KR20220005002A - 재조합 인플루엔자 항원 - Google Patents
재조합 인플루엔자 항원 Download PDFInfo
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- KR20220005002A KR20220005002A KR1020217036388A KR20217036388A KR20220005002A KR 20220005002 A KR20220005002 A KR 20220005002A KR 1020217036388 A KR1020217036388 A KR 1020217036388A KR 20217036388 A KR20217036388 A KR 20217036388A KR 20220005002 A KR20220005002 A KR 20220005002A
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
본 발명은 재조합 인플루엔자 A 적혈구응집소(HA) 폴리펩티드로서, 인플루엔자 A 바이러스 HA의 HA1 및 HA2 도메인을 포함하고,
(a) 위치 355의 아미노산은 W이고;
(b) 위치 432의 아미노산은 I이고/이거나, 위치 380의 아미노산은 I인 아미노산 서열을 포함하고;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따르는 것인 폴리펩티드, 이의 면역원성 단편, 상기 폴리펩티드 또는 면역원성 단편을 암호화하는 핵산 분자, 및 이들의 용도를 제공한다.
(a) 위치 355의 아미노산은 W이고;
(b) 위치 432의 아미노산은 I이고/이거나, 위치 380의 아미노산은 I인 아미노산 서열을 포함하고;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따르는 것인 폴리펩티드, 이의 면역원성 단편, 상기 폴리펩티드 또는 면역원성 단편을 암호화하는 핵산 분자, 및 이들의 용도를 제공한다.
Description
본 발명은 적어도 부분적으로, HHS가 수여한 계약 HHSO100201700018C에 따른 정부 지원으로 이루어졌다. 정부는 본 발명에 있어서 소정의 권리를 갖는다.
본 발명은 의약 분야에 관한 것이다. 재조합 인플루엔자 A 적혈구응집소(HA) 폴리펩티드, 상기 폴리펩티드를 암호화하는 핵산, 이를 포함하는 약제학적 조성물, 및 이들의 사용 방법이 본 명세서에서 제공된다.
인플루엔자 바이러스는 중증도가 준임상적 감염으로부터 사망을 야기할 수 있는 원발성 바이러스성 폐렴까지의 범위인 호흡기 질환(통상 "인플루엔자" 또는 "플루"로 지칭)을 야기하는 주요 인간 병원체이다. 감염의 임상 효과는 인플루엔자 균주의 발병력 및 숙주의 노출, 병력, 연령 및 면역 상태에 따라 달라진다. 매년 전 세계적으로 대략 10억 명의 사람들이 인플루엔자 바이러스로 감염을 겪어, 3백만 내지 5백만의 사례에서 중증의 병이 야기되고, 300,000 내지 500,000건의 인플루엔자 관련 사망이 발생하는 것으로 추정된다. 대부분의 이들 감염은 기여도가 보다 적은 인플루엔자 B 바이러스와 함께, H1 또는 H3 적혈구응집소 서브타입을 지니는 인플루엔자 A 바이러스에 기인할 수 있으며, 이에 따라, 대표적인 3가지 모두가 계절성 백신에 포함된다. 현행의 면역화 관행은 효과적인 계절성 인플루엔자 백신의 시기 적절한 생성을 가능하게 하기 위하여, 유행하는 인플루엔자 바이러스의 조기 확인에 의존한다. 다음 계절 동안 우세적일 균주를 예측함에 있어서 내재적인 어려움과 별개로, 항바이러스 내성 및 면역 회피 또한 현행의 백신이 이환 및 사망을 예방하지 못하게 한다. 이에 더하여, 동물 병원소로부터 비롯되고, 인간 대 인간 확산을 증가시키도록 재배열된(reassorted) 고도의 발병력의 바이러스 균주에 의해 야기되는 범유행의 가능성은 세계의 보건에 대하여 상당한 현실적인 위협을 제기한다.
인플루엔자 A 바이러스는 자연에서 널리 분포되어 있으며, 다양한 조류 및 포유동물을 감염시킬 수 있다. 인플루엔자 바이러스는 오르토믹소비리대(Orthomyxoviridae)의 과에 속하는 외피(enveloped) RNA 바이러스이다. 이들의 게놈은 11종의 상이한 단백질, 1종의 핵단백질(NP), 3종의 중합효소 단백질(PA, PB1 및 PB2), 2종의 기질 단백질(M1 및 M2), 3종의 비 구조 단백질(NS1, NS2 및 PB1-F2) 및 2종의 외부 당단백질: 적혈구응집소(HA) 및 뉴라미니다제(NA)를 암호화하는 8개의 단일 가닥 RNA 절편으로 구성된다. 바이러스는 HA 및 NA 단백질의 항원 구조의 차이를 기반으로 하여 분류되며, 이들의 상이한 조합은 특정 인플루엔자 바이러스 균주로 더 분류되는 독특한 바이러스 서브타입을 나타낸다. 모든 공지되어 있는 서브타입이 조류에서 관찰될 수 있지만, 현재 유행하는 인간 인플루엔자 A 서브타입은 H1N1 및 H3N2이다. 계통발생 분석에 의해, 다음과 같은 2개의 주요 그룹으로의 적혈구응집소의 세분이 증명되었다: 특히, 계통발생 그룹 1에서 H1, H2, H5 및 H9 서브타입, 그리고 특히, 계통발생 그룹 2에서 H3, H4 및 H7 서브타입.
인플루엔자 B형 바이러스 균주는 엄격하게 인간형이다. 인플루엔자 B형 바이러스 균주에서의 HA의 항원 변이는 A형 균주에서 관찰되는 것보다 더 작다. B/야마가타/16/88(B/야마가타로도 지칭) 및 B/빅토리아/2/87(B/빅토리아) 계통에 의해 대표되는 인플루엔자 B 바이러스의 2종의 유전적 및 항원적으로 별개의 계통이 인간에서 유행하고 있다. 인플루엔자 B 바이러스에 의해 야기되는 질환의 스펙트럼은 일반적으로 인플루엔자 A 바이러스에 의해 야기되는 것보다 더 경증이지만, 입원을 필요로 하는 중증의 병이 인플루엔자 B 감염에서 여전히 빈번하게 관찰된다.
인플루엔자 바이러스를 중화시키는 항체가 주로 적혈구응집소(HA)에 대해 유도되는 것이 알려져 있다. 적혈구응집소는 바이러스 코트에 고정되는 삼량체 당단백질이며 이중의 기능을 갖는다: 세포 표면 수용체 시알산으로의 결합을 담당하는 기능, 및 이것이 흡수 후에 바이러스 및 엔도솜 막의 융합을 매개하여 세포의 세포질 내의 바이러스 RNA의 방출을 야기하는 기능. HA는 소위 헤드 도메인(head domain) 및 스템 도메인(stem domain)을 포함한다. 바이러스 막으로의 부착은 스템 도메인에 연결된 C-말단 고정(anchoring) 서열(막횡단 도메인으로도 알려져 있음)에 의해 매개된다. 단백질은 지정된 루프에서 번역 후 절단되어, 2가지 폴리펩티드, HA1 및 HA2를 생성한다(전체 서열은 HA0으로 지칭). 막 원위 헤드 도메인은 주로 HA1으로부터 유래되며, 막 근위 스템 도메인은 주로 HA2로부터 유래된다(도 1).
인플루엔자 바이러스는 어디에나 존재하기 때문에, 바이러스에 의한 감염을 방지하는 것은 거의 불가능하다. 백신접종은 인플루엔자 유행병 및 범유행을 통제하는 데 중요한 역할을 한다. 많은 인플루엔자 백신은 달걀에서 바이러스의 재배열, 적응 및 성장을 포함하는 방법에 의해 제조된다. 그러나 이러한 기존 방법에는 한계가 있다. 모든 인플루엔자 바이러스 균주가 달걀에서 잘 자라는 것은 아니므로, 적응하거나 바이러스 재배열체를 구성해야 한다. 제조 중 HA의 변화는 순환하는 균주와 다르고 최적이 아닌 보호 수준을 제공할 수 있는 균주로 이어질 수 있다. 또 다른 단점은 달걀 알레르기가 있는 사람들이 달걀 기반 백신의 잔류 달걀 단백질에 과민반응을 보일 수 있다는 점이다. 더욱이, 달걀 기반 방법은 연속된 달걀의 공급에 의존하며, 이는 가금류의 질환의 경우에서와 같이 공급의 중단에 취약할 수 있다. 달걀 공급에 의존하지 않고 백신 단백질 제조가 달걀 기반 방법보다 더욱 엄격하게 통제되는 방법을 사용하여 백신을 제조할 필요가 있다. 세포 배양에서 제조된 HA의 재조합 형태(rHA)는 달걀에서 얻은 것에 대한 인플루엔자 백신의 대안적인 항원 공급원으로 사용된다. 그러나 이러한 방법을 사용하여 rHA의 면역원성 및 규칙적인 4차 구조를 유지하고 높은 수율의 삼량체 rHA를 보장하는 문제가 발생하였다. 따라서, 기존의 문제를 해결하는 인플루엔자 백신 또는 진단을 위한 대안적인 항원 공급 방법이 여전히 필요하다.
본 발명의 일부 양태가 아래에 요약된다. 추가 양태는 본 특허 출원의 발명의 상세한 설명, 실시예, 도면, 및 청구범위 섹션에 기술되어 있다.
제1 양태에서, 본 발명은 재조합 인플루엔자 A 적혈구응집소(HA) 폴리펩티드에 관한 것으로, 인플루엔자 A 바이러스 HA의 HA1 및 HA2 도메인을 포함하고,
(a) 위치 355의 아미노산은 트립토판(W)이고;
(b) 위치 432의 아미노산은 이소류신(I)이고/이거나, 위치 380의 아미노산은 I인 아미노산 서열을 포함하며;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따르는 것인 HA 폴리펩티드에 관한 것이다.
추가 양태에서, 본 발명은 적어도 2개의 HA 폴리펩티드를 포함하는 다량체 폴리펩티드, 특히 본원에 기술된 바와 같은 3개의 HA 폴리펩티드를 포함하는 삼량체 폴리펩티드에 관한 것이다.
본 발명에 따르면 놀랍게도 재조합 인플루엔자 HA 폴리펩티드, 특히 재조합 삼량체 HA 폴리펩티드가 야생형 HA 폴리펩티드와 비교하여 높은 수준으로 수득될 수 있고, 이종 아미노산 서열, 예컨대, 이종 삼량체화 도메인의 추가 없이 더 큰 안정성을 나타내는 증가된 용융 온도를 갖는 것으로 나타났다. 또한, 본 발명의 HA 폴리펩티드는 항체 CR9114, CR8020 및/또는 CR6261과 같은(그러나 이에 한정되지 않음) HA 폴리펩티드에 대한 항-HA 항체의 결합에 의해 제시된 바와 같이 정확하게 접힌다. 따라서, 폴리펩티드는 대상체, 특히 인간 대상체에게 투여되는 경우 HA에 대한 면역 반응을 유도할 수 있다. 삼량체 폴리펩티드는 야생형 천연 HA의 4차 구조를 포함하고, 따라서 HA 분자의 막 근위 줄기의 보존된 에피토프를 포함하는 천연 에피토프를 면역계에 제시한다.
추가의 양태에서, 본 발명은 재조합 인플루엔자 HA 폴리펩티드를 암호화하는 핵산 분자를 제공한다.
또 다른 양태에서, 본 발명은 인플루엔자 HA 폴리펩티드를 암호화하는 핵산 분자를 포함하는 벡터, 특히 재조합 아데노바이러스 벡터를 제공한다.
또 다른 양태에서, 본 발명은 본 발명에 따른 인플루엔자 HA 폴리펩티드, 핵산 분자 및/또는 벡터, 및 약제학적으로 허용 가능한 담체를 포함하는 면역원성 조성물을 제공한다.
추가의 양태에서, 본 발명은 의약으로서 사용하기 위한, 특히 인플루엔자 질환, 특히 계통발생 그룹 1 및/또는 2로부터의 인플루엔자 바이러스 A 균주에 의해 유발된 질환 또는 병태의 예방 및/또는 치료를 위한 백신으로서 사용하기 위한 인플루엔자 HA 폴리펩티드, 상기 인플루엔자 HA 폴리펩티드를 암호화하는 핵산 분자, 및/또는 상기 핵산 분자를 포함하는 벡터를 제공한다.
본 발명은 또한 필요로 하는 대상체에서 인플루엔자 HA에 대한 면역 반응의 유도 방법을 제공하며, 방법은 대상체에게 본 발명에 따른 인플루엔자 HA 폴리펩티드, 핵산 분자, 및/또는 벡터를 투여하는 단계를 포함한다. 또 다른 양태에서, 인플루엔자 질환 감염 위험이 있는 것으로 확인된 사람과 같이 필요로 하는 사람에게 전술한 바와 같은 폴리펩티드 또는 면역원성 조성물을 투여하는 단계를 포함하는 인플루엔자 질환에 대한 예방 및/또는 백신접종 방법이 제공된다.
또 다른 양태에서, 위에서 정의된 바와 같은 재조합 HA 폴리펩티드를 제조하는 방법으로서, 원핵생물 또는 진핵생물 세포, 예컨대, 포유동물 세포, 예를 들어, CHO 세포, 또는 곤충 세포에서 전술한 핵산 분자를 발현시키는 단계를 포함하고, 상기 세포로부터 rHA를 정제/단리하는 단계를 선택적으로 추가로 포함하는 방법이 제공된다.
또 다른 양태에서, 본 발명은 연구 도구 또는 진단 도구로서, 또는 항체의 인플루엔자 억제제 생산을 위한 표적으로서 HA 폴리펩티드의 용도를 제공한다.
도 1.
A. 표시된 돌연변이의 위치를 갖는 본 발명의 폴리펩티드 단량체의 3차원 표현. 암회색의 적혈구응집소(HA)의 헤드와 연회색의 스템; B. 표시된 돌연변이의 위치를 갖는 본 발명의 폴리펩티드 단량체의 개략도(검은색: 헤드; 연회색: 스템).
도 2. 도 2a. 인플루엔자 HA의 계통수. 인플루엔자 A 그룹 1 및 그룹 2 및 인플루엔자 B의 상이한 서브타입이 표시되어 있다; 도 2b. OCTET(항-His2 센서)에 의해 결정된 단백질 발현 수준. 마지막 컬럼은 야생형(WT) HA와 비교하여 본 발명의 안정화된 가용성 HA 삼량체의 발현 수준의 배수 증가를 나타낸다; 도 2c. 크기 배제 크로마토그래피(SEC) 프로파일 - 점선은 WT HA를 나타내고, 검은색의 실선은 본 발명에 따른 안정화된 HA 폴리펩티드를 나타낸다.
도 3. 도 3a. 본 발명의 정제된 삼량체(T) 폴리펩티드(검은색 선) 및 폴드온(Foldon) 삼량체화 도메인을 갖는 WT 폴리펩티드(회색 선)의 SEC 분석(UFV4239는 폴드온 도메인을 포함하지 않음에 유의함). WT-폴드온 정제된 폴리펩티드는 -80℃에서 보관 후 피크의 폭 넓어짐 및 다량체 형성(*)을 나타낸다. UFV4239의 누락된 삼량체화 도메인으로 인해, 단량체(M)만 발현되고 정제되었다; 도 3b 및 도 3c. 시차 주사 형광측정법(DSF)에 의한 정제된 폴리펩티드의 온도 안정성 분석(℃ 단위의 Tm50 값); 도 3d. 단일클론 항체(mAb) CR6261, CR8020, CT149, CR9114 및 다중도메인 항체 MD3606의 결합(ELISA, EC50 값).
도 4. A. OCTET(항-His2 센서)에 의해 결정된 단백질 발현 수준; B. EXPI-293 세포 배양 상청액의 SEC 분석. UFV181007은 돌연변이 K380I 및 E432I(검은색의 점선)를 포함한다. UFV181005은 돌연변이 H355W 및 M478I(회색의 점선)를 포함한다. 안정화 돌연변이의 조합(UFV1810009, 검은색 선).
도 5. A. OCTET(항-His2 센서)에 의해 결정된 단백질 발현 수준. 마지막 컬럼은 야생형(WT) HA와 비교하여 본 발명의 안정화된 가용성 HA 삼량체의 발현 수준의 배수 증가를 나타낸다; B. 크기 배제 크로마토그래피(SEC) 프로파일- 점선은 WT HA를 나타내고, 검은색의 실선은 본 발명에 따른 추가의 안정화된 HA 폴리펩티드를 나타낸다.
도 6. 온도 스트레스 전후의 정제된 안정화 HA의 크기 배제 크로마토그래피(SEC) 프로파일. 실험 전(점선)과 4℃(검은색 실선) 및 37℃(회색 실선)에서 60일의 인큐베이션 후 폴리펩티드에 대한 프로파일을 나타내었다.
도 2. 도 2a. 인플루엔자 HA의 계통수. 인플루엔자 A 그룹 1 및 그룹 2 및 인플루엔자 B의 상이한 서브타입이 표시되어 있다; 도 2b. OCTET(항-His2 센서)에 의해 결정된 단백질 발현 수준. 마지막 컬럼은 야생형(WT) HA와 비교하여 본 발명의 안정화된 가용성 HA 삼량체의 발현 수준의 배수 증가를 나타낸다; 도 2c. 크기 배제 크로마토그래피(SEC) 프로파일 - 점선은 WT HA를 나타내고, 검은색의 실선은 본 발명에 따른 안정화된 HA 폴리펩티드를 나타낸다.
도 3. 도 3a. 본 발명의 정제된 삼량체(T) 폴리펩티드(검은색 선) 및 폴드온(Foldon) 삼량체화 도메인을 갖는 WT 폴리펩티드(회색 선)의 SEC 분석(UFV4239는 폴드온 도메인을 포함하지 않음에 유의함). WT-폴드온 정제된 폴리펩티드는 -80℃에서 보관 후 피크의 폭 넓어짐 및 다량체 형성(*)을 나타낸다. UFV4239의 누락된 삼량체화 도메인으로 인해, 단량체(M)만 발현되고 정제되었다; 도 3b 및 도 3c. 시차 주사 형광측정법(DSF)에 의한 정제된 폴리펩티드의 온도 안정성 분석(℃ 단위의 Tm50 값); 도 3d. 단일클론 항체(mAb) CR6261, CR8020, CT149, CR9114 및 다중도메인 항체 MD3606의 결합(ELISA, EC50 값).
도 4. A. OCTET(항-His2 센서)에 의해 결정된 단백질 발현 수준; B. EXPI-293 세포 배양 상청액의 SEC 분석. UFV181007은 돌연변이 K380I 및 E432I(검은색의 점선)를 포함한다. UFV181005은 돌연변이 H355W 및 M478I(회색의 점선)를 포함한다. 안정화 돌연변이의 조합(UFV1810009, 검은색 선).
도 5. A. OCTET(항-His2 센서)에 의해 결정된 단백질 발현 수준. 마지막 컬럼은 야생형(WT) HA와 비교하여 본 발명의 안정화된 가용성 HA 삼량체의 발현 수준의 배수 증가를 나타낸다; B. 크기 배제 크로마토그래피(SEC) 프로파일- 점선은 WT HA를 나타내고, 검은색의 실선은 본 발명에 따른 추가의 안정화된 HA 폴리펩티드를 나타낸다.
도 6. 온도 스트레스 전후의 정제된 안정화 HA의 크기 배제 크로마토그래피(SEC) 프로파일. 실험 전(점선)과 4℃(검은색 실선) 및 37℃(회색 실선)에서 60일의 인큐베이션 후 폴리펩티드에 대한 프로파일을 나타내었다.
정의
본 발명에서 사용되는 용어의 정의가 아래에 제공된다.
본 발명에 따른 아미노산은 20개의 자연 발생(또는 '표준' 아미노산) 또는 이의 변이체, 예를 들어, D-프롤린(프롤린의 D-거울상 이성질체), 또는 자연적으로 단백질에서 관찰되지 않는 임의의 변이체, 예를 들어, 노르류신 중 임의의 것일 수 있다. 표준 아미노산은 이들의 특성을 기반으로 몇 개의 군으로 분류될 수 있다. 중요한 인자는 전하, 친수성 또는 소수성, 크기 및 기능기이다. 이들 특성은 단백질 구조 및 단백질-단백질 상호작용에 중요하다. 다른 시스테인 잔기와 공유적 이황화 결합(또는 이황화 가교)을 형성할 수 있는 시스테인, 폴리펩티드 백본에 대하여 고리를 형성하는 프롤린, 및 다른 아미노산보다 더 유연한 글리신과 같이, 일부 아미노산은 특별한 특성을 갖는다. 표 3은 표준 아미노산의 약어 및 특성을 나타낸다.
본 명세서에서 사용되는 용어 "포함되는" 또는 "포함하는"은 용어 "제한 없이"가 뒤따르는 것으로 간주된다.
본 명세서에서 사용되는 용어 "감염"은 세포 또는 대상체 내의 바이러스의 증식 및/또는 존재에 의한 침습을 의미한다. 일 구현예에서, 감염은 "활성" 감염, 즉, 바이러스가 세포 또는 대상체에서 복제하고 있는 것이다. 이러한 감염은 바이러스에 의해 처음 감염된 세포, 조직 및/또는 기관으로부터 다른 세포, 조직 및/또는 기관으로의 바이러스의 확산을 특징으로 한다. 감염은 또한 잠복 감염, 즉, 바이러스가 복제하고 있지 않은 것일 수도 있다. 특정 구현예에서, 감염은 세포 또는 대상체에서의 바이러스의 존재로부터 야기되거나, 또는 바이러스에 의한 세포 또는 대상체의 침습에 의해 야기되는 병리적 상태를 지칭한다.
인플루엔자 바이러스는 전형적으로 속 A, B 및 C의 인플루엔자 바이러스 타입으로 분류된다. 본 명세서에서 사용되는 용어 "인플루엔자 바이러스 서브타입"은 헤마글루티닌(H) 및 뉴라미다제(N) 바이러스 표면 단백질의 조합을 특징으로 하는 인플루엔자 A 바이러스 변이체를 말한다. 본 발명에 따르면, 인플루엔자 바이러스 서브타입은 이들의 H 번호에 의해, 예를 들어, "H3 서브타입의 HA를 포함하는 인플루엔자 바이러스", "H3 서브타입의 인플루엔자 바이러스" 또는 "H3 인플루엔자"에 의해, 또는 H 번호 및 N 번호의 조합, 예를 들어, "인플루엔자 바이러스 서브타입 H3N2" 또는 "H3N2"에 의해 지칭될 수 있다. 용어 "서브타입"은 구체적으로 보통 돌연변이로부터 야기되는 각각의 서브타입 내의 모든 개별 "균주"를 포함하며, 천연 분리주 및 인공 돌연변이체 또는 재배열체 등을 포함하여, 상이한 병리학적 프로파일을 보인다. 이러한 균주는 또한 바이러스 서브타입의 다양한 "분리주"로 지칭될 수도 있다. 따라서, 본 명세서에서 사용되는 용어 "균주" 및 "분리주"는 상호교환 가능하게 사용될 수 있다. 인간 인플루엔자 바이러스 균주 또는 분리주에 대한 현행의 명명법은 예를 들어, A/모스크바/10/2000(H3N2)과 같이, 보통 괄호 안에 제공되는 HA 및 NA의 항원 설명과 함께, 바이러스의 타입(속), 즉, A, B 또는 C, 처음 분리한 지리학적 위치, 균주 번호 및 분리한 해를 포함한다. 비 인간 균주는 또한 명명에 있어 기원하는 숙주를 포함한다.
인플루엔자 A 바이러스 서브타입은 이들의 계통발생 그룹을 참조하여 더 분류될 수 있다. 계통발생 분석에 의해, 다음과 같은 2개의 주요 그룹으로의 적혈구응집소의 세분이 증명되었다: 특히, 계통발생 그룹 1("그룹 1" 인플루엔자 바이러스)의 H1, H2, H5 및 H9 서브타입 및 특히, 계통발생 그룹 2("그룹 2" 인플루엔자 바이러스)의 H3, H4, H7 및 H10 서브타입.
본 명세서에서 사용되는 용어 "인플루엔자 바이러스 질환" 또는 "인플루엔자"는 대상체에서의 인플루엔자 바이러스, 예를 들어, 인플루엔자 A 또는 B 바이러스의 존재로부터 야기되는 병리학적 상태를 지칭한다. 본 명세서에서 사용되는 용어 "질환" 및 "장애"는 상호교환 가능하게 사용된다. 특정 구현예에서, 이 용어는 인플루엔자 바이러스에 의한 대상체의 감염에 의해 야기되는 호흡기 병을 지칭한다.
본 명세서에서 사용되는 용어 "핵산" 또는 "핵산 분자"는 폴리뉴클레오티드, 예컨대, DNA 분자(예를 들어, cDNA 또는 게놈 DNA) 및 RNA 분자(예를 들어, mRNA) 및 뉴클레오티드 유사체를 사용하여 생성된 DNA 또는 RNA의 유사체를 포함하고자 한 것이다. 핵산은 단일 가닥 또는 이중 가닥일 수 있다. 핵산 분자는 화학적 또는 생화학적으로 변형될 수 있거나, 또는 당업자에 의해 용이하게 인정될 바와 같이, 비 천연 또는 유도체화된 뉴클레오티드 염기를 함유할 수 있다. 이러한 변형은 예를 들어, 표지, 메틸화, 하나 이상의 자연 발생 뉴클레오티드의 유사체로의 치환, 뉴클레오티드 사이 변형, 예를 들어, 비 하전 연결기(예를 들어, 메틸 포스포네이트, 포스포트리에스테르, 포스포르아미데이트, 카르바메이트 등), 하전된 연결기(예를 들어, 포스포로티오에이트, 포스포로디티오에이트 등), 펜덴트 부분(예를 들어, 폴리펩티드), 인터칼레이터(intercalator)(예를 들어, 아크리딘, 소랄렌 등), 킬레이터, 알킬레이터 및 변형된 연결기(예를 들어, 알파 아노머 핵산 등)를 포함한다. 핵산 서열에 대한 언급은 달리 명시되지 않는 한 이의 상보체를 포함한다. 따라서, 특정 서열을 갖는 핵산 분자에 대한 언급은 이의 상보 서열을 지니는 이의 상보 가닥을 포함하는 것으로 이해되어야 한다. 상보 가닥은 또한 예를 들어, 안티-센스 치료법, 혼성화 프로브 및 PCR 프라이머에 유용하다.
본 명세서에서 사용되는 바와 같이, HA에서 아미노산의 넘버링은 문헌[Winter et al. (Nature 292: 72-75, 1981)]에 기술된 바와 같이, H3 넘버링에 기반한다. 따라서, 본 발명의 폴리펩티드의 아미노산 잔기 또는 아미노산 위치의 넘버링은 문헌[Winter et al. (1981)]의 도 2에서 기술되고 도시된 바와 같이, H3 HA의 아미노산의 넘버링(특히, A/아이치/2/68의 HA의 아미노산 위치의 넘버링)에 해당한다. 특히, 넘버링은 서열번호 1의 아미노산 위치의 넘버링에 해당한다. 예를 들어, 표현 "위치 355의 아미노산"은 문헌[Winter et al. (1981)]의 H3 넘버링에 따른 위치 355에 있는 아미노산 잔기, 즉, 서열번호 1의 위치 355에 있는 아미노산 잔기를 지칭한다. 다른 인플루엔자 바이러스 균주 및/또는 서브타입, 예컨대, 예를 들어, H1, H5, 또는 H7 HA에서 균등한 아미노산이 서열 정렬에 의해 결정될 수 있음을 당업자는 이해할 것이다. 따라서, 예를 들어, 서열번호 1과 비교하여 추가의 아미노산 잔기가 부가되거나 제거된 경우, 상이한 HA 서열이 상이한 넘버링 시스템을 가질 수 있음을 주목해야 하고, 이를 당업자는 이해할 것이다. 이와 같이, 특정 아미노산 잔기가 그 번호로 지칭될 경우, 이러한 설명은 주어진 아미노산 서열의 시작부터 셀 때 정확히 해당 번호가 매겨진 위치에 위치한 아미노산에만 한정되지 않고, 오히려 해당 잔기가 동일한 정확한 번호가 매겨진 위치에 있지 않더라도, 예를 들어, HA 서열이 서열번호 1보다 짧거다 긴 경우, 또는 서열번호 1과 비교하여 삽입 또는 결실이 있는 경우, 모든 HA 서열의 균등한/상응하는 아미노산 잔기가 의도되는 것으로 이해된다. 당업자는 예를 들어 주어진 HA 서열을 서열번호 1에 정렬함으로써, 본원에 인용된 특정 번호가 매겨진 잔기 중 임의의 것과 상응하는/동등한 아미노산 위치가 무엇인지 쉽게 결정할 수 있다. 따라서, 인플루엔자 HA 단백질의 특정 아미노산 잔기가 지칭되는 구현예에서, 본 발명은 오로지 정확한 번호가 매겨진 아미노산 위치에서만의 소정의 아미노산 잔기(예를 들어, 위치 355의 트립토판(W) 및/또는 위치 432 및/또는 380의 이소류신(I)의 존재)를 갖는 서열들로 한정되지 않음이 이해된다.
"폴리펩티드"는 당업자에게 공지된 바와 같이 아미드 결합에 의해 연결된 아미노산의 중합체를 지칭한다. 본 명세서에서 사용되는 용어는 공유적 아미드 결합에 의해 연결된 단일 폴리펩티드 사슬을 지칭할 수 있다. 이 용어는 또한 비 공유 상호작용, 예를 들어, 이온 접촉, 수소 결합, 반데르발스 접촉 및 소수성 접촉에 의해 회합된 다수의 폴리펩티드 사슬을 지칭할 수 있다. 당업자는 이 용어가 예를 들어, 번역 후 가공, 예를 들어, 신호 펩티드 절단, 이황화 결합 형성, 글리코실화(예를 들어, N-연결 및 O-연결 글리코실화), 프로테아제 절단 및 지질 변형(예를 들어, S-팔미토일화)에 의해 변형된 폴리펩티드를 포함함을 인식할 것이다.
본 명세서에서 사용되는 용어 "야생형"은 자연적으로 순환하는 인플루엔자 바이러스로부터의 HA를 지칭한다.
발명의 상세한 설명
인플루엔자 바이러스는 세계 공중 보건에 상당한 영향을 미치며, 매년 수백만의 중증의 병의 사례, 수천의 사망 및 상당한 경제적 손실을 야기한다. 현행의 3가 인플루엔자 백신은 백신 균주 및 밀접하게 관련된 분리주에 대하여 강력한 중화 항체 반응을 유도하나, 서브타입 내의 더욱 분기된 균주 또는 다른 서브타입으로 드물게 확대된다. 또한, 적절한 백신 균주의 선택은 많은 시험감염을 제시하며, 준최적의 보호를 빈번하게 야기한다. 추가로, 다음의 유행성 바이러스가 발생할 시기와 장소를 비롯하여 이의 서브타입을 예측하는 것은 현재 불가능하다.
적혈구응집소(HA)는 중화 항체의 주요 표적인 인플루엔자 A 바이러스 유래의 주요 외피 당단백질이다. 적혈구응집소는 유입 과정 동안 2가지 주요 기능을 갖는다. 먼저, 적혈구응집소는 시알산 수용체와의 상호작용을 통하여 표적 세포의 표면으로의 바이러스의 부착을 매개한다. 두 번째로, 바이러스의 세포내 이입 후에, 적혈구응집소는 이후에 바이러스 및 엔도솜 막의 융합을 매개하여, 이의 게놈을 표적 세포의 세포질로 방출시킨다.
HA는 단량체당 약 500개 아미노산의 엑토도메인을 포함하는 삼량체 단백질이고, 각각이 이황화 결합에 의해 연결되는 2개의 폴리펩티드 HA1 및 HA2를 함유하는 세 개의 동일한 서브유닛(단량체)을 포함한다. 각각의 단량체는 처음에는 HA0으로 발현되고, 이후에 상기 이황화 결합을 통해 연결되는 HA1 및 HA2 도메인으로 숙주 프로테아제에 의해 절단된다.
대다수의 N 말단 도메인(HA1 도메인, 약 320 내지 330개 아미노산 길이)은 바이러스 중화 항체에 의해 인식되는 대부분의 결정기 및 수용체 결합 부위를 함유하는 막-원위 구형 도메인(헤드 도메인)을 형성한다. 보다 작은 C 말단 도메인(HA2 도메인, 약 180개 아미노산 길이)은 세포 또는 바이러스 막에 구형 도메인을 고정시키는 스템 유사 구조(스템 도메인)를 형성한다. 가장 보존된 영역 중 하나는 절단 부위 근처의 서열, 특히 HA2 N 말단의 23개 아미노산(융합 단백질)이며, 이는 모든 인플루엔자 A 바이러스 서브타입 간에 보존된다. 이러한 영역의 부분은 HA 전구체 분자(HA0)에서 표면 루프로서 노출되나, HA0이 HA1 및 HA2로 절단되는 경우 접근 불가능하게 된다.
위에서 언급한 바와 같이, 인플루엔자 HA 단백질은 바이러스의 표면에서 발견되는 주요한 단백질이다. 비리온의 표면에서 발견되는 HA는 삼량체 형태이다. 삼량체는 3개의 단량체 각각의 카복시 말단 단부의 막횡단 스패닝 서열에 의해 바이러스 막에 고정되어 있다. 인플루엔자 백신의 주요 보호 효능은 HA 단백질에 대한 항-헤마글루티닌 항체에 기인한다. 이는 구조적으로 관련 있는 HA 단백질에 대한 면역 반응을 높이는 것의 중요성을 강조한다.
인플루엔자 A 바이러스 적혈구응집소(HA0)의 엑토도메인을 나타내는 가용성 폴리펩티드를 제조하기 위하여, HA는 고유의 막횡단 및 세포질 도메인 없이 발현될 필요가 있다. 안정적인 삼량체형 가용성 야생형(WT) HA의 발현은 종종 포유동물 세포에서 매우 빈약하다. 적어도 삼량체화 수준을 개선하기 위하여, 이종 삼량체화 도메인(예를 들어, 폴드온 삼량체화 도메인; 문헌[Stevens et al. Science 303(5665):1866-1870, 2004])은 종종 폴리펩티드의 C 말단에 유전적으로 융합된다. 불행히도, 이종 삼량체화 도메인의 부가는 원치 않는 네오에피토프를 도입하고 종종 발현 수준을 감소시키거나, 또는 폴리펩티드의 4차 구조를 변경할 수 있다.
본 발명은 안정적인 재조합 인플루엔자 A 적혈구응집소(HA) 폴리펩티드로서, 인플루엔자 A 바이러스 HA의 HA1 및 HA2 도메인을 포함하고,
(a) 위치 355의 아미노산은 W이고;
(b) 위치 432의 아미노산은 I이고/이거나, 위치 380의 아미노산은 I인 아미노산 서열을 포함하고;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따르는 것인 HA 폴리펩티드를 제공한다.
본 발명에 따르면, 놀랍게도 안정적인 재조합 HA 폴리펩티드, 특히 가용성 HA 삼량체 폴리펩티드가 HA 폴리펩티드의 코어에서 특정 아미노산 돌연변이의 존재에 의해, 폴드온 도메인 또는 임의의 다른 이종 삼량체화 도메인의 부가 없이, 수득될 수 있음이 밝혀졌다.
따라서, 특정 양태에서, 본 발명은 재조합 인플루엔자 A 적혈구응집소(HA) 폴리펩티드에 관한 것으로, 인플루엔자 A 바이러스 HA의 HA1 및 HA2 도메인을 포함하고,
(a) 위치 355의 아미노산은 W로 돌연변이되고;
(b) 위치 432의 아미노산은 I로 돌연변이되고/되거나, 위치 380의 아미노산은 I로 돌연변이된 아미노산 서열을 포함하고;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따르는 것인 HA 폴리펩티드를 제공한다. 돌연변이는 "매립형" 돌연변이이기 때문에, 즉 이들 잔기의 측쇄가 단백질 표면에 노출되지 않으므로, HA 폴리펩티드의 항원성은 바뀌지 않을 것이다.
특정 구현예에서, 폴리펩티드는 위치 355의 아미노산, 특히 히스티딘(H)의 트립토판(W)으로의 돌연변이 및 위치 432 및/또는 380의 아미노산의 이소류신(I)으로의 돌연변이를 포함한다.
예를 들어, 위치 432의 아미노산의 I로의 돌연변이를 도입함으로써 위치 432의 아미노산 I와 조합하여, 예를 들어, 위치 355의 아미노산, 특히 H의 W로의 돌연변이를 도입함으로써 위치 355에 아미노산 잔기 W를 갖거나; 또는 예를 들어, 위치 432 및 380에서 I로의 돌연변이를 도입함으로써 위치 432의 I와 위치 380의 I의 조합을 갖는 본 발명의 HA 폴리펩티드는 이러한 아미노산 돌연변이가 없는 HA 폴리펩티드와 비교하여 포유동물 세포에서 증가된 발현 수준, 삼량체화 경향 증가(예를 들어, AlphaLISA, Octet 및 SEC에 의해 측정됨), 및/또는 증가된 수준의 열 안정성(예를 들어, 동적 주사 형광측정법/열량측정법(DSF/DSC)에 의해 측정됨)을 보여준다. 또한, 본 발명의 폴리펩티드에 대한 시험한 모든 항체의 결합 강도는 (Octet 및 ELISA에 의해 측정 시) 5 nM 미만이다. 이는 폴리펩티드가 (1차, 2차, 3차 및 4차 구조와 관련하여) 천연의 야생형 HA와 구조적으로 동일함을 분명히 보여준다. 신규한 HA 폴리펩티드는 나아가 링커, 태그, 또는 삼량체화 도메인 서열과 같은 임의의 인공 (이종) 서열의 존재를 필요로 하지 않는다.
특정 구현예에서, 폴리펩티드는 위치 355의 아미노산, 특히 히스티딘(H)의 트립토판(W)으로의 돌연변이 및 위치 432 및/또는 380의 아미노산의 이소류신(I)으로의 돌연변이를 포함한다.
특정 구현예에서, HA 폴리펩티드는
(a)
위치 388의 아미노산은 M이고/이거나;
(b)
위치 478의 아미노산은 I인 아미노산 서열을 포함한다.
이러한 돌연변이는 적어도 특정 HA 서브타입에서 HA 폴리펩티드의 안정성을 추가로 증가시키는 것으로 나타났다.
특정 구현예에서, 상기 HA 단량체는 프로테아제 절단 부위를 포함하지 않는다. 위에 기술된 바와 같이, (HA1 및 HA2의) 인플루엔자 HA0 단백질의 절단은 이의 활성에 요구되어, 숙주 엔도솜 막의 바이러스 막과의 융합을 초래하여 바이러스 게놈의 표적 세포로의 진입을 촉진한다. 특정 구현예에서, 본 발명의 폴리펩티드는 천연 프로테아제 절단 부위를 포함한다. 따라서, HA1 및 HA2에 걸쳐 이어지는 Arg(R) 내지 Gly(G) 서열(즉, 아미노산 위치 329 및 330)은 트립신 및 트립신 유사 프로테아제의 인식 부위이고, 전형적으로 적혈구응집소 활성화를 위하여 절단된다(도 1의 A). 특정 구현예에서, 프로테아제 절단 부위는 위치 329의 아미노산 잔기의 아르기닌(R) 또는 리신(K) 이외의 임의의 아미노산으로의 돌연변이에 의해 제거되었다. 특정 구현예에서, 위치 329의 아미노산 잔기는 아르기닌(R)이 아니다. 바람직한 구현예에서, 폴리펩티드는 위치 329의 아미노산의 글루타민(Q)으로의 돌연변이를 포함한다. 따라서, 특정 구현예에서, 본 발명의 폴리펩티드는 투여 후에 시험관 내 또는 생체 내에서 생산되는 동안 또는 그 이후에 분자의 추정상의 절단을 방지하기 위하여 절단 부위 녹아웃 돌연변이 R329Q를 포함한다. 이로써, 절단 부위 녹아웃 돌연변이, 예를 들어, R329Q 돌연변이는 낮은 pH에 의해 유발된 구조 변화에 대한 무감응을 보장하고 HA의 융합 전 구조를 보존한다.
본 발명에 따르면, HA1 및/또는 HA2 도메인은 인플루엔자 HA 폴리펩티드의 완전한(즉, 전장) HA1 및/또는 HA2 도메인을 포함할 수 있거나, 또는 이들은 HA1 및/또는 HA2 도메인의 적어도 일부를 포함할 수 있다.
분비된 (가용성) HA 폴리펩티드를 생성하기 위하여, 특정 구현예에서 HA 단량체는 절단된 HA2 도메인을 포함한다. 따라서, 특정 구현예에서 본 발명의 폴리펩티드 내의 HA 단량체는 막횡단 및 세포질 도메인을 포함하지 않는다. 특히, 특정 구현예에서, 폴리펩티드 단량체는 C 말단 단부에서 절단된 HA2 도메인을 포함한다. 따라서, 본 발명에 따른 절단된 HA2 도메인은 HA2 도메인의 C 말단 및/또는 N 말단 단부에서 하나 이상의 아미노산 잔기의 결실에 의해 전장 HA2 서열보다 짧다. 따라서, 본 발명은 또한 HA의 세포외 도메인(엑토도메인, ECD)을 포함하거나 이로 구성된 재조합 HA 폴리펩티드를 제공한다.
특정 구현예에서, 위치 515의 아미노산에 해당하는 아미노산으로 시작하는 HA2 도메인의 C 말단 부분이 결실되어, 실질적으로 전체 막횡단 및 세포질 도메인이 제거되었다.
특정 구현예에서, 엑토도메인의 C 말단의 하나 이상의 아미노산이 또한 결실되었다. 본 발명에 따르면, HA2 도메인의 더 큰 부분이 결실되는 경우에도, 안정적인 가용성 및 삼량체 HA 폴리펩티드가 제공될 수 있음이 밝혀졌다. 따라서, 특정 구현예에서, 위치 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, 512, 513, 또는 514의 아미노산 서열에서 시작하는 HA2 도메인의 C 말단 부분이 결실되어(위의 문헌[Winter et al.]에서 기술된 바와 같은 H3 넘버링을 따름) 세포에서 발현된 후 가용성 폴리펩티드를 생성하였다.
유사하게, HA1 도메인은 완전할 수 있거나(즉, 전장 HA1 도메인), 또는 이의 적어도 일부일 수 있다. 특정 구현예에서, 폴리펩티드는 절단된 HA1 도메인을 포함한다. HA1 도메인은 HA1 도메인의 N 및/또는 C 말단 단부에서 절단될 수 있다.
특정 구현예에서, HA 폴리펩티드는 신호 서열을 포함하지 않는다. 신호 서열(때때로 신호 펩티드, 표적화 신호, 국소화 신호, 국소화 서열, 전달 펩티드, 리더 서열 또는 리더 펩티드라고도 함)은 분비 경로로 향하는 새로 합성된 단백질의 대부분의 N 말단에 존재하는 짧은 펩티드(보통 16 내지 30개 아미노산의 길이)이다. 신호 서열은 세포가 단백질, 일반적으로 세포막으로 전위하도록 자극하는 기능을 한다. 많은 경우에, 신호 펩티드를 포함하는 아미노산은 최종 목적지에 도달하면 단백질로부터 절단된다. 인플루엔자 HA에서, 신호 서열은 전형적으로 (H3 넘버링에 따라 위치 -6에서 위치 10까지의 아미노산에 해당하는) 전장 HA0의 아미노산 서열의 처음 16개의 아미노산을 포함한다.
또한, 본 발명은 HA 폴리펩티드의 면역원성 단편을 제공한다. 특정 구현예에서, 헤드 도메인을 구성하는 HA1 도메인의 적어도 일부가 결실되어 본 발명의 HA 폴리펩티드의 면역원성 단편, 예컨대, 무 헤드 HA 폴리펩티드(즉, 스템 단독 폴리펩티드)를 제공할 수 있다.
본 발명의 폴리펩티드는 인플루엔자 A 바이러스의 인플루엔자 바이러스 적혈구응집소(HA)를 나타낸다(그로부터 유래된다). 전술한 바와 같이, 인플루엔자 A는 그룹 1과 그룹 2의 두 개의 주요 그룹으로 나눌 수 있는 여러 서브타입의 HA를 함유한다(도 2a). 본 발명의 폴리펩티드의 안정화 돌연변이는 인플루엔자 A의 모든 적혈구응집소 유형에 적용될 수 있다.
특정 구현예에서, HA1 및 HA2 도메인은 그룹 1 또는 그룹 2 인플루엔자 A 바이러스로부터 유래된다. 특정 구현예에서, HA1 및 HA2 도메인은 동일한 그룹 1 또는 그룹 2 바이러스로부터 유래된다. 특정한 다른 구현예에서, HA1 및 HA2 도메인은 상이한 그룹 1 또는 상이한 그룹 2 바이러스로부터 유래되거나, 또는 HA1 및 HA2 도메인은 상이한 그룹으로부터의 인플루엔자 A 바이러스로부터 유래된다. 예를 들어, HA2 도메인은 그룹 1 바이러스로부터 유래되고, HA2 도메인은 그룹 2 바이러스로부터 유래되거나, 또는 그 반대의 경우도 마찬가지이다. 특정 구현예에서, 헤드 도메인(즉, 헤드 도메인을 형성하는 HA1 도메인의 적어도 일부는 스템 도메인과 상이한 인플루엔자 바이러스(즉, 인플루엔자 HA 폴리펩티드의 스템 도메인을 형성하는 HA2 도메인의 일부)로부터 유래된다.
일부 특정 구현예에서, HA1 및/또는 HA2 도메인은 다음으로 구성된 군으로부터 선택되는 인플루엔자 A 바이러스로부터 유래된다: H1 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/캘리포니아/07/2009 또는 A/미시간/45/2015; H2 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/Env/MPU3156/2005; H5 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/붉은머리오리(Eurasian Wigeon)/MPF461/2007; H9 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/홍콩/1073/1999; H3 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 H/홍콩/1/1968 또는 A/파나마/2007/1999; H14 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/청둥오리(Mallard)/아스트라칸/263/1982; H7 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/청둥오리/네덜란드/12/2000; 및 H10 서브타입의 HA를 포함하는 인플루엔자 바이러스, 예를 들어, 인플루엔자 바이러스 A/닭/독일/N/1949. 당업자는 본 발명의 폴리펩티드가 또한 그룹 1 또는 그룹 2로부터의 다른 인플루엔자 A 바이러스 균주의 HA로부터 유래될 수 있음을 이해할 것이다.
특정 바람직한 구현예에서, HA 서브타입(즉, 그룹 1 또는 그룹 2)에 따라 HA 폴리펩티드 또는 이의 면역원성 단편은 결합 분자 CR9114, CR6261, CR8020 및/또는 MD3606에 결합한다. 따라서, (서열번호 2의 중쇄 가변 영역 및 서열번호 3의 경쇄 가변 영역을 포함하는) 항체 CR6261 및/또는 (서열번호 6의 중쇄 가변 영역 및 서열번호 7의 경쇄 가변 영역을 포함하는) 항체 CR9114, 및/또는 (서열번호 4의 중쇄 가변 영역 및 서열번호 5의 경쇄 가변 영역을 포함하는) 항체 CR8020 및/또는 다중도메인 항체 MD3606(서열번호 8)의 특정 에피토프를 제시하는 신규한 HA 폴리펩티드가 제공된다. 본 발명의 폴리펩티드는 단독으로 또는 다른 예방적 및/또는 치료적 처치와 조합하여 생체 내 투여될 때 인플루엔자 바이러스 중화 항체를 유도하는 데 사용될 수 있다.
특정 구현예에서, 본 발명의 HA 폴리펩티드 또는 이의 면역원성 단편은 예를 들어, 중합체, 리포솜, 바이로솜, 바이러스 유사 입자, 또는 자가 조립 나노입자와 같은 나노입자에 연결된다. 폴리펩티드는 나노입자와 조합되거나, 나노입자 내에 캡시드화되거나 나노입자에 접합(예를 들어, 공유적으로 연결되거나 흡착)될 수 있다.
본 발명은 추가로 전술한 바와 같이 적어도 2개의 HA 폴리펩티드 또는 이의 면역원성 단편을 포함하는 다량체 폴리펩티드를 제공한다.
특정 바람직한 구현예에서, 다량체 폴리펩티드는 삼량체이고, 전술한 바와 같이 3개의 HA 폴리펩티드 또는 이의 면역원성 단편을 포함한다.
따라서, 특정 구현예에서, 본 발명은 안정화된 재조합 안정화된 삼량체 인플루엔자 A 적혈구응집소(HA) 폴리펩티드 또는 이의 면역원성 단편을 제공하며, 상기 폴리펩티드는 3개의 HA 단량체를 포함하고, 상기 HA 단량체는 각각 인플루엔자 A 바이러스 HA의 HA1 및 HA2 도메인을 포함하고,
(a) 위치 355의 아미노산은 W이고;
(b) 위치 432의 아미노산은 I이거나, 또는 위치 432의 아미노산이 I이고 위치 380의 아미노산이 I인 아미노산 서열을 포함하며;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따른다.
위에 언급한 바와 같이, 본 발명에 따르면, 예를 들어, 위치 432의 아미노산의 I로의 돌연변이를 도입함으로써 위치 432의 아미노산 I와 조합하여, 예를 들어, 위치 355의 아미노산의 W로의 돌연변이를 도입함으로써 위치 355에 아미노산 잔기 W를 가짐으로써; 또는 예를 들어, 위치 432 및 380에서 I로의 돌연변이를 도입함으로써 위치 432의 I와 위치 380의 I의 조합을 가짐으로써, 안정적인 HA 삼량체의 발현 수준 및 삼량체화 둘 다가 증가될 수 있음이 밝혀졌다. 따라서, 본 발명의 HA 폴리펩티드는 이러한 아미노산 돌연변이가 없는 HA 폴리펩티드와 비교하여 포유동물 세포에서 증가된 발현 수준, 삼량체화 경향 증가(예를 들어, AlphaLISA, Octet 및 SEC에 의해 측정됨), 및/또는 증가된 수준의 열 안정성(예를 들어, 동적 주사 형광측정법/열량측정법(DSF/DSC)에 의해 측정됨)을 보여준다.
특정 구현예에서, 본 발명의 HA 폴리펩티드는 40℃에서 적어도 3일 동안 안정적이다.
본 발명은 또한 본 발명의 인플루엔자 HA 폴리펩티드 또는 이의 면역원성 단편을 암호화하는 핵산 분자를 제공한다. 유전자 코드의 축중(degeneracy)의 결과로 다수의 상이한 핵산 분자가 동일한 폴리펩티드를 암호화할 수 있다는 점은 당업자에 의해 이해된다. 당업자는 폴리펩티드가 발현될 임의의 특정한 숙주 유기체의 코돈 활용을 반영하기 위하여 통상적인 기법을 이용하여 기재된 폴리뉴클레오티드에 의하여 암호화된 폴리펩티드 서열에 영향을 주지 않는 뉴클레오티드 치환을 제조할 수 있다는 것도 알고 있다. 따라서, 달리 명시되지 않는 한, "아미노산 서열을 암호화하는 핵산 분자"는 서로의 축중 형태이면서 동일한 아미노산 서열을 암호화하는 모든 뉴클레오티드 서열을 포함한다.
특정 구현예에서, 인플루엔자 HA 폴리펩티드 또는 이의 면역원성 단편을 암호화하는 핵산 분자는 인간 세포와 같은 포유동물 세포에서의 발현을 위하여 코돈 최적화된다. 코돈 최적화의 방법은 공지되어 있고 이전에 기술된 바 있다(예를 들어, WO 96/09378).
본 발명은 추가로 위에서 정의된 바와 같은 재조합 HA 폴리펩티드 또는 이의 면역원성 단편을 제조하는 방법을 제공하며, 이는 원핵생물(예를 들어, 대장균) 또는 진핵생물 세포(예를 들어, 포유동물 세포, 예컨대, CHO 또는 PER.C6)에서 전술한 핵산 분자를 발현시키는 단계를 포함하고, 상기 방법은 상기 세포로부터 재조합 HA 폴리펩티드 또는 이의 면역원성 단편을 정제/단리하는 단계를 선택적으로 포함한다. 재조합 인플루엔자 HA 폴리펩티드 또는 이의 면역원성 단편은 본원에 기술된 기술을 포함하여, 당업자에게 재조합 폴리펩티드를 제조하기에 적합한 것으로 간주되는 임의의 기술에 따라 제조될 수 있다. 따라서, 본 발명의 폴리펩티드는 당해 분야에 공지되어 있는 표준 방법에 의해 DNA 서열로서 합성되고, 적합한 제한 효소와 당해 분야에 공지되어 있는 방법을 사용하여 클로닝되고, 이후에 시험관 내 또는 생체 내에서 발현될 수 있다. 본 발명의 HA 폴리펩티드 또는 이의 면역원성 단편을 암호화하는 뉴클레오티드 서열은 당업자에게 잘 알려진 기술에 따라 합성 및/또는 클로닝 및 발현될 수 있다. 예를 들어, 문헌[Sambrook, et al, Molecular Cloning, A Laboratory Manual, Vols. 1-3, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989)] 참조. 인플루엔자 백신을 제조하기 위한 재조합 DNA 기술의 사용은 몇 가지 이점을 제공한다. 여기에는 달걀에서 감염성 바이러스의 적응 및 통과 단계를 피하는 것과 더 안전하고 엄격하게 통제되는 조건 하에서 더 고도로 정제된 단백질의 생산이 포함된다. 더욱이, 바이러스 불활성화 단계를 포함할 필요가 없다. 임의의 적합한 클로닝 및 발현 시스템을 사용하여 본 발명의 HA 폴리펩티드를 재조합적으로 생산할 수 있다.
바람직한 구현예에서, 폴리펩티드 또는 이의 면역원성 단편은 포유동물 세포에서 생산된다. 특정 구현예에서, 폴리펩티드는 적합한 세포(예를 들어, 포유류 세포)에서 발현될 때 글리코실화된다. 따라서, 폴리펩티드는 1개 이상의 천연 및/또는 도입된(즉, 비 천연) 글리코실화 모티프를 함유할 수 있다.
적혈구응집소 서열은 중합효소 연쇄 반응(PCR) 또는 역전사효소 PCR, 역공학과 같은 당해 분야에 공지된 표준 재조합 방법에 의해 생산될 수 있거나, 또는 DNA가 합성될 수 있다. PCR의 경우, 공개적으로 이용 가능한 데이터베이스에서 이용할 수 있는 적혈구응집소 뉴클레오티드 서열을 사용하여 프라이머를 준비할 수 있다. 폴리뉴클레오티드 구축물은 PCR 카세트로부터 조립될 수 있고 숙주 세포에서 증식을 위한 선택 가능한 마커를 함유하는 벡터로 순차적으로 클로닝될 수 있다. 그런 다음, 재조합 벡터는 주사, 형질감염 또는 전기천공 또는 기타 방법(예를 들어, 인산칼슘 형질감염, DEAE-덱스트란 매개 형질감염, 양이온성 지질 매개 형질감염, 전기천공)에 의해 숙주 세포 내로 도입될 수 있다. 리포펙타민(Invitrogen, 미국 캘리포니아 주 칼스배드 소재)과 같은 상업적인 형질감염 시약도 이용 가능하다.
HA 폴리펩티드 또는 이의 면역원성 단편은 음이온 및/또는 양이온 교환 크로마토그래피, 친화도 크로마토그래피를 포함하는 당해 분야에 공지된 방법에 의해 재조합 세포 배양물로부터 회수 및 단리/정제될 수 있다. SDS-PAGE와 같은 기술을 사용하여 이러한 분리/정제 기술에서 용리된 단백질 분획을 분석할 수 있다. 이러한 방법은 당업자에게 잘 알려져 있으므로, 여기에서는 상세하게 제시하지 않을 것이다. 또한, 정제된 폴리펩티드를 당해 분야에 공지된 분광학적 방법(예를 들어, 원편광 분광법, 푸리에 변환 적외 분광법 및 NMR 분광법 또는 X-선 결정학)에 의해 분석하여, 헬릭스 및 베타 시트와 같은 요망되는 구조의 존재를 조사할 수 있다. ELISA, AlphaLISA, 바이오층 간섭법(Octet) 및 FACS 등을 사용하여, CR6261 및/또는 CR9114와 같은 광범위 중화 항체에 대한 본 발명의 폴리펩티드의 결합을 조사할 수 있다. 따라서, 올바른 형태를 갖는 본 발명에 따른 폴리펩티드가 선택될 수 있다. 삼량체 함량은 예를 들어, 비 환원 조건 하에서의 SDS 겔 전기영동, CR6261 및/또는 CR9114와 같은 광범위 중화 항체의 항체 Fab 단편의 존재 하에서의 크기 배제 크로마토그래피 및 차등 표지된 항체를 이용한 AlphaLISA를 이용하여 분석할 수 있다. 폴리펩티드의 안정성은 온도 스트레스, 냉동-해동 주기, 증가된 단백질 농도, 또는 교반 후에 위에 기술된 바와 같이 평가할 수 있다. 폴리펩티드의 용융 온도는 시차 주사 형광측정법(DSF)에 의해 추가로 평가할 수 있다.
일부 구현예에서 본 발명은 서열번호 10, 서열번호 12, 서열번호 14, 서열번호 16, 서열번호 18, 서열번호 20, 서열번호 22, 서열번호 24, 서열번호 26, 서열번호 28, 서열번호 33, 서열번호 34, 서열번호 35, 서열번호 36, 서열번호 38, 서열번호 40, 서열번호 42, 서열번호 44, 서열번호 47, 서열번호 50, 서열번호 51 및 서열번호 52로 구성된 군으로부터 선택되는 인플루엔자 HA 아미노산 서열 중 임의의 하나 또는 이러한 아미노산 서열과 적어도 약 40% 또는 50% 또는 60% 또는 65% 또는 70% 또는 75% 또는 80% 또는 85% 또는 90% 또는 95% 또는 98% 또는 99%의 동일성을 갖는 이의 임의의 변이체 또는 단편으로부터 유래되거나, 이를 포함하거나, 이로 구성되는 재조합 인플루엔자 HA 폴리펩티드를 제공하며, 여기서, 인플루엔자 HA 폴리펩티드는 위치 355에 트립토판(W) 및 위치 432 및/또는 380에 이소류신(I)을 포함하고, 아미노산 넘버링은 서열번호 1의 서열에 기초하거나, 예를 들어 서열번호 1에 대한 HA 아미노산 서열의 정렬에 의해 결정된 바와 같이, 이러한 아미노산 위치에 상응하는 아미노산 위치에서 이루어진다.
특정 구현예에서 본 발명은 서열번호 10의 아미노산 잔기 18 내지 518, 서열번호 12의 아미노산 잔기 18 내지 518, 서열번호 14의 아미노산 잔기 16 내지 514, 서열번호 16의 아미노산 잔기 17 내지 516, 서열번호 18의 아미노산 잔기 19 내지 512, 서열번호 20의 아미노산 잔기 17 내지 521, 서열번호 22의 아미노산 잔기 17 내지 521, 서열번호 24의 아미노산 잔기 18 내지 523, 서열번호 26의 아미노산 잔기 19 내지 515, 서열번호 28의 아미노산 잔기 17 내지 515, 서열번호 33의 아미노산 잔기 17 내지 521, 서열번호 34의 아미노산 18 내지 518, 서열번호 35의 아미노산 18 내지 518, 서열번호 36의 아미노산 18 내지 517, 서열번호 38의 아미노산 18 내지 518, 서열번호 40의 아미노산 17 내지 521, 서열번호 42의 아미노산 17 내지 521, 서열번호 44의 아미노산 17 내지 521, 서열번호 47의 아미노산 17 내지 519, 서열번호 50의 아미노산 17 내지 521, 서열번호 51의 아미노산 19 내지 515, 또는 서열번호 52의 아미노산 17 내지 514로부터 유래되거나, 이를 포함하거나, 이로 구성되는 재조합 인플루엔자 HA 폴리펩티드를 제공한다.
특정 구현예에서, HA 폴리펩티드는 서열번호 10, 서열번호 12, 서열번호 14, 서열번호 16, 서열번호 18, 서열번호 20, 서열번호 22, 서열번호 24, 서열번호 26, 서열번호 28, 서열번호 33, 서열번호 34 or 서열번호 35, 서열번호 36, 서열번호 38, 서열번호 40, 서열번호 42, 서열번호 44, 서열번호 47, 서열번호 50, 서열번호 51 및 서열번호 52로부터 유래되거나, 이를 포함하거나, 이로 구성된 아미노산 서열을 포함한다.
본 발명은 추가로 본 발명의 HA 폴리펩티드 또는 이의 면역원성 단편을 암호화하는 핵산 분자를 포함하는 벡터에 관한 것이다.
특정 구현예에서, 벡터는 인간 재조합 아데노바이러스이다. 따라서, 본 발명은 본 발명에 따른 HA 폴리펩티드 또는 이의 면역원성 단편을 암호화하는 핵산 분자를 포함하는 재조합 아데노바이러스 벡터를 제공한다. 재조합 아데노바이러스 벡터는 막결합 HA를 암호화할 수 있고, 따라서 HA2 도메인을 포함하고, 막횡단 및 세포질 도메인을 포함하는 HA 폴리펩티드를 암호화할 수 있다. 아데노벡터는 또한 가용성 폴리펩티드를 암호화할 수 있고, 따라서 절단된 HA2 도메인을 포함하는 HA 폴리펩티드를 암호화할 수 있다.
재조합 아데노바이러스 벡터의 제조는 당해 분야에 잘 알려져 있다. 본 명세서에서 사용되는 바와 같이, 아데노바이러스에 대한 '재조합'이라는 용어는 아데노바이러스가 인간의 손에 의해 변형되었고, 예컨대, 아데노바이러스가 그 안에 활발하게 클로닝된 변경된 말단부를 가지고/가지거나 이종 유전자를 포함함, 즉, 자연 발생적인 야생형 아데노바이러스가 아님을 내포한다. 특정 구현예에서, 본 발명에 따른 아데노바이러스 벡터는 바이러스 복제에 요구되는 아데노바이러스 게놈의 E1 영역, 예컨대, E1a 영역 및/또는 E1b 영역의 적어도 하나의 필수적인 유전자 기능이 결핍되어 있다. 특정 구현예에서, 본 발명에 따른 아데노바이러스 벡터는 필수적이지 않은 E3 영역의 적어도 일부가 결핍되어 있다. 특정 구현예에서, 벡터는 E1 영역의 적어도 하나의 필수 유전자 기능 및 필수적이지 않은 E3 영역의 적어도 일부가 결핍되어 있다. 아데노바이러스 벡터는 "다중적으로 결핍"될 수 있는데, 이는 아데노바이러스 벡터가 아데노바이러스 게놈의 둘 이상의 영역들 각각에서 하나 이상의 필수적인 유전자 기능이 결핍되어 있음을 의미한다. 예를 들면, 위에서 언급된 E1-결핍된 또는 E1-, E3-결핍된 아데노바이러스 벡터는 E4 영역의 적어도 하나의 필수적인 유전자 및/또는 E2 영역(예컨대, E2A 영역 및/또는 E2B 영역)의 적어도 하나의 필수적인 유전자가 더 결핍되어 있을 수 있다. 아데노바이러스 벡터, 이의 구축 방법 및 이의 증식 방법은 당해 분야에 잘 알려져 있고, 예를 들어, 미국 특허 번호 5,559,099, 5,837,511, 5,846,782, 5,851,806, 5,994,106, 5,994,128, 5,965,541, 5,981,225, 6,040,174, 6,020,191, 및 6,113,913에 기술되어 있다.
특정 구현예에서, 아데노바이러스는 혈청형 26의 인간 아데노바이러스이다.
본 발명은 본 발명에 따른 HA 폴리펩티드, 이의 면역원성 단편, 핵산, 및/또는 벡터, 및 약제학적으로 허용 가능한 담체를 포함하는 면역원성 조성물을 추가로 제공한다. 본 발명은 특히 치료적 유효량의 본 발명의 폴리펩티드, 면역원성 단편, 핵산, 및/또는 벡터를 포함하는 약제학적 조성물에 관한 것이다. 약제학적 조성물은 약제학적으로 허용 가능한 담체를 더 포함한다. 본 발명의 문맥에서, 용어 "약제학적으로 허용 가능한"은 담체가, 사용되는 투여량 및 농도에서, 이들이 투여되는 대상체에게 원치 않는 또는 유해한 효과를 야기하지 않을 것을 의미한다. 이러한 약제학적으로 허용 가능한 담체 및 부형제는 당해 분야에 잘 알려져 있다(문헌[Remington's Pharmaceutical Sciences, 18th edition, A. R. Gennaro, Ed., Mack Publishing Company [1990]]; 문헌[Pharmaceutical Formulation Development of Peptides and Proteins, S. Frokjaer and L. Hovgaard, Eds., Taylor & Francis [2000]]; 및 문헌[Handbook of Pharmaceutical Excipients, 3rd edition, A. Kibbe, Ed., Pharmaceutical Press [2000]] 참조). 용어 "담체"는 폴리펩티드, 핵산, 및/또는 벡터와 함께 투여되는 희석제, 부형제, 또는 비히클을 지칭한다. 염수 용액 및 덱스트로스 및 글리세롤 수용액이 특히 주사 가능한 용액을 위한 액체 담체로서 사용될 수 있다.
본 발명은 또한 의약으로서의 사용을 위한, 본 명세서에서 기술된 HA 폴리펩티드, 면역원성 단편, 핵산, 및/또는 벡터에 관한 것이다. 본 발명은 특히 인플루엔자 바이러스에 대해, 특히 인플루엔자 바이러스의 HA 분자에 대해, 바람직하게는 중화 항체를 이끌어내는 것을 포함하여, 면역 반응을 유도하는 데 사용하기 위한, 본 명세서에서 기술된 HA 폴리펩티드, 핵산, 및/또는 벡터에 관한 것이다. 바람직한 구현예에서, 본 발명은 인플루엔자 백신으로서의 사용을 위한, 본 명세서에서 기술된 HA 폴리펩티드, 면역원성 단편, 핵산, 및/또는 벡터에 관한 것이다.
본 발명은 또한 필요로 하는 대상체에서 인플루엔자 A 바이러스에 대한 면역 반응의 유도 방법, 특히 항체를 이끌어내는 방법에 관한 것으로, 방법은 상기 대상체에게 위에서 기술된 HA 폴리펩티드, 면역원성 단편, 핵산 분자 및/또는 벡터를 투여하는 단계를 포함한다. 본 발명에 따른 대상체는 바람직하게는 인플루엔자 바이러스에 감염될 수 있거나, 다르게는 인플루엔자 바이러스에 대한 면역 반응의 유도로부터 이익을 얻을 수 있는 포유류이며, 예를 들어, 이러한 대상체는 설치류, 예를 들어, 마우스, 페럿, 애완동물 또는 가축, 또는 비 인간 영장류 또는 인간이다. 바람직하게는, 대상체는 인간 대상체, 예컨대, 인플루엔자 질환 감염 위험이 있는 것으로 확인된 사람이다.
특정 구현예에서, 본 발명의 HA 폴리펩티드, 면역원성 단편, 핵산 분자 및/또는 벡터는 보강제와 조합하여 투여된다. 보강제는 본 발명의 폴리펩티드, 핵산 분자 및/또는 벡터의 투여 전에, 투여와 동시에, 또는 투여 후에 투여될 수 있다. 적합한 보강제의 예에는 알루미늄 염, 예를 들어, 수산화알루미늄 및/또는 인산알루미늄; 스쿠알렌-물 에멀젼, 예를 들어, MF59를 비롯한 오일-에멀젼 조성물(또는 수중유 조성물)(예를 들어, WO 90/14837 참조); 사포닌 제형, 예를 들어, QS21 및 면역자극 복합체(ISCOMS)(예를 들어, US 5,057,540; WO 90/03184, WO 96/11711, WO 2004/004762, WO 2005/002620 참조); 예로서, 모노포스포릴 지질 A(MPL), 3-O-데아실화 MPL(3dMPL), CpG-모티프 함유 올리고뉴클레오티드, ADP-리보실화 박테리아 독소 또는 이의 돌연변이체, 예를 들어, E. 콜라이 이열성 장독소 LT, 콜레라 독소 CT, 백일해 독소 PT 또는 파상풍 톡소이드 TT, 매트릭스 M, 또는 이의 조합이 있는, 박테리아 또는 미생물 유도체가 포함된다. 또한, 공지되어 있는 면역증강 기술, 예를 들어, 본 발명의 폴리펩티드를 면역 반응을 향상시키는 것으로 당해 분야에 공지되어 있는 단백질(예를 들어, 파상풍 톡소이드, CRM197, rCTB, 박테리아 플라젤린 또는 기타의 것)에 융합시키는 것, 또는 폴리펩티드를 바이로솜에 포함시키는 것, 또는 이의 조합이 사용될 수 있다. 또한, 예를 들어, 동일한 아데노벡터에 의해 공동 전달되거나 암호화되는 유전적 보강제가 사용될 수 있다.
본 발명에 따른 HA 폴리펩티드, 면역원성 단편, 핵산 분자, 및/또는 벡터의 투여는 표준 투여 경로를 사용하여 수행될 수 있다. 비 제한적인 예에는 비경구 투여, 예를 들어, 정맥내, 피내, 경피, 근육내, 피하 등 또는 점막 투여, 예를 들어, 비내, 구강 등이 포함된다. 당업자는 면역 반응을 유도하기 위하여 본 발명에 따른 폴리펩티드, 핵산 분자 및/또는 벡터를 투여하는 다양한 가능성을 결정할 수 있을 것이다.
본 발명은 추가로 필요로 하는 대상체에서 인플루엔자 바이러스 질환의 예방 및/또는 치료, 바람직하게는 예방 방법을 제공하며, 방법은 상기 대상체에게 본 명세서에 기술된 치료적 유효량의 HA 폴리펩티드, 면역원성 단편, 핵산 분자 및/또는 벡터를 투여하는 단계를 포함한다. 치료적 유효량은 인플루엔자 바이러스에 의한 감염으로부터 야기되는 질환 또는 병태를 예방, 완화 및/또는 치료하는 데 유효한 폴리펩티드, 면역원성 단편, 핵산 분자, 및/또는 벡터의 양을 지칭한다. 예방은 인플루엔자 바이러스의 확산의 억제 또는 감소, 또는 인플루엔자 바이러스에 의한 감염과 관련된 증상 중 하나 이상의 발병, 발생 또는 진행의 억제 또는 감소를 포함한다. 본 명세서에서 사용되는 바와 같이, 개선은 인플루엔자 감염의 가시적이거나 또는 감지 가능한 질환 증상, 바이러스 혈증, 또는 임의의 다른 측정 가능한 징후의 감소를 의미할 수 있다.
치료를 필요로 하는 대상체는 인플루엔자 바이러스 감염에 기인한 병태로 이미 고통 받는 대상체뿐만 아니라, 인플루엔자 바이러스 감염이 예방되어야 하는 대상체도 포함한다. 따라서, 본 발명의 폴리펩티드, 면역원성 단편, 핵산 및/또는 벡터는 미경험 대상체, 즉, 인플루엔자 바이러스 감염에 의해 야기되는 질환을 갖지 않거나, 인플루엔자 바이러스 감염으로 감염된 적이 없으며 현재 감염되지 않은 대상체, 또는 인플루엔자 바이러스에 이미 감염된 적이 있는 대상체에게 투여될 수 있다.
일 구현예에서, 예방 및/또는 치료는 인플루엔자 바이러스 감염에 민감한 환자군에서 표적화될 수 있다. 이러한 환자군은, 예를 들어 노인(예를 들어, 50세 이상, 60세 이상, 그리고 바람직하게는 65세 이상), 어린이(예를 들어, 5세 이하, 1세 이하), 입원 환자, 면역손상 대상체, 및 항바이러스 화합물 치료를 받았으나 부적절한 항바이러스 반응을 나타낸 환자를 포함하나, 이에 한정되지 않는다.
본 발명의 폴리펩티드, 면역원성 단편, 핵산 분자 및/또는 벡터는 하나 이상의 다른 활성제, 예컨대, 대안적인 인플루엔자 백신, 단일클론 항체, 항바이러스제, 항박테리아제, 및/또는 면역조절제와 병용하여 대상체에게 투여될 수 있다. 하나 이상의 다른 활성제는 인플루엔자 바이러스 질환의 치료 및/또는 예방에 유리할 수 있거나 인플루엔자 바이러스 질환과 관련된 증상 또는 병태를 개선할 수 있다. 일부 구현예에서, 하나 이상의 다른 활성제는 진통제, 해열 약제 또는 호흡을 완화하거나 보조하는 치료제이다.
또한, 본 발명의 HA 폴리펩티드 또는 이의 단편은 연구 도구로서, 진단 도구로서, 또는 항체 시약 또는 치료 항체의 생산을 위한 표적으로서 사용될 수 있다. 예를 들어, 일부 구현예에서 HA 폴리펩티드는 예를 들어, ELISA 분석, 비아코어(Biacore)/SPR 결합 분석 및/또는 당해 분야에 공지된 항체 결합에 대한 임의의 기타 분석에서 항-HA 항체의 분석, 이의 결합 및/또는 이의 역가의 측정을 위한 분석물로서 유용할 수 있다. 또 다른 예로서, 본 발명의 HA 폴리펩티드는 항-HA 항체의 효능을 분석 및/또는 비교하는 데 사용될 수 있다.
본 발명의 HA 폴리펩티드 또는 이의 단편은 또한 치료 항체 및/또는 연구 도구로서 또는 임의의 기타 원하는 용도에 사용될 수 있는 항체의 생성에 유용할 수 있다. 예를 들어, 본 발명의 HA 폴리펩티드는 연구 도구 및/또는 치료제로서 사용하기 위한 HA 단백질에 대한 항체를 얻기 위하여 비 인간 동물의 면역화에 사용될 수 있다. 그러면 단일클론성 또는 다클론성일 수 있는 이러한 항체 및/또는 이러한 항체를 생성하는 세포를 이러한 동물로부터 얻을 수 있다.
진단 도구로서 사용하기 위한 본 발명의 폴리펩티드는 주어진 분석에 적합한 임의의 검출 기술에 유용한 태그를 포함할 수 있다. 사용된 태그는 사용된 특정 검출/분석/진단 기술 및/또는 방법에 따라 달라질 것이다. 방법은 용액에서 수행될 수 있거나, 또는 본 발명의 폴리펩티드(들)는 담체 또는 기질, 예를 들어, 미세역가 플레이트(예를 들어, ELISA용), 막 및 비드 등에 결합되거나 부착될 수 있다.
본 발명을 다음의 실시예 및 도면에서 추가로 설명한다. 실시예는 본 발명의 범주를 어떠한 방식으로든 제한하고자 하지 않는다.
실시예
실시예 1:
가용성 HA 폴리펩티드 - 본 발명의 폴리펩티드의 구조 및 설계 요소
인플루엔자 A 바이러스 적혈구응집소(HA0)의 엑토도메인을 나타내는 가용성 폴리펩티드를 제조하기 위하여, HA는 고유의 막횡단 및 세포질 도메인 없이 발현될 필요가 있다. 안정적인 삼량체형 가용성 야생형(WT) HA의 발현은 종종 포유동물 세포에서 매우 빈약하다. 적어도 삼량체화 수준을 개선하기 위하여, 폴드온 삼량체화 도메인은 종종 폴리펩티드의 C 말단에 유전적으로 융합된다. 불행히도, 폴드온 도메인의 부가는 원치 않는 네오에피토프를 도입하고 종종 발현 수준을 감소시키거나, 또는 폴리펩티드의 구조를 변경할 수 있다. 본 발명에 따르면, 가용성의 안정적인 HA 삼량체의 발현 및 삼량체화 수준은 폴드온 또는 임의의 기타 비 천연 삼량체화 서열의 부가 없이 HA 폴리펩티드의 코어에, 특히 아미노산 위치 355 및 432에서, 또는 아미노산 위치 355 및 380 및 432에서, 특정 아미노산 돌연변이를 도입함으로써 증가될 수 있음이 밝혀졌다. 본 발명의 HA 단량체에서 아미노산 위치의 넘버링에 대해서는 문헌[Winter et al. 1981]에 의한 H3 넘버링이 사용됨을 유의한다(위 참조). 따라서, 본 발명의 HA 폴리펩티드 단량체에서 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 (서열번호 1의 아미노산 서열을 갖는) 기준 H3N2 균주 A/아이치/2/68로부터의 HA 내 아미노산 위치의 넘버링에 따른다.
본 발명에 따른 주요 돌연변이의 주요 구조적 요소 및 위치는 인플루엔자 A H1A/캘리포니아/07/2009 균주의 HA에서 도 1의 A에 도시되어 있다(도 1의 A). 도시된 바와 같이, HA 단량체는 절단된 HA2 도메인(특히 HA2 도메인은 아미노산 위치 514 다음에 절단됨(즉, HA2 도메인의 C 말단 부분이 위치 515의 아미노산으로부터 시작하여 결실됨)을 포함하여, 막횡단 및 세포질 도메인을 결실시키고 HA의 가용성 엑토도메인을 생성한다(도 1의 B).
본 발명의 폴리펩티드는 위치 329(도 1의 B)에서 천연 일염기성 절단 부위인 아미노산 아르기닌(R)의 예를 들어, 글루타민(Q)으로의 돌연변이에 의해 프로테아제 절단에 대해 저항성이 되도록 만들어질 수 있다. 천연의 전장 HA와 대조적으로, R329Q 돌연변이를 포함하는 폴리펩티드는 세린 프로테아제(예를 들어, 트립신)에 의해 절단될 수 없다. HA의 절단은 단백질이 막 융합 및 바이러스 진입에 필요한 구조적 변화를 겪을 수 있도록 한다.
실시예 2:
상이한 서브타입에서 야생형 HA와 비교하여 가용성의 안정화된 HA의 발현
이 실시예에서, 그룹 1 및 그룹 2 둘 다로부터의 인플루엔자 바이러스로부터의 몇몇 HA를 선택하고 안정화된 가용성 삼량체 HA 폴리펩티드로서 발현시키고, 이들의 각각의 야생형 가용성 HA 엑토도메인(즉, 막횡단 및 세포질내 도메인이 없음)과 비교하였다. 본 발명에 따르면, 위치 355의 트립토판(W) 및 위치 380 및 432의 이소류신(I)은, 이들 아미노산이 HA 아미노 서열에 아직 존재하지 않은 경우, 인간에서의 8개의 가장 많은 순환 서브타입(도 2a)을 포함한 5개의 상이한 그룹 1 균주 및 5개의 상이한 그룹 2 균주의 HA의 아미노산 서열에 도입되었다. 또한, 일부 폴리펩티드의 위치 388에서 A-나선의 상단에 메티오닌(M)이 도입되었다. A/청둥오리/네덜란드/12/200(UFV181146) 및 A/닭/독일/N/1949(UFV181147)로부터 유래된 폴리펩티드를 제외하고 위치 478에서 이소류신이 도입되거나 WT 서열에 이미 존재하는 경우 유지되었다. Expi293F 배양 상청액에서 본 발명의 폴리펩티드의 발현 수준 및 삼량체화를 본 발명의 돌연변이가 없는 각각의 가용성 WT 폴리펩티드와 비교하였다.
표 1은 제조된 본 발명에 따른 폴리펩티드를 나타낸다.
[표 1]
본 발명의 폴리펩티드
+는 상기 위치에서 상기 아미노산의 존재를 의미하고; 빈 세포는 상기 아미노산의 부재(즉, 야생형 아미노산 잔기의 존재)를 의미한다.
도 2 및 표 1에 열거된 폴리펩티드를 암호화하는 DNA 단편을 합성하고(Genscript) pcDNA2004 발현 벡터(증진된 CMV 프로모터가 있는 변형된 pcDNA3 플라스미드)에 클로닝하였다. 본 발명의 폴리펩티드는 부위 특이적 비오틴화, 스크리닝 및 정제 목적을 위한 C 말단 링커-소르타제(Sortase)-링커-His 태그를 포함하였고, 진핵생물 Expi293F 현탁 세포주에서 마이크로 규모(200 μL)로 생산되었다. 야생형(WT) 전장(FL) HA 폴리펩티드는 스크리닝 목적을 위한 링커-His 태그를 함유하였다.
세포를 ExpiFectamine 293 형질감염 키트(Gibco , ThermoFisher Scientific)를 이용하여 2.5E+06vc/mL의 세포 밀도에서 96-하프 딥 웰 플레이트(System Duetz)에서 산업 등급의 DNA(0.01 EU/μg 이하의 내독소 수준 및 90% 이상의 초나선 함량)로 일시적으로 형질감염시키고, 37℃, 250 rpm, 8% CO2 및 75%의 습도에서 Expi293 발현 배지(Gibco, ThermoFisher Scientific)를 함유하는 진탕 플라스크에서 인큐베이션하였다. 분비된 폴리펩티드를 함유하는 세포 배양 상청액을 제3일에 수확하고, 원심분리(400xg에서 10분) 후 여과(96웰 필터 플레이트, 0.22 μm PVDF 막, Corning)하여 정화하였다.
수확된 배양 상청액의 발현된 가용성 HA 폴리펩티드의 수준을 OCTET 플랫폼(ForteBio)을 이용한 바이오층 간섭법에 의해 평가하였다. 요컨대, 항-HIS(HIS2) 바이오센서(ForteBio)를 사용하여, 정제된 폴리펩티드 UFV180436의 잘 정의된 기준 배치의 희석물 계열의 결합 이동을 측정하여 표준 곡선을 확립하였다. 이어서, 본 발명의 폴리펩티드를 함유하는 (동역학 완충액(ForteBio)에) 사전 희석된 세포 배양 상청액의 결합 이동을 측정하고, 폴리펩티드의 농도를 확립된 표준 곡선을 사용하여 계산하였다.
Expi293F 세포 배양 수확물에서 발현된 폴리펩티드의 존재 및 이의 4차 구조(폴리펩티드가 단량체, 삼량체 또는 다량체인지를 나타냄)는 BEH 200A 컬럼(Waters, 주입 부피 40 μL, 유속 0.35 mL/분)을 구비한 Vanquish 시스템(ThermoFisher Scientific)을 사용하여 초고성능 액체 크로마토그래피(UHPLC)에서 분석적 크기 배제 크로마토그래피(SEC)에 의해 평가하였다. 용리는 Helios 광산란 검출기(Wyatt Technologies)에 의해 모니터링하였다. SEC 프로파일을 Astra 6 소프트웨어 패키지(Wyatt Technology)로 분석하였다.
결과 및 결론
상이한 균주의 야생형 HA에서 위치 355의 트립토판 및 위치 380 및/또는 432의 이소류신의 도입은 OCTET에 의해 결정된 바와 같이 본 발명의 테스트한 모든 폴리펩티드에 대해 발현의 증가를 초래하였다(도 2b).
조 세포 배양 상청액의 SEC 분석은 본 발명의 폴리펩티드에 안정화 돌연변이를 도입할 때 모든 가용성의 안정화된 HA에 대해, 각각의 야생형 HA 엑토도메인에 대해 관찰된 삼량체 피크보다 더 긴 6분에서 7분 사이의 체류 시간에 뚜렷한 삼량체(T) 피크가 나타남을 보여주었다(도 2c). 상이한 인플루엔자 HA 서브타입 간의 체류 시간의 차이는 글리코실화 수준 및 복잡성의 차이로 인한 것일 수 있음을 주목한다.
종합하면, 데이터는 본 발명의 HA 폴리펩티드로의 돌연변이 355W, 380I 및/또는 432I의 도입은 안정적인 가용성 삼량체 HA의 증가된 발현 및 형성을 초래함을 확인시켜 준다.
실시예 3:
폴드온 삼량체화 도메인을 함유하는 야생형 HA와 비교한 정제된 삼량체 전장 HA의 시험관 내 특성화
중요한 안정화 돌연변이 355W, 380I 및/또는 432I의 기여를 추가로 특성화하기 위하여, 돌연변이를 H1 균주 A/캘리포니아/07/2009(UFV181009), A/미시간/45/2015((UFV181091), 및 H3 균주 A/홍콩/1/1968(UFV180660) 및 A/인디애나/11/2011(UFV181099)로부터 유래된 HA 엑토도메인 폴리펩티드(즉, TM 및 IC 도메인 제외)에 도입하고, 폴드온 삼량체화 도메인을 함유하는 야생형(WT) HA 엑토도메인과 비교하였다(폴드온 삼량체화 도메인이 결여된 UFV4239(서열번호 29)는 제외). 폴리펩티드는 표 2에 나타낸 바와 같은 아미노산을 포함하였다. 모든 폴리펩티드는 정제 및 스크리닝 목적을 위한 His 태그를 추가로 포함하고, ExpiCHO 세포에서 생산된 후, 정제 및 특성화되었다.
본 발명의 폴리펩티드를 암호화하는 DNA 단편을 실시예 2에 기술된 바와 같이 합성하였다. 폴리펩티드를 ExpiCHO 현탁 세포(350 mL 규모)에서 생산하였고, 제조업체의 프로토콜에 따라 ExpiFectamine 형질감염 시약(Gibco, ThermoFisher Scientific)을 사용하여 각각의 산업 등급의 DNA로 일시적 형질감염에 의해 ExpiCHO 발현 배지에서 배양하였다. ExpiFectamine CHO 증진제 및 ExpiCHO 피드(Gibco, ThermoFisher Scientific)를 제조업체의 프로토콜에 따라 형질감염 1일 후 세포 배양물에 첨가하였다. ExpiCHO 형질감염된 세포 현탁액을 32℃, 5% CO2에서 인큐베이션하고, 분비된 폴리펩티드를 함유하는 배양 상청액을 7 내지 11일 사이에 수확하였다. 배양 상청액을 원심분리에 이어, 0.2 μm 보틀 탑 필터(Corning)를 이용한 여과에 의해 정화하였다.
수확된 배양 상청액으로부터, 본 발명의 his 태그가 붙은 폴리펩티드 및 폴드온 삼량체화 도메인을 함유하는 각각의 야생형 균주를 
KTA Avant 25 시스템(GE Healthcare Life Sciences)을 사용하여 2단계 프로토콜에 따라 정제하였다. 먼저, 사전 패킹된 cOmplete His-태그 정제 컬럼(Roche)을 사용하여 고정화된 금속 친화도 크로마토그래피를 수행하고, 1 mM의 이미다졸로 세척하고, 300 mM의 이미다졸로 용리시켰다. 두 번째로, HiLoad Superdex 200 pg 26/600 컬럼(GE Healthcare Life Sciences)을 사용하여 크기 배제 크로마토그래피를 수행하였다. 삼량체 피크 분획을 모아 냉동하고 -80℃에서 보관하였다(1개월 및 6개월).
본 발명의 정제된 폴리펩티드의 삼량체 함량을 실시예 2에 기술된 바와 같이 초고성능 액체 크로마토그래피(UHPLC)에서 분석적 SEC에 의해 평가하였다. 각각의 정제된 폴리펩티드 20 μg을 주입하고, 컬럼에 흘려보냈다.
정제된 폴리펩티드의 열 안정성을 6 μg의 폴리펩티드 용액에 첨가된 Sypro Orange 염료(ThermoFisher Scientific)의 형광 발광을 모니터링함으로써 시차 주사 형광측정법(DSF)에 의해 결정하였다. 온도가 25℃에서 95℃(시간당 60℃)로 점진적으로 증가하면, 폴리펩티드가 펼쳐지고 형광 염료가 노출된 소수성 잔기에 결합하여 특징적인 발광 변화를 초래한다. 용융 곡선을 ViiA7 실시간 PCR 장치(Applied BioSystems)를 사용하여 측정하고, Tm50 값을 Spotfire suite(Tibco Software Inc.)에 의해 계산하였다. Tm50 값은 단백질의 50%가 펼쳐지는 온도를 나타내며, 따라서 폴리펩티드의 온도 안정성에 대한 척도이다.
정제된 폴리펩티드의 3차원 입체형태를 ELISA에서 항원성을 테스트함으로써 평가하였다(항체 결합의 EC50 값). 이를 위하여, 폴리펩티드를 10 nM의 농도로 코팅하고, 출발 농도로서 70 nM을 사용하여, 일련의 단일클론 항체(mAb) 희석액: 특히 CR6261(그룹 1 특이적), CR8020(그룹 2 특이적), CR9114(그룹 1 및 2 모두 특이적), 및 MD3606(그룹 1 및 2 특이적 다중도메인 항체)과 인큐베이션하였다. 항체 결합을 2차 항체 항-인간 Fc HRP(마우스 항 인간 IgG, Jackson ImmunoResearch)와의 인큐베이션에 의해 결정하였고, POD 기질의 첨가에 의해 가시화하였다. EnSight™ 다중모드 플레이트 판독기(PerkinElmer)를 사용하여 판독을 수행하였다. 두 가지 독립적인 실험의 EC50 값을 Spotfire 제품군(Tibco Software Inc.)과 도 3d에 열거된 평균 및 표준 편차를 이용하여 계산하였다.
결과 및 결론
[표 2]
본 발명의 폴리펩티드
SEC 분석 결과는 상이한 인플루엔자 HA 균주의 본 발명의 폴리펩티드 내로의 (안정화) 아미노산의 존재(또는 동시 도입)가 고도로 순수하고 안정적인 가용성 삼량체 HA 폴리펩티드의 정제를 가능하게 함을 확인시켜 주었다. 아미노산의 안정화 효과는 H1A/캘리포니아/07/2009(UFV181009)에서 유래된 정제된 폴리펩티드의 경우 가장 잘 관찰되었으며, 여기서 상응하는 야생형 구축물(UFV4239, 서열번호 29)은 폴드온 삼량체화 도메인을 보유하지 않았고 단량체 피크만을 생성하였으나, 본 발명의 안정화된 폴리펩티드는 고순도 삼량체 피크를 보여준다(도 3a). 다른 H1 균주 A/미시간/45/2015의 야생형 HA 분자와 H3 균주 A/홍콩/1/1968 및 A/인디애나/11/2011은 추가적인 C 말단 폴드온 도메인을 가지고 발현되었고 삼량체 HA를 형성하였다. 그러나 본 발명의 각각의 안정화된 폴리펩티드와 달리, 폴드온 도메인을 갖는 야생형 HA의 삼량체 피크는 더 넓고 비대칭적이며, 바람직하지 않은 형태(*) 또는 덜 컴팩트한 접힘의 대안적인 고분자량 및/또는 저분자량의 폴리펩티드의 존재를 시사하는 숄더(shoulder)를 보여주었다(Seok et al., Sci. Cell Rep. 8;7:1-7540, 2017]).
모든 폴리펩티드의 추가 특성화는 안정화 아미노산을 포함하는 본 발명의 폴리펩티드가 폴드온 삼량체화 도메인이 있거나 없는(UFV4239, 서열번호 29) WT 폴리펩티드와 비교하여 상당히 더 높은 열 안정성을 나타냄을 보여주었다(도 3b 및 3c).
도입된 안정화 돌연변이는 매립형 돌연변이(즉, 표면이 아니라 HA 폴리펩티드 내부에 있음)이므로, 단량체 또는 삼량체 HA의 표면에 영향을 미치지 않을 것이다. HA 표면의 무결성을 확인하기 위하여, 널리 알려진 폭넓게 중화하는 항체 패널의 폴리펩티드에 대한 결합을 ELISA로 평가하였다. 본 발명의 야생형 및 안정화된 폴리펩티드는 이들의 예상되는 결합 폭에 따라 모든 항체에 대해 낮은 nM 범위의 EC50 값으로 비슷한 결합을 나타내었다. 본 발명의 H3 A/홍콩/1/1968 및 H3 A/인디애나/11/2011 유래 HA 폴리펩티드에 대한 CR9114 결합의 개선(약 4 내지 8배)이 관찰되었다(도 3d).
결론적으로, 본 실시예에 기술된 본 발명의 폴리펩티드는 고순도 삼량체 폴리펩티드로서 세포 배양 상청액으로부터 정제되었고, (폴드온 삼량체화 도메인이 있거나 없는) WT HA와 비교하여 개선된 열 안정성을 나타내었고 적절하게 접혔다.
실시예 4:
안정화 돌연변이의 조합의 특성화
본 발명의 폴리펩티드에서 안정화 돌연변이를 조합하는 것의 유익한 효과를 평가하기 위하여, 조합 355W + 478I 및 조합 380I + 432I를 H1 균주 A/캘리포니아/07/2009의 HA 엑토도메인에 단계적으로 도입하였다(도 4의 A, '.'는 첫 번째 줄에 열거된 H1 야생형(WT) 잔기의 변경되지 않은 존재를 나타낸다).
본 발명의 폴리펩티드를 암호화하는 DNA 단편을 실시예 2에 기술된 바와 같이 합성하였다. 부위 특이적 비오틴화 및 스크리닝 및 정제 목적을 위한 C 말단 링커-소르타제-링커-His 태그를 포함하는 폴리펩티드를 실시예 2에 기술된 바와 같이 진핵생물 Expi293F 세포에서 마이크로 규모(200 μL)로 생산하였다. 발현된 폴리펩티드의 수준을 OCTET에 의해 결정하였고, 삼량체 함량을 실시예 2에 기술된 바와 같이 분석적 SEC에 의해 분석하였다.
결과 및 결론
안정화 돌연변이의 상이한 조합을 갖는 본 발명의 폴리펩티드의 발현 수준의 평가는 UFV181007(서열번호 35)에 존재하는 돌연변이 380I 및 432I가 발현에 영향을 미치지 않았지만 WT 구축물과 비교하여 유의하게 삼량체의 수준을 증가시켰음을 보여주었다(도 4의 B). 돌연변이 355W 및 478I(예를 들어, UFV181005: 서열번호 34)의 부가는 현저한 발현의 증가를 초래하지만(도 4의 A), 삼량체의 형성은 관찰되지 않았다(도 4의 B). 355W, 478I, 380I 및 432I(예를 들어, UFV181009: 서열번호 10)를 조합할 때 발현 수준이 모두 증가되었고(도 4의 A), 삼량체 함량은 세포 배양 상청액에서 유의하게 개선되었다(도 4의 B).
결론적으로, 돌연변이 355W 및 478I는 본 발명의 폴리펩티드의 발현 수준을 증가시킨 반면, 돌연변이 380I 및 432I는 삼량체 형성을 개선하였다. 안정화 돌연변이의 조합은 본 발명의 폴리펩티드의 발현 및 삼량체 수준을 상승적으로 증가시켰다.
실시예 5: 다양한 HA 서브타입에서 야생형 HA와 비교하여 추가적인 가용성의 안정화된 HA의 발현
이 실시예에서, 추가로 안정화된 HA를 발현시키고, 각각의 야생형 가용성 HA 엑토도메인과 비교하였다(도 5의 A). 위치 355의 트립토판(W) 및 위치 380 및 432의 이소류신(I)이 2개의 추가 그룹 1 균주 및 4개의 추가 그룹 2 균주의 HA의 아미노산 서열에 도입되었다. 형질감염 3일 후 Expi293F 배양 상청액에서 폴리펩티드의 발현 수준 및 삼량체화를 본 발명의 돌연변이가 없는 각각의 가용성 WT 폴리펩티드와 비교하였다. 표 4는 제조된 본 발명에 따른 추가의 폴리펩티드를 나타낸다.
본 발명의 폴리펩티드를 암호화하는 DNA 단편을 실시예 2에 기술된 바와 같이 합성하였다. 플라스미드를 실시예 2에 기술된 바와 같이 진핵생물 Expi293F 세포에서 마이크로 규모(200 μL)로 형질감염시켰다. 스크리닝 및 정제 목적을 위한 C 말단 링커 His-태그를 포함한 모든 폴리펩티드가 발현된 반면, 안정화된 폴리펩티드는 부위 특이적 비오틴화를 위한 His 태그 앞에 추가의 소르타제-링커 서열을 포함한다. 발현된 폴리펩티드의 수준을 OCTET에 의해 결정하였고, 삼량체 함량을 실시예 2에 기술된 바와 같이 분석적 SEC에 의해 분석하였다.
결과 및 결론
실시예 2에서 관찰된 바와 같이, 상이한 균주의 야생형 HA에서 위치 355의 트립토판 및 위치 380 및 432의 이소류신의 도입은 OCTET 측정에 기초하여 추가로 테스트한 이러한 모든 폴리펩티드의 발현의 증가를 초래하였다. 하나의 예외는 H1 A/사우스캐롤라이나/1/1918 (UFV181084) 유래 HA에 대해 관찰되었는데, 이는 OCTET에 의해 결정된 바와 같이 약간의 감소를 보였지만(도 5의 A), SEC의 곡선 아래 면적을 기초로 하지 않았다(도 5의 B).
조 세포 배양 상청액의 SEC 분석은 모든 추가적인 가용성의 안정화된 HA에 안정화 돌연변이를 도입할 때, 각각의 야생형 HA 엑토도메인과 비교하여 더 많은 삼량체 폴리펩티드(T) 및 더 적은 단량체 폴리펩티드(M) 및 고분자량 종이 관찰되었음을 보여주었다(도 5의 B). 실시예 2에서 주목한 바와 같이, 상이한 인플루엔자 HA 서브타입 간의 체류 시간의 차이는 글리코실화 수준 및 복잡성의 차이로 인한 것일 가능성이 높다.
종합하면, 데이터는 본 발명의 추가적인 HA 폴리펩티드로의 돌연변이 355W, 380I 및/또는 432I의 도입이 안정적인 가용성 삼량체 HA의 증가된 발현 및 형성을 초래함을 확인시켜 준다.
실시예 6: 정제된 삼량체 전장 HA의 시험관 내 특성화(추가 데이터)
이 실시예에서는 추가적인 안정화된 HA를 발현시키고, 정제하고, 장기간 온도 스트레스에 노출시켰다. 이들 HA, UFV190839(서열번호 50), UFV190068(서열번호 51) 및 UFV190841(서열번호 52)은 각각 H3 A/홍콩/1/1968 H7 A/청둥오리/NL/12/2000, 및 H10 A/닭/독일/N/1949에서 유래되었다. 요컨대, 정제된 삼량체 폴리펩티드를 4℃(냉장고) 및 37℃(인큐베이터)에서 60일 동안 보관한 후, 분석적 SEC에 의해 단백질 무결성을 평가하였다.
본 발명에 따르면, 위치 355의 트립토판(W) 및 위치 380 및 432의 이소류신(I)이 3개의 상이한 그룹 2 균주의 HA의 아미노산 서열에 도입되었다.
본 발명의 폴리펩티드를 암호화하는 DNA 단편을 실시예 2에 기술된 바와 같이 합성하였다. 폴리펩티드를 실시예 3에 기술된 바와 같이 진핵생물 ExpiCHO 세포에서 중간 규모(30 mL)로 생산하였고, 5일째에 수확하였다. 부위 특이적 비오틴화, 스크리닝 및 정제 목적을 위한 C 말단 링커-소르타제-링커 His-태그를 포함하는 모든 폴리펩티드를 발현시켰다. 단백질을 실시예 3에 기술된 바와 같이 2단계 공정에 의해 정제하였으나, 이제는 HiLoad Superdex 200 16/600 컬럼이 사용되었다(GE Healthcare Life Sciences). 발현된 폴리펩티드의 수준을 OCTET에 의해 결정하였고, 현재 Unix-C 300 A 컬럼(Sepax Technologies)이 사용되었다는 일탈과 함께 실시예 2에 기술된 바와 같이 삼량체 함량을 분석적 SEC에 의해 분석하였다.
결과 및 결론
SEC 분석 결과는 정제 후 수득된 안정화 아미노산을 포함하는 본 발명의 폴리펩티드가 고도로 순수하고 삼량체임을 나타내었다. 나아가, 가용성 HA 폴리펩티드는 온도 스트레스에 저항적이었다. 4℃ 및 37℃에서 60일의 인큐베이션은 스트레스 전 물질에 대해 관찰된 것과 비교하여 단백질의 양 및 삼량체 상태에 영향을 미치지 않았으며(도 6), 37℃에서 인큐베이션 후 H10 유래 HA에 대해 오로지 소량의 삼량체 이외의 폴리펩티드가 관찰되었다(약 4.75분의 체류 시간).
실시예 2에서 주목한 바와 같이, 상이한 인플루엔자 HA 서브타입 간의 체류 시간의 차이는 글리코실화 수준 및 복잡성의 차이로 인한 것일 가능성이 높다. 또한, 60일 동안 스트레스를 가한 물질과 비교하여 출발 물질에 대해 관찰된 체류 시간의 작은 차이는 컬럼 노화로 인한 것일 가능성이 높다(즉, 유사한 이동이 내부 대조군에 대해 관찰됨).
결론적으로, 본 실시예에 기술된 본 발명의 폴리펩티드는 고순도 삼량체 폴리펩티드로서 배양 상청액으로부터 정제되었고, 60일의 기간 동안 온도 스트레스에 대해 고도로 불활성인 것으로 나타났다.
[표 3]
표준 아미노산, 약어 및 특성
서열
서열번호 1 CAA24269.1 적혈구응집소(인플루엔자 A 바이러스(A/아이치/2/1968(H3N2) (신호 서열 제외))
QDLPGNDNST ATLCLGHHAV PNGTLVKTIT DDQIEVTNAT ELVQSSSTGK 50
ICNNPHRILD GIDCTLIDAL LGDPHCDVFQ NETWDLFVER SKAFSNCYPY 100
DVPDYASLRS LVASSGTLEF ITEGFTWTGV TQNGGSNACK RGPGSGFFSR 150
LNWLTKSGST YPVLNVTMPN NDNFDKLYIW GIHHPSTNQE QTSLYVQASG 200
RVTVSTRRSQ QTIIPNIGSR PWVRGLSSRI SIYWTIVKPG DVLVINSNGN 250
LIAPRGYFKM RTGKSSIMRS DAPIDTCISE CITPNGSIPN DKPFQNVNKI 300
TYGACPKYVK QNTLKLATGM RNVPEKQTRG LFGAIAGFIE NGWEGMIDGW 350
YGFRHQNSEG TGQAADLKST QAAIDQINGK LNRVIEKTNE KFHQIEKEFS 400
EVEGRIQDLE KYVEDTKIDL WSYNAELLVA LENQHTIDLT DSEMNKLFEK 450
TRRQLRENAE EMGNGCFKIY HKCDNACIES IRNGTYDHDV YRDEALNNRF 500
QIKGVELKSG YKDWILWISF AISCFLLCVV LLGFIMWACQ RGNIRCNICI 550
CR6261 VH 단백질(서열번호 2)
EVQLVESGAEVKKPGSSVKVSCKASGGPFRSYAISWVRQAPGQGPEWMGGIIPIFGTTKYAPKFQGRVTITADDFAGTVYMELSSLRSEDTAMYYCAKHMGYQVRETMDVWGKGTTVTVSS
CR6261 VL 단백질(서열번호 3)
QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNDYVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEANYYCATWDRRPTAYVVFGGGTKLTVL
CR8020 VH 단백질(서열번호 4)
QVQLQQSGAEVKTPGASVKVSCKASGYTFTSFGVSWIRQAPGQGLEWIGWISAYNGDTYYAQKFQARVTMTTDTSTTTAYMEMRSLRSDDTAVYYCAREPPLFYSSWSLDNWGQGTLVTVSS
CR8020 VL 단백질(서열번호 5)
EIVLTQSPGTLSLSPGERATLSCRASQSVSMNYLAWFQQKPGQAPRLLIYGASRRATGIPDRISGSGSGTDFTLTISRLEPADFAVYYCQQYGTSPRTFGQGAKVEIK
CR9114 VH 단백질(서열번호 6)
QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTISADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDVWGQGTTVTVSS
CR9114 VL 단백질(서열번호 7)
SYVLTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVVPDRFSGSKSGTSASLAISGLQSEDEAEYYCAAWDDSLKGAVFGGGTQLTVL
MD3606 단백질(서열번호 8)
EVQLVESGGGLVQPGGSLRLSCAVSISIFDIYAMDWYRQAPGKQRDLVATSFRDGSTNYADSVKGRFTISRDNAKNTLYLQMNSLKPEDTAVYLCHVSLYRDPLGVAGGMGVYWGKGALVTVSSGGGGSGGGGSEVQLVESGGGLVQAGGSLKLSCAASGRTYAMGWFRQAPGKEREFVAHINALGTRTYYSDSVKGRFTISRDNAKNTEYLEMNNLKPEDTAVYYCTAQGQWRAAPVAVAAEYEFWGQGTQVTVSSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAATGFTLENKAIGWFRQTPGSEREGVLCISKSGSWTYYTDSMRGRFTISRDNAENTVYLQMDSLKPEDTAVYYCATTTAGGGLCWDGTTFSRLASSWGQGTQVTVSSGGGGSGGGGSEVQLVESGGGLVQPGGSLKLSCAASGFTFSTSWMYWLRQAPGKGLEWVSVINTDGGTYYADSVKDRFTISRDNAKDTLYLQMSSLKSEDTAVYYCAKDWGGPEPTRGQGTQVTVSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
서열번호 9: UFV181157(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFATANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETPSSDNGTCYPGDFIDYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLSKSYINDKGKEVLVLWGIHHPSTSADQQSLYQNADAYVFVGSSRYSKKFKPEIAIRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFAMERNAGSGIIISDTPVHDCNTTCQTPKGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNIPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQNAIDEITNKVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCMESVKNGTYDYPKYSEEAKLNREEIDGSHHHHHH
서열번호 10: UFV181009(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFATANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETPSSDNGTCYPGDFIDYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLSKSYINDKGKEVLVLWGIHHPSTSADQQSLYQNADAYVFVGSSRYSKKFKPEIAIRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFAMERNAGSGIIISDTPVHDCNTTCQTPKGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNIPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHWQNEQGSGYAADLKSTQNAIDEITNIVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLINERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCIESVKNGTYDYPKYSEEAKLNREEIDSGSLPETGGGSHHHHHH
서열번호 11: UFV181134(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFTTANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETSNSDNGTCYPGDFINYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLNQSYINDKGKEVLVLWGIHHPSTTADQQSLYQNADAYVFVGTSRYSKKFKPEIATRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFTMERNAGSGIIISDTPVHDCNTTCQTPEGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNVPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQNAIDKITNKVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDYHDSNVKNLYEKVRNQLKNNAKEIGNGCFEFYHKCDNTCMESVKNGTYDYPKYSEEAKLNREKIDGSHHHHHH
서열번호 12: UFV181091(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFTTANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETSNSDNGTCYPGDFINYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLNQSYINDKGKEVLVLWGIHHPSTTADQQSLYQNADAYVFVGTSRYSKKFKPEIATRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFTMERNAGSGIIISDTPVHDCNTTCQTPEGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNVPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHWQNEQGSGYAADLKSTQNAIDKITNIVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLINERTLDYHDSNVKNLYEKVRNQLKNNAKEIGNGCFEFYHKCDNTCIESVKNGTYDYPKYSEEAKLNREKIDSGSLPETGGGSHHHHHH
서열번호 13: UFV181153(신호 펩티드 및 태그는 밑줄 표시됨)
MAIIYLILLFAAVRGDQICIGYHSNNSTEKVDTILERNVTVTHAQDILEKTHNGKLCKLNGIPPLELGDCSIAGWLLGNPECDRLLTVPEWSYIMEKENPRNGLCYPGSFNDYEELKHLLSSVTHFEKVKILPRDRWTQHTTTGGSRACAVSGNPSFFRNMVWLTKKGSNYPIAKGSYNNTSGEQMLIIWGVHHPNDDAEQRTLYQNVGTYVSVGTSTLNKRSVPEIATRPKVNGQGGRMEFSWTILDMLDTINFESTGNLIAPEYGFRISKRGSSGIMKTEGTLENCETKCQTPLGAINTTLPFHNIHPLTIGECPKYVKSERLVLATGLRNVPQIESRGLFGAIAGFIEGGWQGMVDGWYGYHHSNDQGSGYAADKESTQRAIDGITNKVNSVIEKMNTQFEAVGKEFNNLEKRLENLNKKMEDGFLDVWTYNAELLVLMENERTLDFHDSNVKNLYDKVRMQLRDNAKELGNGCFEFYHKCDDECMNSVKNGTYDYPKYEEESKLNRNEIKGSHHHHHH
서열번호 14: UFV181154(신호 펩티드 및 태그는 밑줄 표시됨)
MAIIYLILLFAAVRGDQICIGYHSNNSTEKVDTILERNVTVTHAQDILEKTHNGKLCKLNGIPPLELGDCSIAGWLLGNPECDRLLTVPEWSYIMEKENPRNGLCYPGSFNDYEELKHLLSSVTHFEKVKILPRDRWTQHTTTGGSRACAVSGNPSFFRNMVWLTKKGSNYPIAKGSYNNTSGEQMLIIWGVHHPNDDAEQRTLYQNVGTYVSVGTSTLNKRSVPEIATRPKVNGQGGRMEFSWTILDMLDTINFESTGNLIAPEYGFRISKRGSSGIMKTEGTLENCETKCQTPLGAINTTLPFHNIHPLTIGECPKYVKSERLVLATGLRNVPQIESRGLFGAIAGFIEGGWQGMVDGWYGYHWSNDQGSGYAADKESTQRAIDGITNIVNSVIEKMNTQFEAVGKEFNNLEKRLENLNKKMEDGFLDVWTYNAELLVLMINERTLDFHDSNVKNLYDKVRMQLRDNAKELGNGCFEFYHKCDDECINSVKNGTYDYPKYEEESKLNRNEIKSGSLPETGGGSHHHHHH
서열번호 15: UFV181158(신호 펩티드 및 태그는 밑줄 표시됨)
MEKIVLLFAIVSLVQSDQICIGYHANNSTEQVDTIMEKNVTVTHAQDILEKTHNGKLCSLNGVKPLILRDCSVAGWLLGNPMCDEFLNVPEWSYIVEKDSPINGLCYPGDFNDYEELKHLLSSTNHFEKIQIIPRSSWSNHDASSGVSSACPYNGRSSFFRNVVWLIKKNNAYPTIKRSYNNTNQEDLLVLWGIHHPNDAAEQTKLYQNPTTYVSVGTSTLNQRSVPEIATRPKVNGQSGRMEFFWTILKPNDAINFESNGNFIAPEYAYKIVKKGDSAIMKSGLEYGNCNTKCQTPMGAINSSMPFHNIHPLTIGECPKYVKSDRLVLATGLRNVPQRETRGLFGAIAGFIEGGWQGMVDGWYGYLHSNEQGSGYAADKESTQKAIDGITNKINSIIDKMNTQFEAVGKEFNNLERRIENLNKKMEDGFLDVWTYNAELLVLMENERTLDFHDSNVKNLYDKVRLQLRDNAKELGNGCFEFYHKCDDECMESVRNGTYDYPQYSEEARLNREEISGSHHHHHH
서열번호 16: UFV181159(신호 펩티드 및 태그는 밑줄 표시됨)
MEKIVLLFAIVSLVQSDQICIGYHANNSTEQVDTIMEKNVTVTHAQDILEKTHNGKLCSLNGVKPLILRDCSVAGWLLGNPMCDEFLNVPEWSYIVEKDSPINGLCYPGDFNDYEELKHLLSSTNHFEKIQIIPRSSWSNHDASSGVSSACPYNGRSSFFRNVVWLIKKNNAYPTIKRSYNNTNQEDLLVLWGIHHPNDAAEQTKLYQNPTTYVSVGTSTLNQRSVPEIATRPKVNGQSGRMEFFWTILKPNDAINFESNGNFIAPEYAYKIVKKGDSAIMKSGLEYGNCNTKCQTPMGAINSSMPFHNIHPLTIGECPKYVKSDRLVLATGLRNVPQRETRGLFGAIAGFIEGGWQGMVDGWYGYLWSNEQGSGYAADKESTQKAIDGITNIINSIIDKMNTQFEAVGKEFNNLERRIENLNKKMEDGFLDVWTYNAELLVLMINERTLDFHDSNVKNLYDKVRLQLRDNAKELGNGCFEFYHKCDDECIESVRNGTYDYPQYSEEARLNREEISSGSLPETGGGSHHHHHH
서열번호 17: UFV181155(신호 펩티드 및 태그는 밑줄 표시됨)
METISLITILLVVTASNADKICIGHQSTNSTETVDTLTETNVPVTHAKELLHTEHNGMLCATSLGHPLILDTCTIEGLVYGNPSCDLLLGGREWSYIVERSSAVNGTCYPGNVENLEELRTLFSSASSYQRIQIFPDTTWNVTYTGTSRACSGSFYRSMRWLTQKSGFYPVQDAQYTNNRGKSILFVWGIHHPPTYTEQTNLYIRNDTTTSVTTEDLNRTFKPVIGPRPLVNGLQGRIDYYWSVLKPGQTLRVRSNGNLIAPWYGHVLSGGSHGRILKTDLKGGNCVVQCQTEKGGLNSTLPFHNISKYAFGTCPKYVRVNSLKLAVGLRNVPARSSRGLFGAIAGFIEGGWPGLVAGWYGFQHSNDQGVGMAADRDSTQKAIDKITSKVNNIVDKMNKQYEIIDHEFSEVETRLNMINNKIDDQIQDVWAYNAELLVLLENQKTLDEHDANVNNLYNKVKRALGSNAMEDGKGCFELYHKCDDQCMETIRNGTYNRRKYREESRLERQKIEGSHHHHHH
서열번호 18: UFV181156(신호 펩티드 및 태그는 밑줄 표시됨)
METISLITILLVVTASNADKICIGHQSTNSTETVDTLTETNVPVTHAKELLHTEHNGMLCATSLGHPLILDTCTIEGLVYGNPSCDLLLGGREWSYIVERSSAVNGTCYPGNVENLEELRTLFSSASSYQRIQIFPDTTWNVTYTGTSRACSGSFYRSMRWLTQKSGFYPVQDAQYTNNRGKSILFVWGIHHPPTYTEQTNLYIRNDTTTSVTTEDLNRTFKPVIGPRPLVNGLQGRIDYYWSVLKPGQTLRVRSNGNLIAPWYGHVLSGGSHGRILKTDLKGGNCVVQCQTEKGGLNSTLPFHNISKYAFGTCPKYVRVNSLKLAVGLRNVPARSSRGLFGAIAGFIEGGWPGLVAGWYGFQWSNDQGVGMAADRDSTQKAIDKITSIVNNIVDKMNKQYEIIDHEFSEVETRLNMINNKIDDQIQDVWAYNAELLVLLINQKTLDEHDANVNNLYNKVKRALGSNAMEDGKGCFELYHKCDDQCIETIRNGTYNRRKYREESRLERQKIESGSLPETGGGSHHHHHH
서열번호 19: UFV181141(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYIFCLALGQDLPGNDNSTATLCLGHHAVPNGTLVKTITDDQIEVTNATELVQSSSTGKICNNPHRILDGIDCTLIDALLGDPHCDVFQNETWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFITEGFTWTGVTQNGGSNACKRGPGSGFFSRLNWLTKSGSTYPVLNVTMPNNDNFDKLYIWGVHHPSTNQEQTSLYVQASGRVTVSTRRSQQTIIPNIGSRPWVRGLSSRISIYWTIVKPGDVLVINSNGNLIAPRGYFKMRTGKSSIMRSDAPIDTCISECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTRGLFGAIAGFIENGWEGMIDGWYGFRHQNSEGTGQAADLKSTQAAIDQINGKLNRVIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFEKTRRQLRENAEDMGNGCFKIYHKCDNACIESIRNGTYDHDVYRDEALNNRFQIKGVGSHHHHHH
서열번호 20: UFV180660(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYIFCLALGQDLPGNDNSTATLCLGHHAVPNGTLVKTITDDQIEVTNATELVQSSSTGKICNNPHRILDGIDCTLIDALLGDPHCDVFQNETWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFITEGFTWTGVTQNGGSNACKRGPGSGFFSRLNWLTKSGSTYPVLNVTMPNNDNFDKLYIWGVHHPSTNQEQTSLYVQASGRVTVSTRRSQQTIIPNIWSRPWVRGLSSRISIYWTIVKPGDVLVINSNGNLIAPRGYFKMRTGKSSIMRSDAPIDTCISECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTRGLFGAIAGFIENGWEGMIDGWYGFRWQNSEGTGQAADLKSTQAAIDQINGILNRVIEKMNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFEKTRRQLRENAEDMGNGCFKIYHKCDNACIESIRNGNYDHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 21: UFV181137(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVSNGTLVKTITNDQIEVTNATELVQSSSTGRICDSPHQILDGENCTLIDALLGDPHCDGFQNKEWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNNESFNWTGVAQNGTSSACKRRSNKSFFSRLNWLHQLKYKYPALNVTMPNNEKFDKLYIWGVHHPSTDSDQISIYAQASGRVTVSTKRSQQTVIPNIGSSPWVRGVSSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGKSSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRHQNSEGTGQAADLKSTQAAINQINGKLNRLIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFERTKKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVGSHHHHHH
서열번호 22: UFV181096(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVSNGTLVKTITNDQIEVTNATELVQSSSTGRICDSPHQILDGENCTLIDALLGDPHCDGFQNKEWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNNESFNWTGVAQNGTSSACKRRSNKSFFSRLNWLHQLKYKYPALNVTMPNNEKFDKLYIWGVHHPSTDSDQISIYAQASGRVTVSTKRSQQTVIPNIGSSPWVRGVSSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGKSSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRWQNSEGTGQAADLKSTQAAINQINGILNRLIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFERTKKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 23: UFV181145(신호 펩티드 및 태그는 밑줄 표시됨)
MIALILVALALSHTAYSQITNGTTGNPIICLGHHAVENGTSVKTLTDNHVEVVSAKELVETNHTDELCPSPLKLVDGQDCDLINGALGSPGCDRLQDTTWDVFIERPTAVDTCYPFDVPDYQSLRSILASSGSLEFIAEQFTWNGVKVDGSSSACLRGGRNSFFSRLNWLTKETNGNYGPINVTKENTGSYVRLYLWGVHHPSSDNEQTDLYKVATGRVTVSTRSDQISIVPNIGSRPRVRNQSGRISIYWTLVNPGDSIIFNSIGNLIAPRGHYKISKSTKSTVLKSDKRIGSCTSPCLTDKGSIQSDKPFQNVSRIAIGNCPKYVKQGSLMLATGMRNIPGKQAKGLFGAIAGFIENGWQGLIDGWYGFRHQNAEGTGTAADLKSTQAAIDQINGKLNRLIEKTNEKYHQIEKEFEQVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDVTDSEMNKLFERVRRQLRENAEDQGNGCFEIFHQCDNNCIESIRNGTYDHNIYRDEAINNRIKINPVGSHHHHHH
서열번호 24: UFV180661(신호 펩티드 및 태그는 밑줄 표시됨)
MIALILVALALSHTAYSQITNGTTGNPIICLGHHAVENGTSVKTLTDNHVEVVSAKELVETNHTDELCPSPLKLVDGQDCDLINGALGSPGCDRLQDTTWDVFIERPTAVDTCYPFDVPDYQSLRSILASSGSLEFIAEQFTWNGVKVDGSSSACLRGGRNSFFSRLNWLTKETNGNYGPINVTKENTGSYVRLYLWGVHHPSSDNEQTDLYKVATGRVTVSTRSDQISIVPNIGSRPRVRNQSGRISIYWTLVNPGDSIIFNSIGNLIAPRGHYKISKSTKSTVLKSDKRIGSCTSPCLTDKGSIQSDKPFQNVSRIAIGNCPKYVKQGSLMLATGMRNIPGKQAKGLFGAIAGFIENGWQGLIDGWYGFRWQNAEGTGTAADLKSTQAAIDQINGILNRLIEKMNEKYHQIEKEFEQVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDVTDSEMNKLFERVRRQLRENAEDQGNGCFEIFHQCDNNCIESIRNGTYDHNIYRDEAINNRIKINPVSGSLPETGGGSHHHHHH
서열번호 25: UFV181146(신호 펩티드 및 태그는 밑줄 표시됨)
MNTQILVFALMAIIPTNADKICLGHHAVSNGTKVNTLTERGVEVVNATETVERTNVPRICSKGKRTVDLGQCGLLGTITGPPQCDQFLEFSADLIIERREGSDVCYPGKFVNEEALRQILRESGGIDKETMGFTYSGIRTNGATSACRRSGSSFYAEMKWLLSNTDNAAFPQMTKSYKNTRKDPALIIWGIHHSGSTTEQTKLYGSGNKLITVGSSNYQQSFVPSPGARPQVNGQSGRIDFHWLILNPNDTVTFSFNGAFIAPDRASFLRGKSMGIQSGVQVDANCEGDCYHSGGTIISNLPFQNINSRAVGKCPRYVKQESLLLATGMKNVPEIPKGRGLFGAIAGFIENGWEGLIDGWYGFRHQNAQGEGTAADYKSTQSAIDQITGKLNRLIEKTNQQFELIDNEFTEVEKQIGNVINWTRDSMTEVWSYNAELLVAMENQHTIDLADSEMNKLYERVKRQLRENAEEDGTGCFEIFHKCDDDCMASIRNNTYDHSKYREEAMQNRIQIDPVGSHHHHHH
서열번호 26: UFV180664(신호 펩티드 및 태그는 밑줄 표시됨)
MNTQILVFALMAIIPTNADKICLGHHAVSNGTKVNTLTERGVEVVNATETVERTNVPRICSKGKRTVDLGQCGLLGTITGPPQCDQFLEFSADLIIERREGSDVCYPGKFVNEEALRQILRESGGIDKETMGFTYSGIRTNGATSACRRSGSSFYAEMKWLLSNTDNAAFPQMTKSYKNTRKDPALIIWGIHHSGSTTEQTKLYGSGNKLITVGSSNYQQSFVPSPGARPQVNGQSGRIDFHWLILNPNDTVTFSFNGAFIAPDRASFLRGKSMGIQSGVQVDANCEGDCYHSGGTIISNLPFQNINSRAVGKCPRYVKQESLLLATGMKNVPEIPKGRGLFGAIAGFIENGWEGLIDGWYGFRWQNAQGEGTAADYKSTQSAIDQITGILNRLIEKMNQQFELIDNEFTEVEKQIGNVINWTRDSMTEVWSYNAELLVAMINQHTIDLADSEMNKLYERVKRQLRENAEEDGTGCFEIFHKCDDDCMASIRNNTYDHSKYREEAMQNRIQIDPVSGSLPETGGGSHHHHHH
서열번호 27: UFV181147(신호 펩티드 및 태그는 밑줄 표시됨)
MYKVVVIIALLGAVKGLDRICLGHHAVANGTIVKTLTNEQEEVTNATETVESTNLNKLCMKGRSYKDLGNCHPVGMLIGTPVCDPHLTGTWDTLIERENAIAHCYPGATINEEALRQKIMESGGISKMSTGFTYGSSINSAGTTKACMRNGGDSFYAELKWLVSKTKGQNFPQTTNTYRNTDTAEHLIIWGIHHPSSTQEKNDLYGTQSLSISVESSTYQNNFVPVVGARPQVNGQSGRIDFHWTLVQPGDNITFSHNGGLIAPSRVSKLTGRGLGIQSEALIDNSCESKCFWRGGSINTKLPFQNLSPRTVGQCPKYVNQRSLLLATGMRNVPEVVQGRGLFGAIAGFIENGWEGMVDGWYGFRHQNAQGTGQAADYKSTQAAIDQITGKLNRLIEKTNTEFESIESEFSETEHQIGNVINWTKDSITDIWTYQAELLVAMENQHTIDMADSEMLNLYERVRKQLRQNAEEDGKGCFEIYHTCDDSCMESIRNNTYDHSQYREEALLNRLNINSVGSHHHHHH
서열번호 28: UFV180662(신호 펩티드 및 태그는 밑줄 표시됨)
MYKVVVIIALLGAVKGLDRICLGHHAVANGTIVKTLTNEQEEVTNATETVESTNLNKLCMKGRSYKDLGNCHPVGMLIGTPVCDPHLTGTWDTLIERENAIAHCYPGATINEEALRQKIMESGGISKMSTGFTYGSSINSAGTTKACMRNGGDSFYAELKWLVSKTKGQNFPQTTNTYRNTDTAEHLIIWGIHHPSSTQEKNDLYGTQSLSISVESSTYQNNFVPVVGARPQVNGQSGRIDFHWTLVQPGDNITFSHNGGLIAPSRVSKLTGRGLGIQSEALIDNSCESKCFWRGGSINTKLPFQNLSPRTVGQCPKYVNQRSLLLATGMRNVPEVVQGRGLFGAIAGFIENGWEGMVDGWYGFRWQNAQGTGQAADYKSTQAAIDQITGILNRLIEKMNTEFESIESEFSETEHQIGNVINWTKDSITDIWTYQAELLVAMINQHTIDMADSEMLNLYERVRKQLRQNAEEDGKGCFEIYHTCDDSCMESIRNNTYDHSQYREEALLNRLNINSSGSLPETGGGSHHHHHH
서열번호 29: UFV4239(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFATANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETPSSDNGTCYPGDFIDYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLSKSYINDKGKEVLVLWGIHHPSTSADQQSLYQNADAYVFVGSSRYSKKFKPEIAIRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFAMERNAGSGIIISDTPVHDCNTTCQTPKGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNIPSIQSQGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQNAIDEITNKVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCMESVKNGTYDYPKYSEEAKLNREEIDGRSLVPRGSGHHHHHH
서열번호 30: UFV180843(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFTTANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETSNSDNGTCYPGDFINYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLNQSYINDKGKEVLVLWGIHHPSTTADQQSLYQNADAYVFVGTSRYSKKFKPEIATRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFTMERNAGSGIIISDTPVHDCNTTCQTPEGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNVPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQNAIDKITNKVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDYHDSNVKNLYEKVRNQLKNNAKEIGNGCFEFYHKCDNTCMESVKNGTYDYPKYSEEAKLNREKIDSGSLVPSGSPGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGSLPETGGGSHHHHHH
서열번호 31: UFV180436(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYIFCLALGQDLPGNDNSTATLCLGHHAVPNGTLVKTITDDQIEVTNATELVQSSSTGKICNNPHRILDGIDCTLIDALLGDPHCDVFQNETWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFITEGFTWTGVTQNGGSNACKRGPGSGFFSRLNWLTKSGSTYPVLNVTMPNNDNFDKLYIWGVHHPSTNQEQTSLYVQASGRVTVSTRRSQQTIIPNIGSRPWVRGLSSRISIYWTIVKPGDVLVINSNGNLIAPRGYFKMRTGKSSIMRSDAPIDTCISECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTRGLFGAIAGFIENGWEGMIDGWYGFRHQNSEGTGQAADLKSTQAAIDQINGKLNRVIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFEKTRRQLRENAEDMGNGCFKIYHKCDNACIESIRNGTYDHDVYRDEALNNRFQSGSLVPSGSPGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGSLPETGGGSHHHHHH
서열번호 32: UFV170466(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIVALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTIVKTITNDQIEVTNATELVQSSSTGEICDSPHQILDGENCTLIDALLGDPQCDGFQNKKWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNNESFNWTGVTQNGTSSACIRRSNSSFFSRLNWLTHLNFKYPALNVTMPNNEQFDKLYIWGVHHPGTDKDQIFLYAQSSGRITVSTKRSQQAVIPNIGSRPRIRNIPSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGKSSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQSTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRHQNSEGRGQAADLKSTQAAIDQINGKLNRLIGKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFEKTKKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYNHDVYRDEALNNRFQSGSLVPRGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGSEPEA
서열번호 33: UFV181099(신호 펩티드 및 태그는 밑줄 표시됨)
MKTIVALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTIVKTITNDQIEVTNATELVQSSSTGEICDSPHQILDGENCTLIDALLGDPQCDGFQNKKWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNNESFNWTGVTQNGTSSACIRRSNSSFFSRLNWLTHLNFKYPALNVTMPNNEQFDKLYIWGVHHPGTDKDQIFLYAQSSGRITVSTKRSQQAVIPNIGSRPRIRNIPSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGKSSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQSTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRWQNSEGRGQAADLKSTQAAIDQINGILNRLIGKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFEKTKKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYNHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 34: UFV181005(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFATANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETPSSDNGTCYPGDFIDYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLSKSYINDKGKEVLVLWGIHHPSTSADQQSLYQNADAYVFVGSSRYSKKFKPEIAIRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFAMERNAGSGIIISDTPVHDCNTTCQTPKGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNIPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHWQNEQGSGYAADLKSTQNAIDEITNKVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCIESVKNGTYDYPKYSEEAKLNREEIDSGSLPETGGGSHHHHHH
서열번호 35: UFV181007(신호 펩티드 및 태그는 밑줄 표시됨)
MKAILVVLLYTFATANADTLCIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDKHNGKLCKLRGVAPLHLGKCNIAGWILGNPECESLSTASSWSYIVETPSSDNGTCYPGDFIDYEELREQLSSVSSFERFEIFPKTSSWPNHDSNKGVTAACPHAGAKSFYKNLIWLVKKGNSYPKLSKSYINDKGKEVLVLWGIHHPSTSADQQSLYQNADAYVFVGSSRYSKKFKPEIAIRPKVRDQEGRMNYYWTLVEPGDKITFEATGNLVVPRYAFAMERNAGSGIIISDTPVHDCNTTCQTPKGAINTSLPFQNIHPITIGKCPKYVKSTKLRLATGLRNIPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADLKSTQNAIDEITNIVNSVIEKMNTQFTAVGKEFNHLEKRIENLNKKVDDGFLDIWTYNAELLVLLINERTLDYHDSNVKNLYEKVRSQLKNNAKEIGNGCFEFYHKCDNTCMESVKNGTYDYPKYSEEAKLNREEIDSGSLPETGGGSHHHHHH
서열번호 36: UFV181090 (신호 펩티드 및 태그는 밑줄 표시됨)
MKVKLLVLLCTFTATYADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENSHNGKLCLLKGIAPLQLGNCSVAGWILGNPECELLISKESWSYIVEKPNPENGTCYPGHFADYEELREQLSSVSSFERFEIFPKESSWPNHTVTGVSASCSHNGESSFYRNLLWLTGKNGLYPNLSKSYANNKEKEVLVLWGVHHPPNIGDQKALYHTENAYVSVVSSHYSRKFTPEIAKRPKVRDQEGRINYYWTLLEPGDTIIFEANGNLIAPRYAFALSRGFGSGIINSNAPMDKCDAKCQTPQGAINSSLPFQNVHPVTIGECPKYVRSAKLRMVTGLRNIPSIQSRGLFGAIAGFIEGGWTGMVDGWYGYHWQNEQGSGYAADQKSTQNAINGITNIVNSVIEKMNTQFTAVGKEFNKLERRMENLNKKVDDGFIDIWTYNAELLVLLINERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNDECIESVKNGTYDYPKYSEESKLNREKIDSGSLPETGGGSHHHHHH
서열번호 37: UFV181135 (신호 펩티드 및 태그는 밑줄 표시됨)
MKVKLLVLLCTFTATYADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENSHNGKLCLLKGIAPLQLGNCSVAGWILGNPECELLISKESWSYIVEKPNPENGTCYPGHFADYEELREQLSSVSSFERFEIFPKESSWPNHTVTGVSASCSHNGESSFYRNLLWLTGKNGLYPNLSKSYANNKEKEVLVLWGVHHPPNIGDQKALYHTENAYVSVVSSHYSRKFTPEIAKRPKVRDQEGRINYYWTLLEPGDTIIFEANGNLIAPRYAFALSRGFGSGIINSNAPMDKCDAKCQTPQGAINSSLPFQNVHPVTIGECPKYVRSAKLRMVTGLRNIPSIQSQGLFGAIAGFIEGGWTGMVDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKMNTQFTAVGKEFNKLERRMENLNKKVDDGFIDIWTYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGSHHHHHH
서열번호 38: UFV181084 (신호 펩티드 및 태그는 밑줄 표시됨)
MEARLLVLLCAFAATNADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCKLKGIAPLQLGKCNIAGWLLGNPECDLLLTASSWSYIVETSNSENGTCYPGDFIDYEELREQLSSVSSFEKFEIFPKTSSWPNHETTKGVTAACSYAGASSFYRNLLWLTKKGSSYPKLSKSYVNNKGKEVLVLWGVHHPPTGTDQQSLYQNADAYVSVGSSKYNRRFTPEIAARPKVRDQAGRMNYYWTLLEPGDTITFEATGNLIAPWYAFALNRGSGSGIITSDAPVHDCNTKCQTPHGAINSSLPFQNIHPVTIGECPKYVRSTKLRMATGLRNIPSIQSRGLFGAIAGFIEGGWTGMIDGWYGYHWQNEQGSGYAADQKSTQNAIDGITNIVNSVIEKMNTQFTAVGKEFNNLERRIENLNKKVDDGFLDIWTYNAELLVLLINERTLDFHDSNVRNLYEKVKSQLKNNAKEIGNGCFEFYHKCDDACIESVRNGTYDYPKYSEESKLNREEIDSGSLPETGGGSHHHHHH
서열번호 39: UFV181131 (신호 펩티드 및 태그는 밑줄 표시됨)
MEARLLVLLCAFAATNADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDSHNGKLCKLKGIAPLQLGKCNIAGWLLGNPECDLLLTASSWSYIVETSNSENGTCYPGDFIDYEELREQLSSVSSFEKFEIFPKTSSWPNHETTKGVTAACSYAGASSFYRNLLWLTKKGSSYPKLSKSYVNNKGKEVLVLWGVHHPPTGTDQQSLYQNADAYVSVGSSKYNRRFTPEIAARPKVRDQAGRMNYYWTLLEPGDTITFEATGNLIAPWYAFALNRGSGSGIITSDAPVHDCNTKCQTPHGAINSSLPFQNIHPVTIGECPKYVRSTKLRMATGLRNIPSIQSRGLFGAIAGFIEGGWTGMIDGWYGYHHQNEQGSGYAADQKSTQNAIDGITNKVNSVIEKMNTQFTAVGKEFNNLERRIENLNKKVDDGFLDIWTYNAELLVLLENERTLDFHDSNVRNLYEKVKSQLKNNAKEIGNGCFEFYHKCDDACMESVRNGTYDYPKYSEESKLNREEIDGSHHHHHH
서열번호 40: UFV181095 (신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTLVKTITNDQIEVTNATELVQSSSTGRICDSPHRILDGKNCTLIDALLGDPHCDGFQNKEWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFINEDFNWTGVAQDGKSYTCKRGSVNSFFSRLNWLHKLEYKYPALNVTMPNNGKFDKLYIWGVHHPSTDSDQTSLYVRASGRVTVSTKRSQQTVIPNIGSRPWVRGLSSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRNGKSSIMRSDAPIGNCSSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRWQNSEGTGQAADLKSTQAAIDQINGILNRLIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFERTRKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 41: UFV181140 (신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTLVKTITNDQIEVTNATELVQSSSTGRICDSPHRILDGKNCTLIDALLGDPHCDGFQNKEWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFINEDFNWTGVAQDGKSYTCKRGSVNSFFSRLNWLHKLEYKYPALNVTMPNNGKFDKLYIWGVHHPSTDSDQTSLYVRASGRVTVSTKRSQQTVIPNIGSRPWVRGLSSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRNGKSSIMRSDAPIGNCSSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRHQNSEGTGQAADLKSTQAAIDQINGKLNRLIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFERTRKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVGSHHHHHH
서열번호 42: UFV181093 (신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYIFCQVLAQNLPGNDNSTATLCLGHHAVPNGTLVKTITNDQIEVTNATELVQSSSTGRICDSPHRILDGKNCTLIDALLGDPHCDGFQNEKWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFINEGFNWTGVTQNGGSYACKRGPDKSFFSRLNWLYESESTYPVLNVTMPNNDNFDKLYIWGVHHPSTDKEQTNLYVQASGRVTVSTKRSQQTIIPNVGSRPWVRGLSSRISIYWTIVKPGDILLINSNGNLIAPRGYFKIRTGKSSIMRSDAPIGTCSSECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMIDGWYGFRWQNSEGTGQAADLKSTQAAIDQINGILNRVIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFEKTRRQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 43: UFV181136 (신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYIFCQVLAQNLPGNDNSTATLCLGHHAVPNGTLVKTITNDQIEVTNATELVQSSSTGRICDSPHRILDGKNCTLIDALLGDPHCDGFQNEKWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFINEGFNWTGVTQNGGSYACKRGPDKSFFSRLNWLYESESTYPVLNVTMPNNDNFDKLYIWGVHHPSTDKEQTNLYVQASGRVTVSTKRSQQTIIPNVGSRPWVRGLSSRISIYWTIVKPGDILLINSNGNLIAPRGYFKIRTGKSSIMRSDAPIGTCSSECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMIDGWYGFRHQNSEGTGQAADLKSTQAAIDQINGKLNRVIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFEKTRRQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVGSHHHHHH
서열번호 44: UFV181097 (신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTIVKTITNDQIEVTNATELVQSSSTGGICDSPHQILDGENCTLIDALLGDPQCDGFQNKKWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNDESFNWTGVTQNGTSSSCKRRSNNSFFSRLNWLTHLKFKYPALNVTMPNNEKFDKLYIWGVHHPVTDNDQIFLYAQASGRITVSTKRSQQTVIPNIGSRPRIRNIPSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGKSSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRWQNSEGIGQAADLKSTQAAINQINGILNRLIGKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFERTKKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 45: UFV181138 (신호 펩티드 및 태그는 밑줄 표시됨)
MKTIIALSYILCLVFAQKLPGNDNSTATLCLGHHAVPNGTIVKTITNDQIEVTNATELVQSSSTGGICDSPHQILDGENCTLIDALLGDPQCDGFQNKKWDLFVERSKAYSNCYPYDVPDYASLRSLVASSGTLEFNDESFNWTGVTQNGTSSSCKRRSNNSFFSRLNWLTHLKFKYPALNVTMPNNEKFDKLYIWGVHHPVTDNDQIFLYAQASGRITVSTKRSQQTVIPNIGSRPRIRNIPSRISIYWTIVKPGDILLINSTGNLIAPRGYFKIRSGKSSIMRSDAPIGKCNSECITPNGSIPNDKPFQNVNRITYGACPRYVKQNTLKLATGMRNVPEKQTRGIFGAIAGFIENGWEGMVDGWYGFRHQNSEGIGQAADLKSTQAAINQINGKLNRLIGKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDLTDSEMNKLFERTKKQLRENAEDMGNGCFKIYHKCDNACIGSIRNGTYDHDVYRDEALNNRFQIKGVGSHHHHHH
서열번호 46: UFV181148 (신호 펩티드 및 태그는 밑줄 표시됨)
MLSIVILFLLVAENSSQNYTGNPVICMGHHAVANGTMVKTLTDDQVEVVTAQELVESQNLPELCPSPLRLVDGQTCDIINGALGSPGCDHLNGAEWDVFIERPNAMDTCYPFDVPDYQSLRSILANNGKFEFIAEEFQWTTVKQNGKSGACKRANVNDFFRRLNWLVKSDRNAYPLQNLTKVNNGDYARLYIWGVHHPSTDTEQTNLYKNNPGRVTVSTKTSQTSVIPNIGSRPWVRGQSGRISFYWTIVEPGDLIVFNTIGNLIAPRGHYKLNNQKKGTILNTAIPIGSCVSKCHTDKGSLSTTKPFQNISRIAIGDCPKYVKQGSLKLATGMRNIPEKASRGLFGAIAGFIENGWQGLIDGWYGFRHQNAEGTGTAADLKSTQAAIDQINGKLNRLIEKTNEKYHQIEKEFEQVEGRIQDLEKYVEDTKIDLWSYNAELLVALENQHTIDVTDSEMNKLFERVRRQLRENAEDKGNGCFEIFHKCDNNCIESIRNGTYDHDIYRDEAINNRFQIQGVGSHHHHHH
서열번호 47: UFV181149 (신호 펩티드 및 태그는 밑줄 표시됨)
MLSIVILFLLVAENSSQNYTGNPVICMGHHAVANGTMVKTLTDDQVEVVTAQELVESQNLPELCPSPLRLVDGQTCDIINGALGSPGCDHLNGAEWDVFIERPNAMDTCYPFDVPDYQSLRSILANNGKFEFIAEEFQWTTVKQNGKSGACKRANVNDFFRRLNWLVKSDRNAYPLQNLTKVNNGDYARLYIWGVHHPSTDTEQTNLYKNNPGRVTVSTKTSQTSVIPNIGSRPWVRGQSGRISFYWTIVEPGDLIVFNTIGNLIAPRGHYKLNNQKKGTILNTAIPIGSCVSKCHTDKGSLSTTKPFQNISRIAIGDCPKYVKQGSLKLATGMRNIPEKASRGLFGAIAGFIENGWQGLIDGWYGFRWQNAEGTGTAADLKSTQAAIDQINGILNRLIEKTNEKYHQIEKEFEQVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDVTDSEMNKLFERVRRQLRENAEDKGNGCFEIFHKCDNNCIESIRNGTYDHDIYRDEAINNRFQIQGVSGSLPETGGGSHHHHHH
서열번호 50: UFV190839 (신호 펩티드 및 태그는 밑줄 표시됨
MKTIIALSYIFCLALGQDLPGNDNSTATLCLGHHAVPNGTLVKTITDDQIEVTNATELVQSSSTGKICNNPHRILDGIDCTLIDALLGDPHCDVFQNETWDLFVERSKAFSNCYPYDVPDYASLRSLVASSGTLEFITEGFTWTGVTQNGGSNACKRGPGSGFFSRLNWLTKSGSTYPVLNVTMPNNDNFDKLYIWGVHHPSTNQEQTSLYVQASGRVTVSTRRSQQTIIPNIGSRPWVRGLSSRISIYWTIVKPGDVLVINSNGNLIAPRGYFKMRTGKSSIMRSDAPIDTCISECITPNGSIPNDKPFQNVNKITYGACPKYVKQNTLKLATGMRNVPEKQTRGLFGAIAGFIENGWEGMIDGWYGFRWQNSEGTGQAADLKSTQAAIDQINGILNRVIEKTNEKFHQIEKEFSEVEGRIQDLEKYVEDTKIDLWSYNAELLVALINQHTIDLTDSEMNKLFEKTRRQLRENAEDMGNGCFKIYHKCDNACIESIRNGTYDHDVYRDEALNNRFQIKGVSGSLPETGGGSHHHHHH
서열번호 51: UFV190068 (신호 펩티드 및 태그는 밑줄 표시됨) MNTQILVFALMAIIPTNADKICLGHHAVSNGTKVNTLTERGVEVVNATETVERTNVPRICSKGKRTVDLGQCGLLGTITGPPQCDQFLEFSADLIIERREGSDVCYPGKFVNEEALRQILRESGGIDKETMGFTYSGIRTNGATSACRRSGSSFYAEMKWLLSNTDNAAFPQMTKSYKNTRKDPALIIWGIHHSGSTTEQTKLYGSGNKLITVGSSNYQQSFVPSPGARPQVNGQSGRIDFHWLILNPNDTVTFSFNGAFIAPDRASFLRGKSMGIQSGVQVDANCEGDCYHSGGTIISNLPFQNINSRAVGKCPRYVKQESLLLATGMKNVPEIPKGRGLFGAIAGFIENGWEGLIDGWYGFRWQNAQGEGTAADYKSTQSAIDQITGILNRLIEKTNQQFELIDNEFTEVEKQIGNVINWTRDSMTEVWSYNAELLVAMINQHTIDLADSEMNKLYERVKRQLRENAEEDGTGCFEIFHKCDDDCMASIRNNTYDHSKYREEAMQNRIQIDPVSGSLPETGGGSHHHHHH
서열번호 52: UFV190841 (신호 펩티드 및 태그는 밑줄 표시됨)
MYKVVVIIALLGAVKGDRICLGHHAVANGTIVKTLTNEQEEVTNATETVESTNLNKLCMKGRSYKDLGNCHPVGMLIGTPVCDPHLTGTWDTLIERENAIAHCYPGATINEEALRQKIMESGGISKMSTGFTYGSSINSAGTTKACMRNGGDSFYAELKWLVSKTKGQNFPQTTNTYRNTDTAEHLIIWGIHHPSSTQEKNDLYGTQSLSISVESSTYQNNFVPVVGARPQVNGQSGRIDFHWTLVQPGDNITFSHNGGLIAPSRVSKLTGRGLGIQSEALIDNSCESKCFWRGGSINTKLPFQNLSPRTVGQCPKYVNQRSLLLATGMRNVPEVVQGRGLFGAIAGFIENGWEGMVDGWYGFRWQNAQGTGQAADYKSTQAAIDQITGILNRLIEKTNTEFESIESEFSETEHQIGNVINWTKDSITDIWTYQAELLVAMINQHTIDMADSEMLNLYERVRKQLRQNAEEDGKGCFEIYHTCDDSCMESIRNNTYDHSQYREEALLNRLNINSSGSLPETGGGSHHHHHH
SEQUENCE LISTING
<110> Janssen Vaccines & Prevention B.V.
<120> RECOMBINANT INFLUENZA ANTIGENS
<130> CRU6016WOPCT1
<140> PCT/EP2020/061335
<141> 2020-04-23
<150> US 62/838,690
<151> 2019-04-25
<160> 52
<170> PatentIn version 3.5
<210> 1
<211> 550
<212> PRT
<213> Artificial Sequence
<220>
<223> CAA24269.1 haemagglutinin (Influenza A virus
(A/Aichi/2/1968(H3N2) (excluding signal sequence)
<400> 1
Gln Asp Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
1 5 10 15
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asp Asp
20 25 30
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
35 40 45
Gly Lys Ile Cys Asn Asn Pro His Arg Ile Leu Asp Gly Ile Asp Cys
50 55 60
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Val Phe Gln
65 70 75 80
Asn Glu Thr Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
85 90 95
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
100 105 110
Ala Ser Ser Gly Thr Leu Glu Phe Ile Thr Glu Gly Phe Thr Trp Thr
115 120 125
Gly Val Thr Gln Asn Gly Gly Ser Asn Ala Cys Lys Arg Gly Pro Gly
130 135 140
Ser Gly Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Ser Gly Ser Thr
145 150 155 160
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
165 170 175
Leu Tyr Ile Trp Gly Ile His His Pro Ser Thr Asn Gln Glu Gln Thr
180 185 190
Ser Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Arg Arg
195 200 205
Ser Gln Gln Thr Ile Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg
210 215 220
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
225 230 235 240
Asp Val Leu Val Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
245 250 255
Tyr Phe Lys Met Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
260 265 270
Pro Ile Asp Thr Cys Ile Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
275 280 285
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
290 295 300
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
305 310 315 320
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Leu Phe Gly Ala Ile Ala
325 330 335
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
340 345 350
Phe Arg His Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
355 360 365
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Val
370 375 380
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
385 390 395 400
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
405 410 415
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
420 425 430
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
435 440 445
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Glu Met Gly Asn
450 455 460
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Glu Ser
465 470 475 480
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
485 490 495
Asn Asn Arg Phe Gln Ile Lys Gly Val Glu Leu Lys Ser Gly Tyr Lys
500 505 510
Asp Trp Ile Leu Trp Ile Ser Phe Ala Ile Ser Cys Phe Leu Leu Cys
515 520 525
Val Val Leu Leu Gly Phe Ile Met Trp Ala Cys Gln Arg Gly Asn Ile
530 535 540
Arg Cys Asn Ile Cys Ile
545 550
<210> 2
<211> 121
<212> PRT
<213> Artificial Sequence
<220>
<223> CR6261 VH
<400> 2
Glu Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Pro Phe Arg Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Pro Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Thr Lys Tyr Ala Pro Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Asp Phe Ala Gly Thr Val Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Lys His Met Gly Tyr Gln Val Arg Glu Thr Met Asp Val Trp Gly
100 105 110
Lys Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 3
<211> 111
<212> PRT
<213> Artificial Sequence
<220>
<223> CR6261 VL
<400> 3
Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln
1 5 10 15
Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asp
20 25 30
Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln
65 70 75 80
Thr Gly Asp Glu Ala Asn Tyr Tyr Cys Ala Thr Trp Asp Arg Arg Pro
85 90 95
Thr Ala Tyr Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 4
<211> 122
<212> PRT
<213> Artificial Sequence
<220>
<223> CR8020 VH
<400> 4
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys Thr Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Phe
20 25 30
Gly Val Ser Trp Ile Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Trp Ile Ser Ala Tyr Asn Gly Asp Thr Tyr Tyr Ala Gln Lys Phe
50 55 60
Gln Ala Arg Val Thr Met Thr Thr Asp Thr Ser Thr Thr Thr Ala Tyr
65 70 75 80
Met Glu Met Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Pro Pro Leu Phe Tyr Ser Ser Trp Ser Leu Asp Asn Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 5
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> CR8020 VL
<400> 5
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Met Asn
20 25 30
Tyr Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Arg Arg Ala Thr Gly Ile Pro Asp Arg Ile Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Ala Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Thr Ser Pro
85 90 95
Arg Thr Phe Gly Gln Gly Ala Lys Val Glu Ile Lys
100 105
<210> 6
<211> 121
<212> PRT
<213> Artificial Sequence
<220>
<223> CR9114 VH
<400> 6
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ser Ser Gly Gly Thr Ser Asn Asn Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Asp Trp Met
35 40 45
Gly Gly Ile Ser Pro Ile Phe Gly Ser Thr Ala Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Ser Ala Asp Ile Phe Ser Asn Thr Ala Tyr
65 70 75 80
Met Glu Leu Asn Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg His Gly Asn Tyr Tyr Tyr Tyr Ser Gly Met Asp Val Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 7
<211> 110
<212> PRT
<213> Artificial Sequence
<220>
<223> CR9114 VL
<400> 7
Ser Tyr Val Leu Thr Gln Pro Pro Ala Val Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Ser Gly Ser Asp Ser Asn Ile Gly Arg Arg
20 25 30
Ser Val Asn Trp Tyr Gln Gln Phe Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Ser Asn Asp Gln Arg Pro Ser Val Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu
85 90 95
Lys Gly Ala Val Phe Gly Gly Gly Thr Gln Leu Thr Val Leu
100 105 110
<210> 8
<211> 749
<212> PRT
<213> Artificial Sequence
<220>
<223> MD3606
<400> 8
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 Val Ser Ile Ser Ile Phe Asp Ile Tyr
20 25 30
Ala Met Asp Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Asp Leu Val
35 40 45
Ala Thr Ser Phe Arg Asp Gly Ser Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Leu Cys His
85 90 95
Val Ser Leu Tyr Arg Asp Pro Leu Gly Val Ala Gly Gly Met Gly Val
100 105 110
Tyr Trp Gly Lys Gly Ala Leu Val Thr Val Ser Ser Gly Gly Gly Gly
115 120 125
Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly
130 135 140
Leu Val Gln Ala Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly
145 150 155 160
Arg Thr Tyr Ala Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg
165 170 175
Glu Phe Val Ala His Ile Asn Ala Leu Gly Thr Arg Thr Tyr Tyr Ser
180 185 190
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn
195 200 205
Thr Glu Tyr Leu Glu Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val
210 215 220
Tyr Tyr Cys Thr Ala Gln Gly Gln Trp Arg Ala Ala Pro Val Ala Val
225 230 235 240
Ala Ala Glu Tyr Glu Phe Trp Gly Gln Gly Thr Gln Val Thr Val Ser
245 250 255
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val
260 265 270
Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser
275 280 285
Cys Ala Ala Thr Gly Phe Thr Leu Glu Asn Lys Ala Ile Gly Trp Phe
290 295 300
Arg Gln Thr Pro Gly Ser Glu Arg Glu Gly Val Leu Cys Ile Ser Lys
305 310 315 320
Ser Gly Ser Trp Thr Tyr Tyr Thr Asp Ser Met Arg Gly Arg Phe Thr
325 330 335
Ile Ser Arg Asp Asn Ala Glu Asn Thr Val Tyr Leu Gln Met Asp Ser
340 345 350
Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Thr Thr Thr Ala
355 360 365
Gly Gly Gly Leu Cys Trp Asp Gly Thr Thr Phe Ser Arg Leu Ala Ser
370 375 380
Ser Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser Gly Gly Gly Gly
385 390 395 400
Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly
405 410 415
Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly
420 425 430
Phe Thr Phe Ser Thr Ser Trp Met Tyr Trp Leu Arg Gln Ala Pro Gly
435 440 445
Lys Gly Leu Glu Trp Val Ser Val Ile Asn Thr Asp Gly Gly Thr Tyr
450 455 460
Tyr Ala Asp Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ala
465 470 475 480
Lys Asp Thr Leu Tyr Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr
485 490 495
Ala Val Tyr Tyr Cys Ala Lys Asp Trp Gly Gly Pro Glu Pro Thr Arg
500 505 510
Gly Gln Gly Thr Gln Val Thr Val Ser Ser Asp Lys Thr His Thr Cys
515 520 525
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
530 535 540
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
545 550 555 560
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
565 570 575
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
580 585 590
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
595 600 605
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
610 615 620
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
625 630 635 640
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
645 650 655
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
660 665 670
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
675 680 685
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
690 695 700
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
705 710 715 720
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
725 730 735
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
740 745
<210> 9
<211> 526
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181157
<400> 9
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Pro Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Ser Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Ile Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Ala Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Lys Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Glu Ile Thr Asn Lys Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Gly Ser His His His His His His
515 520 525
<210> 10
<211> 535
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181009
<400> 10
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Pro Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Ser Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Ile Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Ala Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Lys Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His Trp Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Glu Ile Thr Asn Ile Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Ile Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Ile Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly
515 520 525
Ser His His His His His His
530 535
<210> 11
<211> 526
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181134
<400> 11
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Thr Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Ser Asn Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asn Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Asn Gln Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Thr Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Thr Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Thr Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Thr Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Glu Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Val Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Lys Ile Thr Asn Lys Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Asn Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Lys Ile Asp Gly Ser His His His His His His
515 520 525
<210> 12
<211> 535
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181091
<400> 12
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Thr Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Ser Asn Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asn Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Asn Gln Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Thr Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Thr Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Thr Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Thr Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Glu Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Val Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His Trp Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Lys Ile Thr Asn Ile Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Ile Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Asn Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Ile Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Lys Ile Asp Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly
515 520 525
Ser His His His His His His
530 535
<210> 13
<211> 522
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181153
<400> 13
Met Ala Ile Ile Tyr Leu Ile Leu Leu Phe Ala Ala Val Arg Gly Asp
1 5 10 15
Gln Ile Cys Ile Gly Tyr His Ser Asn Asn Ser Thr Glu Lys Val Asp
20 25 30
Thr Ile Leu Glu Arg Asn Val Thr Val Thr His Ala Gln Asp Ile Leu
35 40 45
Glu Lys Thr His Asn Gly Lys Leu Cys Lys Leu Asn Gly Ile Pro Pro
50 55 60
Leu Glu Leu Gly Asp Cys Ser Ile Ala Gly Trp Leu Leu Gly Asn Pro
65 70 75 80
Glu Cys Asp Arg Leu Leu Thr Val Pro Glu Trp Ser Tyr Ile Met Glu
85 90 95
Lys Glu Asn Pro Arg Asn Gly Leu Cys Tyr Pro Gly Ser Phe Asn Asp
100 105 110
Tyr Glu Glu Leu Lys His Leu Leu Ser Ser Val Thr His Phe Glu Lys
115 120 125
Val Lys Ile Leu Pro Arg Asp Arg Trp Thr Gln His Thr Thr Thr Gly
130 135 140
Gly Ser Arg Ala Cys Ala Val Ser Gly Asn Pro Ser Phe Phe Arg Asn
145 150 155 160
Met Val Trp Leu Thr Lys Lys Gly Ser Asn Tyr Pro Ile Ala Lys Gly
165 170 175
Ser Tyr Asn Asn Thr Ser Gly Glu Gln Met Leu Ile Ile Trp Gly Val
180 185 190
His His Pro Asn Asp Asp Ala Glu Gln Arg Thr Leu Tyr Gln Asn Val
195 200 205
Gly Thr Tyr Val Ser Val Gly Thr Ser Thr Leu Asn Lys Arg Ser Val
210 215 220
Pro Glu Ile Ala Thr Arg Pro Lys Val Asn Gly Gln Gly Gly Arg Met
225 230 235 240
Glu Phe Ser Trp Thr Ile Leu Asp Met Leu Asp Thr Ile Asn Phe Glu
245 250 255
Ser Thr Gly Asn Leu Ile Ala Pro Glu Tyr Gly Phe Arg Ile Ser Lys
260 265 270
Arg Gly Ser Ser Gly Ile Met Lys Thr Glu Gly Thr Leu Glu Asn Cys
275 280 285
Glu Thr Lys Cys Gln Thr Pro Leu Gly Ala Ile Asn Thr Thr Leu Pro
290 295 300
Phe His Asn Ile His Pro Leu Thr Ile Gly Glu Cys Pro Lys Tyr Val
305 310 315 320
Lys Ser Glu Arg Leu Val Leu Ala Thr Gly Leu Arg Asn Val Pro Gln
325 330 335
Ile Glu Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly
340 345 350
Gly Trp Gln Gly Met Val Asp Gly Trp Tyr Gly Tyr His His Ser Asn
355 360 365
Asp Gln Gly Ser Gly Tyr Ala Ala Asp Lys Glu Ser Thr Gln Arg Ala
370 375 380
Ile Asp Gly Ile Thr Asn Lys Val Asn Ser Val Ile Glu Lys Met Asn
385 390 395 400
Thr Gln Phe Glu Ala Val Gly Lys Glu Phe Asn Asn Leu Glu Lys Arg
405 410 415
Leu Glu Asn Leu Asn Lys Lys Met Glu Asp Gly Phe Leu Asp Val Trp
420 425 430
Thr Tyr Asn Ala Glu Leu Leu Val Leu Met Glu Asn Glu Arg Thr Leu
435 440 445
Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Asp Lys Val Arg Met
450 455 460
Gln Leu Arg Asp Asn Ala Lys Glu Leu Gly Asn Gly Cys Phe Glu Phe
465 470 475 480
Tyr His Lys Cys Asp Asp Glu Cys Met Asn Ser Val Lys Asn Gly Thr
485 490 495
Tyr Asp Tyr Pro Lys Tyr Glu Glu Glu Ser Lys Leu Asn Arg Asn Glu
500 505 510
Ile Lys Gly Ser His His His His His His
515 520
<210> 14
<211> 531
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181154
<400> 14
Met Ala Ile Ile Tyr Leu Ile Leu Leu Phe Ala Ala Val Arg Gly Asp
1 5 10 15
Gln Ile Cys Ile Gly Tyr His Ser Asn Asn Ser Thr Glu Lys Val Asp
20 25 30
Thr Ile Leu Glu Arg Asn Val Thr Val Thr His Ala Gln Asp Ile Leu
35 40 45
Glu Lys Thr His Asn Gly Lys Leu Cys Lys Leu Asn Gly Ile Pro Pro
50 55 60
Leu Glu Leu Gly Asp Cys Ser Ile Ala Gly Trp Leu Leu Gly Asn Pro
65 70 75 80
Glu Cys Asp Arg Leu Leu Thr Val Pro Glu Trp Ser Tyr Ile Met Glu
85 90 95
Lys Glu Asn Pro Arg Asn Gly Leu Cys Tyr Pro Gly Ser Phe Asn Asp
100 105 110
Tyr Glu Glu Leu Lys His Leu Leu Ser Ser Val Thr His Phe Glu Lys
115 120 125
Val Lys Ile Leu Pro Arg Asp Arg Trp Thr Gln His Thr Thr Thr Gly
130 135 140
Gly Ser Arg Ala Cys Ala Val Ser Gly Asn Pro Ser Phe Phe Arg Asn
145 150 155 160
Met Val Trp Leu Thr Lys Lys Gly Ser Asn Tyr Pro Ile Ala Lys Gly
165 170 175
Ser Tyr Asn Asn Thr Ser Gly Glu Gln Met Leu Ile Ile Trp Gly Val
180 185 190
His His Pro Asn Asp Asp Ala Glu Gln Arg Thr Leu Tyr Gln Asn Val
195 200 205
Gly Thr Tyr Val Ser Val Gly Thr Ser Thr Leu Asn Lys Arg Ser Val
210 215 220
Pro Glu Ile Ala Thr Arg Pro Lys Val Asn Gly Gln Gly Gly Arg Met
225 230 235 240
Glu Phe Ser Trp Thr Ile Leu Asp Met Leu Asp Thr Ile Asn Phe Glu
245 250 255
Ser Thr Gly Asn Leu Ile Ala Pro Glu Tyr Gly Phe Arg Ile Ser Lys
260 265 270
Arg Gly Ser Ser Gly Ile Met Lys Thr Glu Gly Thr Leu Glu Asn Cys
275 280 285
Glu Thr Lys Cys Gln Thr Pro Leu Gly Ala Ile Asn Thr Thr Leu Pro
290 295 300
Phe His Asn Ile His Pro Leu Thr Ile Gly Glu Cys Pro Lys Tyr Val
305 310 315 320
Lys Ser Glu Arg Leu Val Leu Ala Thr Gly Leu Arg Asn Val Pro Gln
325 330 335
Ile Glu Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly
340 345 350
Gly Trp Gln Gly Met Val Asp Gly Trp Tyr Gly Tyr His Trp Ser Asn
355 360 365
Asp Gln Gly Ser Gly Tyr Ala Ala Asp Lys Glu Ser Thr Gln Arg Ala
370 375 380
Ile Asp Gly Ile Thr Asn Ile Val Asn Ser Val Ile Glu Lys Met Asn
385 390 395 400
Thr Gln Phe Glu Ala Val Gly Lys Glu Phe Asn Asn Leu Glu Lys Arg
405 410 415
Leu Glu Asn Leu Asn Lys Lys Met Glu Asp Gly Phe Leu Asp Val Trp
420 425 430
Thr Tyr Asn Ala Glu Leu Leu Val Leu Met Ile Asn Glu Arg Thr Leu
435 440 445
Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Asp Lys Val Arg Met
450 455 460
Gln Leu Arg Asp Asn Ala Lys Glu Leu Gly Asn Gly Cys Phe Glu Phe
465 470 475 480
Tyr His Lys Cys Asp Asp Glu Cys Ile Asn Ser Val Lys Asn Gly Thr
485 490 495
Tyr Asp Tyr Pro Lys Tyr Glu Glu Glu Ser Lys Leu Asn Arg Asn Glu
500 505 510
Ile Lys Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His His His
515 520 525
His His His
530
<210> 15
<211> 524
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181158
<400> 15
Met Glu Lys Ile Val Leu Leu Phe Ala Ile Val Ser Leu Val Gln Ser
1 5 10 15
Asp Gln Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Glu Gln Val
20 25 30
Asp Thr Ile Met Glu Lys Asn Val Thr Val Thr His Ala Gln Asp Ile
35 40 45
Leu Glu Lys Thr His Asn Gly Lys Leu Cys Ser Leu Asn Gly Val Lys
50 55 60
Pro Leu Ile Leu Arg Asp Cys Ser Val Ala Gly Trp Leu Leu Gly Asn
65 70 75 80
Pro Met Cys Asp Glu Phe Leu Asn Val Pro Glu Trp Ser Tyr Ile Val
85 90 95
Glu Lys Asp Ser Pro Ile Asn Gly Leu Cys Tyr Pro Gly Asp Phe Asn
100 105 110
Asp Tyr Glu Glu Leu Lys His Leu Leu Ser Ser Thr Asn His Phe Glu
115 120 125
Lys Ile Gln Ile Ile Pro Arg Ser Ser Trp Ser Asn His Asp Ala Ser
130 135 140
Ser Gly Val Ser Ser Ala Cys Pro Tyr Asn Gly Arg Ser Ser Phe Phe
145 150 155 160
Arg Asn Val Val Trp Leu Ile Lys Lys Asn Asn Ala Tyr Pro Thr Ile
165 170 175
Lys Arg Ser Tyr Asn Asn Thr Asn Gln Glu Asp Leu Leu Val Leu Trp
180 185 190
Gly Ile His His Pro Asn Asp Ala Ala Glu Gln Thr Lys Leu Tyr Gln
195 200 205
Asn Pro Thr Thr Tyr Val Ser Val Gly Thr Ser Thr Leu Asn Gln Arg
210 215 220
Ser Val Pro Glu Ile Ala Thr Arg Pro Lys Val Asn Gly Gln Ser Gly
225 230 235 240
Arg Met Glu Phe Phe Trp Thr Ile Leu Lys Pro Asn Asp Ala Ile Asn
245 250 255
Phe Glu Ser Asn Gly Asn Phe Ile Ala Pro Glu Tyr Ala Tyr Lys Ile
260 265 270
Val Lys Lys Gly Asp Ser Ala Ile Met Lys Ser Gly Leu Glu Tyr Gly
275 280 285
Asn Cys Asn Thr Lys Cys Gln Thr Pro Met Gly Ala Ile Asn Ser Ser
290 295 300
Met Pro Phe His Asn Ile His Pro Leu Thr Ile Gly Glu Cys Pro Lys
305 310 315 320
Tyr Val Lys Ser Asp Arg Leu Val Leu Ala Thr Gly Leu Arg Asn Val
325 330 335
Pro Gln Arg Glu Thr Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile
340 345 350
Glu Gly Gly Trp Gln Gly Met Val Asp Gly Trp Tyr Gly Tyr Leu His
355 360 365
Ser Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Lys Glu Ser Thr Gln
370 375 380
Lys Ala Ile Asp Gly Ile Thr Asn Lys Ile Asn Ser Ile Ile Asp Lys
385 390 395 400
Met Asn Thr Gln Phe Glu Ala Val Gly Lys Glu Phe Asn Asn Leu Glu
405 410 415
Arg Arg Ile Glu Asn Leu Asn Lys Lys Met Glu Asp Gly Phe Leu Asp
420 425 430
Val Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Met Glu Asn Glu Arg
435 440 445
Thr Leu Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Asp Lys Val
450 455 460
Arg Leu Gln Leu Arg Asp Asn Ala Lys Glu Leu Gly Asn Gly Cys Phe
465 470 475 480
Glu Phe Tyr His Lys Cys Asp Asp Glu Cys Met Glu Ser Val Arg Asn
485 490 495
Gly Thr Tyr Asp Tyr Pro Gln Tyr Ser Glu Glu Ala Arg Leu Asn Arg
500 505 510
Glu Glu Ile Ser Gly Ser His His His His His His
515 520
<210> 16
<211> 533
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181159
<400> 16
Met Glu Lys Ile Val Leu Leu Phe Ala Ile Val Ser Leu Val Gln Ser
1 5 10 15
Asp Gln Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Glu Gln Val
20 25 30
Asp Thr Ile Met Glu Lys Asn Val Thr Val Thr His Ala Gln Asp Ile
35 40 45
Leu Glu Lys Thr His Asn Gly Lys Leu Cys Ser Leu Asn Gly Val Lys
50 55 60
Pro Leu Ile Leu Arg Asp Cys Ser Val Ala Gly Trp Leu Leu Gly Asn
65 70 75 80
Pro Met Cys Asp Glu Phe Leu Asn Val Pro Glu Trp Ser Tyr Ile Val
85 90 95
Glu Lys Asp Ser Pro Ile Asn Gly Leu Cys Tyr Pro Gly Asp Phe Asn
100 105 110
Asp Tyr Glu Glu Leu Lys His Leu Leu Ser Ser Thr Asn His Phe Glu
115 120 125
Lys Ile Gln Ile Ile Pro Arg Ser Ser Trp Ser Asn His Asp Ala Ser
130 135 140
Ser Gly Val Ser Ser Ala Cys Pro Tyr Asn Gly Arg Ser Ser Phe Phe
145 150 155 160
Arg Asn Val Val Trp Leu Ile Lys Lys Asn Asn Ala Tyr Pro Thr Ile
165 170 175
Lys Arg Ser Tyr Asn Asn Thr Asn Gln Glu Asp Leu Leu Val Leu Trp
180 185 190
Gly Ile His His Pro Asn Asp Ala Ala Glu Gln Thr Lys Leu Tyr Gln
195 200 205
Asn Pro Thr Thr Tyr Val Ser Val Gly Thr Ser Thr Leu Asn Gln Arg
210 215 220
Ser Val Pro Glu Ile Ala Thr Arg Pro Lys Val Asn Gly Gln Ser Gly
225 230 235 240
Arg Met Glu Phe Phe Trp Thr Ile Leu Lys Pro Asn Asp Ala Ile Asn
245 250 255
Phe Glu Ser Asn Gly Asn Phe Ile Ala Pro Glu Tyr Ala Tyr Lys Ile
260 265 270
Val Lys Lys Gly Asp Ser Ala Ile Met Lys Ser Gly Leu Glu Tyr Gly
275 280 285
Asn Cys Asn Thr Lys Cys Gln Thr Pro Met Gly Ala Ile Asn Ser Ser
290 295 300
Met Pro Phe His Asn Ile His Pro Leu Thr Ile Gly Glu Cys Pro Lys
305 310 315 320
Tyr Val Lys Ser Asp Arg Leu Val Leu Ala Thr Gly Leu Arg Asn Val
325 330 335
Pro Gln Arg Glu Thr Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile
340 345 350
Glu Gly Gly Trp Gln Gly Met Val Asp Gly Trp Tyr Gly Tyr Leu Trp
355 360 365
Ser Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Lys Glu Ser Thr Gln
370 375 380
Lys Ala Ile Asp Gly Ile Thr Asn Ile Ile Asn Ser Ile Ile Asp Lys
385 390 395 400
Met Asn Thr Gln Phe Glu Ala Val Gly Lys Glu Phe Asn Asn Leu Glu
405 410 415
Arg Arg Ile Glu Asn Leu Asn Lys Lys Met Glu Asp Gly Phe Leu Asp
420 425 430
Val Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Met Ile Asn Glu Arg
435 440 445
Thr Leu Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Asp Lys Val
450 455 460
Arg Leu Gln Leu Arg Asp Asn Ala Lys Glu Leu Gly Asn Gly Cys Phe
465 470 475 480
Glu Phe Tyr His Lys Cys Asp Asp Glu Cys Ile Glu Ser Val Arg Asn
485 490 495
Gly Thr Tyr Asp Tyr Pro Gln Tyr Ser Glu Glu Ala Arg Leu Asn Arg
500 505 510
Glu Glu Ile Ser Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His
515 520 525
His His His His His
530
<210> 17
<211> 520
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181155
<400> 17
Met Glu Thr Ile Ser Leu Ile Thr Ile Leu Leu Val Val Thr Ala Ser
1 5 10 15
Asn Ala Asp Lys Ile Cys Ile Gly His Gln Ser Thr Asn Ser Thr Glu
20 25 30
Thr Val Asp Thr Leu Thr Glu Thr Asn Val Pro Val Thr His Ala Lys
35 40 45
Glu Leu Leu His Thr Glu His Asn Gly Met Leu Cys Ala Thr Ser Leu
50 55 60
Gly His Pro Leu Ile Leu Asp Thr Cys Thr Ile Glu Gly Leu Val Tyr
65 70 75 80
Gly Asn Pro Ser Cys Asp Leu Leu Leu Gly Gly Arg Glu Trp Ser Tyr
85 90 95
Ile Val Glu Arg Ser Ser Ala Val Asn Gly Thr Cys Tyr Pro Gly Asn
100 105 110
Val Glu Asn Leu Glu Glu Leu Arg Thr Leu Phe Ser Ser Ala Ser Ser
115 120 125
Tyr Gln Arg Ile Gln Ile Phe Pro Asp Thr Thr Trp Asn Val Thr Tyr
130 135 140
Thr Gly Thr Ser Arg Ala Cys Ser Gly Ser Phe Tyr Arg Ser Met Arg
145 150 155 160
Trp Leu Thr Gln Lys Ser Gly Phe Tyr Pro Val Gln Asp Ala Gln Tyr
165 170 175
Thr Asn Asn Arg Gly Lys Ser Ile Leu Phe Val Trp Gly Ile His His
180 185 190
Pro Pro Thr Tyr Thr Glu Gln Thr Asn Leu Tyr Ile Arg Asn Asp Thr
195 200 205
Thr Thr Ser Val Thr Thr Glu Asp Leu Asn Arg Thr Phe Lys Pro Val
210 215 220
Ile Gly Pro Arg Pro Leu Val Asn Gly Leu Gln Gly Arg Ile Asp Tyr
225 230 235 240
Tyr Trp Ser Val Leu Lys Pro Gly Gln Thr Leu Arg Val Arg Ser Asn
245 250 255
Gly Asn Leu Ile Ala Pro Trp Tyr Gly His Val Leu Ser Gly Gly Ser
260 265 270
His Gly Arg Ile Leu Lys Thr Asp Leu Lys Gly Gly Asn Cys Val Val
275 280 285
Gln Cys Gln Thr Glu Lys Gly Gly Leu Asn Ser Thr Leu Pro Phe His
290 295 300
Asn Ile Ser Lys Tyr Ala Phe Gly Thr Cys Pro Lys Tyr Val Arg Val
305 310 315 320
Asn Ser Leu Lys Leu Ala Val Gly Leu Arg Asn Val Pro Ala Arg Ser
325 330 335
Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly Gly Trp
340 345 350
Pro Gly Leu Val Ala Gly Trp Tyr Gly Phe Gln His Ser Asn Asp Gln
355 360 365
Gly Val Gly Met Ala Ala Asp Arg Asp Ser Thr Gln Lys Ala Ile Asp
370 375 380
Lys Ile Thr Ser Lys Val Asn Asn Ile Val Asp Lys Met Asn Lys Gln
385 390 395 400
Tyr Glu Ile Ile Asp His Glu Phe Ser Glu Val Glu Thr Arg Leu Asn
405 410 415
Met Ile Asn Asn Lys Ile Asp Asp Gln Ile Gln Asp Val Trp Ala Tyr
420 425 430
Asn Ala Glu Leu Leu Val Leu Leu Glu Asn Gln Lys Thr Leu Asp Glu
435 440 445
His Asp Ala Asn Val Asn Asn Leu Tyr Asn Lys Val Lys Arg Ala Leu
450 455 460
Gly Ser Asn Ala Met Glu Asp Gly Lys Gly Cys Phe Glu Leu Tyr His
465 470 475 480
Lys Cys Asp Asp Gln Cys Met Glu Thr Ile Arg Asn Gly Thr Tyr Asn
485 490 495
Arg Arg Lys Tyr Arg Glu Glu Ser Arg Leu Glu Arg Gln Lys Ile Glu
500 505 510
Gly Ser His His His His His His
515 520
<210> 18
<211> 529
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181156
<400> 18
Met Glu Thr Ile Ser Leu Ile Thr Ile Leu Leu Val Val Thr Ala Ser
1 5 10 15
Asn Ala Asp Lys Ile Cys Ile Gly His Gln Ser Thr Asn Ser Thr Glu
20 25 30
Thr Val Asp Thr Leu Thr Glu Thr Asn Val Pro Val Thr His Ala Lys
35 40 45
Glu Leu Leu His Thr Glu His Asn Gly Met Leu Cys Ala Thr Ser Leu
50 55 60
Gly His Pro Leu Ile Leu Asp Thr Cys Thr Ile Glu Gly Leu Val Tyr
65 70 75 80
Gly Asn Pro Ser Cys Asp Leu Leu Leu Gly Gly Arg Glu Trp Ser Tyr
85 90 95
Ile Val Glu Arg Ser Ser Ala Val Asn Gly Thr Cys Tyr Pro Gly Asn
100 105 110
Val Glu Asn Leu Glu Glu Leu Arg Thr Leu Phe Ser Ser Ala Ser Ser
115 120 125
Tyr Gln Arg Ile Gln Ile Phe Pro Asp Thr Thr Trp Asn Val Thr Tyr
130 135 140
Thr Gly Thr Ser Arg Ala Cys Ser Gly Ser Phe Tyr Arg Ser Met Arg
145 150 155 160
Trp Leu Thr Gln Lys Ser Gly Phe Tyr Pro Val Gln Asp Ala Gln Tyr
165 170 175
Thr Asn Asn Arg Gly Lys Ser Ile Leu Phe Val Trp Gly Ile His His
180 185 190
Pro Pro Thr Tyr Thr Glu Gln Thr Asn Leu Tyr Ile Arg Asn Asp Thr
195 200 205
Thr Thr Ser Val Thr Thr Glu Asp Leu Asn Arg Thr Phe Lys Pro Val
210 215 220
Ile Gly Pro Arg Pro Leu Val Asn Gly Leu Gln Gly Arg Ile Asp Tyr
225 230 235 240
Tyr Trp Ser Val Leu Lys Pro Gly Gln Thr Leu Arg Val Arg Ser Asn
245 250 255
Gly Asn Leu Ile Ala Pro Trp Tyr Gly His Val Leu Ser Gly Gly Ser
260 265 270
His Gly Arg Ile Leu Lys Thr Asp Leu Lys Gly Gly Asn Cys Val Val
275 280 285
Gln Cys Gln Thr Glu Lys Gly Gly Leu Asn Ser Thr Leu Pro Phe His
290 295 300
Asn Ile Ser Lys Tyr Ala Phe Gly Thr Cys Pro Lys Tyr Val Arg Val
305 310 315 320
Asn Ser Leu Lys Leu Ala Val Gly Leu Arg Asn Val Pro Ala Arg Ser
325 330 335
Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly Gly Trp
340 345 350
Pro Gly Leu Val Ala Gly Trp Tyr Gly Phe Gln Trp Ser Asn Asp Gln
355 360 365
Gly Val Gly Met Ala Ala Asp Arg Asp Ser Thr Gln Lys Ala Ile Asp
370 375 380
Lys Ile Thr Ser Ile Val Asn Asn Ile Val Asp Lys Met Asn Lys Gln
385 390 395 400
Tyr Glu Ile Ile Asp His Glu Phe Ser Glu Val Glu Thr Arg Leu Asn
405 410 415
Met Ile Asn Asn Lys Ile Asp Asp Gln Ile Gln Asp Val Trp Ala Tyr
420 425 430
Asn Ala Glu Leu Leu Val Leu Leu Ile Asn Gln Lys Thr Leu Asp Glu
435 440 445
His Asp Ala Asn Val Asn Asn Leu Tyr Asn Lys Val Lys Arg Ala Leu
450 455 460
Gly Ser Asn Ala Met Glu Asp Gly Lys Gly Cys Phe Glu Leu Tyr His
465 470 475 480
Lys Cys Asp Asp Gln Cys Ile Glu Thr Ile Arg Asn Gly Thr Tyr Asn
485 490 495
Arg Arg Lys Tyr Arg Glu Glu Ser Arg Leu Glu Arg Gln Lys Ile Glu
500 505 510
Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His His His His His
515 520 525
His
<210> 19
<211> 529
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181141
<400> 19
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Phe Cys Leu Ala Leu Gly
1 5 10 15
Gln Asp Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asp Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Lys Ile Cys Asn Asn Pro His Arg Ile Leu Asp Gly Ile Asp Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Val Phe Gln
85 90 95
Asn Glu Thr Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Thr Glu Gly Phe Thr Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Gly Ser Asn Ala Cys Lys Arg Gly Pro Gly
145 150 155 160
Ser Gly Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Ser Gly Ser Thr
165 170 175
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asn Gln Glu Gln Thr
195 200 205
Ser Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Arg Arg
210 215 220
Ser Gln Gln Thr Ile Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Val Leu Val Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Met Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Asp Thr Cys Ile Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Leu Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Val
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Glu Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Gly Ser His His His His His
515 520 525
His
<210> 20
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV180660
<400> 20
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Phe Cys Leu Ala Leu Gly
1 5 10 15
Gln Asp Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asp Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Lys Ile Cys Asn Asn Pro His Arg Ile Leu Asp Gly Ile Asp Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Val Phe Gln
85 90 95
Asn Glu Thr Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Thr Glu Gly Phe Thr Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Gly Ser Asn Ala Cys Lys Arg Gly Pro Gly
145 150 155 160
Ser Gly Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Ser Gly Ser Thr
165 170 175
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asn Gln Glu Gln Thr
195 200 205
Ser Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Arg Arg
210 215 220
Ser Gln Gln Thr Ile Ile Pro Asn Ile Trp Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Val Leu Val Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Met Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Asp Thr Cys Ile Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Leu Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn Arg Val
385 390 395 400
Ile Glu Lys Met Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Glu Ser
485 490 495
Ile Arg Asn Gly Asn Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 21
<211> 529
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181137
<400> 21
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Ser Asn Gly Thr Leu Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Arg Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Gly Phe Gln
85 90 95
Asn Lys Glu Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Asn Asn Glu Ser Phe Asn Trp Thr
130 135 140
Gly Val Ala Gln Asn Gly Thr Ser Ser Ala Cys Lys Arg Arg Ser Asn
145 150 155 160
Lys Ser Phe Phe Ser Arg Leu Asn Trp Leu His Gln Leu Lys Tyr Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Glu Lys Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asp Ser Asp Gln Ile
195 200 205
Ser Ile Tyr Ala Gln Ala Ser Gly Arg Val Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Ser Pro Trp Val Arg
225 230 235 240
Gly Val Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Ser Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Lys Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asn Gln Ile Asn Gly Lys Leu Asn Arg Leu
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Arg Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Gly Ser His His His His His
515 520 525
His
<210> 22
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181096
<400> 22
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Ser Asn Gly Thr Leu Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Arg Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Gly Phe Gln
85 90 95
Asn Lys Glu Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Asn Asn Glu Ser Phe Asn Trp Thr
130 135 140
Gly Val Ala Gln Asn Gly Thr Ser Ser Ala Cys Lys Arg Arg Ser Asn
145 150 155 160
Lys Ser Phe Phe Ser Arg Leu Asn Trp Leu His Gln Leu Lys Tyr Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Glu Lys Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asp Ser Asp Gln Ile
195 200 205
Ser Ile Tyr Ala Gln Ala Ser Gly Arg Val Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Ser Pro Trp Val Arg
225 230 235 240
Gly Val Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Ser Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Lys Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asn Gln Ile Asn Gly Ile Leu Asn Arg Leu
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Arg Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 23
<211> 531
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181145
<400> 23
Met Ile Ala Leu Ile Leu Val Ala Leu Ala Leu Ser His Thr Ala Tyr
1 5 10 15
Ser Gln Ile Thr Asn Gly Thr Thr Gly Asn Pro Ile Ile Cys Leu Gly
20 25 30
His His Ala Val Glu Asn Gly Thr Ser Val Lys Thr Leu Thr Asp Asn
35 40 45
His Val Glu Val Val Ser Ala Lys Glu Leu Val Glu Thr Asn His Thr
50 55 60
Asp Glu Leu Cys Pro Ser Pro Leu Lys Leu Val Asp Gly Gln Asp Cys
65 70 75 80
Asp Leu Ile Asn Gly Ala Leu Gly Ser Pro Gly Cys Asp Arg Leu Gln
85 90 95
Asp Thr Thr Trp Asp Val Phe Ile Glu Arg Pro Thr Ala Val Asp Thr
100 105 110
Cys Tyr Pro Phe Asp Val Pro Asp Tyr Gln Ser Leu Arg Ser Ile Leu
115 120 125
Ala Ser Ser Gly Ser Leu Glu Phe Ile Ala Glu Gln Phe Thr Trp Asn
130 135 140
Gly Val Lys Val Asp Gly Ser Ser Ser Ala Cys Leu Arg Gly Gly Arg
145 150 155 160
Asn Ser Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Glu Thr Asn Gly
165 170 175
Asn Tyr Gly Pro Ile Asn Val Thr Lys Glu Asn Thr Gly Ser Tyr Val
180 185 190
Arg Leu Tyr Leu Trp Gly Val His His Pro Ser Ser Asp Asn Glu Gln
195 200 205
Thr Asp Leu Tyr Lys Val Ala Thr Gly Arg Val Thr Val Ser Thr Arg
210 215 220
Ser Asp Gln Ile Ser Ile Val Pro Asn Ile Gly Ser Arg Pro Arg Val
225 230 235 240
Arg Asn Gln Ser Gly Arg Ile Ser Ile Tyr Trp Thr Leu Val Asn Pro
245 250 255
Gly Asp Ser Ile Ile Phe Asn Ser Ile Gly Asn Leu Ile Ala Pro Arg
260 265 270
Gly His Tyr Lys Ile Ser Lys Ser Thr Lys Ser Thr Val Leu Lys Ser
275 280 285
Asp Lys Arg Ile Gly Ser Cys Thr Ser Pro Cys Leu Thr Asp Lys Gly
290 295 300
Ser Ile Gln Ser Asp Lys Pro Phe Gln Asn Val Ser Arg Ile Ala Ile
305 310 315 320
Gly Asn Cys Pro Lys Tyr Val Lys Gln Gly Ser Leu Met Leu Ala Thr
325 330 335
Gly Met Arg Asn Ile Pro Gly Lys Gln Ala Lys Gly Leu Phe Gly Ala
340 345 350
Ile Ala Gly Phe Ile Glu Asn Gly Trp Gln Gly Leu Ile Asp Gly Trp
355 360 365
Tyr Gly Phe Arg His Gln Asn Ala Glu Gly Thr Gly Thr Ala Ala Asp
370 375 380
Leu Lys Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn
385 390 395 400
Arg Leu Ile Glu Lys Thr Asn Glu Lys Tyr His Gln Ile Glu Lys Glu
405 410 415
Phe Glu Gln Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu
420 425 430
Asp Thr Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala
435 440 445
Leu Glu Asn Gln His Thr Ile Asp Val Thr Asp Ser Glu Met Asn Lys
450 455 460
Leu Phe Glu Arg Val Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Gln
465 470 475 480
Gly Asn Gly Cys Phe Glu Ile Phe His Gln Cys Asp Asn Asn Cys Ile
485 490 495
Glu Ser Ile Arg Asn Gly Thr Tyr Asp His Asn Ile Tyr Arg Asp Glu
500 505 510
Ala Ile Asn Asn Arg Ile Lys Ile Asn Pro Val Gly Ser His His His
515 520 525
His His His
530
<210> 24
<211> 540
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV180661
<400> 24
Met Ile Ala Leu Ile Leu Val Ala Leu Ala Leu Ser His Thr Ala Tyr
1 5 10 15
Ser Gln Ile Thr Asn Gly Thr Thr Gly Asn Pro Ile Ile Cys Leu Gly
20 25 30
His His Ala Val Glu Asn Gly Thr Ser Val Lys Thr Leu Thr Asp Asn
35 40 45
His Val Glu Val Val Ser Ala Lys Glu Leu Val Glu Thr Asn His Thr
50 55 60
Asp Glu Leu Cys Pro Ser Pro Leu Lys Leu Val Asp Gly Gln Asp Cys
65 70 75 80
Asp Leu Ile Asn Gly Ala Leu Gly Ser Pro Gly Cys Asp Arg Leu Gln
85 90 95
Asp Thr Thr Trp Asp Val Phe Ile Glu Arg Pro Thr Ala Val Asp Thr
100 105 110
Cys Tyr Pro Phe Asp Val Pro Asp Tyr Gln Ser Leu Arg Ser Ile Leu
115 120 125
Ala Ser Ser Gly Ser Leu Glu Phe Ile Ala Glu Gln Phe Thr Trp Asn
130 135 140
Gly Val Lys Val Asp Gly Ser Ser Ser Ala Cys Leu Arg Gly Gly Arg
145 150 155 160
Asn Ser Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Glu Thr Asn Gly
165 170 175
Asn Tyr Gly Pro Ile Asn Val Thr Lys Glu Asn Thr Gly Ser Tyr Val
180 185 190
Arg Leu Tyr Leu Trp Gly Val His His Pro Ser Ser Asp Asn Glu Gln
195 200 205
Thr Asp Leu Tyr Lys Val Ala Thr Gly Arg Val Thr Val Ser Thr Arg
210 215 220
Ser Asp Gln Ile Ser Ile Val Pro Asn Ile Gly Ser Arg Pro Arg Val
225 230 235 240
Arg Asn Gln Ser Gly Arg Ile Ser Ile Tyr Trp Thr Leu Val Asn Pro
245 250 255
Gly Asp Ser Ile Ile Phe Asn Ser Ile Gly Asn Leu Ile Ala Pro Arg
260 265 270
Gly His Tyr Lys Ile Ser Lys Ser Thr Lys Ser Thr Val Leu Lys Ser
275 280 285
Asp Lys Arg Ile Gly Ser Cys Thr Ser Pro Cys Leu Thr Asp Lys Gly
290 295 300
Ser Ile Gln Ser Asp Lys Pro Phe Gln Asn Val Ser Arg Ile Ala Ile
305 310 315 320
Gly Asn Cys Pro Lys Tyr Val Lys Gln Gly Ser Leu Met Leu Ala Thr
325 330 335
Gly Met Arg Asn Ile Pro Gly Lys Gln Ala Lys Gly Leu Phe Gly Ala
340 345 350
Ile Ala Gly Phe Ile Glu Asn Gly Trp Gln Gly Leu Ile Asp Gly Trp
355 360 365
Tyr Gly Phe Arg Trp Gln Asn Ala Glu Gly Thr Gly Thr Ala Ala Asp
370 375 380
Leu Lys Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn
385 390 395 400
Arg Leu Ile Glu Lys Met Asn Glu Lys Tyr His Gln Ile Glu Lys Glu
405 410 415
Phe Glu Gln Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu
420 425 430
Asp Thr Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala
435 440 445
Leu Ile Asn Gln His Thr Ile Asp Val Thr Asp Ser Glu Met Asn Lys
450 455 460
Leu Phe Glu Arg Val Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Gln
465 470 475 480
Gly Asn Gly Cys Phe Glu Ile Phe His Gln Cys Asp Asn Asn Cys Ile
485 490 495
Glu Ser Ile Arg Asn Gly Thr Tyr Asp His Asn Ile Tyr Arg Asp Glu
500 505 510
Ala Ile Asn Asn Arg Ile Lys Ile Asn Pro Val Ser Gly Ser Leu Pro
515 520 525
Glu Thr Gly Gly Gly Ser His His His His His His
530 535 540
<210> 25
<211> 523
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181146
<400> 25
Met Asn Thr Gln Ile Leu Val Phe Ala Leu Met Ala Ile Ile Pro Thr
1 5 10 15
Asn Ala Asp Lys Ile Cys Leu Gly His His Ala Val Ser Asn Gly Thr
20 25 30
Lys Val Asn Thr Leu Thr Glu Arg Gly Val Glu Val Val Asn Ala Thr
35 40 45
Glu Thr Val Glu Arg Thr Asn Val Pro Arg Ile Cys Ser Lys Gly Lys
50 55 60
Arg Thr Val Asp Leu Gly Gln Cys Gly Leu Leu Gly Thr Ile Thr Gly
65 70 75 80
Pro Pro Gln Cys Asp Gln Phe Leu Glu Phe Ser Ala Asp Leu Ile Ile
85 90 95
Glu Arg Arg Glu Gly Ser Asp Val Cys Tyr Pro Gly Lys Phe Val Asn
100 105 110
Glu Glu Ala Leu Arg Gln Ile Leu Arg Glu Ser Gly Gly Ile Asp Lys
115 120 125
Glu Thr Met Gly Phe Thr Tyr Ser Gly Ile Arg Thr Asn Gly Ala Thr
130 135 140
Ser Ala Cys Arg Arg Ser Gly Ser Ser Phe Tyr Ala Glu Met Lys Trp
145 150 155 160
Leu Leu Ser Asn Thr Asp Asn Ala Ala Phe Pro Gln Met Thr Lys Ser
165 170 175
Tyr Lys Asn Thr Arg Lys Asp Pro Ala Leu Ile Ile Trp Gly Ile His
180 185 190
His Ser Gly Ser Thr Thr Glu Gln Thr Lys Leu Tyr Gly Ser Gly Asn
195 200 205
Lys Leu Ile Thr Val Gly Ser Ser Asn Tyr Gln Gln Ser Phe Val Pro
210 215 220
Ser Pro Gly Ala Arg Pro Gln Val Asn Gly Gln Ser Gly Arg Ile Asp
225 230 235 240
Phe His Trp Leu Ile Leu Asn Pro Asn Asp Thr Val Thr Phe Ser Phe
245 250 255
Asn Gly Ala Phe Ile Ala Pro Asp Arg Ala Ser Phe Leu Arg Gly Lys
260 265 270
Ser Met Gly Ile Gln Ser Gly Val Gln Val Asp Ala Asn Cys Glu Gly
275 280 285
Asp Cys Tyr His Ser Gly Gly Thr Ile Ile Ser Asn Leu Pro Phe Gln
290 295 300
Asn Ile Asn Ser Arg Ala Val Gly Lys Cys Pro Arg Tyr Val Lys Gln
305 310 315 320
Glu Ser Leu Leu Leu Ala Thr Gly Met Lys Asn Val Pro Glu Ile Pro
325 330 335
Lys Gly Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Asn Gly
340 345 350
Trp Glu Gly Leu Ile Asp Gly Trp Tyr Gly Phe Arg His Gln Asn Ala
355 360 365
Gln Gly Glu Gly Thr Ala Ala Asp Tyr Lys Ser Thr Gln Ser Ala Ile
370 375 380
Asp Gln Ile Thr Gly Lys Leu Asn Arg Leu Ile Glu Lys Thr Asn Gln
385 390 395 400
Gln Phe Glu Leu Ile Asp Asn Glu Phe Thr Glu Val Glu Lys Gln Ile
405 410 415
Gly Asn Val Ile Asn Trp Thr Arg Asp Ser Met Thr Glu Val Trp Ser
420 425 430
Tyr Asn Ala Glu Leu Leu Val Ala Met Glu Asn Gln His Thr Ile Asp
435 440 445
Leu Ala Asp Ser Glu Met Asn Lys Leu Tyr Glu Arg Val Lys Arg Gln
450 455 460
Leu Arg Glu Asn Ala Glu Glu Asp Gly Thr Gly Cys Phe Glu Ile Phe
465 470 475 480
His Lys Cys Asp Asp Asp Cys Met Ala Ser Ile Arg Asn Asn Thr Tyr
485 490 495
Asp His Ser Lys Tyr Arg Glu Glu Ala Met Gln Asn Arg Ile Gln Ile
500 505 510
Asp Pro Val Gly Ser His His His His His His
515 520
<210> 26
<211> 532
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV180664
<400> 26
Met Asn Thr Gln Ile Leu Val Phe Ala Leu Met Ala Ile Ile Pro Thr
1 5 10 15
Asn Ala Asp Lys Ile Cys Leu Gly His His Ala Val Ser Asn Gly Thr
20 25 30
Lys Val Asn Thr Leu Thr Glu Arg Gly Val Glu Val Val Asn Ala Thr
35 40 45
Glu Thr Val Glu Arg Thr Asn Val Pro Arg Ile Cys Ser Lys Gly Lys
50 55 60
Arg Thr Val Asp Leu Gly Gln Cys Gly Leu Leu Gly Thr Ile Thr Gly
65 70 75 80
Pro Pro Gln Cys Asp Gln Phe Leu Glu Phe Ser Ala Asp Leu Ile Ile
85 90 95
Glu Arg Arg Glu Gly Ser Asp Val Cys Tyr Pro Gly Lys Phe Val Asn
100 105 110
Glu Glu Ala Leu Arg Gln Ile Leu Arg Glu Ser Gly Gly Ile Asp Lys
115 120 125
Glu Thr Met Gly Phe Thr Tyr Ser Gly Ile Arg Thr Asn Gly Ala Thr
130 135 140
Ser Ala Cys Arg Arg Ser Gly Ser Ser Phe Tyr Ala Glu Met Lys Trp
145 150 155 160
Leu Leu Ser Asn Thr Asp Asn Ala Ala Phe Pro Gln Met Thr Lys Ser
165 170 175
Tyr Lys Asn Thr Arg Lys Asp Pro Ala Leu Ile Ile Trp Gly Ile His
180 185 190
His Ser Gly Ser Thr Thr Glu Gln Thr Lys Leu Tyr Gly Ser Gly Asn
195 200 205
Lys Leu Ile Thr Val Gly Ser Ser Asn Tyr Gln Gln Ser Phe Val Pro
210 215 220
Ser Pro Gly Ala Arg Pro Gln Val Asn Gly Gln Ser Gly Arg Ile Asp
225 230 235 240
Phe His Trp Leu Ile Leu Asn Pro Asn Asp Thr Val Thr Phe Ser Phe
245 250 255
Asn Gly Ala Phe Ile Ala Pro Asp Arg Ala Ser Phe Leu Arg Gly Lys
260 265 270
Ser Met Gly Ile Gln Ser Gly Val Gln Val Asp Ala Asn Cys Glu Gly
275 280 285
Asp Cys Tyr His Ser Gly Gly Thr Ile Ile Ser Asn Leu Pro Phe Gln
290 295 300
Asn Ile Asn Ser Arg Ala Val Gly Lys Cys Pro Arg Tyr Val Lys Gln
305 310 315 320
Glu Ser Leu Leu Leu Ala Thr Gly Met Lys Asn Val Pro Glu Ile Pro
325 330 335
Lys Gly Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Asn Gly
340 345 350
Trp Glu Gly Leu Ile Asp Gly Trp Tyr Gly Phe Arg Trp Gln Asn Ala
355 360 365
Gln Gly Glu Gly Thr Ala Ala Asp Tyr Lys Ser Thr Gln Ser Ala Ile
370 375 380
Asp Gln Ile Thr Gly Ile Leu Asn Arg Leu Ile Glu Lys Met Asn Gln
385 390 395 400
Gln Phe Glu Leu Ile Asp Asn Glu Phe Thr Glu Val Glu Lys Gln Ile
405 410 415
Gly Asn Val Ile Asn Trp Thr Arg Asp Ser Met Thr Glu Val Trp Ser
420 425 430
Tyr Asn Ala Glu Leu Leu Val Ala Met Ile Asn Gln His Thr Ile Asp
435 440 445
Leu Ala Asp Ser Glu Met Asn Lys Leu Tyr Glu Arg Val Lys Arg Gln
450 455 460
Leu Arg Glu Asn Ala Glu Glu Asp Gly Thr Gly Cys Phe Glu Ile Phe
465 470 475 480
His Lys Cys Asp Asp Asp Cys Met Ala Ser Ile Arg Asn Asn Thr Tyr
485 490 495
Asp His Ser Lys Tyr Arg Glu Glu Ala Met Gln Asn Arg Ile Gln Ile
500 505 510
Asp Pro Val Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His His
515 520 525
His His His His
530
<210> 27
<211> 524
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181147
<400> 27
Met Tyr Lys Val Val Val Ile Ile Ala Leu Leu Gly Ala Val Lys Gly
1 5 10 15
Leu Asp Arg Ile Cys Leu Gly His His Ala Val Ala Asn Gly Thr Ile
20 25 30
Val Lys Thr Leu Thr Asn Glu Gln Glu Glu Val Thr Asn Ala Thr Glu
35 40 45
Thr Val Glu Ser Thr Asn Leu Asn Lys Leu Cys Met Lys Gly Arg Ser
50 55 60
Tyr Lys Asp Leu Gly Asn Cys His Pro Val Gly Met Leu Ile Gly Thr
65 70 75 80
Pro Val Cys Asp Pro His Leu Thr Gly Thr Trp Asp Thr Leu Ile Glu
85 90 95
Arg Glu Asn Ala Ile Ala His Cys Tyr Pro Gly Ala Thr Ile Asn Glu
100 105 110
Glu Ala Leu Arg Gln Lys Ile Met Glu Ser Gly Gly Ile Ser Lys Met
115 120 125
Ser Thr Gly Phe Thr Tyr Gly Ser Ser Ile Asn Ser Ala Gly Thr Thr
130 135 140
Lys Ala Cys Met Arg Asn Gly Gly Asp Ser Phe Tyr Ala Glu Leu Lys
145 150 155 160
Trp Leu Val Ser Lys Thr Lys Gly Gln Asn Phe Pro Gln Thr Thr Asn
165 170 175
Thr Tyr Arg Asn Thr Asp Thr Ala Glu His Leu Ile Ile Trp Gly Ile
180 185 190
His His Pro Ser Ser Thr Gln Glu Lys Asn Asp Leu Tyr Gly Thr Gln
195 200 205
Ser Leu Ser Ile Ser Val Glu Ser Ser Thr Tyr Gln Asn Asn Phe Val
210 215 220
Pro Val Val Gly Ala Arg Pro Gln Val Asn Gly Gln Ser Gly Arg Ile
225 230 235 240
Asp Phe His Trp Thr Leu Val Gln Pro Gly Asp Asn Ile Thr Phe Ser
245 250 255
His Asn Gly Gly Leu Ile Ala Pro Ser Arg Val Ser Lys Leu Thr Gly
260 265 270
Arg Gly Leu Gly Ile Gln Ser Glu Ala Leu Ile Asp Asn Ser Cys Glu
275 280 285
Ser Lys Cys Phe Trp Arg Gly Gly Ser Ile Asn Thr Lys Leu Pro Phe
290 295 300
Gln Asn Leu Ser Pro Arg Thr Val Gly Gln Cys Pro Lys Tyr Val Asn
305 310 315 320
Gln Arg Ser Leu Leu Leu Ala Thr Gly Met Arg Asn Val Pro Glu Val
325 330 335
Val Gln Gly Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Asn
340 345 350
Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly Phe Arg His Gln Asn
355 360 365
Ala Gln Gly Thr Gly Gln Ala Ala Asp Tyr Lys Ser Thr Gln Ala Ala
370 375 380
Ile Asp Gln Ile Thr Gly Lys Leu Asn Arg Leu Ile Glu Lys Thr Asn
385 390 395 400
Thr Glu Phe Glu Ser Ile Glu Ser Glu Phe Ser Glu Thr Glu His Gln
405 410 415
Ile Gly Asn Val Ile Asn Trp Thr Lys Asp Ser Ile Thr Asp Ile Trp
420 425 430
Thr Tyr Gln Ala Glu Leu Leu Val Ala Met Glu Asn Gln His Thr Ile
435 440 445
Asp Met Ala Asp Ser Glu Met Leu Asn Leu Tyr Glu Arg Val Arg Lys
450 455 460
Gln Leu Arg Gln Asn Ala Glu Glu Asp Gly Lys Gly Cys Phe Glu Ile
465 470 475 480
Tyr His Thr Cys Asp Asp Ser Cys Met Glu Ser Ile Arg Asn Asn Thr
485 490 495
Tyr Asp His Ser Gln Tyr Arg Glu Glu Ala Leu Leu Asn Arg Leu Asn
500 505 510
Ile Asn Ser Val Gly Ser His His His His His His
515 520
<210> 28
<211> 532
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV180662
<400> 28
Met Tyr Lys Val Val Val Ile Ile Ala Leu Leu Gly Ala Val Lys Gly
1 5 10 15
Leu Asp Arg Ile Cys Leu Gly His His Ala Val Ala Asn Gly Thr Ile
20 25 30
Val Lys Thr Leu Thr Asn Glu Gln Glu Glu Val Thr Asn Ala Thr Glu
35 40 45
Thr Val Glu Ser Thr Asn Leu Asn Lys Leu Cys Met Lys Gly Arg Ser
50 55 60
Tyr Lys Asp Leu Gly Asn Cys His Pro Val Gly Met Leu Ile Gly Thr
65 70 75 80
Pro Val Cys Asp Pro His Leu Thr Gly Thr Trp Asp Thr Leu Ile Glu
85 90 95
Arg Glu Asn Ala Ile Ala His Cys Tyr Pro Gly Ala Thr Ile Asn Glu
100 105 110
Glu Ala Leu Arg Gln Lys Ile Met Glu Ser Gly Gly Ile Ser Lys Met
115 120 125
Ser Thr Gly Phe Thr Tyr Gly Ser Ser Ile Asn Ser Ala Gly Thr Thr
130 135 140
Lys Ala Cys Met Arg Asn Gly Gly Asp Ser Phe Tyr Ala Glu Leu Lys
145 150 155 160
Trp Leu Val Ser Lys Thr Lys Gly Gln Asn Phe Pro Gln Thr Thr Asn
165 170 175
Thr Tyr Arg Asn Thr Asp Thr Ala Glu His Leu Ile Ile Trp Gly Ile
180 185 190
His His Pro Ser Ser Thr Gln Glu Lys Asn Asp Leu Tyr Gly Thr Gln
195 200 205
Ser Leu Ser Ile Ser Val Glu Ser Ser Thr Tyr Gln Asn Asn Phe Val
210 215 220
Pro Val Val Gly Ala Arg Pro Gln Val Asn Gly Gln Ser Gly Arg Ile
225 230 235 240
Asp Phe His Trp Thr Leu Val Gln Pro Gly Asp Asn Ile Thr Phe Ser
245 250 255
His Asn Gly Gly Leu Ile Ala Pro Ser Arg Val Ser Lys Leu Thr Gly
260 265 270
Arg Gly Leu Gly Ile Gln Ser Glu Ala Leu Ile Asp Asn Ser Cys Glu
275 280 285
Ser Lys Cys Phe Trp Arg Gly Gly Ser Ile Asn Thr Lys Leu Pro Phe
290 295 300
Gln Asn Leu Ser Pro Arg Thr Val Gly Gln Cys Pro Lys Tyr Val Asn
305 310 315 320
Gln Arg Ser Leu Leu Leu Ala Thr Gly Met Arg Asn Val Pro Glu Val
325 330 335
Val Gln Gly Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Asn
340 345 350
Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly Phe Arg Trp Gln Asn
355 360 365
Ala Gln Gly Thr Gly Gln Ala Ala Asp Tyr Lys Ser Thr Gln Ala Ala
370 375 380
Ile Asp Gln Ile Thr Gly Ile Leu Asn Arg Leu Ile Glu Lys Met Asn
385 390 395 400
Thr Glu Phe Glu Ser Ile Glu Ser Glu Phe Ser Glu Thr Glu His Gln
405 410 415
Ile Gly Asn Val Ile Asn Trp Thr Lys Asp Ser Ile Thr Asp Ile Trp
420 425 430
Thr Tyr Gln Ala Glu Leu Leu Val Ala Met Ile Asn Gln His Thr Ile
435 440 445
Asp Met Ala Asp Ser Glu Met Leu Asn Leu Tyr Glu Arg Val Arg Lys
450 455 460
Gln Leu Arg Gln Asn Ala Glu Glu Asp Gly Lys Gly Cys Phe Glu Ile
465 470 475 480
Tyr His Thr Cys Asp Asp Ser Cys Met Glu Ser Ile Arg Asn Asn Thr
485 490 495
Tyr Asp His Ser Gln Tyr Arg Glu Glu Ala Leu Leu Asn Arg Leu Asn
500 505 510
Ile Asn Ser Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His His
515 520 525
His His His His
530
<210> 29
<211> 534
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV4239
<400> 29
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Pro Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Ser Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Ile Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Ala Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Lys Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Gln Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Glu Ile Thr Asn Lys Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Gly Arg Ser Leu Val Pro Arg Gly Ser Gly
515 520 525
His His His His His His
530
<210> 30
<211> 574
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV180843
<400> 30
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Thr Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Ser Asn Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asn Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Asn Gln Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Thr Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Thr Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Thr Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Thr Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Glu Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Val Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Lys Ile Thr Asn Lys Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Asn Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Lys Ile Asp Ser Gly Ser Leu Val Pro Ser Gly Ser Pro
515 520 525
Gly Ser Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val
530 535 540
Arg Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu Gly Gly Ser
545 550 555 560
Leu Pro Glu Thr Gly Gly Gly Ser His His His His His His
565 570
<210> 31
<211> 573
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV180436
<400> 31
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Phe Cys Leu Ala Leu Gly
1 5 10 15
Gln Asp Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asp Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Lys Ile Cys Asn Asn Pro His Arg Ile Leu Asp Gly Ile Asp Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Val Phe Gln
85 90 95
Asn Glu Thr Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Thr Glu Gly Phe Thr Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Gly Ser Asn Ala Cys Lys Arg Gly Pro Gly
145 150 155 160
Ser Gly Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Ser Gly Ser Thr
165 170 175
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asn Gln Glu Gln Thr
195 200 205
Ser Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Arg Arg
210 215 220
Ser Gln Gln Thr Ile Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Val Leu Val Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Met Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Asp Thr Cys Ile Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Leu Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Val
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Glu Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ser Gly Ser Leu Val Pro Ser Gly Ser Pro Gly
515 520 525
Ser Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg
530 535 540
Lys Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu Gly Gly Ser Leu
545 550 555 560
Pro Glu Thr Gly Gly Gly Ser His His His His His His
565 570
<210> 32
<211> 562
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV170466
<400> 32
Met Lys Thr Ile Val Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Ile Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Glu Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro Gln Cys Asp Gly Phe Gln
85 90 95
Asn Lys Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Asn Asn Glu Ser Phe Asn Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Thr Ser Ser Ala Cys Ile Arg Arg Ser Asn
145 150 155 160
Ser Ser Phe Phe Ser Arg Leu Asn Trp Leu Thr His Leu Asn Phe Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Glu Gln Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Gly Thr Asp Lys Asp Gln Ile
195 200 205
Phe Leu Tyr Ala Gln Ser Ser Gly Arg Ile Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Ala Val Ile Pro Asn Ile Gly Ser Arg Pro Arg Ile Arg
225 230 235 240
Asn Ile Pro Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Ser Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Lys Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Ser Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Arg Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Leu
385 390 395 400
Ile Gly Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asn His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ser Gly Ser Leu Val Pro Arg Gly Ser Gly Ser
515 520 525
Gly Tyr Ile Pro Glu Ala Pro Arg Asp Gly Gln Ala Tyr Val Arg Lys
530 535 540
Asp Gly Glu Trp Val Leu Leu Ser Thr Phe Leu Gly Gly Ser Glu Pro
545 550 555 560
Glu Ala
<210> 33
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181099
<400> 33
Met Lys Thr Ile Val Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Ile Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Glu Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro Gln Cys Asp Gly Phe Gln
85 90 95
Asn Lys Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Asn Asn Glu Ser Phe Asn Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Thr Ser Ser Ala Cys Ile Arg Arg Ser Asn
145 150 155 160
Ser Ser Phe Phe Ser Arg Leu Asn Trp Leu Thr His Leu Asn Phe Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Glu Gln Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Gly Thr Asp Lys Asp Gln Ile
195 200 205
Phe Leu Tyr Ala Gln Ser Ser Gly Arg Ile Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Ala Val Ile Pro Asn Ile Gly Ser Arg Pro Arg Ile Arg
225 230 235 240
Asn Ile Pro Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Ser Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Lys Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Ser Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Arg Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn Arg Leu
385 390 395 400
Ile Gly Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asn His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 34
<211> 535
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181005
<400> 34
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Pro Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Ser Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Ile Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Ala Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Lys Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His Trp Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Glu Ile Thr Asn Lys Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Ile Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly
515 520 525
Ser His His His His His His
530 535
<210> 35
<211> 535
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181007
<400> 35
Met Lys Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn
1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu Arg Gly Val
50 55 60
Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Ser Leu Ser Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Pro Ser Ser Asp Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp
130 135 140
Ser Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser
145 150 155 160
Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Ile Asn Asp Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Ile His His Pro Ser Thr Ser Ala Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser Arg Tyr Ser
210 215 220
Lys Lys Phe Lys Pro Glu Ile Ala Ile Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Glu Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe
260 265 270
Ala Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro
275 280 285
Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Lys Gly Ala Ile Asn
290 295 300
Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile Gly Lys Cys
305 310 315 320
Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Leu Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr
355 360 365
His His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Glu Ile Thr Asn Ile Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415
Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Ile Asn
435 440 445
Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn Leu Tyr Glu
450 455 460
Lys Val Arg Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asn Thr Cys Met Glu Ser Val
485 490 495
Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly
515 520 525
Ser His His His His His His
530 535
<210> 36
<211> 534
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181090
<400> 36
Met Lys Val Lys Leu Leu Val Leu Leu Cys Thr Phe Thr Ala Thr Tyr
1 5 10 15
Ala Asp Thr Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asn Ser His Asn Gly Lys Leu Cys Leu Leu Lys Gly Ile
50 55 60
Ala Pro Leu Gln Leu Gly Asn Cys Ser Val Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Leu Leu Ile Ser Lys Glu Ser Trp Ser Tyr Ile
85 90 95
Val Glu Lys Pro Asn Pro Glu Asn Gly Thr Cys Tyr Pro Gly His Phe
100 105 110
Ala Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn His Thr
130 135 140
Val Thr Gly Val Ser Ala Ser Cys Ser His Asn Gly Glu Ser Ser Phe
145 150 155 160
Tyr Arg Asn Leu Leu Trp Leu Thr Gly Lys Asn Gly Leu Tyr Pro Asn
165 170 175
Leu Ser Lys Ser Tyr Ala Asn Asn Lys Glu Lys Glu Val Leu Val Leu
180 185 190
Trp Gly Val His His Pro Pro Asn Ile Gly Asp Gln Lys Ala Leu Tyr
195 200 205
His Thr Glu Asn Ala Tyr Val Ser Val Val Ser Ser His Tyr Ser Arg
210 215 220
Lys Phe Thr Pro Glu Ile Ala Lys Arg Pro Lys Val Arg Asp Gln Glu
225 230 235 240
Gly Arg Ile Asn Tyr Tyr Trp Thr Leu Leu Glu Pro Gly Asp Thr Ile
245 250 255
Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala
260 265 270
Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Asn Ser Asn Ala Pro Met
275 280 285
Asp Lys Cys Asp Ala Lys Cys Gln Thr Pro Gln Gly Ala Ile Asn Ser
290 295 300
Ser Leu Pro Phe Gln Asn Val His Pro Val Thr Ile Gly Glu Cys Pro
305 310 315 320
Lys Tyr Val Arg Ser Ala Lys Leu Arg Met Val Thr Gly Leu Arg Asn
325 330 335
Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe
340 345 350
Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr His
355 360 365
Trp Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Gln Lys Ser Thr
370 375 380
Gln Asn Ala Ile Asn Gly Ile Thr Asn Ile Val Asn Ser Val Ile Glu
385 390 395 400
Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn Lys Leu
405 410 415
Glu Arg Arg Met Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe Ile
420 425 430
Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Ile Asn Glu
435 440 445
Arg Thr Leu Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Glu Lys
450 455 460
Val Lys Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly Cys
465 470 475 480
Phe Glu Phe Tyr His Lys Cys Asn Asp Glu Cys Ile Glu Ser Val Lys
485 490 495
Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ser Lys Leu Asn
500 505 510
Arg Glu Lys Ile Asp Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser
515 520 525
His His His His His His
530
<210> 37
<211> 525
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181135
<400> 37
Met Lys Val Lys Leu Leu Val Leu Leu Cys Thr Phe Thr Ala Thr Tyr
1 5 10 15
Ala Asp Thr Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asn Ser His Asn Gly Lys Leu Cys Leu Leu Lys Gly Ile
50 55 60
Ala Pro Leu Gln Leu Gly Asn Cys Ser Val Ala Gly Trp Ile Leu Gly
65 70 75 80
Asn Pro Glu Cys Glu Leu Leu Ile Ser Lys Glu Ser Trp Ser Tyr Ile
85 90 95
Val Glu Lys Pro Asn Pro Glu Asn Gly Thr Cys Tyr Pro Gly His Phe
100 105 110
Ala Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn His Thr
130 135 140
Val Thr Gly Val Ser Ala Ser Cys Ser His Asn Gly Glu Ser Ser Phe
145 150 155 160
Tyr Arg Asn Leu Leu Trp Leu Thr Gly Lys Asn Gly Leu Tyr Pro Asn
165 170 175
Leu Ser Lys Ser Tyr Ala Asn Asn Lys Glu Lys Glu Val Leu Val Leu
180 185 190
Trp Gly Val His His Pro Pro Asn Ile Gly Asp Gln Lys Ala Leu Tyr
195 200 205
His Thr Glu Asn Ala Tyr Val Ser Val Val Ser Ser His Tyr Ser Arg
210 215 220
Lys Phe Thr Pro Glu Ile Ala Lys Arg Pro Lys Val Arg Asp Gln Glu
225 230 235 240
Gly Arg Ile Asn Tyr Tyr Trp Thr Leu Leu Glu Pro Gly Asp Thr Ile
245 250 255
Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala
260 265 270
Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Asn Ser Asn Ala Pro Met
275 280 285
Asp Lys Cys Asp Ala Lys Cys Gln Thr Pro Gln Gly Ala Ile Asn Ser
290 295 300
Ser Leu Pro Phe Gln Asn Val His Pro Val Thr Ile Gly Glu Cys Pro
305 310 315 320
Lys Tyr Val Arg Ser Ala Lys Leu Arg Met Val Thr Gly Leu Arg Asn
325 330 335
Ile Pro Ser Ile Gln Ser Gln Gly Leu Phe Gly Ala Ile Ala Gly Phe
340 345 350
Ile Glu Gly Gly Trp Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr His
355 360 365
His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Gln Lys Ser Thr
370 375 380
Gln Asn Ala Ile Asn Gly Ile Thr Asn Lys Val Asn Ser Val Ile Glu
385 390 395 400
Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn Lys Leu
405 410 415
Glu Arg Arg Met Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe Ile
420 425 430
Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn Glu
435 440 445
Arg Thr Leu Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Glu Lys
450 455 460
Val Lys Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly Cys
465 470 475 480
Phe Glu Phe Tyr His Lys Cys Asn Asp Glu Cys Met Glu Ser Val Lys
485 490 495
Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ser Lys Leu Asn
500 505 510
Arg Glu Lys Ile Asp Gly Ser His His His His His His
515 520 525
<210> 38
<211> 535
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181084
<400> 38
Met Glu Ala Arg Leu Leu Val Leu Leu Cys Ala Phe Ala Ala Thr Asn
1 5 10 15
Ala Asp Thr Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Ser His Asn Gly Lys Leu Cys Lys Leu Lys Gly Ile
50 55 60
Ala Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly
65 70 75 80
Asn Pro Glu Cys Asp Leu Leu Leu Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Ser Asn Ser Glu Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Lys Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Glu
130 135 140
Thr Thr Lys Gly Val Thr Ala Ala Cys Ser Tyr Ala Gly Ala Ser Ser
145 150 155 160
Phe Tyr Arg Asn Leu Leu Trp Leu Thr Lys Lys Gly Ser Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Val Asn Asn Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Val His His Pro Pro Thr Gly Thr Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Ser Val Gly Ser Ser Lys Tyr Asn
210 215 220
Arg Arg Phe Thr Pro Glu Ile Ala Ala Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Ala Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Glu Pro Gly Asp Thr
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Ile Ala Pro Trp Tyr Ala Phe
260 265 270
Ala Leu Asn Arg Gly Ser Gly Ser Gly Ile Ile Thr Ser Asp Ala Pro
275 280 285
Val His Asp Cys Asn Thr Lys Cys Gln Thr Pro His Gly Ala Ile Asn
290 295 300
Ser Ser Leu Pro Phe Gln Asn Ile His Pro Val Thr Ile Gly Glu Cys
305 310 315 320
Pro Lys Tyr Val Arg Ser Thr Lys Leu Arg Met Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Ile Asp Gly Trp Tyr Gly Tyr
355 360 365
His Trp Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Gln Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Gly Ile Thr Asn Ile Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn Asn
405 410 415
Leu Glu Arg Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Ile Asn
435 440 445
Glu Arg Thr Leu Asp Phe His Asp Ser Asn Val Arg Asn Leu Tyr Glu
450 455 460
Lys Val Lys Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asp Ala Cys Ile Glu Ser Val
485 490 495
Arg Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ser Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly
515 520 525
Ser His His His His His His
530 535
<210> 39
<211> 526
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181131
<400> 39
Met Glu Ala Arg Leu Leu Val Leu Leu Cys Ala Phe Ala Ala Thr Asn
1 5 10 15
Ala Asp Thr Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30
Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val Asn
35 40 45
Leu Leu Glu Asp Ser His Asn Gly Lys Leu Cys Lys Leu Lys Gly Ile
50 55 60
Ala Pro Leu Gln Leu Gly Lys Cys Asn Ile Ala Gly Trp Leu Leu Gly
65 70 75 80
Asn Pro Glu Cys Asp Leu Leu Leu Thr Ala Ser Ser Trp Ser Tyr Ile
85 90 95
Val Glu Thr Ser Asn Ser Glu Asn Gly Thr Cys Tyr Pro Gly Asp Phe
100 105 110
Ile Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe
115 120 125
Glu Lys Phe Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Glu
130 135 140
Thr Thr Lys Gly Val Thr Ala Ala Cys Ser Tyr Ala Gly Ala Ser Ser
145 150 155 160
Phe Tyr Arg Asn Leu Leu Trp Leu Thr Lys Lys Gly Ser Ser Tyr Pro
165 170 175
Lys Leu Ser Lys Ser Tyr Val Asn Asn Lys Gly Lys Glu Val Leu Val
180 185 190
Leu Trp Gly Val His His Pro Pro Thr Gly Thr Asp Gln Gln Ser Leu
195 200 205
Tyr Gln Asn Ala Asp Ala Tyr Val Ser Val Gly Ser Ser Lys Tyr Asn
210 215 220
Arg Arg Phe Thr Pro Glu Ile Ala Ala Arg Pro Lys Val Arg Asp Gln
225 230 235 240
Ala Gly Arg Met Asn Tyr Tyr Trp Thr Leu Leu Glu Pro Gly Asp Thr
245 250 255
Ile Thr Phe Glu Ala Thr Gly Asn Leu Ile Ala Pro Trp Tyr Ala Phe
260 265 270
Ala Leu Asn Arg Gly Ser Gly Ser Gly Ile Ile Thr Ser Asp Ala Pro
275 280 285
Val His Asp Cys Asn Thr Lys Cys Gln Thr Pro His Gly Ala Ile Asn
290 295 300
Ser Ser Leu Pro Phe Gln Asn Ile His Pro Val Thr Ile Gly Glu Cys
305 310 315 320
Pro Lys Tyr Val Arg Ser Thr Lys Leu Arg Met Ala Thr Gly Leu Arg
325 330 335
Asn Ile Pro Ser Ile Gln Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly
340 345 350
Phe Ile Glu Gly Gly Trp Thr Gly Met Ile Asp Gly Trp Tyr Gly Tyr
355 360 365
His His Gln Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Gln Lys Ser
370 375 380
Thr Gln Asn Ala Ile Asp Gly Ile Thr Asn Lys Val Asn Ser Val Ile
385 390 395 400
Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn Asn
405 410 415
Leu Glu Arg Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp Gly Phe
420 425 430
Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu Leu Glu Asn
435 440 445
Glu Arg Thr Leu Asp Phe His Asp Ser Asn Val Arg Asn Leu Tyr Glu
450 455 460
Lys Val Lys Ser Gln Leu Lys Asn Asn Ala Lys Glu Ile Gly Asn Gly
465 470 475 480
Cys Phe Glu Phe Tyr His Lys Cys Asp Asp Ala Cys Met Glu Ser Val
485 490 495
Arg Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ser Lys Leu
500 505 510
Asn Arg Glu Glu Ile Asp Gly Ser His His His His His His
515 520 525
<210> 40
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181095
<400> 40
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Arg Ile Cys Asp Ser Pro His Arg Ile Leu Asp Gly Lys Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Gly Phe Gln
85 90 95
Asn Lys Glu Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Asn Glu Asp Phe Asn Trp Thr
130 135 140
Gly Val Ala Gln Asp Gly Lys Ser Tyr Thr Cys Lys Arg Gly Ser Val
145 150 155 160
Asn Ser Phe Phe Ser Arg Leu Asn Trp Leu His Lys Leu Glu Tyr Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Gly Lys Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asp Ser Asp Gln Thr
195 200 205
Ser Leu Tyr Val Arg Ala Ser Gly Arg Val Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Asn Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Asn Cys Ser Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn Arg Leu
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Arg Thr Arg Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 41
<211> 529
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181140
<400> 41
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Arg Ile Cys Asp Ser Pro His Arg Ile Leu Asp Gly Lys Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Gly Phe Gln
85 90 95
Asn Lys Glu Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Asn Glu Asp Phe Asn Trp Thr
130 135 140
Gly Val Ala Gln Asp Gly Lys Ser Tyr Thr Cys Lys Arg Gly Ser Val
145 150 155 160
Asn Ser Phe Phe Ser Arg Leu Asn Trp Leu His Lys Leu Glu Tyr Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Gly Lys Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asp Ser Asp Gln Thr
195 200 205
Ser Leu Tyr Val Arg Ala Ser Gly Arg Val Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Asn Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Asn Cys Ser Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Leu
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Arg Thr Arg Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Gly Ser His His His His His
515 520 525
His
<210> 42
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181093
<400> 42
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Phe Cys Gln Val Leu Ala
1 5 10 15
Gln Asn Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Arg Ile Cys Asp Ser Pro His Arg Ile Leu Asp Gly Lys Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Gly Phe Gln
85 90 95
Asn Glu Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Asn Glu Gly Phe Asn Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Gly Ser Tyr Ala Cys Lys Arg Gly Pro Asp
145 150 155 160
Lys Ser Phe Phe Ser Arg Leu Asn Trp Leu Tyr Glu Ser Glu Ser Thr
165 170 175
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asp Lys Glu Gln Thr
195 200 205
Asn Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Ile Ile Pro Asn Val Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Thr Cys Ser Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn Arg Val
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 43
<211> 529
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181136
<400> 43
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Phe Cys Gln Val Leu Ala
1 5 10 15
Gln Asn Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Arg Ile Cys Asp Ser Pro His Arg Ile Leu Asp Gly Lys Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Gly Phe Gln
85 90 95
Asn Glu Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Asn Glu Gly Phe Asn Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Gly Ser Tyr Ala Cys Lys Arg Gly Pro Asp
145 150 155 160
Lys Ser Phe Phe Ser Arg Leu Asn Trp Leu Tyr Glu Ser Glu Ser Thr
165 170 175
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asp Lys Glu Gln Thr
195 200 205
Asn Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Ile Ile Pro Asn Val Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Thr Cys Ser Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Val
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Gly Ser His His His His His
515 520 525
His
<210> 44
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181097
<400> 44
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Ile Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Gly Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro Gln Cys Asp Gly Phe Gln
85 90 95
Asn Lys Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Asn Asp Glu Ser Phe Asn Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Thr Ser Ser Ser Cys Lys Arg Arg Ser Asn
145 150 155 160
Asn Ser Phe Phe Ser Arg Leu Asn Trp Leu Thr His Leu Lys Phe Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Glu Lys Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Val Thr Asp Asn Asp Gln Ile
195 200 205
Phe Leu Tyr Ala Gln Ala Ser Gly Arg Ile Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Arg Pro Arg Ile Arg
225 230 235 240
Asn Ile Pro Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Ser Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Lys Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Ile Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asn Gln Ile Asn Gly Ile Leu Asn Arg Leu
385 390 395 400
Ile Gly Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Arg Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 45
<211> 529
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181138
<400> 45
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Leu Cys Leu Val Phe Ala
1 5 10 15
Gln Lys Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Ile Val Lys Thr Ile Thr Asn Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Gly Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro Gln Cys Asp Gly Phe Gln
85 90 95
Asn Lys Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Tyr Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Asn Asp Glu Ser Phe Asn Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Thr Ser Ser Ser Cys Lys Arg Arg Ser Asn
145 150 155 160
Asn Ser Phe Phe Ser Arg Leu Asn Trp Leu Thr His Leu Lys Phe Lys
165 170 175
Tyr Pro Ala Leu Asn Val Thr Met Pro Asn Asn Glu Lys Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Val Thr Asp Asn Asp Gln Ile
195 200 205
Phe Leu Tyr Ala Gln Ala Ser Gly Arg Ile Thr Val Ser Thr Lys Arg
210 215 220
Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Arg Pro Arg Ile Arg
225 230 235 240
Asn Ile Pro Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Ile Leu Leu Ile Asn Ser Thr Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Ile Arg Ser Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Gly Lys Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp Gly Trp Tyr Gly
355 360 365
Phe Arg His Gln Asn Ser Glu Gly Ile Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asn Gln Ile Asn Gly Lys Leu Asn Arg Leu
385 390 395 400
Ile Gly Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Arg Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Gly Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Gly Ser His His His His His
515 520 525
His
<210> 46
<211> 527
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181148
<400> 46
Met Leu Ser Ile Val Ile Leu Phe Leu Leu Val Ala Glu Asn Ser Ser
1 5 10 15
Gln Asn Tyr Thr Gly Asn Pro Val Ile Cys Met Gly His His Ala Val
20 25 30
Ala Asn Gly Thr Met Val Lys Thr Leu Thr Asp Asp Gln Val Glu Val
35 40 45
Val Thr Ala Gln Glu Leu Val Glu Ser Gln Asn Leu Pro Glu Leu Cys
50 55 60
Pro Ser Pro Leu Arg Leu Val Asp Gly Gln Thr Cys Asp Ile Ile Asn
65 70 75 80
Gly Ala Leu Gly Ser Pro Gly Cys Asp His Leu Asn Gly Ala Glu Trp
85 90 95
Asp Val Phe Ile Glu Arg Pro Asn Ala Met Asp Thr Cys Tyr Pro Phe
100 105 110
Asp Val Pro Asp Tyr Gln Ser Leu Arg Ser Ile Leu Ala Asn Asn Gly
115 120 125
Lys Phe Glu Phe Ile Ala Glu Glu Phe Gln Trp Thr Thr Val Lys Gln
130 135 140
Asn Gly Lys Ser Gly Ala Cys Lys Arg Ala Asn Val Asn Asp Phe Phe
145 150 155 160
Arg Arg Leu Asn Trp Leu Val Lys Ser Asp Arg Asn Ala Tyr Pro Leu
165 170 175
Gln Asn Leu Thr Lys Val Asn Asn Gly Asp Tyr Ala Arg Leu Tyr Ile
180 185 190
Trp Gly Val His His Pro Ser Thr Asp Thr Glu Gln Thr Asn Leu Tyr
195 200 205
Lys Asn Asn Pro Gly Arg Val Thr Val Ser Thr Lys Thr Ser Gln Thr
210 215 220
Ser Val Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg Gly Gln Ser
225 230 235 240
Gly Arg Ile Ser Phe Tyr Trp Thr Ile Val Glu Pro Gly Asp Leu Ile
245 250 255
Val Phe Asn Thr Ile Gly Asn Leu Ile Ala Pro Arg Gly His Tyr Lys
260 265 270
Leu Asn Asn Gln Lys Lys Gly Thr Ile Leu Asn Thr Ala Ile Pro Ile
275 280 285
Gly Ser Cys Val Ser Lys Cys His Thr Asp Lys Gly Ser Leu Ser Thr
290 295 300
Thr Lys Pro Phe Gln Asn Ile Ser Arg Ile Ala Ile Gly Asp Cys Pro
305 310 315 320
Lys Tyr Val Lys Gln Gly Ser Leu Lys Leu Ala Thr Gly Met Arg Asn
325 330 335
Ile Pro Glu Lys Ala Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe
340 345 350
Ile Glu Asn Gly Trp Gln Gly Leu Ile Asp Gly Trp Tyr Gly Phe Arg
355 360 365
His Gln Asn Ala Glu Gly Thr Gly Thr Ala Ala Asp Leu Lys Ser Thr
370 375 380
Gln Ala Ala Ile Asp Gln Ile Asn Gly Lys Leu Asn Arg Leu Ile Glu
385 390 395 400
Lys Thr Asn Glu Lys Tyr His Gln Ile Glu Lys Glu Phe Glu Gln Val
405 410 415
Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr Lys Ile
420 425 430
Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu Asn Gln
435 440 445
His Thr Ile Asp Val Thr Asp Ser Glu Met Asn Lys Leu Phe Glu Arg
450 455 460
Val Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Lys Gly Asn Gly Cys
465 470 475 480
Phe Glu Ile Phe His Lys Cys Asp Asn Asn Cys Ile Glu Ser Ile Arg
485 490 495
Asn Gly Thr Tyr Asp His Asp Ile Tyr Arg Asp Glu Ala Ile Asn Asn
500 505 510
Arg Phe Gln Ile Gln Gly Val Gly Ser His His His His His His
515 520 525
<210> 47
<211> 536
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV181149
<400> 47
Met Leu Ser Ile Val Ile Leu Phe Leu Leu Val Ala Glu Asn Ser Ser
1 5 10 15
Gln Asn Tyr Thr Gly Asn Pro Val Ile Cys Met Gly His His Ala Val
20 25 30
Ala Asn Gly Thr Met Val Lys Thr Leu Thr Asp Asp Gln Val Glu Val
35 40 45
Val Thr Ala Gln Glu Leu Val Glu Ser Gln Asn Leu Pro Glu Leu Cys
50 55 60
Pro Ser Pro Leu Arg Leu Val Asp Gly Gln Thr Cys Asp Ile Ile Asn
65 70 75 80
Gly Ala Leu Gly Ser Pro Gly Cys Asp His Leu Asn Gly Ala Glu Trp
85 90 95
Asp Val Phe Ile Glu Arg Pro Asn Ala Met Asp Thr Cys Tyr Pro Phe
100 105 110
Asp Val Pro Asp Tyr Gln Ser Leu Arg Ser Ile Leu Ala Asn Asn Gly
115 120 125
Lys Phe Glu Phe Ile Ala Glu Glu Phe Gln Trp Thr Thr Val Lys Gln
130 135 140
Asn Gly Lys Ser Gly Ala Cys Lys Arg Ala Asn Val Asn Asp Phe Phe
145 150 155 160
Arg Arg Leu Asn Trp Leu Val Lys Ser Asp Arg Asn Ala Tyr Pro Leu
165 170 175
Gln Asn Leu Thr Lys Val Asn Asn Gly Asp Tyr Ala Arg Leu Tyr Ile
180 185 190
Trp Gly Val His His Pro Ser Thr Asp Thr Glu Gln Thr Asn Leu Tyr
195 200 205
Lys Asn Asn Pro Gly Arg Val Thr Val Ser Thr Lys Thr Ser Gln Thr
210 215 220
Ser Val Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg Gly Gln Ser
225 230 235 240
Gly Arg Ile Ser Phe Tyr Trp Thr Ile Val Glu Pro Gly Asp Leu Ile
245 250 255
Val Phe Asn Thr Ile Gly Asn Leu Ile Ala Pro Arg Gly His Tyr Lys
260 265 270
Leu Asn Asn Gln Lys Lys Gly Thr Ile Leu Asn Thr Ala Ile Pro Ile
275 280 285
Gly Ser Cys Val Ser Lys Cys His Thr Asp Lys Gly Ser Leu Ser Thr
290 295 300
Thr Lys Pro Phe Gln Asn Ile Ser Arg Ile Ala Ile Gly Asp Cys Pro
305 310 315 320
Lys Tyr Val Lys Gln Gly Ser Leu Lys Leu Ala Thr Gly Met Arg Asn
325 330 335
Ile Pro Glu Lys Ala Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe
340 345 350
Ile Glu Asn Gly Trp Gln Gly Leu Ile Asp Gly Trp Tyr Gly Phe Arg
355 360 365
Trp Gln Asn Ala Glu Gly Thr Gly Thr Ala Ala Asp Leu Lys Ser Thr
370 375 380
Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn Arg Leu Ile Glu
385 390 395 400
Lys Thr Asn Glu Lys Tyr His Gln Ile Glu Lys Glu Phe Glu Gln Val
405 410 415
Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr Lys Ile
420 425 430
Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile Asn Gln
435 440 445
His Thr Ile Asp Val Thr Asp Ser Glu Met Asn Lys Leu Phe Glu Arg
450 455 460
Val Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Lys Gly Asn Gly Cys
465 470 475 480
Phe Glu Ile Phe His Lys Cys Asp Asn Asn Cys Ile Glu Ser Ile Arg
485 490 495
Asn Gly Thr Tyr Asp His Asp Ile Tyr Arg Asp Glu Ala Ile Asn Asn
500 505 510
Arg Phe Gln Ile Gln Gly Val Ser Gly Ser Leu Pro Glu Thr Gly Gly
515 520 525
Gly Ser His His His His His His
530 535
<210> 48
<400> 48
000
<210> 49
<400> 49
000
<210> 50
<211> 538
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV190839
<400> 50
Met Lys Thr Ile Ile Ala Leu Ser Tyr Ile Phe Cys Leu Ala Leu Gly
1 5 10 15
Gln Asp Leu Pro Gly Asn Asp Asn Ser Thr Ala Thr Leu Cys Leu Gly
20 25 30
His His Ala Val Pro Asn Gly Thr Leu Val Lys Thr Ile Thr Asp Asp
35 40 45
Gln Ile Glu Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr
50 55 60
Gly Lys Ile Cys Asn Asn Pro His Arg Ile Leu Asp Gly Ile Asp Cys
65 70 75 80
Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro His Cys Asp Val Phe Gln
85 90 95
Asn Glu Thr Trp Asp Leu Phe Val Glu Arg Ser Lys Ala Phe Ser Asn
100 105 110
Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val
115 120 125
Ala Ser Ser Gly Thr Leu Glu Phe Ile Thr Glu Gly Phe Thr Trp Thr
130 135 140
Gly Val Thr Gln Asn Gly Gly Ser Asn Ala Cys Lys Arg Gly Pro Gly
145 150 155 160
Ser Gly Phe Phe Ser Arg Leu Asn Trp Leu Thr Lys Ser Gly Ser Thr
165 170 175
Tyr Pro Val Leu Asn Val Thr Met Pro Asn Asn Asp Asn Phe Asp Lys
180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Ser Thr Asn Gln Glu Gln Thr
195 200 205
Ser Leu Tyr Val Gln Ala Ser Gly Arg Val Thr Val Ser Thr Arg Arg
210 215 220
Ser Gln Gln Thr Ile Ile Pro Asn Ile Gly Ser Arg Pro Trp Val Arg
225 230 235 240
Gly Leu Ser Ser Arg Ile Ser Ile Tyr Trp Thr Ile Val Lys Pro Gly
245 250 255
Asp Val Leu Val Ile Asn Ser Asn Gly Asn Leu Ile Ala Pro Arg Gly
260 265 270
Tyr Phe Lys Met Arg Thr Gly Lys Ser Ser Ile Met Arg Ser Asp Ala
275 280 285
Pro Ile Asp Thr Cys Ile Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile
290 295 300
Pro Asn Asp Lys Pro Phe Gln Asn Val Asn Lys Ile Thr Tyr Gly Ala
305 310 315 320
Cys Pro Lys Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335
Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Leu Phe Gly Ala Ile Ala
340 345 350
Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Ile Asp Gly Trp Tyr Gly
355 360 365
Phe Arg Trp Gln Asn Ser Glu Gly Thr Gly Gln Ala Ala Asp Leu Lys
370 375 380
Ser Thr Gln Ala Ala Ile Asp Gln Ile Asn Gly Ile Leu Asn Arg Val
385 390 395 400
Ile Glu Lys Thr Asn Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser
405 410 415
Glu Val Glu Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr
420 425 430
Lys Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Ile
435 440 445
Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460
Glu Lys Thr Arg Arg Gln Leu Arg Glu Asn Ala Glu Asp Met Gly Asn
465 470 475 480
Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn Ala Cys Ile Glu Ser
485 490 495
Ile Arg Asn Gly Thr Tyr Asp His Asp Val Tyr Arg Asp Glu Ala Leu
500 505 510
Asn Asn Arg Phe Gln Ile Lys Gly Val Ser Gly Ser Leu Pro Glu Thr
515 520 525
Gly Gly Gly Ser His His His His His His
530 535
<210> 51
<211> 532
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV190068
<400> 51
Met Asn Thr Gln Ile Leu Val Phe Ala Leu Met Ala Ile Ile Pro Thr
1 5 10 15
Asn Ala Asp Lys Ile Cys Leu Gly His His Ala Val Ser Asn Gly Thr
20 25 30
Lys Val Asn Thr Leu Thr Glu Arg Gly Val Glu Val Val Asn Ala Thr
35 40 45
Glu Thr Val Glu Arg Thr Asn Val Pro Arg Ile Cys Ser Lys Gly Lys
50 55 60
Arg Thr Val Asp Leu Gly Gln Cys Gly Leu Leu Gly Thr Ile Thr Gly
65 70 75 80
Pro Pro Gln Cys Asp Gln Phe Leu Glu Phe Ser Ala Asp Leu Ile Ile
85 90 95
Glu Arg Arg Glu Gly Ser Asp Val Cys Tyr Pro Gly Lys Phe Val Asn
100 105 110
Glu Glu Ala Leu Arg Gln Ile Leu Arg Glu Ser Gly Gly Ile Asp Lys
115 120 125
Glu Thr Met Gly Phe Thr Tyr Ser Gly Ile Arg Thr Asn Gly Ala Thr
130 135 140
Ser Ala Cys Arg Arg Ser Gly Ser Ser Phe Tyr Ala Glu Met Lys Trp
145 150 155 160
Leu Leu Ser Asn Thr Asp Asn Ala Ala Phe Pro Gln Met Thr Lys Ser
165 170 175
Tyr Lys Asn Thr Arg Lys Asp Pro Ala Leu Ile Ile Trp Gly Ile His
180 185 190
His Ser Gly Ser Thr Thr Glu Gln Thr Lys Leu Tyr Gly Ser Gly Asn
195 200 205
Lys Leu Ile Thr Val Gly Ser Ser Asn Tyr Gln Gln Ser Phe Val Pro
210 215 220
Ser Pro Gly Ala Arg Pro Gln Val Asn Gly Gln Ser Gly Arg Ile Asp
225 230 235 240
Phe His Trp Leu Ile Leu Asn Pro Asn Asp Thr Val Thr Phe Ser Phe
245 250 255
Asn Gly Ala Phe Ile Ala Pro Asp Arg Ala Ser Phe Leu Arg Gly Lys
260 265 270
Ser Met Gly Ile Gln Ser Gly Val Gln Val Asp Ala Asn Cys Glu Gly
275 280 285
Asp Cys Tyr His Ser Gly Gly Thr Ile Ile Ser Asn Leu Pro Phe Gln
290 295 300
Asn Ile Asn Ser Arg Ala Val Gly Lys Cys Pro Arg Tyr Val Lys Gln
305 310 315 320
Glu Ser Leu Leu Leu Ala Thr Gly Met Lys Asn Val Pro Glu Ile Pro
325 330 335
Lys Gly Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Asn Gly
340 345 350
Trp Glu Gly Leu Ile Asp Gly Trp Tyr Gly Phe Arg Trp Gln Asn Ala
355 360 365
Gln Gly Glu Gly Thr Ala Ala Asp Tyr Lys Ser Thr Gln Ser Ala Ile
370 375 380
Asp Gln Ile Thr Gly Ile Leu Asn Arg Leu Ile Glu Lys Thr Asn Gln
385 390 395 400
Gln Phe Glu Leu Ile Asp Asn Glu Phe Thr Glu Val Glu Lys Gln Ile
405 410 415
Gly Asn Val Ile Asn Trp Thr Arg Asp Ser Met Thr Glu Val Trp Ser
420 425 430
Tyr Asn Ala Glu Leu Leu Val Ala Met Ile Asn Gln His Thr Ile Asp
435 440 445
Leu Ala Asp Ser Glu Met Asn Lys Leu Tyr Glu Arg Val Lys Arg Gln
450 455 460
Leu Arg Glu Asn Ala Glu Glu Asp Gly Thr Gly Cys Phe Glu Ile Phe
465 470 475 480
His Lys Cys Asp Asp Asp Cys Met Ala Ser Ile Arg Asn Asn Thr Tyr
485 490 495
Asp His Ser Lys Tyr Arg Glu Glu Ala Met Gln Asn Arg Ile Gln Ile
500 505 510
Asp Pro Val Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His His
515 520 525
His His His His
530
<210> 52
<211> 531
<212> PRT
<213> Artificial Sequence
<220>
<223> UFV190841
<400> 52
Met Tyr Lys Val Val Val Ile Ile Ala Leu Leu Gly Ala Val Lys Gly
1 5 10 15
Asp Arg Ile Cys Leu Gly His His Ala Val Ala Asn Gly Thr Ile Val
20 25 30
Lys Thr Leu Thr Asn Glu Gln Glu Glu Val Thr Asn Ala Thr Glu Thr
35 40 45
Val Glu Ser Thr Asn Leu Asn Lys Leu Cys Met Lys Gly Arg Ser Tyr
50 55 60
Lys Asp Leu Gly Asn Cys His Pro Val Gly Met Leu Ile Gly Thr Pro
65 70 75 80
Val Cys Asp Pro His Leu Thr Gly Thr Trp Asp Thr Leu Ile Glu Arg
85 90 95
Glu Asn Ala Ile Ala His Cys Tyr Pro Gly Ala Thr Ile Asn Glu Glu
100 105 110
Ala Leu Arg Gln Lys Ile Met Glu Ser Gly Gly Ile Ser Lys Met Ser
115 120 125
Thr Gly Phe Thr Tyr Gly Ser Ser Ile Asn Ser Ala Gly Thr Thr Lys
130 135 140
Ala Cys Met Arg Asn Gly Gly Asp Ser Phe Tyr Ala Glu Leu Lys Trp
145 150 155 160
Leu Val Ser Lys Thr Lys Gly Gln Asn Phe Pro Gln Thr Thr Asn Thr
165 170 175
Tyr Arg Asn Thr Asp Thr Ala Glu His Leu Ile Ile Trp Gly Ile His
180 185 190
His Pro Ser Ser Thr Gln Glu Lys Asn Asp Leu Tyr Gly Thr Gln Ser
195 200 205
Leu Ser Ile Ser Val Glu Ser Ser Thr Tyr Gln Asn Asn Phe Val Pro
210 215 220
Val Val Gly Ala Arg Pro Gln Val Asn Gly Gln Ser Gly Arg Ile Asp
225 230 235 240
Phe His Trp Thr Leu Val Gln Pro Gly Asp Asn Ile Thr Phe Ser His
245 250 255
Asn Gly Gly Leu Ile Ala Pro Ser Arg Val Ser Lys Leu Thr Gly Arg
260 265 270
Gly Leu Gly Ile Gln Ser Glu Ala Leu Ile Asp Asn Ser Cys Glu Ser
275 280 285
Lys Cys Phe Trp Arg Gly Gly Ser Ile Asn Thr Lys Leu Pro Phe Gln
290 295 300
Asn Leu Ser Pro Arg Thr Val Gly Gln Cys Pro Lys Tyr Val Asn Gln
305 310 315 320
Arg Ser Leu Leu Leu Ala Thr Gly Met Arg Asn Val Pro Glu Val Val
325 330 335
Gln Gly Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Asn Gly
340 345 350
Trp Glu Gly Met Val Asp Gly Trp Tyr Gly Phe Arg Trp Gln Asn Ala
355 360 365
Gln Gly Thr Gly Gln Ala Ala Asp Tyr Lys Ser Thr Gln Ala Ala Ile
370 375 380
Asp Gln Ile Thr Gly Ile Leu Asn Arg Leu Ile Glu Lys Thr Asn Thr
385 390 395 400
Glu Phe Glu Ser Ile Glu Ser Glu Phe Ser Glu Thr Glu His Gln Ile
405 410 415
Gly Asn Val Ile Asn Trp Thr Lys Asp Ser Ile Thr Asp Ile Trp Thr
420 425 430
Tyr Gln Ala Glu Leu Leu Val Ala Met Ile Asn Gln His Thr Ile Asp
435 440 445
Met Ala Asp Ser Glu Met Leu Asn Leu Tyr Glu Arg Val Arg Lys Gln
450 455 460
Leu Arg Gln Asn Ala Glu Glu Asp Gly Lys Gly Cys Phe Glu Ile Tyr
465 470 475 480
His Thr Cys Asp Asp Ser Cys Met Glu Ser Ile Arg Asn Asn Thr Tyr
485 490 495
Asp His Ser Gln Tyr Arg Glu Glu Ala Leu Leu Asn Arg Leu Asn Ile
500 505 510
Asn Ser Ser Gly Ser Leu Pro Glu Thr Gly Gly Gly Ser His His His
515 520 525
His His His
530
Claims (18)
- 재조합 인플루엔자 A 적혈구응집소(HA) 폴리펩티드로서, 인플루엔자 A 바이러스 HA의 HA1 및 HA2 도메인을 포함하고,
(a) 위치 355의 아미노산은 W이고;
(b) 위치 432의 아미노산은 I이고/이거나, 위치 380의 아미노산은 I인 아미노산 서열을 포함하고;
HA 폴리펩티드의 아미노산 서열의 아미노산 위치의 넘버링은 기준 H3N2 인플루엔자 균주, 특히 기준 균주 H3N2 A/아이치/2/68(서열번호 1)로부터의 HA의 아미노산 서열의 아미노산의 넘버링에 따르는 것인 HA 폴리펩티드. - 제1항에 있어서,
(a) 위치 388의 아미노산은 M이고/이거나;
(b) 위치 478의 아미노산은 I인 아미노산 서열을 포함하는 것인 HA 폴리펩티드. - 제1항 또는 제2항에 있어서, HA1 및 HA2 도메인 사이에 프로테아제 절단 부위를 포함하지 않는 것인 HA 폴리펩티드.
- 제1항 내지 제3항 중 어느 한 항에 있어서, HA1 및 HA2 도메인은 그룹 1 및/또는 그룹 2 인플루엔자 A 바이러스로부터 유래되는 것인 HA 폴리펩티드.
- 제1항 내지 제4항 중 어느 한 항에 있어서, 절단된 HA1 및/또는 HA2 도메인을 포함하는 것인 HA 폴리펩티드.
- 제5항에 있어서, 막횡단 및 세포질내 도메인이 HA2 도메인으로부터 결실된 것인 HA 폴리펩티드.
- 제5항 또는 제6항에 있어서, 위치 515의 아미노산에 해당하는 아미노산으로 시작하는 HA2 도메인의 적어도 C 말단 부분은 결실된 것인 HA 폴리펩티드.
- 제1항 내지 제7항 중 어느 한 항에 있어서, (절단된) HA2 도메인의 C 말단에 위치한 검출 및/또는 정제 태그를 포함하는 것인 HA 폴리펩티드.
- 제1항 내지 제8항 중 어느 한 항의 폴리펩티드의 면역원성 단편.
- 제1항 내지 제8항 중 어느 한 항의 적어도 두 개의 HA 폴리펩티드, 또는 제9항의 면역원성 단편을 포함하는 다량체 폴리펩티드.
- 제10항에 있어서, 폴리펩티드는 삼량체이고, 제1항 내지 제8항 중 어느 한 항의 세 개의 HA 폴리펩티드를 포함하는 것인 다량체 폴리펩티드.
- 제1항 내지 제8항, 제10항, 또는 제11항 중 어느 한 항의 HA 폴리펩티드, 또는 제9항의 면역원성 단편을 암호화하는 핵산.
- 제11항의 핵산 분자를 포함하는 벡터.
- 제12항에 있어서, 재조합 아데노바이러스 벡터인 것인, 벡터.
- 제1항 내지 제8항, 제10항, 또는 제11항 중 어느 한 항의 재조합 HA 폴리펩티드, 또는 제9항의 면역원성 단편을 제조하는 방법으로서, 원핵생물 또는 진핵생물 세포에서 제12항의 핵산 분자를 발현시키는 단계를 포함하고, 상기 세포로부터 HA 폴리펩티드 또는 이의 단편을 단리하는 단계를 추가로 선택적으로 포함하는 방법.
- 제1항 내지 제8항, 제10항 또는 제11항 중 어느 한 항의 HA 폴리펩티드, 제9항의 면역원성 단편, 제12항의 핵산, 및/또는 제13항 또는 제14항의 벡터, 및 약제학적으로 허용 가능한 담체를 포함하는 면역원성 조성물.
- 인플루엔자 바이러스에 대한 면역 반응을 유도하는 데 사용하기 위한, 제1항 내지 제8항, 제10항 또는 제11항 중 어느 한 항의 HA 폴리펩티드, 제9항의 면역원성 단편, 제12항의 핵산, 및/또는 제13항 또는 제14항의 벡터.
- 백신으로서 사용하기 위한, 제1항 내지 제8항, 제10항 또는 제11항 중 어느 한 항의 HA 폴리펩티드, 제9항의 면역원성 단편, 제12항의 핵산, 및/또는 제13항 또는 제14항의 벡터.
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| US201962838690P | 2019-04-25 | 2019-04-25 | |
| US62/838,690 | 2019-04-25 | ||
| PCT/EP2020/061335 WO2020216844A1 (en) | 2019-04-25 | 2020-04-23 | Recombinant influenza antigens |
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| US10456462B2 (en) | 2015-07-07 | 2019-10-29 | Janssen Vaccines & Preventions B.V. | Vaccine against RSV |
| WO2017174564A1 (en) | 2016-04-05 | 2017-10-12 | Janssen Vaccines & Prevention B.V. | Vaccine against rsv |
| PL3464331T3 (pl) | 2016-05-30 | 2021-04-19 | Janssen Vaccines & Prevention B.V. | Stabilizowane przedfuzyjne białka F RSV |
| CN117693359A (zh) * | 2021-06-10 | 2024-03-12 | 乔治亚大学研究基金会股份有限公司 | 作为免疫原的广泛反应性病毒抗原、其组合物和使用方法 |
| WO2023126982A1 (en) * | 2021-12-31 | 2023-07-06 | Mynvax Private Limited | Polypeptide fragments, immunogenic composition against influenza virus, and implementations thereof |
| TW202408567A (zh) * | 2022-05-06 | 2024-03-01 | 法商賽諾菲公司 | 用於核酸疫苗之訊息序列 |
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| US5057540A (en) | 1987-05-29 | 1991-10-15 | Cambridge Biotech Corporation | Saponin adjuvant |
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| JPH0832638B2 (ja) | 1989-05-25 | 1996-03-29 | カイロン コーポレイション | サブミクロン油滴乳剤を含んで成るアジュバント製剤 |
| FR2705686B1 (fr) | 1993-05-28 | 1995-08-18 | Transgene Sa | Nouveaux adénovirus défectifs et lignées de complémentation correspondantes. |
| US5851806A (en) | 1994-06-10 | 1998-12-22 | Genvec, Inc. | Complementary adenoviral systems and cell lines |
| EP0784690B1 (en) | 1994-06-10 | 2006-08-16 | Genvec, Inc. | Complementary adenoviral vector systems and cell lines |
| US5846782A (en) | 1995-11-28 | 1998-12-08 | Genvec, Inc. | Targeting adenovirus with use of constrained peptide motifs |
| US5965541A (en) | 1995-11-28 | 1999-10-12 | Genvec, Inc. | Vectors and methods for gene transfer to cells |
| US5559099A (en) | 1994-09-08 | 1996-09-24 | Genvec, Inc. | Penton base protein and methods of using same |
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| SI0833934T2 (sl) | 1995-06-15 | 2013-04-30 | Crucell Holland B.V. | Pakirni sistemi za humani rekombinantni adenovirus za uporabo v genski terapiji |
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| US6020191A (en) | 1997-04-14 | 2000-02-01 | Genzyme Corporation | Adenoviral vectors capable of facilitating increased persistence of transgene expression |
| US5981225A (en) | 1998-04-16 | 1999-11-09 | Baylor College Of Medicine | Gene transfer vector, recombinant adenovirus particles containing the same, method for producing the same and method of use of the same |
| US6113913A (en) | 1998-06-26 | 2000-09-05 | Genvec, Inc. | Recombinant adenovirus |
| SE0202110D0 (sv) | 2002-07-05 | 2002-07-05 | Isconova Ab | Iscom preparation and use thereof |
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| SG11201402633UA (en) * | 2011-11-28 | 2014-09-26 | Crucell Holland Bv | Influenza virus vaccines and uses thereof |
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