을 일컬으며, 초가변영역으로도 불리우며, 면역 글로불린 분자마다 특히 아미노산 서열의 변화가 큰 부분을 일컫는다. CDR에는 경쇄 및 중쇄 각각 3개의 CDR(CDR-L1, CDR-L2, CDR-L3, 및 CDR-H1, CDR-H2, CDR-H3)이 있다. 본원에서는, 면역 글로불린 분자의 CDR은 카밧(Kabat)의 번호붙임시스템(Kabat 등, 1987, Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH, USA)에 따라 결정된다.
또한, 본 발명에서의, 경쇄 및 중쇄의 아미노산 서열로 규정되는 항체에 있어서는, CDR의 아미노산 서열이 소정의 서열로부터 불변이며, CDR 이외가 변이 등에 의해 변화하고 있어도 된다. CDR 이외에 변이 등이 있을 때, 상동성은 90% 이상인 것이 바람직하다.
또한, RGMa항체는, 서열번호 5, 6, 8, 9, 11, 12, 14, 15 및 상기 서열의 1개와 적어도 50%의 서열 동일성을 가지는 수식 CDR 아미노산 서열로 이루어지는 군으로부터 선택되는 아미노산 서열을 포함하는 적어도 1개의 CDR을 더 포함하는 것이 바람직하다. 상기한 서열 동일성은, 바람직하게는 80% 이상이며, 보다 바람직하게는 90% 이상이다.
RGMa항체는, 표 1에 나타낸 가변 도메인 CDR 세트, 또는 상기 3개의 CDR 중 적어도 1개가, 상위 서열과 적어도 50%, 바람직하게는 80%, 보다 바람직하게는 90%의 서열 동일성을 가지는 수식 CDR 아미노산 서열인 가변 도메인 세트로부터 선택되는 적어도 3개의 CDR을 포함하는 것이 보다 바람직하다. 또한, RGMa항체는, 표 1에 나타낸 가변 도메인 CDR 세트를 적어도 2개 포함하는 것이 더욱 바람직하다. 또한, 적어도 2개의 가변 도메인 CDR 세트는, VH5F9 세트 및 VL5F9 세트의 조합, 또는, VH8D1 세트 및 VL8D1 세트의 조합인 것이 바람직하다.
[표 1]
RGMa항체는, 프레임워크 영역을 포함해도 된다. 프레임워크 영역이 포함하는 아미노산 서열로서는, 서열번호 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 및 40을 예로 들 수 있다. 이들 아미노산 서열은, 1종 단독이라도 되고, 2종 이상의 조합이라도 된다. 또한, RGMa항체는, 중쇄 가변 도메인 및 경쇄 가변 도메인으로서, 서열번호 41, 42, 43, 44, 45, 46, 47, 48 및 49로부터 선택되는 적어도 1개의 중쇄 가변 도메인; 및/또는 서열번호 50, 51 및 52로부터 선택되는 적어도 1개의 경쇄 가변 도메인을 포함하는 것이 바람직하다.
또한, RGMa항체는, RGMa 저해를 일으키는 부위이면 RGMa에 결합할 때의 결합 부위는 특별히 한정되지 않지만, 예를 들면, 인간 RGMa에서 언급하면,
EEVVNAVEDWDSQG(서열번호 53)
NQQIDFQAFHTNAE(서열번호 54)
PTAPETFPYET(서열번호 55)
KLPVEDLYYQA(서열번호 56)
LYERTRDLPGRAAAGL(서열번호 57)
로 표시되는 아미노산 서열을 가지는 하나 또는 복수의 펩티드에 결합하는 것이 바람직하다.
RGMa항체는, 서열번호 53 및/또는 서열번호 54로 표시되는 아미노산 서열을 가지는 펩티드에 결합하는 것이 보다 바람직하고, 서열번호 53 및/또는 서열번호 54로 표시되는 아미노산 서열을 가지는 펩티드와, 서열번호 55 및/또는 서열번호 56으로 표시되는 아미노산 서열을 가지는 펩티드에 결합하는 것이 더욱 바람직하다.
RGMa항체는, 인간 RGMa의 아미노산 서열에서의 250번째 이후에 결합하는 항체인 것이 바람직하다.
RGMa항체는, 서열번호 53과 서열번호 54, 그리고 서열번호 55 또는 서열번호 56로 표시되는 아미노산 서열을 가지는 펩티드에 결합하는 것이 보다 바람직하다.
RGMa항체는, RGMa 단백질 또는 그의 부분 단편(예를 들면, 서열번호 53∼57 중 하나 이상을 포함하는 단편)을 항원으로 하여, 해당 항원을 마우스 등의 포유동물에 면역하여 얻어지는 폴리클로날 항체 및 모노클로날 항체, 유전자재조합 기술을 사용하여 제조되는 키메라 항체 및 인간화 항체, 및 인간 항체 생산 트랜스제닉 동물 등을 사용하여 제조되는 인간 항체 등이라도 된다.
본 발명에 있어서, RGMa항체는, 의약으로서 인간에 투여하는 경우, 부작용의 관점에서, 인간화 항체 또는 인간 항체가 바람직하다.
RGMa항체는, RGMa 단백질 또는 그의 부분 단편(예를 들면, 서열번호 53∼57 중 하나 이상을 포함하는 단편)을 항원으로서 인식하는 폴리클로날 항체 및/또는 모노클로날 항체의 부분 단편으로서 사용해도 되며, 저분자 항체라도 된다.
RGMa항체로서는, 표 1에 나타낸 것에 더하여, 경쇄의 상보성 결정 영역 1(LCDR1), 경쇄의 상보성 결정 영역 2(LCDR2), 경쇄의 상보성 결정 영역 3(LCDR3), 중쇄의 상보성 결정 영역 1(HCDR1), 중쇄의 상보성 결정 영역 2(HCDR2) 및 중쇄의 상보성 결정 영역 3(HCDR3)의 각각의 아미노산 서열이,
LCDR1: RASQDISSYLN(서열번호 58)
LCDR2: YTSRLHS(서열번호 59)
LCDR3: QQLNTLP(서열번호 60)
HCDR1: DAWMD(서열번호 61)
HCDR2: EIRSKANNHATYYAESVKG(서열번호 62) 및
HCDR3: RDGAY(서열번호 63)을 포함하는,
또는,
LCDR1: RSSQSLVHSNGNTYLH(서열번호 64)
LCDR2: KVSNRFS(서열번호 65)
LCDR3: SQSTHVP(서열번호 66)
HCDR1: TSYYWN(서열번호 67)
HCDR2: YISYDGTNNYNPSLKN(서열번호 68) 및
HCDR3: SFG을 포함하는,
단리된 RGMa항체, 또는 그의 항원 결합 단편이라도 된다.
각 CDR서열에 있어서는, 1개 또는 수개의 아미노산이 치환, 결실, 및/또는 부가되어 있어도 되고, 예를 들면, 1개 또는 2개의 아미노산이 치환, 결실, 및/또는 부가되어 있어도 된다.
RGMa항체로서는, 경쇄에 서열번호 73의 아미노산 서열을, 중쇄에 서열번호 74의 아미노산 서열을 가지는 항체가 예시된다. 이들 서열번호로 표시되는 아미노산 서열에 있어서, 1개 또는 수개의 아미노산(1∼20 개, 1∼10 개 또는 1∼5 개)의 치환, 결실, 부가 또는 삽입이 있어도 된다. 이와 같은 치환, 결실, 부가는 CDR에 도입되어도 되지만, CDR 이외의 영역에 도입되는 것이 바람직하다.
정상 영역을 인간 유래로 한 마우스/인간 키메라 항체라도 되고, 마우스/인간 키메라 항체로서는, 경쇄에 서열번호 77의 아미노산 서열(가변 영역은 1∼107)을, 중쇄에 서열번호 78의 아미노산 서열(가변 영역은 1∼116)을 가지는 항체가 예시된다. 이들 서열번호로 표시되는 아미노산 서열에 있어서, 1개 또는 수개의 아미노산(1∼20 개, 1∼10 개 또는 1∼5 개)의 치환, 결실, 부가 또는 삽입이 있어도 된다. 이와 같은 치환, 결실, 부가는 CDR에 도입되어도 되지만, CDR 이외의 영역에 도입되는 것이 바람직하다.
CDR 이외를 인간 유래로 한 인간화 항체라도 된다. 인간화 항체로서는, 중쇄에 서열번호 70∼87(가변 영역은 N말단측 116잔기까지) 중 어떤 아미노산 서열을 가지고, 경쇄에 서열번호 88∼94(가변 영역은 N말단측 1∼107 잔기까지) 중 어떤 아미노산 서열을 가지는 항체가 예시된다. 이들 서열번호로 표시되는 아미노산 서열은, 1개 또는 수개의 아미노산(1∼20 개, 1∼10 개 또는 1∼5 개)의 치환, 결실, 부가 또는 삽입이 있어도 된다. 이와 같은 치환, 결실, 부가는 CDR에 도입되어도 되지만, CDR 이외의 영역에 도입되는 것이 바람직하다.
중쇄 아미노산 서열과 경쇄 아미노산 서열은 이들의 임의의 조합이면 되지만, 바람직하게는, 중쇄에 서열번호 84의 아미노산 서열, 경쇄에 서열번호 88의 아미노산 서열을 가지는 항체이다. 서열번호 84의 아미노산 서열 중, 중쇄 가변 영역에 상당하는 아미노산 서열은, 서열번호 95로 표시되고, 경쇄 가변 영역에 상당하는 아미노산 서열은, 서열번호 96로 나타내진다.
RGMa항체로서는, 중쇄 가변 영역(VH)이
EVQLVESGGGLVQPGRSLRLSCTASGFTFSDAWMDWVRQAPGKGLEWVAEIRSKANNHATYYAESVKGRFTISRDDSKSIVYLQMNSLRTEDTALYYCTRRDGAYWGKGTTVTVSS(서열번호 95)
또는 해당 아미노산 서열에 적어도 90%의 동일성이 있는 아미노산 서열을 포함하고,
경쇄 가변 영역(VL)이
DIQMTQSPSSVSASVGDRVTITCRASQDISSYLNWYQQKPGKAPKLLIYYTSRLHSGVPSRFSGSGSGTDFTLTISSLQPEDFASYFCQQLNTLPWTFGGGTKVEME(서열번호 96)
또는 해당 아미노산 서열에 적어도 90%의 동일성이 있는 아미노산 서열을 포함하는, 단리된 RGMa항체, 또는 그의 항원 결합 단편이 바람직하다.
RGMa항체로서는, 경쇄에 서열번호 75의 아미노산 서열을, 중쇄에 서열번호 76의 아미노산 서열을 가지는 항체가 예시된다. 이들 서열번호로 표시되는 아미노산 서열에 있어서, 1개 또는 수개의 아미노산(1∼20 개, 1∼10 개 또는 1∼5 개)의 치환, 결실, 부가 또는 삽입이 있어도 된다. 이와 같은 치환, 결실, 부가는 CDR에 도입되어도 되지만, CDR 이외의 영역에 도입되는 것이 바람직하다.
정상 영역을 인간 유래로 한 마우스/인간 키메라 항체라도 되고, CDR 이외를 인간 유래로 한 인간화 항체라도 된다.
또한, 항RGMa항체로서는, 하기 (a1)∼(h1)로부터 선택되는, 단리된 RGMa항체, 또는 그의 항원 결합 단편이라도 된다.
(a1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 99에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편,
(b1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 103에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편,
(c1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 104에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편,
(d1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 105에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편,
(e1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 106에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편,
(f1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 107에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편,
(g1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 108에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편, 및
(h1) 서열번호 97에 기재된 아미노산 서열을 포함하는 LCDR1, 서열번호 98에 기재된 아미노산 서열을 포함하는 LCDR2 및 서열번호 109에 기재된 아미노산 서열을 포함하는 LCDR3를 포함하는 경쇄 가변 영역, 및 서열번호 100에 기재된 아미노산 서열을 포함하는 HCDR1, 서열번호 101에 기재된 아미노산 서열을 포함하는 HCDR2 및 서열번호 102에 기재된 아미노산 서열을 포함하는 HCDR3를 포함하는 중쇄 가변 영역을 포함하는 항RGMa항체, 또는 그의 항원 결합 단편.
RGMa항체는, 전술한 아미노산 서열을 가지는 항체이며, 인간화 항체인 것이 바람직하고, 또한, 인간 IgG의 정상 영역을 가지는 것이 바람직하다.
본 발명에서의 RGMa 저해 물질에 의해, HAM을 치료 또는 예방할 수 있다.
후술하는 실시예 및 도 13에 나타낸 바와 같이, HAM환자에서는 HAM환자 유래 세포에 의해 신경세포사, 즉 척수 조직의 손상 및 변성의 유도가 일어나고 있다.
본 발명자들이 검토한 결과, 후술하는 실시예에 나타낸 바와 같이, HAM환자의 주요 HTLV-1 감염 세포인 CD4양성 T세포에 있어서 RGMa가 현저하게 발현하고 있어, HAM에는 RGMa의 발현이 관여하는 것을 알 수 있다. 또한, RGMa항체를 사용하여 RGMa의 저해를 행하는 시험의 결과, RGMa가 CXCL10, IL-2, 및 IL-10의 생산에 관여하는 것을 알 수 있다.
CXCL10은, IFN-γ에 응답하여 HAM병원성의 HTLV-1 감염 T세포로부터 생산되는 단백질이며, HAM의 병태를 유도하는 것이 알려져 있고(Brain 2013), 또한 HAM의 증상의 진행 속도와도 강하게 상관관계가 있는 것이 알려져 있다(PLoS Negl Trop Dis 2013). HAM환자의 세포에 RGMa항체를 작용시키면, 이 CXCL10의 생산이 억제되었다. 또한, IL-10은 사이토카인의 하나이며, 염증을 억제하도록 작용한다. HAM환자의 세포에 RGMa항체를 작용시키면, IL-10의 생산이 현저하게 증가했다.
HAM의 참된 치료 효과를 부여하기 위해서는, 신경 세포의 상해를 억누르는 것이 요구된다. HAM환자의 HTLV-1 감염 세포에서는 HTLV-1-tax의 발현 레벨이 높고(Blood 2002), HTLV-1-tax가 병태 형성에 중요한 것이 나타나 있다(J Clin Invest 2014). 본 발명에서는, HTLV-1-tax가 RGMa의 발현을 유도하고, RGMa 발현 레벨이 높은 HAM환자 유래 세포가 신경세포사를 유도하는 것이 나타나 있고, 중요한 점은, HTLV-1-tax 발현세포에 의한 신경세포사를 RGMa 저해 물질이 억제하는 것으로 나타났다.
이상과 같이, RGMa 저해 물질은, HAM의 염증 병태의 유도를 억제하고, 또한, HAM환자 세포에 의한 염증반응을 억제할 수 있으므로, HAM을 치료 또는 예방할 수 있다. 또한 RGMa 저해 물질은 HAM 특유의 염증반응을 억제할 뿐만 아니라, HAM환자의 병원적인 세포인 HTLV-1-tax 발현세포에 의한 신경세포사를 억제할 수 있으므로, HAM을 치료 또는 예방할 수 있다.
전술한 바와 같이 RGMa 저해 물질은 HAM을 치료 또는 예방할 수 있고, 본 발명은, HAM의 치료에 사용하기 위한 RGMa 저해 물질; HAM의 치료에 사용하기 위한 의약조성물; HAM을 치료하기 위한 RGMa 저해 물질의 사용; HAM의 치료용 의약의 제조에서의 RGMa 저해 물질의 사용; HAM의 치료용 의약의 제조에 사용하기 위한 RGMa 저해 물질; 유효량의 RGMa 저해 물질을, 그 필요가 있는 대상에 투여하는 것을 포함하는, HAM의 치료 또는 예방 방법을 제공한다.
본 발명의 HAM의 치료 또는 예방제는, RGMa 저해 물질을 포함하고, 나아가서는 약학적으로 허용되는 담체 및/또는 첨가제를 적절하게 배합하여 제제화해도 된다.
제제화의 형태로서는, 예를 들면, 정제, 피복 정제, 환제, 산제(散劑), 과립제, 캡슐제, 액제, 현탁제, 유제 등의 경구제; 주사제, 수액, 좌제, 연고, 패치제 등의 비경구제가 있다.
담체 또는 첨가제의 배합 비율에 대해서는, 의약품 분야에 있어서 통상 채용되고 있는 범위에 기초하여 적절하게 설정하면 된다.
상기 담체로서는, 특별히 제한되지 않지만, 예를 들면, 물, 생리식염수,그 외의 수성 용매, 및 수성 또는 유성 기제 등이 있다. 상기 첨가제로서는, 특별히 제한되지 않지만, 부형제(賦形劑), 결합제, pH조정제, 붕괴제, 흡수 촉진제, 활택제, 착색제, 교미제(矯味劑), 및 향료 등이 있다.
본 발명에서의 RGMa 저해 물질이 RGMa를 인식하는 항체일 때, 상기 항체를 약학적으로 허용되는 담체와 함께 제제화된 주사제 또는 수액으로 만들어, 비경구 투여 경로, 예를 들면, 정맥내, 근육내, 피부내, 복강내, 피하 또는 국소에 투여하는 것이 바람직하다.
RGMa항체를 포함하는 주사제 또는 수액은, 용액, 현탁액 또는 유탁액으로서 사용할 수 있다. 그 용제로서는, 예를 들면, 주사용 증류수, 생리식염수, 포도당 용액, 및 등장액(예를 들면, 염화나트륨, 염화칼륨, 글리세린, 만니톨, 소르비톨, 붕산, 붕사, 프로필렌글리콜 등의 용액) 등을 사용할 수 있다. 이들 용제는 1종 단독으로 사용해도 되고, 2종 이상을 조합하여 사용할 수도 있다.
또한, 상기 주사제 또는 수액은, 안정제, 용해 보조제, 현탁화제, 유화제, 무통화제, 완충제, 보존제, 방부제, pH조정제 등의 첨가제를 포함해도 된다.
안정제로서는, 예를 들면, 알부민, 글로불린, 젤라틴, 만니톨, 글루코오스, 덱스트란, 에틸렌글리콜, 프로필렌글리콜, 아스코르브산, 아황산수소 나트륨, 티오황산나트륨, EDTA 나트륨, 시트르산 나트륨, 디부틸하이드록시톨루엔 등을 사용할 수 있다.
용해 보조제로서는, 예를 들면, 알코올(예를 들면, 에탄올 등), 폴리알코올(예를 들면, 프로필렌글리콜, 폴리에틸렌글리콜 등), 비이온 계면활성제(예를 들면, 폴리소르베이트 80(등록상표), HCO-50 등) 등을 사용할 수 있다.
현탁화제로서는, 예를 들면, 모노스테아르산 글리세린, 모노스테아르산 알루미늄, 메틸셀룰로오스, 카르복시메틸셀룰로오스, 하이드록시메틸셀룰로오스, 라우릴 황산 나트륨 등을 사용할 수 있다.
유화제로서는, 예를 들면, 아라비아검, 알긴산 나트륨, 트라가칸트 등을 사용할 수 있다.
무통화제로서는, 예를 들면, 벤질알코올, 클로로부탄올, 소르비톨 등을 사용할 수 있다.
완충제로서는, 예를 들면, 인산 완충액, 아세트산 완충액, 붕산 완충액, 탄산 완충액, 시트르산 완충액, 트리스 완충액 등을 사용할 수 있다.
보존제로서는, 예를 들면, 파라옥시벤조산 메틸, 파라옥시벤조산 에틸, 파라옥시벤조산 프로필, 파라옥시벤조산 부틸, 클로로부탄올, 벤질알코올, 염화 벤잘코늄, 데하이드로아세트산 나트륨, 에데트산 나트륨, 붕산, 붕사 등을 사용할 수 있다.
방부제로서는, 예를 들면, 염화 벤잘코늄, 파라옥시벤조산, 클로로부탄올 등을 사용할 수 있다.
pH조정제로서는, 예를 들면, 염산, 수산화 나트륨, 인산, 아세트산 등을 사용할 수 있다.
본 발명에서의 RGMa 저해 물질이, RGMa를 발현하는 유전자의 siRNA(short interfering RNA), shRNA(short hairpin RNA), 및 안티센스올리고뉴클레오티드 등의 RGMa의 발현을 억제하는 물질일 때, 비바이러스 벡터 또는 바이러스 벡터의 형태로 투여할 수 있다.
RGMa 저해 물질이 비바이러스 벡터 형태일 때, 투여의 방법으로서는, 리포솜을 사용하여 핵산 분자를 도입하는 방법(리포솜법, HVJ-리포솜법, 카티오닉리포솜법, 리포펙숀법, 리포펙트아민법 등), 마이크로인젝션법, 유전자총(Gene Gun)으로 캐리어(금속 입자)와 함께 핵산 분자를 세포에 이입(移入)하는 방법 등을 예로 들 수 있다.
siRNA 또는 shRNA를, 바이러스 벡터를 사용하여 생체에 투여하는 경우, 재조함 아데노바이러스, 레트로바이러스 등의 바이러스 벡터를 이용할 수 있다. 무독화한 레트로바이러스, 아데노바이러스, 아데노 수반 바이러스, 헤르페스바이러스, 백시니어 바이러스, 폭스바이러스, 폴리오바이러스, 신드비스바이러스, 센다이바이러스, SV40 등의 DNA바이러스 또는 RNA바이러스에, siRNA 또는 shRNA를 발현하는 DNA를 도입하고, 세포 또는 조직에 이 재조합 바이러스를 감염시킴으로써, 세포 또는 조직 내에 유전자를 도입할 수 있다.
이와 같이 하여 얻어지는 제제는, 예를 들면, 인간, 및 래트, 마우스, 토끼, 양, 돼지, 소, 고양이, 개, 및 원숭이 등의 다른 포유동물에 대하여, 그 유효량을 투여함으로써, HAM을 예방 또는 치료할 수 있다. 투여량은, 목적, 질환의 중증도, 환자의 연령, 체중, 성별, 기왕력, 유효성분의 종류 등을 고려하여, 적절하게 설정된다.
[실시예]
[실시예 1: HAM 및 성인 T세포 백혈병 림프종(ALT; adult T-cell leukemia/lymphoma)의 발증 기구 및 병태의 규명을 위한 망라적 비교 해석]
도 1에 나타낸 바와 같이, HTLV-1 감염 미발증자의 검체에 대해서는, CD4양성 세포 중, HTLV-1 감염 세포의 지표인 CADM1과, CD7에 의해 분류된 군 P, D, N에 대하여, mRNA의 발현 분석을 행하였다.
또한 HAM은, HAM환자 말초혈단핵세포로부터, 자기(磁氣) 비즈로 농축한 CD4 양성 세포와 CD4 음성 세포도 4예씩 분석했다. HTLV-1 비감염의 건강한 사람 (Normal)에 대해서도 동일하게 분리했다. 말초혈단핵세포는, PBMC로도 기재한다. ATL(smodering, chronic, acute의 각 병형)는, 주로 CD4 양성 세포로 이루어지는PBM를 사용했다.
아질렌트테크놀로지가부시키가이샤에서 제조한 1색마이크로어레이 유전자 발현분석법을 행하였다.
신경과 관련된 분자에 대하여 망라하여 비교 분석한 바, HAM환자의 CD4 양성 세포에 있어서 RGMa가 현저하게 발현되어 있는 것을 알 수 있다.
정상 T세포 4예(Normal.CD4), HAM 환자 유래 CD4양성 T세포 4예(HAM.CD4), 건강한 사람 유래 CD4양성/CADM1음성/CD7양성 T세포 3예(Normal.P), HTLV-1 감염자 CD4양성/CADM1음성/CD7양성 T세포 5예(P군), HTLV-1 감염자 CD4양성/CADM1양성/CD7양성 T세포 5예(D군), HTLV-1 감염자 CD4양성/CADM1양성/CD7음성 T세포 5예(N군), 급성형 ATL환자 CD4양성/CADM1양성/CD7음성 T세포 3예(Acute.N), 건강한 사람 CD4양성 T세포 21예(Normal.CD4.1), 무증후성 ATL환자 유래 PBMC 3예(Smoldering), 만성형 ATL환자 유래 PBMC 20예(Chronic), 급성형 ATL환자 유래 PBMC 26예(Acute)에 대하여, 아질렌트테크놀로지가부시키가이샤의 Human Gene Expression 4x44K Microarray를 사용하여 전체유전자 발현 데이터를 취득하고, 중앙값으로 표준화한 후, RGMa 유전자 레벨을 그래프화하였다.
그래프에서는 어레이의 형광강도의 Log2값을 플롯했다. HAM 환자 유래 CD4양성 T세포군은,그 외의 모든 군과 유의차를 가지고 상승하고 있었다(P<0.05). 그래프를 도 2에 나타낸다.
[실시예 2: CD4양성 HAM환자에서의 유전자 발현 어세이]
HAM환자 5명의 말초혈로부터 분리한 PBMC로부터 human CD4+ isolation kit(Miltenyi Biotec)를 사용하여 CD4양성 T세포를 분리하고, HTLV-1 감염 세포가 다수 포함되는 세포집단으로 했다. 마찬가지로 건강한 사람 4명의 PBMC로부터 CD4양성 T세포를 분리하고, 컨트롤 군으로 했다.
분리한 CD4양성 T세포로부터 전체 RNA를 회수하고, ReverTra Ace(도요보)를 사용하여 cDNA를 제작했다. 제작된 cDNA를 사용하여 HAM환자 CD4양성 T세포와 건강한 사람 CD4양성 T세포 사이에서의 RGMa의 발현량의 차이를 Real time PCR에 의해 분석했다. 인터널(internal) 컨트롤에는 18srRNA를 사용했다.
분석 결과의 그래프를 도 3에 나타낸다. 그래프에서의 HD-CD4+는, 컨트롤 군을 나타내고, HAM-CD4+는, HAM환자의 군을 나타낸다.
[실시예 3: PBMC에서의 RGMa 발현 세포의 분석]
HAM환자 PBMC에 Clear Back(MBL)을 사용하여 Fc block 처리한 후, 항CD3-PECy7(TONBO), CD4-FITC(eBioscience), CD14-PE(eBioscience) 항체를 가하여 4℃로 30분간 염색을 했다.
항체 염색된 PBMC를 세정한 후에 AriaIIIu(BD)를 사용하여 FACS소팅을 행하고, CD3양성 CD4음성 세포(CD3+CD4-), CD3양 성CD4양성 세포(CD3+CD4+), CD3음성 CD14음성 세포(CD3-CD14-) 및 CD3음성 CD14양성 세포(CD3-CD14+)의 분리 회수를 행하였다.
회수된 각 세포로부터 전체 RNA를 회수하고, ReverTra Ace(도요보)를 사용하여 cDNA를 제작했다. 제작된 cDNA를 사용하여 세포종간에서의 RGMa의 발현량의 차이를 Real time PCR에 의해 분석했다. 인터널 컨트롤에는 18s rRNA를 사용했다. 분석 결과의 그래프를 도 4에 나타낸다.
HAM-PBMC에서는, 감염 세포가 많이 존재하는 CD3양성 CD4양성 세포(CD3+CD4+)에 있어서 RGMa의 발현이 가장 높은 것을 알았다.
[실시예 4: HAM환자 PBMC의 배양에 따른 RGMa 발현 변화]
HAM환자 2명의 PBMC를 배지(10% FBS(GIBCO)를 포함하는 RPMI1640 배지(wako))에 현탁하고 1e5 cells씩 96 well round bottom plate에 10well분 파종 한 후에 1, 3, 5, 7 일간 배양했다.
배양을 행하지 않은 Day0의 PBMC와 함께 각 기간 배양한 PBMC로부터 전체 RNA를 추출하고 ReverTra Ace(도요보)를 사용하여 cDNA를 제작했다. HAM환자 PBMC는 배양하면 HTLV-1 바이러스가 과잉으로 발현하는 것이 알려져 있고, 제작된 cDNA를 사용하여 HAM환자 PBMC의 배양에 따른, 즉 바이러스의 발현에 따른 RGMa의 발현변화에 대하여, Real time PCR에 의해 분석했다. 인터널 컨트롤에는 18s rRNA를 사용했다. 분석 결과를 나타낸 그래프를 도 5에 나타낸다.
[실시예 5: 전체 프로모터 상의 H3K27me3 레벨의 분석]
정상 T세포 3예(Normal T cell), HAM 환자 유래 CD4양성 T세포 4예(HAM), 급성형 ATL환자 유래 PBMC 3예(ATL)에 대하여, Agilent Technologies사의 SurePrint G3 Human Promoter 2x400K Microarray를 사용하여 전체 프로모터 상의 H3K27me3 레벨을 취득하고, 표준화한 후, RGMa유전자의 전사개시점으로부터 -2,916bp 상류 부근의 H3K27me3 레벨을 그래프화하였다.
그래프를 도 6에 나타낸다. 그래프에서는 어레이의 형광강도의 Log2값을 플롯했다. 그리고, 도면 중의 HAM50 및 HAM123은, 각각 CD4양성 T세포를 얻은 HAM환자를 지칭한다. HAM환자 유래의 CD4양성 T세포에서는, RGMa의 유전자 발현 억제가 해제되어 있는 것이 시사되었다.
[실시예 6: RGMa유전자 mRNA 레벨의 정량(定量)]
인간 CD4양성 T세포 백혈병세포주 Jurkat에 대하여, HTLV-1 Tax를 코딩하는 cDNA를 삽입하는 렌티바이러스 벡터를 도입하고, 도입 후 3일간의 계시적(繼時的)인 RGMa유전자 mRNA의 레벨을 정량적 RT-PCR로 측정했다. RPL19유전자 mRNA도 측정하여, 내부 표준으로서 사용했다.
정량의 결과를 나타낸 그래프를 도 7에 나타낸다. HTLV-1 바이러스에 의해 RGMa 발현이 야기되는 것이 시사되었다.
[실시예 7: HTLV-1-tax 발현 유도 세포주 JPX-9 세포에서의 Tax 의존적인 RGMa의 발현 유도]
JPX9 세포를 배지(10% FBS를 포함하는 RPMI1640 배지)에서 24시간 배양하고, HTLV-1-tax 발현을 유도하는 염화카드뮴(CdCl2; 나카라이테스크)을 최종농도 20μM로 되도록 첨가한 후, 1, 2, 및 3 일간 배양했다. 염화 카드뮴 처리 및 미처리의 JPX9 세포에 대하여, Tax 및 RGMa의 단백질 발현에 대하여 FACS 분석했다.
염화 카드뮴 처리 및 미처리의 JPX9 세포를 각각 세정하고, Foxp3/Transcription Factor Staining Buffer Kit(eBioscience)를 사용하여 세포 투과 처리를 행하였다. 그 후, 항Tax-FITC항체(Lt-4: 류큐(琉球)대학다나카선생님으로부터 분여)를 첨가하고 4℃에서 1시간 처리함으로써 세포 내에 발현되는 Tax 단백질을 염색했다. 염색된 Tax 단백질은 CantoII를 사용한 FACS 분석에 의해 검출했다.
염화 카드뮴 처리 및 미처리의 JPX9세 포를 각각 세정하고, 항RGMa항체(가부시키가이샤면역생물연구소(IBL) 제조)를 첨가하고 4℃에서 30분간 처리했다. 그 후, 세포를 세정하고, 항mouse IgG-PE항체(BioLegend)를 첨가하고 4℃에서 30분간 반응시켜, JPX9에 발현되는 RGMa 단백질을 염색했다. 염색된 RGMa 단백질은 CantoII를 사용한 FACS 분석에 의해 검출했다. 도 8에 Tax 및 RGMa의 단백질 발현의 분석 결과를 나타낸다.
[실시예 8: HAM환자 PBMC에 대한 RGMa항체의 작용 검토]
(자발적 증식 활성에 대한 RGMa항체의 작용)
4예의 HAM환자 PBMC를 배지(10% FBS를 포함하는 RPMI1640 배지)에 현탁하고, 1e5 cells씩 96 well round bottom plate에 파종하고, 최종농도 10μg/ml로 되도록 RGMa항체(R&D Systems 제조)를 가하여 합계 0.1ml의 배양액에 있어서 37℃, 5% CO2 조건 하에서 7일간 배양했다.
아무 것도 첨가하지 않은 군(Medium), 동일 농도의 Normal Goat IgG(Santa Cruz Biotechnology) 첨가 군(Normal IgG), 및 1μg/ml 프레드니솔론(PSL)(후나코시) 첨가 군을 컨트롤로 했다.
배양 개시로부터 6일 후, 각 웰에 1μ Ci 3H-Thymidine를 첨가하고, 37℃, 5% CO2 조건 하에서 16시간 배양을 행하였다. 그 후, 배양 세포를 셀 하베스터(Tomtec MH3 PerkinElmer)를 사용하여 유리 필터(Printed Filtermat A PerkinElmer)에 흡착시키고, 건조시킨 후 고체 신틸레이터 Meltilex-A(PerkinElmer)를 배어들게 하고, MicroBeta(WALLAC MicroBeta TriLux 1450-021)를 사용하여 세포에 받아들여진 3H-Thymidine량의 측정을 행하였다. 각 HAM환자 PBMC의 Medium군에서의 3H-Thymidine의 카운트의 평균을 100%로 하고, 각 군의 상대값을 산출하고, HAM환자 4예에 3H-Thymidine 흡수율의 평균값을 구했다. 결과를 도 9에 나타낸다.
(HTLV-1 프로바이러스량 변화에 대한 RGMa항체의 작용)
4예의 HAM환자 PBMC를 배지(10% FBS를 포함하는 RPMI1640 배지)에 현탁하고 1e5 cells씩 96 well round bottom plate에 파종하고, 최종농도 10μg/ml로 되도록RGMa항체(R&D Systems 제조)를 가하여 합계 0.1ml의 배양액에 있어서 37℃, 5% CO2 조건 하에서 7일간 배양했다.
아무 것도 첨가하지 않은 군(Medium), 동일 농도의 Normal Goat IgG (Santa Cruz Biotechnology) 첨가 군(Normal IgG), 1μg/ml 프레드니솔론(PSL)(후나코시) 첨가 군을 컨트롤로 했다. 배양 개시로부터 7일 후, 원심분리하여, 상청액을 제외한 세포괴(細胞塊)로부터 게놈 DNA 추출을 행하였다. 추출된 게놈 DNA를 사용하여, 실시간 PCR에 의해 HTLV-1 프로바이러스량(감염 세포율)의 측정을 행하였다.
각 HAM환자 PBMC의 Medium군 있어서의 HTLV-1 프로바이러스량을 100%로 하여, 각 군에서의 HTLV-1 프로바이러스량의 상대값을 산출하고, HAM환자 4예의 HTLV-1 프로바이러스량의 평균값을 구했다. 결과를 도 10에 나타낸다.
(CXCL10 생산에 대한 RGMa항체의 작용)
HAM환자 PBMC으로부터의 CXCL10 생산에 대한 RGMa항체의 작용에 대하여 분석하기 위하여, 4예의 HAM환자 PBMC를 배지(10% FBS를 포함하는 RPMI1640 배지)에 현탁하고 1e5 cells씩 96 well round bottom plate에 파종하고, 최종농도 10μg/ml로 되도록 RGMa항체(R&D Systems 제조)를 가하여 합계 0.1ml의 배양액에 있어서 37℃, 5% CO2 조건 하에서 7일간 배양했다.
아무 것도 첨가하지 않은 군(Medium), 동일 농도의 Normal Goat IgG(Santa Cruz Biotechnology) 첨가 군(Normal IgG), 1μg/ml 프레드니솔론(PSL)(후나코시) 첨가 군을 컨트롤로 했다.
배양 개시로부터 7일 후, 배양액을 원심분리하여 배양 상청액만을 회수했다. 배양 상청액 중의 CXCL10 농도는 Cytokine Beads Array kit(BD Biosciences)을 사용하여, 플로우사이토미터 FACSCantoII(BD Biosciences)에 의해 측정했다.
Medium군의 배양액 중에서의 CXCL10 농도를 100%로 하고, 각 군에서의 각 군의 배양액 중 CXCL10 농도의 상대값을 산출하고, HAM환자 4예의 CXCL10 농도의 평균값을 구했다. 결과를 도 11에 나타낸다.
(HAM환자 PBMC의 사이토카인 생산에 대한 RGMa항체의 작용)
HAM환자 PBMC에서의 각종 사이토카인의 생산에 대한 RGMa항체의 작용에 대하여 분석하기 위하여, 4예의 HAM환자 PBMC를 배지(10% FBS를 포함하는 RPMI1640 배지)에 현탁하고 1e5 cells씩 98 well round bottom plate에 파종하고, 최종농도 10μg/ml로 되도록 RGMa항체(R&D Systems 제조)를 가하고 합계 0.1ml의 배양액에 있어서 37℃, 5% CO2 조건 하에서 7일간 배양했다.
아무 것도 첨가하지 않은 군(Medium), 동일 농도의 Normal Goat IgG(Santa Cruz Biotechnology) 첨가 군(Normal IgG), 1μg/ml 프레드니솔론(PSL)(후나코시) 첨가 군을 컨트롤로 했다.
배양 개시로부터 7일 후, 배양액을 원심분리하여 배양 상청액만을 회수했다. 배양 상청액 중의 IFNγ, TNF, IL-2, IL-10 농도는 Cytokine Beads Array kit(BD Biosciences)을 사용하여, 플로우사이토미터 FACSCantoII(BD Biosciences)에 의해 측정했다.
Medium군의 배양액 중에서의 각 사이토카인 농도를 100%로 하고, 각 배양 조건에서의 사이토카인 농도의 상대값을 산출하고, HAM환자 4예의 평균값을 구했다.
[실시예 9: HAM-PBMC에 의한 신경 세포주의 아포토시스 유도]
신경 세포주 NB-1 또는 SK-N-AS를 6well plate에 파종하고 24시간 배양 후, 건강한 사람(HD) 또는 HAM환자 PBMC를 첨가하여 공배양을 행하였다. 공배양 개시로부터 48시간 후에 배지와 함께 첨가한 PBMC를 제거하고, PBS로 세정 후에 신경 세포주를 회수했다.
회수된 각 신경 세포주에 대하여, 아포토시스에 의해 DNA 단편화를 일으키고 있는 세포를 특이적으로 검출하는 TUNEL법(MEBSTAIN Apoptosis TUNEL Kit Direct(MBL))에 의해 분석을 행하였다. 도 13에 분석 결과를 나타낸다. 히스토그램에서의 X축이, DNA 단편화 양성의 강도를 나타낸다. HAM유래 세포는 HD유래 세포와 비교하여, 신경 세포주에 아포토시스를 더욱 강하게 유도했다.
구체적으로는, 하기 <실험 수순>에 따라, 세포사의 분석을 행하였다.
<실험 수순>
신경 세포주 NB-1, SK-N-AS를 각각 6well plate에 파종하고, 24시간 배양했다.
다음으로, HD 또는 HAM-PBMC를 첨가(신경 세포주의 파종수의 2배량)하고, 48시간 배양했다.
그 후, 4% 파라포름알데히드를 사용하여 세포 고정하고, 70% 에탄올에 의해 세포 투과 처리를 행하였다.
DNA nick end labeling을 행하기 위해, MEBSTAIN Apoptosis TUNEL Kit Direct(MBL)의 TdT solution 20uL(TdT buffer II : TdT : FITC-dUTP = 18:1:1)에 세포를 현탁시키고, 37℃에서 60분간 반응시킨 후, FACS 분석을 행하였다.
[실시예 10: HTLV-1 Tax 발현 T세포주에 의해 유도되는 신경 세포주의 아포토시스에 대한 RGMa항체의 억제 작용]
신경 세포주 NB-1을 6well plate에 파종하고 24시간 배양 후, 미자극 JPX9(JPX9(-)) 또는 20μM 염화 카드뮴을 24시간 첨가하여 Tax 발현을 유도한 JPX9(JPX9(+CdCl2))을 가하여 공배양을 행하였다. 또한, NB-1 세포와 (JPX9(+CdCl2)의 공배양에 최종농도 10μg/ml로 되도록 Normal Mouse IgG2b(MBL) 또는 RGMa항체(IBL)를 가하였다. 공배양 개시로부터 48시간 후에 배지와 함께 첨가한 JPX9을 제거하고, PBS로 세정 후에 신경 세포주를 회수했다. 회수된 각 신경 세포주에 대하여, 아포토시스에 의해 DNA 단편화를 일으키고 있는 세포를 특이적으로 검출하는TUNEL법(MEBSTAIN Apoptosis TUNEL Kit Direct(MBL))에 의해 분석을 행하였다. 도 14에 분석 결과를 나타낸다. 히스토그램에서의 X축이, DNA 단편화 양성의 강도를 나타낸다.
구체적으로는, 하기 <실험 수순>에 따라, 세포사의 분석을 행하였다.
<실험 수순>
신경 세포주(NB-1)를 6well plate에 파종하고, 24시간 배양했다.
다음으로, JPX9(-) 또는 JPX9(+CdCl2)을 NB-1 세포에 첨가하여 공배양했다. 그리고, JPX9(-) 또는 JPX9(+CdCl2)의 세포수는 NB-1 세포의 파종수의 2배량으로 했다. JPX9(+CdCl2)에 대해서는, 염화 카드뮴을 제거하기 위해 배지 10ml에 3회 세정하여 NB-1 세포에 가하였다.
계속해서, NB-1 세포와 (JPX9(+CdCl2)의 공배양에 Normal Mouse IgG2b(MBL) 또는 항RGMa항체(IBL)를 최종농도 10μg/ml로 되도록 가하고, 48시간 배양했다.
그 후, 4% 파라포름알데히드를 사용하여 세포 고정하고, 70% 에탄올에 의해 세포 투과 처리를 행하고, 항CD45-V450항체를 가하고 염색했다.
DNA nick end labeling을 행하기 위해, MEBSTAIN Apoptosis TUNEL Kit Direct(MBL)의 TdT solution 20uL(TdT buffer II : TdT : FITC-dUTP = 18:1:1)에 세포를 현탁하고, 37℃에서 60분간 반응시킨 후, FACS 분석을 행하였다.
본 명세서에 있어서 인용하여 기재하는 간행물, 특허문헌 및 비특허문헌에 기재된 모든 내용은, 그대로 참고로서 본명세서에 있어서 원용된다.
SEQUENCE LISTING
<110> The University of Tokyo
St. Marianna University School of Medicine
<120> Therapeutic agent or prophylactic agent for HAM
<130> T0529AKP0016
<160> 109
<170> PatentIn version 3.5
<210> 1
<211> 1299
<212> DNA
<213> Homo sapiens
<400> 1
atggggagag gggcaggacg ttcagccctg ggattctggc cgaccctcgc cttccttctc 60
tgcagcttcc ccgcagccac ctccccgtgc aagatcctca agtgcaactc tgagttctgg 120
agcgccacgt cgggcagcca cgccccagcc tcagacgaca cccccgagtt ctgtgcagcc 180
ttgcgcagct acgccctgtg cacgcggcgg acggcccgca cctgccgggg tgacctggcc 240
taccactcgg ccgtccatgg catagaggac ctcatgagcc agcacaactg ctccaaggat 300
ggccccacct cgcagccacg cctgcgcacg ctcccaccgg ccggagacag ccaggagcgc 360
tcggacagcc ccgagatctg ccattacgag aagagctttc acaagcactc ggccaccccc 420
aactacacgc actgtggcct cttcggggac ccacacctca ggactttcac cgaccgcttc 480
cagacctgca aggtgcaggg cgcctggccg ctcatcgaca ataattacct gaacgtgcag 540
gtcaccaaca cgcctgtgct gcccggctca gcggccactg ccaccagcaa gctcaccatc 600
atcttcaaga acttccagga gtgtgtggac cagaaggtgt accaggctga gatggacgag 660
ctcccggccg ccttcgtgga tggctctaag aacggtgggg acaagcacgg ggccaacagc 720
ctgaagatca ctgagaaggt gtcaggccag cacgtggaga tccaggccaa gtacatcggc 780
accaccatcg tggtgcgcca ggtgggccgc tacctgacct ttgccgtccg catgccagag 840
gaagtggtca atgctgtgga ggactgggac agccagggtc tctacctctg cctgcggggc 900
tgccccctca accagcagat cgacttccag gccttccaca ccaatgctga gggcaccggt 960
gcccgcaggc tggcagccgc cagccctgca cccacagccc ccgagacctt cccatacgag 1020
acagccgtgg ccaagtgcaa ggagaagctg ccggtggagg acctgtacta ccaggcctgc 1080
gtcttcgacc tcctcaccac gggcgacgtg aacttcacac tggccgccta ctacgcgttg 1140
gaggatgtca agatgctcca ctccaacaaa gacaaactgc acctgtatga gaggactcgg 1200
gacctgccag gcagggcggc tgcggggctg cccctggccc cccggcccct cctgggcgcc 1260
ctcgtcccgc tcctggccct gctccctgtg ttctgctag 1299
<210> 2
<211> 1296
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<400> 2
atggggagag gggcaggacg ttcagccctg ggattgtggc cgaccctcgc cttccttctc 60
tgcagctttc ccgcagctat ctctccctgc aagatcctca agtgcaactc tgagttctgg 120
agcgccacgt cgtcaggcag ccacgcccct gcctctgacg acgtgcccga gttctgtgct 180
gccctgcgca cctacgccct gtgcacgcga cggacagccc gcacctgccg gggcgacctg 240
gcttaccact cggctgtcca tggcatagag gacctcatga gccagcacaa ctgctccaag 300
gatggcccca cctcacagcc tcgagtgcgc acgctcccgc cagctgggga cagccaggag 360
cgctcagata gccccgagat ctgccactat gagaagagtt tccacaagca ctcagctgcc 420
cccaactaca ctcactgcgg cctctttggg gacccacacc tcaggacttt cacagaccac 480
ttccagacat gtaaggtgca aggcgcttgg cctctcatcg acaataatta cctgaacgtg 540
caggtcacca atacacctgt gctgcccggc tctgccgcca ctgccaccag caagctcacc 600
atcatcttca agaacttcca agagtgtgtg gaccagaaag tataccaagc cgagatggac 660
gagcttccgt ccgcctttgc cgatggctcc aaaaacggtg gagataaaca cggagccaac 720
agcctgaaga tcacagagaa ggtgtcaggc cagcacgtgg agatccaggc caagtacatc 780
ggcaccacca tcgtggtgag acaggtgggc cgctacctga ccttcgccgt ccggatgccc 840
gaggaggtag tcaacgccgt ggaggaccgt gacagccaag gcctctacct ctgcctgcgg 900
ggctgcccgc tcaaccagca gatcgacttc caggctttcc gtgccaacgc cgagagccct 960
cgcaggccag cagctgccag cccctctcct gtggtccccg agacatttcc gtacgagaca 1020
gctgtggcca agtgcaaaga gaagctgcct gtagaagact tgtactacca ggcctgtgtc 1080
ttcgacctcc tcacgactgg cgacgtgaac ttcacgctgg ccgcctacta tgctttggag 1140
gatggcaaga tgctccactc caacaaggac aagctacacc tgtttgaaag gactcgggag 1200
ctgcctggcg ctgtggccgc tgcagcattt cccttggccc ccgagatgct cccgggcacc 1260
gtcacacttc tggtcctgct gcctctgttc tggtag 1296
<210> 3
<211> 432
<212> PRT
<213> Homo sapiens
<400> 3
Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Phe Trp Pro Thr Leu
1 5 10 15
Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Thr Ser Pro Cys Lys Ile
20 25 30
Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Gly Ser His Ala
35 40 45
Pro Ala Ser Asp Asp Thr Pro Glu Phe Cys Ala Ala Leu Arg Ser Tyr
50 55 60
Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp Leu Ala
65 70 75 80
Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser Gln His Asn
85 90 95
Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Leu Arg Thr Leu Pro
100 105 110
Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu Ile Cys His
115 120 125
Tyr Glu Lys Ser Phe His Lys His Ser Ala Thr Pro Asn Tyr Thr His
130 135 140
Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr Asp Arg Phe
145 150 155 160
Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp Asn Asn Tyr
165 170 175
Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly Ser Ala Ala
180 185 190
Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe Gln Glu Cys
195 200 205
Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu Pro Ala Ala
210 215 220
Phe Val Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly Ala Asn Ser
225 230 235 240
Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu Ile Gln Ala
245 250 255
Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly Arg Tyr Leu
260 265 270
Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala Val Glu Asp
275 280 285
Trp Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys Pro Leu Asn
290 295 300
Gln Gln Ile Asp Phe Gln Ala Phe His Thr Asn Ala Glu Gly Thr Gly
305 310 315 320
Ala Arg Arg Leu Ala Ala Ala Ser Pro Ala Pro Thr Ala Pro Glu Thr
325 330 335
Phe Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu Pro Val
340 345 350
Glu Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr Thr Gly
355 360 365
Asp Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp Val Lys
370 375 380
Met Leu His Ser Asn Lys Asp Lys Leu His Leu Tyr Glu Arg Thr Arg
385 390 395 400
Asp Leu Pro Gly Arg Ala Ala Ala Gly Leu Pro Leu Ala Pro Arg Pro
405 410 415
Leu Leu Gly Ala Leu Val Pro Leu Leu Ala Leu Leu Pro Val Phe Cys
420 425 430
<210> 4
<211> 431
<212> PRT
<213> Rattus norvegicus
<400> 4
Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Leu Trp Pro Thr Leu
1 5 10 15
Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Ile Ser Pro Cys Lys Ile
20 25 30
Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Ser Gly Ser His
35 40 45
Ala Pro Ala Ser Asp Asp Val Pro Glu Phe Cys Ala Ala Leu Arg Thr
50 55 60
Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp Leu
65 70 75 80
Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser Gln His
85 90 95
Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Val Arg Thr Leu
100 105 110
Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu Ile Cys
115 120 125
His Tyr Glu Lys Ser Phe His Lys His Ser Ala Ala Pro Asn Tyr Thr
130 135 140
His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr Asp His
145 150 155 160
Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp Asn Asn
165 170 175
Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly Ser Ala
180 185 190
Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe Gln Glu
195 200 205
Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu Pro Ser
210 215 220
Ala Phe Ala Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly Ala Asn
225 230 235 240
Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu Ile Gln
245 250 255
Ala Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly Arg Tyr
260 265 270
Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala Val Glu
275 280 285
Asp Arg Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys Pro Leu
290 295 300
Asn Gln Gln Ile Asp Phe Gln Ala Phe Arg Ala Asn Ala Glu Ser Pro
305 310 315 320
Arg Arg Pro Ala Ala Ala Ser Pro Ser Pro Val Val Pro Glu Thr Phe
325 330 335
Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu Pro Val Glu
340 345 350
Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr Thr Gly Asp
355 360 365
Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp Gly Lys Met
370 375 380
Leu His Ser Asn Lys Asp Lys Leu His Leu Phe Glu Arg Thr Arg Glu
385 390 395 400
Leu Pro Gly Ala Val Ala Ala Ala Ala Phe Pro Leu Ala Pro Glu Met
405 410 415
Leu Pro Gly Thr Val Thr Leu Leu Val Leu Leu Pro Leu Phe Trp
420 425 430
<210> 5
<211> 5
<212> PRT
<213> Artificial
<220>
<223> VH 5F9 CDR H1
<400> 5
Asn Tyr Gly Met Asn
1 5
<210> 6
<211> 17
<212> PRT
<213> Artificial
<220>
<223> VH 5F9 CDR-H2
<400> 6
Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 7
<211> 6
<212> PRT
<213> Artificial
<220>
<223> VH 5F9 CDR-H3
<400> 7
Gly Thr Thr Pro Asp Tyr
1 5
<210> 8
<211> 16
<212> PRT
<213> Artificial
<220>
<223> VL 5F9 CDR-L1
<400> 8
Arg Ser Ser Gln Ser Leu Glu Tyr Ser Asp Gly Tyr Thr Phe Leu Glu
1 5 10 15
<210> 9
<211> 7
<212> PRT
<213> Artificial
<220>
<223> VL 5F9 CDR-L2
<400> 9
Glu Val Ser Asn Arg Phe Ser
1 5
<210> 10
<211> 9
<212> PRT
<213> Artificial
<220>
<223> VL 5F9 CDR-L3
<400> 10
Phe Gln Ala Thr His Asp Pro Leu Thr
1 5
<210> 11
<211> 5
<212> PRT
<213> Artificial
<220>
<223> VH 8D1 CDR-H1
<400> 11
Ser Tyr Val Met His
1 5
<210> 12
<211> 17
<212> PRT
<213> Artificial
<220>
<223> VH 8D1 CDR-H2
<400> 12
Tyr Ile Ile Pro Tyr Asn Asp Asn Thr Lys Tyr Asn Glu Lys Phe Lys
1 5 10 15
Gly
<210> 13
<211> 14
<212> PRT
<213> Artificial
<220>
<223> VH 8D1 CDR-H3
<400> 13
Ala Arg Arg Asn Glu Tyr Tyr Gly Ser Ser Phe Phe Asp Tyr
1 5 10
<210> 14
<211> 11
<212> PRT
<213> Artificial
<220>
<223> VL 8D1 CDR-L1
<400> 14
Gln Ala Ser Gln Asp Ile Asp Asn Tyr Leu Ala
1 5 10
<210> 15
<211> 7
<212> PRT
<213> Artificial
<220>
<223> VL 8D1 CDR-L2
<400> 15
Gly Ala Thr Asn Leu Ala Asp
1 5
<210> 16
<211> 9
<212> PRT
<213> Artificial
<220>
<223> VL 8D1 CDR-L3
<400> 16
Leu Gln Gly Tyr Ile Pro Pro Arg Thr
1 5
<210> 17
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9
<400> 17
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Ser
1 5 10 15
Ser Leu Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Ile Arg Gln Ala Pro Lys Lys Gly Leu Glu Trp Ile
35 40 45
Gly Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Glu Met Asn Ser Leu Arg Ser Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Lys Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Val Met Val Thr
100 105 110
Val Ser Ser
115
<210> 18
<211> 113
<212> PRT
<213> Artificial
<220>
<223> VL 5F9
<400> 18
Asp Val Val Leu Thr Gln Thr Pro Val Ser Leu Ser Val Thr Leu Gly
1 5 10 15
Asp Gln Ala Ser Met Ser Cys Arg Ser Ser Gln Ser Leu Glu Tyr Ser
20 25 30
Asp Gly Tyr Thr Phe Leu Glu Trp Phe Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Glu Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Pro Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Ala
85 90 95
Thr His Asp Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 19
<211> 121
<212> PRT
<213> Artificial
<220>
<223> VH 8D1
<400> 19
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Thr
1 5 10 15
Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Val Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Tyr Ile Ile Pro Tyr Asn Asp Asn Thr Lys Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ser Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Asn Glu Tyr Tyr Gly Ser Ser Phe Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Thr Leu Thr Val Ser Ser
115 120
<210> 20
<211> 108
<212> PRT
<213> Artificial
<220>
<223> VL 8D1
<400> 20
Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Leu Glu
1 5 10 15
Glu Ile Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Asp Asn Tyr
20 25 30
Leu Ala Trp Tyr His Gln Lys Pro Gly Lys Ser Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Arg Leu Gln Ile
65 70 75 80
Glu Asp Leu Gly Ile Tyr Tyr Cys Leu Gln Gly Tyr Ile Pro Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg
100 105
<210> 21
<211> 5
<212> PRT
<213> Artificial
<220>
<223> VH 5F9 CDR H1
<400> 21
Asn Tyr Gly Met Asn
1 5
<210> 22
<211> 30
<212> PRT
<213> Artificial
<220>
<223> VH3-48/JH3 FR1
<400> 22
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 23
<211> 14
<212> PRT
<213> Artificial
<220>
<223> VH3-48/JH3 FR2
<400> 23
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser
1 5 10
<210> 24
<211> 32
<212> PRT
<213> Artificial
<220>
<223> VH3-48/JH3 FR3
<400> 24
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 25
<211> 11
<212> PRT
<213> Artificial
<220>
<223> VH3-49/JH3 FR4
<400> 25
Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser
1 5 10
<210> 26
<211> 11
<212> PRT
<213> Artificial
<220>
<223> VH3-48/JH4 FR4
<400> 26
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
1 5 10
<210> 27
<211> 11
<212> PRT
<213> Artificial
<220>
<223> VH3-48/JH6 FR4
<400> 27
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
1 5 10
<210> 28
<211> 30
<212> PRT
<213> Artificial
<220>
<223> VH3-33/JH6 FR!
<400> 28
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 29
<211> 14
<212> PRT
<213> Artificial
<220>
<223> VH3-33/JH6 FR2
<400> 29
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala
1 5 10
<210> 30
<211> 32
<212> PRT
<213> Artificial
<220>
<223> VH3-33/JH6 FR3
<400> 30
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 31
<211> 30
<212> PRT
<213> Artificial
<220>
<223> VH3-23/JH3 FR1
<400> 31
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 32
<211> 14
<212> PRT
<213> Artificial
<220>
<223> VH3-23/JH3 FR2
<400> 32
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser
1 5 10
<210> 33
<211> 32
<212> PRT
<213> Artificial
<220>
<223> VH3-23/JH3 FR3
<400> 33
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys
20 25 30
<210> 34
<211> 23
<212> PRT
<213> Artificial
<220>
<223> A18/JK2 FR1
<400> 34
Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys
20
<210> 35
<211> 15
<212> PRT
<213> Artificial
<220>
<223> A18/JK2 FR2
<400> 35
Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr
1 5 10 15
<210> 36
<211> 32
<212> PRT
<213> Artificial
<220>
<223> A18/JK2 FR3
<400> 36
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys
20 25 30
<210> 37
<211> 11
<212> PRT
<213> Artificial
<220>
<223> A18/JK2 FR4
<400> 37
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
1 5 10
<210> 38
<211> 23
<212> PRT
<213> Artificial
<220>
<223> A17/JK2 FR1
<400> 38
Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys
20
<210> 39
<211> 15
<212> PRT
<213> Artificial
<220>
<223> A17/JK2 FR2
<400> 39
Trp Phe Gln Gln Arg Pro Gly Gln Ser Pro Arg Arg Leu Ile Tyr
1 5 10 15
<210> 40
<211> 32
<212> PRT
<213> Artificial
<220>
<223> A17/JK2 FR3
<400> 40
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys
20 25 30
<210> 41
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.1-GL
<400> 41
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Met Val Thr
100 105 110
Val Ser Ser
115
<210> 42
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.2-GL
<400> 42
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 43
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.3-GL
<400> 43
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr
100 105 110
Val Ser Ser
115
<210> 44
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.4-GL
<400> 44
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Met Val Thr
100 105 110
Val Ser Ser
115
<210> 45
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.5-GL
<400> 45
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 46
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.6-GL
<400> 46
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr
100 105 110
Val Ser Ser
115
<210> 47
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.7-GL
<400> 47
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Met Val Thr
100 105 110
Val Ser Ser
115
<210> 48
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.8-GL
<400> 48
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 49
<211> 115
<212> PRT
<213> Artificial
<220>
<223> VH 5F9.9-GL
<400> 49
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Met Ile Tyr Tyr Asp Ser Ser Glu Lys His Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Thr Thr Pro Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr
100 105 110
Val Ser Ser
115
<210> 50
<211> 113
<212> PRT
<213> Artificial
<220>
<223> VL 5F9.1-GL
<400> 50
Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Glu Tyr Ser
20 25 30
Asp Gly Tyr Thr Phe Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Glu Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Ala
85 90 95
Thr His Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 51
<211> 113
<212> PRT
<213> Artificial
<220>
<223> VL 5F9.2-GL
<400> 51
Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Glu Tyr Ser
20 25 30
Asp Gly Tyr Thr Phe Leu Glu Trp Phe Gln Gln Arg Pro Gly Gln Ser
35 40 45
Pro Arg Arg Leu Ile Tyr Glu Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Ala
85 90 95
Thr His Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 52
<211> 113
<212> PRT
<213> Artificial
<220>
<223> VL 5F9.3-GL
<400> 52
Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Glu Tyr Ser
20 25 30
Asp Gly Tyr Thr Phe Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Glu Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Ala
85 90 95
Thr His Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg
<210> 53
<211> 14
<212> PRT
<213> Mus musculus
<400> 53
Glu Glu Val Val Asn Ala Val Glu Asp Trp Asp Ser Gln Gly
1 5 10
<210> 54
<211> 14
<212> PRT
<213> Mus musculus
<400> 54
Asn Gln Gln Ile Asp Phe Gln Ala Phe His Thr Asn Ala Glu
1 5 10
<210> 55
<211> 11
<212> PRT
<213> Mus musculus
<400> 55
Pro Thr Ala Pro Glu Thr Phe Pro Tyr Glu Thr
1 5 10
<210> 56
<211> 11
<212> PRT
<213> Mus musculus
<400> 56
Lys Leu Pro Val Glu Asp Leu Tyr Tyr Gln Ala
1 5 10
<210> 57
<211> 16
<212> PRT
<213> Homo sapiens
<400> 57
Leu Tyr Glu Arg Thr Arg Asp Leu Pro Gly Arg Ala Ala Ala Gly Leu
1 5 10 15
<210> 58
<211> 11
<212> PRT
<213> Mus musculus
<400> 58
Arg Ala Ser Gln Asp Ile Ser Ser Tyr Leu Asn
1 5 10
<210> 59
<211> 7
<212> PRT
<213> Mus musculus
<400> 59
Tyr Thr Ser Arg Leu His Ser
1 5
<210> 60
<211> 7
<212> PRT
<213> Mus musculus
<400> 60
Gln Gln Leu Asn Thr Leu Pro
1 5
<210> 61
<211> 5
<212> PRT
<213> Mus musculus
<400> 61
Asp Ala Trp Met Asp
1 5
<210> 62
<211> 19
<212> PRT
<213> Mus musculus
<400> 62
Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu Ser
1 5 10 15
Val Lys Gly
<210> 63
<211> 5
<212> PRT
<213> Mus musculus
<400> 63
Arg Asp Gly Ala Tyr
1 5
<210> 64
<211> 16
<212> PRT
<213> Mus musculus
<400> 64
Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His
1 5 10 15
<210> 65
<211> 7
<212> PRT
<213> Mus musculus
<400> 65
Lys Val Ser Asn Arg Phe Ser
1 5
<210> 66
<211> 7
<212> PRT
<213> Mus musculus
<400> 66
Ser Gln Ser Thr His Val Pro
1 5
<210> 67
<211> 6
<212> PRT
<213> Mus musculus
<400> 67
Thr Ser Tyr Tyr Trp Asn
1 5
<210> 68
<211> 16
<212> PRT
<213> Mus musculus
<400> 68
Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Asn Pro Ser Leu Lys Asn
1 5 10 15
<210> 69
<400> 69
000
<210> 70
<211> 450
<212> PRT
<213> Homo sapiens
<400> 70
Met Gln Pro Pro Arg Glu Arg Leu Val Val Thr Gly Arg Ala Gly Trp
1 5 10 15
Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Phe Trp Pro
20 25 30
Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Thr Ser Pro Cys
35 40 45
Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Gly Ser
50 55 60
His Ala Pro Ala Ser Asp Asp Thr Pro Glu Phe Cys Ala Ala Leu Arg
65 70 75 80
Ser Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly Asp
85 90 95
Leu Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser Gln
100 105 110
His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Leu Arg Thr
115 120 125
Leu Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu Ile
130 135 140
Cys His Tyr Glu Lys Ser Phe His Lys His Ser Ala Thr Pro Asn Tyr
145 150 155 160
Thr His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr Asp
165 170 175
Arg Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp Asn
180 185 190
Asn Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly Ser
195 200 205
Ala Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe Gln
210 215 220
Glu Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu Pro
225 230 235 240
Ala Ala Phe Val Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly Ala
245 250 255
Asn Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu Ile
260 265 270
Gln Ala Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly Arg
275 280 285
Tyr Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala Val
290 295 300
Glu Asp Trp Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys Pro
305 310 315 320
Leu Asn Gln Gln Ile Asp Phe Gln Ala Phe His Thr Asn Ala Glu Gly
325 330 335
Thr Gly Ala Arg Arg Leu Ala Ala Ala Ser Pro Ala Pro Thr Ala Pro
340 345 350
Glu Thr Phe Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu
355 360 365
Pro Val Glu Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr
370 375 380
Thr Gly Asp Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp
385 390 395 400
Val Lys Met Leu His Ser Asn Lys Asp Lys Leu His Leu Tyr Asp Arg
405 410 415
Thr Arg Asp Leu Pro Gly Arg Ala Ala Ala Gly Leu Pro Leu Ala Pro
420 425 430
Arg Pro Leu Leu Gly Ala Leu Val Pro Leu Leu Ala Leu Leu Pro Val
435 440 445
Phe Cys
450
<210> 71
<211> 454
<212> PRT
<213> Mus musculus
<400> 71
Met Gln Pro Pro Arg Glu Arg Leu Val Val Thr Gly Arg Ala Gly Trp
1 5 10 15
Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Leu Trp Pro
20 25 30
Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Ile Ser Pro Cys
35 40 45
Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Ser Gly
50 55 60
Ser His Ala Pro Ala Ser Asp Asp Val Pro Glu Phe Cys Ala Ala Leu
65 70 75 80
Arg Thr Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly
85 90 95
Asp Leu Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser
100 105 110
Gln His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Val Arg
115 120 125
Thr Leu Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu
130 135 140
Ile Cys His Tyr Glu Lys Ser Phe His Lys His Ser Ala Ala Pro Asn
145 150 155 160
Tyr Thr His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr
165 170 175
Asp His Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp
180 185 190
Asn Asn Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly
195 200 205
Ser Ala Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe
210 215 220
Gln Glu Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu
225 230 235 240
Pro Ser Ala Phe Ala Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly
245 250 255
Ala Asn Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu
260 265 270
Ile Gln Ala Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly
275 280 285
Arg Tyr Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala
290 295 300
Val Glu Asp Arg Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys
305 310 315 320
Pro Leu Asn Gln Gln Ile Asp Phe Gln Ala Phe Arg Ala Asn Ala Glu
325 330 335
Ser Pro Arg Arg Pro Ala Ala Ala Ser Pro Ser Pro Val Val Pro Glu
340 345 350
Thr Phe Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu Pro
355 360 365
Val Glu Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr Thr
370 375 380
Gly Asp Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp Gly
385 390 395 400
Lys Met Leu His Ser Asn Lys Asp Lys Leu His Leu Phe Glu Arg Thr
405 410 415
Arg Glu Leu Pro Gly Ala Val Ala Ala Ala Ala Ala Ala Ala Thr Thr
420 425 430
Phe Pro Leu Ala Pro Gln Ile Leu Leu Gly Thr Ile Pro Leu Leu Val
435 440 445
Leu Leu Pro Val Leu Trp
450
<210> 72
<211> 449
<212> PRT
<213> Rattus norvegicus
<400> 72
Met Gln Pro Pro Arg Glu Arg Leu Val Val Thr Gly Arg Ala Gly Trp
1 5 10 15
Met Gly Met Gly Arg Gly Ala Gly Arg Ser Ala Leu Gly Leu Trp Pro
20 25 30
Thr Leu Ala Phe Leu Leu Cys Ser Phe Pro Ala Ala Ile Ser Pro Cys
35 40 45
Lys Ile Leu Lys Cys Asn Ser Glu Phe Trp Ser Ala Thr Ser Ser Gly
50 55 60
Ser His Ala Pro Ala Ser Asp Asp Val Pro Glu Phe Cys Ala Ala Leu
65 70 75 80
Arg Thr Tyr Ala Leu Cys Thr Arg Arg Thr Ala Arg Thr Cys Arg Gly
85 90 95
Asp Leu Ala Tyr His Ser Ala Val His Gly Ile Glu Asp Leu Met Ser
100 105 110
Gln His Asn Cys Ser Lys Asp Gly Pro Thr Ser Gln Pro Arg Val Arg
115 120 125
Thr Leu Pro Pro Ala Gly Asp Ser Gln Glu Arg Ser Asp Ser Pro Glu
130 135 140
Ile Cys His Tyr Glu Lys Ser Phe His Lys His Ser Ala Ala Pro Asn
145 150 155 160
Tyr Thr His Cys Gly Leu Phe Gly Asp Pro His Leu Arg Thr Phe Thr
165 170 175
Asp His Phe Gln Thr Cys Lys Val Gln Gly Ala Trp Pro Leu Ile Asp
180 185 190
Asn Asn Tyr Leu Asn Val Gln Val Thr Asn Thr Pro Val Leu Pro Gly
195 200 205
Ser Ala Ala Thr Ala Thr Ser Lys Leu Thr Ile Ile Phe Lys Asn Phe
210 215 220
Gln Glu Cys Val Asp Gln Lys Val Tyr Gln Ala Glu Met Asp Glu Leu
225 230 235 240
Pro Ser Ala Phe Ala Asp Gly Ser Lys Asn Gly Gly Asp Lys His Gly
245 250 255
Ala Asn Ser Leu Lys Ile Thr Glu Lys Val Ser Gly Gln His Val Glu
260 265 270
Ile Gln Ala Lys Tyr Ile Gly Thr Thr Ile Val Val Arg Gln Val Gly
275 280 285
Arg Tyr Leu Thr Phe Ala Val Arg Met Pro Glu Glu Val Val Asn Ala
290 295 300
Val Glu Asp Arg Asp Ser Gln Gly Leu Tyr Leu Cys Leu Arg Gly Cys
305 310 315 320
Pro Leu Asn Gln Gln Ile Asp Phe Gln Ala Phe Arg Ala Asn Ala Glu
325 330 335
Ser Pro Arg Arg Pro Ala Ala Ala Ser Pro Ser Pro Val Val Pro Glu
340 345 350
Thr Phe Pro Tyr Glu Thr Ala Val Ala Lys Cys Lys Glu Lys Leu Pro
355 360 365
Val Glu Asp Leu Tyr Tyr Gln Ala Cys Val Phe Asp Leu Leu Thr Thr
370 375 380
Gly Asp Val Asn Phe Thr Leu Ala Ala Tyr Tyr Ala Leu Glu Asp Gly
385 390 395 400
Lys Met Leu His Ser Asn Lys Asp Lys Leu His Leu Phe Glu Arg Thr
405 410 415
Arg Glu Leu Pro Gly Ala Val Ala Ala Ala Ala Phe Pro Leu Ala Pro
420 425 430
Glu Met Leu Pro Gly Thr Val Thr Leu Leu Val Leu Leu Pro Leu Phe
435 440 445
Trp
<210> 73
<211> 234
<212> PRT
<213> Mus musculus
<400> 73
Met Met Ser Ser Ala Gln Phe Leu Gly Leu Leu Leu Leu Cys Phe Gln
1 5 10 15
Gly Thr Arg Cys Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser
20 25 30
Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp
35 40 45
Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val
50 55 60
Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser
85 90 95
Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Leu Asn
100 105 110
Thr Leu Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
115 120 125
Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln
130 135 140
Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr
145 150 155 160
Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln
165 170 175
Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg
195 200 205
His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro
210 215 220
Ile Val Lys Ser Phe Asn Arg Asn Glu Cys
225 230
<210> 74
<211> 471
<212> PRT
<213> Mus musculus
<400> 74
Met Tyr Leu Gly Leu Asn Tyr Val Phe Ile Val Phe Leu Leu Asn Gly
1 5 10 15
Val Gln Ser Glu Val Lys Leu Glu Glu Ser Gly Gly Gly Leu Val Gln
20 25 30
Pro Gly Gly Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
35 40 45
Ser Asp Ala Trp Met Asp Trp Val Arg Gln Ser Pro Glu Lys Gly Leu
50 55 60
Glu Trp Val Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr
65 70 75 80
Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser
85 90 95
Lys Arg Ser Val Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr
100 105 110
Gly Ile Tyr Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Gln Gly
115 120 125
Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser Val Tyr
130 135 140
Pro Leu Ala Pro Gly Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu
145 150 155 160
Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Ser Val Thr Val Thr Trp
165 170 175
Asn Ser Gly Ser Leu Ser Ser Ser Val His Thr Phe Pro Ala Leu Leu
180 185 190
Gln Ser Gly Leu Tyr Thr Met Ser Ser Ser Val Thr Val Pro Ser Ser
195 200 205
Thr Trp Pro Ser Gln Thr Val Thr Cys Ser Val Ala His Pro Ala Ser
210 215 220
Ser Thr Thr Val Asp Lys Lys Leu Glu Pro Ser Gly Pro Ile Ser Thr
225 230 235 240
Ile Asn Pro Cys Pro Pro Cys Lys Glu Cys His Lys Cys Pro Ala Pro
245 250 255
Asn Leu Glu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Ile Lys
260 265 270
Asp Val Leu Met Ile Ser Leu Thr Pro Lys Val Thr Cys Val Val Val
275 280 285
Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn
290 295 300
Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr
305 310 315 320
Asn Ser Thr Ile Arg Val Val Ser Thr Leu Pro Ile Gln His Gln Asp
325 330 335
Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu
340 345 350
Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ile Lys Gly Leu Val Arg
355 360 365
Ala Pro Gln Val Tyr Ile Leu Pro Pro Pro Ala Glu Gln Leu Ser Arg
370 375 380
Lys Asp Val Ser Leu Thr Cys Leu Val Val Gly Phe Asn Pro Gly Asp
385 390 395 400
Ile Ser Val Glu Trp Thr Ser Asn Gly His Thr Glu Glu Asn Tyr Lys
405 410 415
Asp Thr Ala Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Ile Tyr Ser
420 425 430
Lys Leu Asn Met Lys Thr Ser Lys Trp Glu Lys Thr Asp Ser Phe Ser
435 440 445
Cys Asn Val Arg His Glu Gly Leu Lys Asn Tyr Tyr Leu Lys Lys Thr
450 455 460
Ile Ser Arg Ser Pro Gly Lys
465 470
<210> 75
<211> 238
<212> PRT
<213> Mus musculus
<400> 75
Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala
1 5 10 15
Ser Ser Ser Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val
20 25 30
Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
35 40 45
Val His Ser Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Arg Pro
50 55 60
Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser
65 70 75 80
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Leu Tyr Phe Cys
100 105 110
Ser Gln Ser Thr His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu
115 120 125
Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro
130 135 140
Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu
145 150 155 160
Asn Asn Phe Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly
165 170 175
Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser
180 185 190
Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp
195 200 205
Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr
210 215 220
Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys
225 230 235
<210> 76
<211> 466
<212> PRT
<213> Mus musculus
<400> 76
Met Lys Val Leu Ser Leu Leu Tyr Leu Leu Thr Ala Ile Pro Gly Ile
1 5 10 15
Met Ser Asp Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro
20 25 30
Ser Gln Ser Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr
35 40 45
Thr Ser Tyr Tyr Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu
50 55 60
Glu Trp Met Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Asn Pro
65 70 75 80
Ser Leu Lys Asn Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln
85 90 95
Phe Phe Leu Arg Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr
100 105 110
Tyr Cys Ala Gly Ser Phe Gly Tyr Ser Gln Gly Thr Leu Val Thr Val
115 120 125
Ser Ala Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly
130 135 140
Cys Gly Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys
145 150 155 160
Gly Tyr Phe Pro Glu Ser Val Thr Val Thr Trp Asn Ser Gly Ser Leu
165 170 175
Ser Ser Ser Val His Thr Phe Pro Ala Leu Leu Gln Ser Gly Leu Tyr
180 185 190
Thr Met Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser Gln
195 200 205
Thr Val Thr Cys Ser Val Ala His Pro Ala Ser Ser Thr Thr Val Asp
210 215 220
Lys Lys Leu Glu Pro Ser Gly Pro Ile Ser Thr Ile Asn Pro Cys Pro
225 230 235 240
Pro Cys Lys Glu Cys His Lys Cys Pro Ala Pro Asn Leu Glu Gly Gly
245 250 255
Pro Ser Val Phe Ile Phe Pro Pro Asn Ile Lys Asp Val Leu Met Ile
260 265 270
Ser Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Val Ser Glu Asp
275 280 285
Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val Glu Val His
290 295 300
Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser Thr Ile Arg
305 310 315 320
Val Val Ser Thr Leu Pro Ile Gln His Gln Asp Trp Met Ser Gly Lys
325 330 335
Glu Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro Ser Pro Ile Glu
340 345 350
Arg Thr Ile Ser Lys Ile Lys Gly Leu Val Arg Ala Pro Gln Val Tyr
355 360 365
Ile Leu Pro Pro Pro Ala Glu Gln Leu Ser Arg Lys Asp Val Ser Leu
370 375 380
Thr Cys Leu Val Val Gly Phe Asn Pro Gly Asp Ile Ser Val Glu Trp
385 390 395 400
Thr Ser Asn Gly His Thr Glu Glu Asn Tyr Lys Asp Thr Ala Pro Val
405 410 415
Leu Asp Ser Asp Gly Ser Tyr Phe Ile Tyr Ser Lys Leu Asn Met Lys
420 425 430
Thr Ser Lys Trp Glu Lys Thr Asp Ser Phe Ser Cys Asn Val Arg His
435 440 445
Glu Gly Leu Lys Asn Tyr Tyr Leu Lys Lys Thr Ile Ser Arg Ser Pro
450 455 460
Gly Lys
465
<210> 77
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> chimeric antibody light chain
<400> 77
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 78
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> chimeric antibody heavy chain
<400> 78
Glu Val Lys Leu Glu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Met Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Val Arg Gln Ser Pro Glu Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Arg Ser
65 70 75 80
Val Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu Asp Thr Gly Ile Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 79
<211> 1461
<212> PRT
<213> Homo sapiens
<400> 79
Met Ala Ala Glu Arg Gly Ala Arg Arg Leu Leu Ser Thr Pro Ser Phe
1 5 10 15
Trp Leu Tyr Cys Leu Leu Leu Leu Gly Arg Arg Ala Pro Gly Ala Ala
20 25 30
Ala Ala Arg Ser Gly Ser Ala Pro Gln Ser Pro Gly Ala Ser Ile Arg
35 40 45
Thr Phe Thr Pro Phe Tyr Phe Leu Val Glu Pro Val Asp Thr Leu Ser
50 55 60
Val Arg Gly Ser Ser Val Ile Leu Asn Cys Ser Ala Tyr Ser Glu Pro
65 70 75 80
Ser Pro Lys Ile Glu Trp Lys Lys Asp Gly Thr Phe Leu Asn Leu Val
85 90 95
Ser Asp Asp Arg Arg Gln Leu Leu Pro Asp Gly Ser Leu Phe Ile Ser
100 105 110
Asn Val Val His Ser Lys His Asn Lys Pro Asp Glu Gly Tyr Tyr Gln
115 120 125
Cys Val Ala Thr Val Glu Ser Leu Gly Thr Ile Ile Ser Arg Thr Ala
130 135 140
Lys Leu Ile Val Ala Gly Leu Pro Arg Phe Thr Ser Gln Pro Glu Pro
145 150 155 160
Ser Ser Val Tyr Ala Gly Asn Asn Ala Ile Leu Asn Cys Glu Val Asn
165 170 175
Ala Asp Leu Val Pro Phe Val Arg Trp Glu Gln Asn Arg Gln Pro Leu
180 185 190
Leu Leu Asp Asp Arg Val Ile Lys Leu Pro Ser Gly Met Leu Val Ile
195 200 205
Ser Asn Ala Thr Glu Gly Asp Gly Gly Leu Tyr Arg Cys Val Val Glu
210 215 220
Ser Gly Gly Pro Pro Lys Tyr Ser Asp Glu Val Glu Leu Lys Val Leu
225 230 235 240
Pro Asp Pro Glu Val Ile Ser Asp Leu Val Phe Leu Lys Gln Pro Ser
245 250 255
Pro Leu Val Arg Val Ile Gly Gln Asp Val Val Leu Pro Cys Val Ala
260 265 270
Ser Gly Leu Pro Thr Pro Thr Ile Lys Trp Met Lys Asn Glu Glu Ala
275 280 285
Leu Asp Thr Glu Ser Ser Glu Arg Leu Val Leu Leu Ala Gly Gly Ser
290 295 300
Leu Glu Ile Ser Asp Val Thr Glu Asp Asp Ala Gly Thr Tyr Phe Cys
305 310 315 320
Ile Ala Asp Asn Gly Asn Glu Thr Ile Glu Ala Gln Ala Glu Leu Thr
325 330 335
Val Gln Ala Gln Pro Glu Phe Leu Lys Gln Pro Thr Asn Ile Tyr Ala
340 345 350
His Glu Ser Met Asp Ile Val Phe Glu Cys Glu Val Thr Gly Lys Pro
355 360 365
Thr Pro Thr Val Lys Trp Val Lys Asn Gly Asp Met Val Ile Pro Ser
370 375 380
Asp Tyr Phe Lys Ile Val Lys Glu His Asn Leu Gln Val Leu Gly Leu
385 390 395 400
Val Lys Ser Asp Glu Gly Phe Tyr Gln Cys Ile Ala Glu Asn Asp Val
405 410 415
Gly Asn Ala Gln Ala Gly Ala Gln Leu Ile Ile Leu Glu His Ala Pro
420 425 430
Ala Thr Thr Gly Pro Leu Pro Ser Ala Pro Arg Asp Val Val Ala Ser
435 440 445
Leu Val Ser Thr Arg Phe Ile Lys Leu Thr Trp Arg Thr Pro Ala Ser
450 455 460
Asp Pro His Gly Asp Asn Leu Thr Tyr Ser Val Phe Tyr Thr Lys Glu
465 470 475 480
Gly Ile Ala Arg Glu Arg Val Glu Asn Thr Ser His Pro Gly Glu Met
485 490 495
Gln Val Thr Ile Gln Asn Leu Met Pro Ala Thr Val Tyr Ile Phe Arg
500 505 510
Val Met Ala Gln Asn Lys His Gly Ser Gly Glu Ser Ser Ala Pro Leu
515 520 525
Arg Val Glu Thr Gln Pro Glu Val Gln Leu Pro Gly Pro Ala Pro Asn
530 535 540
Leu Arg Ala Tyr Ala Ala Ser Pro Thr Ser Ile Thr Val Thr Trp Glu
545 550 555 560
Thr Pro Val Ser Gly Asn Gly Glu Ile Gln Asn Tyr Lys Leu Tyr Tyr
565 570 575
Met Glu Lys Gly Thr Asp Lys Glu Gln Asp Val Asp Val Ser Ser His
580 585 590
Ser Tyr Thr Ile Asn Gly Leu Lys Lys Tyr Thr Glu Tyr Ser Phe Arg
595 600 605
Val Val Ala Tyr Asn Lys His Gly Pro Gly Val Ser Thr Pro Asp Val
610 615 620
Ala Val Arg Thr Leu Ser Asp Val Pro Ser Ala Ala Pro Gln Asn Leu
625 630 635 640
Ser Leu Glu Val Arg Asn Ser Lys Ser Ile Met Ile His Trp Gln Pro
645 650 655
Pro Ala Pro Ala Thr Gln Asn Gly Gln Ile Thr Gly Tyr Lys Ile Arg
660 665 670
Tyr Arg Lys Ala Ser Arg Lys Ser Asp Val Thr Glu Thr Leu Val Ser
675 680 685
Gly Thr Gln Leu Ser Gln Leu Ile Glu Gly Leu Asp Arg Gly Thr Glu
690 695 700
Tyr Asn Phe Arg Val Ala Ala Leu Thr Ile Asn Gly Thr Gly Pro Ala
705 710 715 720
Thr Asp Trp Leu Ser Ala Glu Thr Phe Glu Ser Asp Leu Asp Glu Thr
725 730 735
Arg Val Pro Glu Val Pro Ser Ser Leu His Val Arg Pro Leu Val Thr
740 745 750
Ser Ile Val Val Ser Trp Thr Pro Pro Glu Asn Gln Asn Ile Val Val
755 760 765
Arg Gly Tyr Ala Ile Gly Tyr Gly Ile Gly Ser Pro His Ala Gln Thr
770 775 780
Ile Lys Val Asp Tyr Lys Gln Arg Tyr Tyr Thr Ile Glu Asn Leu Asp
785 790 795 800
Pro Ser Ser His Tyr Val Ile Thr Leu Lys Ala Phe Asn Asn Val Gly
805 810 815
Glu Gly Ile Pro Leu Tyr Glu Ser Ala Val Thr Arg Pro His Thr Asp
820 825 830
Thr Ser Glu Val Asp Leu Phe Val Ile Asn Ala Pro Tyr Thr Pro Val
835 840 845
Pro Asp Pro Thr Pro Met Met Pro Pro Val Gly Val Gln Ala Ser Ile
850 855 860
Leu Ser His Asp Thr Ile Arg Ile Thr Trp Ala Asp Asn Ser Leu Pro
865 870 875 880
Lys His Gln Lys Ile Thr Asp Ser Arg Tyr Tyr Thr Val Arg Trp Lys
885 890 895
Thr Asn Ile Pro Ala Asn Thr Lys Tyr Lys Asn Ala Asn Ala Thr Thr
900 905 910
Leu Ser Tyr Leu Val Thr Gly Leu Lys Pro Asn Thr Leu Tyr Glu Phe
915 920 925
Ser Val Met Val Thr Lys Gly Arg Arg Ser Ser Thr Trp Ser Met Thr
930 935 940
Ala His Gly Thr Thr Phe Glu Leu Val Pro Thr Ser Pro Pro Lys Asp
945 950 955 960
Val Thr Val Val Ser Lys Glu Gly Lys Pro Lys Thr Ile Ile Val Asn
965 970 975
Trp Gln Pro Pro Ser Glu Ala Asn Gly Lys Ile Thr Gly Tyr Ile Ile
980 985 990
Tyr Tyr Ser Thr Asp Val Asn Ala Glu Ile His Asp Trp Val Ile Glu
995 1000 1005
Pro Val Val Gly Asn Arg Leu Thr His Gln Ile Gln Glu Leu Thr
1010 1015 1020
Leu Asp Thr Pro Tyr Tyr Phe Lys Ile Gln Ala Arg Asn Ser Lys
1025 1030 1035
Gly Met Gly Pro Met Ser Glu Ala Val Gln Phe Arg Thr Pro Lys
1040 1045 1050
Ala Asp Ser Ser Asp Lys Met Pro Asn Asp Gln Ala Ser Gly Ser
1055 1060 1065
Gly Gly Lys Gly Ser Arg Leu Pro Asp Leu Gly Ser Asp Tyr Lys
1070 1075 1080
Pro Pro Met Ser Gly Ser Asn Ser Pro His Gly Ser Pro Thr Ser
1085 1090 1095
Pro Leu Asp Ser Asn Met Leu Leu Val Ile Ile Val Ser Val Gly
1100 1105 1110
Val Ile Thr Ile Val Val Val Val Ile Ile Ala Val Phe Cys Thr
1115 1120 1125
Arg Arg Thr Thr Ser His Gln Lys Lys Lys Arg Ala Ala Cys Lys
1130 1135 1140
Ser Val Asn Gly Ser His Lys Tyr Lys Gly Asn Ser Lys Asp Val
1145 1150 1155
Lys Pro Pro Asp Leu Trp Ile His His Glu Arg Leu Glu Leu Lys
1160 1165 1170
Pro Ile Asp Lys Ser Pro Asp Pro Asn Pro Ile Met Thr Asp Thr
1175 1180 1185
Pro Ile Pro Arg Asn Ser Gln Asp Ile Thr Pro Val Asp Asn Ser
1190 1195 1200
Met Asp Ser Asn Ile His Gln Arg Arg Asn Ser Tyr Arg Gly His
1205 1210 1215
Glu Ser Glu Asp Ser Met Ser Thr Leu Ala Gly Arg Arg Gly Met
1220 1225 1230
Arg Pro Lys Met Met Met Pro Phe Asp Ser Gln Pro Pro Gln Pro
1235 1240 1245
Val Ile Ser Ala His Pro Ile His Ser Leu Asp Asn Pro His His
1250 1255 1260
His Phe His Ser Ser Ser Leu Ala Ser Pro Ala Arg Ser His Leu
1265 1270 1275
Tyr His Pro Gly Ser Pro Trp Pro Ile Gly Thr Ser Met Ser Leu
1280 1285 1290
Ser Asp Arg Ala Asn Ser Thr Glu Ser Val Arg Asn Thr Pro Ser
1295 1300 1305
Thr Asp Thr Met Pro Ala Ser Ser Ser Gln Thr Cys Cys Thr Asp
1310 1315 1320
His Gln Asp Pro Glu Gly Ala Thr Ser Ser Ser Tyr Leu Ala Ser
1325 1330 1335
Ser Gln Glu Glu Asp Ser Gly Gln Ser Leu Pro Thr Ala His Val
1340 1345 1350
Arg Pro Ser His Pro Leu Lys Ser Phe Ala Val Pro Ala Ile Pro
1355 1360 1365
Pro Pro Gly Pro Pro Thr Tyr Asp Pro Ala Leu Pro Ser Thr Pro
1370 1375 1380
Leu Leu Ser Gln Gln Ala Leu Asn His His Ile His Ser Val Lys
1385 1390 1395
Thr Ala Ser Ile Gly Thr Leu Gly Arg Ser Arg Pro Pro Met Pro
1400 1405 1410
Val Val Val Pro Ser Ala Pro Glu Val Gln Glu Thr Thr Arg Met
1415 1420 1425
Leu Glu Asp Ser Glu Ser Ser Tyr Glu Pro Asp Glu Leu Thr Lys
1430 1435 1440
Glu Met Ala His Leu Glu Gly Leu Met Lys Asp Leu Asn Ala Ile
1445 1450 1455
Thr Thr Ala
1460
<210> 80
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HA
<400> 80
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 81
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HB
<400> 81
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 82
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HC
<400> 82
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 83
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HD
<400> 83
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 84
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HE
<400> 84
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 85
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HF
<400> 85
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 86
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HG
<400> 86
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 87
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain HH
<400> 87
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 88
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KA
<400> 88
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Ser Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 89
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KB
<400> 89
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Ser Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 90
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KC
<400> 90
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Ser Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 91
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KD
<400> 91
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 92
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KE
<400> 92
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Ser Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 93
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KF
<400> 93
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 94
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain KG
<400> 94
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 95
<211> 116
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody heavy chain
<400> 95
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Ala
20 25 30
Trp Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Glu Ile Arg Ser Lys Ala Asn Asn His Ala Thr Tyr Tyr Ala Glu
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ile
65 70 75 80
Val Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Leu Tyr
85 90 95
Tyr Cys Thr Arg Arg Asp Gly Ala Tyr Trp Gly Lys Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 96
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> humanized antibody light chain
<400> 96
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Ser Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Ser Tyr Phe Cys Gln Gln Leu Asn Thr Leu Pro Trp
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Met Glu
100 105
<210> 97
<211> 14
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 97
Thr Gly Thr Ser Ser Ser Val Gly Asp Ser Ile Tyr Val Ser
1 5 10
<210> 98
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 98
Asp Val Thr Lys Arg Pro Ser
1 5
<210> 99
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 99
Cys Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 100
<211> 5
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 100
Ser His Gly Ile Ser
1 5
<210> 101
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 101
Trp Ile Ser Pro Tyr Ser Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln
1 5 10 15
Gly
<210> 102
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 102
Val Gly Ser Gly Pro Tyr Tyr Tyr Met Asp Val
1 5 10
<210> 103
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 103
Tyr Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 104
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 104
Phe Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 105
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 105
His Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 106
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 106
Leu Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 107
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 107
Val Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 108
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 108
Ile Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
<210> 109
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Synthetic peptide
<400> 109
Lys Ser Tyr Ala Gly Thr Asp Thr Leu
1 5
Claims (4)
- RGMa 저해 물질을 포함하는, HTLV-1 관련 척수증(HAM)의 치료 또는 예방제.
- 제1항에 있어서,
상기 RGMa 저해 물질이, RGMa를 인식하는 항체인, HAM의 치료 또는 예방제. - 그것을 필요로 하는 HTLV-1 관련 척수증(HAM)환자에 약리학적으로 유효량의 RGMa 저해 물질을 투여하는 것을 포함하는, HAM의 치료 방법.
- 제3항에 있어서,
상기 RGMa 저해 물질이, RGMa를 인식하는 항체인, HAM의 치료 방법.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JPJP-P-2018-135925 | 2018-07-19 | ||
| JP2018135925 | 2018-07-19 | ||
| PCT/JP2019/028431 WO2020017629A1 (ja) | 2018-07-19 | 2019-07-19 | Ｈｔｌｖ－１関連脊髄症（ｈａｍ）治療又は予防剤、及びｈａｍの治療方法 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20210034055A true KR20210034055A (ko) | 2021-03-29 |
Family
ID=69163691
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020217005032A Pending KR20210034055A (ko) | 2018-07-19 | 2019-07-19 | Htlv-1 관련 척수증(ham) 치료 또는 예방제, 및 ham의 치료 방법 |
Country Status (16)
| Country | Link |
|---|---|
| US (1) | US20210269535A1 (ko) |
| EP (1) | EP3824907A4 (ko) |
| JP (3) | JP7150286B2 (ko) |
| KR (1) | KR20210034055A (ko) |
| CN (4) | CN116785439A (ko) |
| AU (1) | AU2019306721B2 (ko) |
| BR (1) | BR112021000949A2 (ko) |
| CA (1) | CA3112511A1 (ko) |
| IL (1) | IL280257A (ko) |
| MX (1) | MX2021000732A (ko) |
| PE (1) | PE20211376A1 (ko) |
| PH (1) | PH12021550134A1 (ko) |
| SG (1) | SG11202100561QA (ko) |
| TW (2) | TWI825131B (ko) |
| WO (1) | WO2020017629A1 (ko) |
| ZA (1) | ZA202101113B (ko) |
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| JP2010100578A (ja) | 2008-10-24 | 2010-05-06 | St Marianna Univ School Of Medicine | Ｈｔｌｖ−ｉ関連脊髄症を治療または予防するための医薬、およびｈｔｌｖ−ｉ関連脊髄症の患者に対する抗体療法の効果を予測する方法 |
| KR20110071059A (ko) | 2008-09-02 | 2011-06-28 | 도레이 토넨 기노우마쿠 고도카이샤 | 미세다공성 고분자막, 이 미세다공성 고분자막의 제조방법 및 전지 세퍼레이터 필름으로서의 이 미세다공성 고분자막의 사용 |
| JP2014138599A (ja) | 2008-02-29 | 2014-07-31 | Abbott Gmbh & Co Kg | Ｒｇｍａタンパク質に対するモノクローナル抗体及びその使用 |
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| EP1442749A1 (en) * | 2003-01-31 | 2004-08-04 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Use of anti-CD100 antibodies for the treatment and the diagnosis of inflammatory disorder affecting the central nervous system |
| JP3981148B2 (ja) * | 2004-03-11 | 2007-09-26 | バイオクルーズ株式会社 | 軸索再生促進剤 |
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| US9133244B2 (en) * | 2011-01-18 | 2015-09-15 | Bioniz, Llc | Compositions and methods for modulating gamma-c-cytokine activity |
| JP5963233B2 (ja) * | 2011-12-07 | 2016-08-03 | 学校法人 聖マリアンナ医科大学 | Ｈｔｌｖ−１関連脊髄症を治療または予防するための医薬および前記医薬を用いた抗体療法の治療効果の確認方法 |
| JP6310845B2 (ja) * | 2012-07-06 | 2018-04-11 | 学校法人 聖マリアンナ医科大学 | Ｈｔｌｖ−１関連脊髄症患者の治療方法および治療剤 |
| CA2983898A1 (en) * | 2015-04-28 | 2016-11-03 | Mitsubishi Tanabe Pharma Corporation | Rgma binding protein and use thereof |
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2019
- 2019-07-19 KR KR1020217005032A patent/KR20210034055A/ko active Pending
- 2019-07-19 CA CA3112511A patent/CA3112511A1/en active Pending
- 2019-07-19 JP JP2020531380A patent/JP7150286B2/ja active Active
- 2019-07-19 AU AU2019306721A patent/AU2019306721B2/en active Active
- 2019-07-19 EP EP19838340.8A patent/EP3824907A4/en active Pending
- 2019-07-19 SG SG11202100561QA patent/SG11202100561QA/en unknown
- 2019-07-19 CN CN202310983277.XA patent/CN116785439A/zh active Pending
- 2019-07-19 BR BR112021000949-3A patent/BR112021000949A2/pt unknown
- 2019-07-19 MX MX2021000732A patent/MX2021000732A/es unknown
- 2019-07-19 PE PE2021000080A patent/PE20211376A1/es unknown
- 2019-07-19 TW TW108125746A patent/TWI825131B/zh active
- 2019-07-19 CN CN201980048012.2A patent/CN112566664B/zh active Active
- 2019-07-19 TW TW112142583A patent/TW202408577A/zh unknown
- 2019-07-19 WO PCT/JP2019/028431 patent/WO2020017629A1/ja active Application Filing
- 2019-07-19 CN CN202310983220.XA patent/CN116942831A/zh active Pending
- 2019-07-19 CN CN202310983521.2A patent/CN116850291A/zh active Pending
- 2019-07-19 US US17/261,480 patent/US20210269535A1/en active Pending
-
2021
- 2021-01-18 IL IL280257A patent/IL280257A/en unknown
- 2021-01-18 PH PH12021550134A patent/PH12021550134A1/en unknown
- 2021-02-18 ZA ZA2021/01113A patent/ZA202101113B/en unknown
-
2022
- 2022-09-16 JP JP2022148180A patent/JP7475011B2/ja active Active
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2024
- 2024-04-08 JP JP2024061895A patent/JP2024095754A/ja active Pending
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Also Published As
| Publication number | Publication date |
|---|---|
| JP7475011B2 (ja) | 2024-04-26 |
| MX2021000732A (es) | 2021-04-28 |
| TW202019482A (zh) | 2020-06-01 |
| PH12021550134A1 (en) | 2021-09-27 |
| CN112566664A (zh) | 2021-03-26 |
| IL280257A (en) | 2021-03-25 |
| CN116850291A (zh) | 2023-10-10 |
| ZA202101113B (en) | 2022-07-27 |
| JPWO2020017629A1 (ja) | 2021-08-02 |
| EP3824907A4 (en) | 2022-05-04 |
| JP7150286B2 (ja) | 2022-10-11 |
| TWI825131B (zh) | 2023-12-11 |
| TW202408577A (zh) | 2024-03-01 |
| EP3824907A1 (en) | 2021-05-26 |
| CN112566664B (zh) | 2023-08-25 |
| CN116942831A (zh) | 2023-10-27 |
| BR112021000949A2 (pt) | 2021-04-27 |
| US20210269535A1 (en) | 2021-09-02 |
| JP2022180487A (ja) | 2022-12-06 |
| JP2024095754A (ja) | 2024-07-10 |
| AU2019306721B2 (en) | 2024-09-05 |
| WO2020017629A1 (ja) | 2020-01-23 |
| SG11202100561QA (en) | 2021-02-25 |
| AU2019306721A1 (en) | 2021-03-11 |
| PE20211376A1 (es) | 2021-07-27 |
| CA3112511A1 (en) | 2020-01-23 |
| CN116785439A (zh) | 2023-09-22 |
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