具体实施方式
下面通过实施例的方式进一步说明本发明,但并不因此将本发明限制在所述的实施例范围之中。下列实施例中未注明具体条件的实验方法,按照常规方法和条件,或按照商品说明书选择。
实施例1.抗B7H4抗体分子的获得
可以利用B7H4重组蛋白或过表达B7H4的细胞对实验动物进行免疫以获得针对B7H4特异性结合的抗体分子,该实验动物可以是小鼠、大鼠、兔、羊、骆驼等。通常,其得到的抗体分子是非人源的。在获得非人源抗体后,需要对这些分子利用抗体工程技术进行人源化改造,以降低免疫原性并提高成药性。然而,抗体的人源化过程有其技术复杂性,经过人源化改造的分子往往会降低对抗原的亲和力。另一方面,转基因技术的进步使得可以培育出基因工程化小鼠,其携带人免疫球蛋白免疫库并使其内源的鼠的免疫库缺失。HarbourH2L2小鼠(Harbour Antibodies BV)是一种携带人免疫球蛋白免疫库的转基因小鼠,这种转基因小鼠产生的抗体具有全人源的序列,因而无需再进一步做人源化改造,大大提高了治疗性抗体开发的效率。
1.1.小鼠免疫
用可溶的重组人B7H4-ECD-mFc融合蛋白(Sino Biological,#10738-H05H)作为抗原对Harbour H2L2小鼠进行多轮免疫。抗原蛋白与免疫佐剂混合成免疫原试剂,然后通过皮下经腹股沟注射或通过腹腔注射。在每一轮免疫中,每只小鼠接受的总注射剂量是100微升。在首轮免疫中,每只小鼠接受用50微克抗原蛋白与完全弗氏佐剂(Sigma,#F5881)以体积比1:1混合配制的免疫原试剂的免疫。在随后的每轮增强免疫中,每只小鼠接受用25微克抗原蛋白与Sigma Adjuvant System佐剂(Sigma,#S6322)混合配制的免疫原试剂的免疫。每轮增强免疫的间隔时间至少为两周,通常为6到7轮增强免疫。免疫时间为第0、14、28、42、56、70、84、98天;并且在第49、77天,检测小鼠血清抗体滴度。在进行H2L2小鼠脾B细胞分离前5天,以每只小鼠25微克抗原蛋白的剂量进行最后一次增强免疫。
或者用过表达人B7H4的CHO-K1细胞(CHO-K1/hu B7H4,和铂医药)转染编码小鼠CD40L的质粒后,与免疫佐剂混合成免疫原试剂后免疫小鼠,每只小鼠每次免疫5×106细胞,免疫过程同蛋白免疫。
1.2.血清滴度检测
在特定的时间点,收取小鼠血清,用ELISA方法检测血清中抗体结合B7H4蛋白的滴度以及用FACS方法检测血清中抗体结合过表达B7H4的细胞的滴度。
在ELISA方法中,用1μg/mL的hB7H4-ECD-his蛋白(Sino Biological,#10738-H08H)100μL/孔包被ELISA板(corning,9018),4℃孵育过夜;漂洗2次后,用含1%BSA的PBST在37℃阻断2小时;加入100μL/孔梯度稀释的血清37℃孵育1小时;漂洗3次后,加入100μL/孔1:5000稀释的anti-rat-HRP(sigma,#A5795),37℃孵育30分钟。漂洗3次后,加入100μL/孔TMB底物孵育约10分钟,加入50μL/孔1NHCl终止显色,读450nm的吸光度(MolecularDevices,Plus 384)。
在FACS方法中,梯度稀释的鼠血清与HEK293-B7H4细胞4℃孵育1小时;细胞洗2次后,加入二抗anti rat IgG(H+L)(Life technologies,A11006)4℃孵育1小时,洗2次后,重悬细胞用流式细胞仪检测(BD,Flibur)。HEK293细胞作为本底对照。
1.3.通过杂交瘤技术筛选抗B7H4的抗体
选择血清效价高的免疫小鼠进行一次终免疫后处死小鼠,取脾细胞和SP2/0骨髓瘤细胞(ATCC,CRL-1581)进行电融合,细胞比例为4:1,电融合参数为V1:50V,t1:15s,V2:600V,t2:20μs,t3:0.5s,n:1,t4:7s,V+/-:+,fade:on。细胞重悬于含20%FBS和HT的DMEM的培液中铺板1×10
5/100μL/孔,24小时后再加100μL/孔含20%FBS和2×HT的DMEM继续培养。后续取上清检测抗体滴度。一般在融合后9-15天,蛋白免疫的小鼠取上清用Acumen进行初筛,检测与CHO-K1/huB7H4细胞的结合;细胞免疫的小鼠取上清用Mirrorball(SPTLabtech,
fluorescence cytometer)进行初筛,检测与HEK-293/huB7H4细胞的结合。阳性的克隆然后用ELISA和FACS进一步确认,检测与过表达人B7H4的CHO-K1细胞株(CHO-K1/huB7H4)、过表达食蟹猴B7H4的CHO-K1细胞株(CHO-K1/cynoB7H4)、过表达小鼠B7H4的CHO-K1细胞株(CHO-K1/mB7H4)的结合能力。阳性的孔用有限稀释法进一步亚克隆,再进一步用ELISA与FACS方法筛选。对人和猴B7H4结合较好的克隆被挑选出来进行测序。
1.4.通过B细胞体外克隆技术筛选抗B7H4的抗体
将小鼠的脾脏取出,研磨后用200目滤网进行过滤,单细胞悬液按照小鼠记忆性B细胞分选试剂盒(Miltenyi,#130-095-838)进行分选。分选得到的细胞进行免疫荧光染色。
在流式细胞分选仪S3e上对B200阳性(BioLegend,#103227)、IgM阴性(BioLegend,#406506)、B7H4特异性阳性细胞(BioLegend,#405207)进行分选。分选获得的细胞按照每孔5个细胞的密度培养在细胞培养96孔板上,细胞培养板上预先铺好辐照过的EL4细胞作为饲养层细胞。
培养14天后收取培养上清进行ELISA检测,对于B7H4蛋白有结合活性的孔,将细胞取出进行RT-PCR(SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing(#634892),I-5TM 2×High-Fidelity Master Mix(#I5HM-5000)。扩增获得的轻、重链通过overlap PCR拼接成scFv,在E.coli中进行表达,表达上清进行ELISA检测,将阳性的克隆进行测序。
1.5.抗B7H4抗体的序列分析和序列优化
利用常规的测序手段获得编码抗体分子可变结构域的核苷酸序列以及对应的氨基酸序列。获得3个单克隆序列。在本实施例中,从免疫的Harbour H2L2小鼠得到的抗B7H4单克隆抗体分子可变结构域的序列是人源抗体序列,表1列出其胚系基因分析和翻译后修饰位点(PTM)分析。
蛋白质或多肽氨基酸链在细胞中翻译合成后有时会引入化学修饰,称为翻译后修饰(PTM)。对于抗体而言,一些PTM的位点是非常保守的,例如,在人的IgG1抗体的恒定结构域的第297位(EU编号)的保守的氨基酸天冬酰胺Asn通常会发生糖基化修饰形成糖链,而该糖链结构对于抗体结构和相关的效应子功能是至关重要的。但是,如果在抗体的可变结构域尤其是抗原结合区域(如CDR)中存在PTM,那么这些PTM的存在有可能会对抗原的结合有较大的影响,也可能对抗体的物理化学性质带来变化。例如,糖基化、脱酰胺、异构化、氧化等都可能增加抗体分子的不稳定性或异质性,从而增加抗体开发的难度和风险。因而避免一些潜在的PTM对于治疗性抗体的开发是非常重要的。随着经验的积累,人们发现一些PTM是和氨基酸序列的组成尤其是相邻氨基酸组成的“模式”是高度相关的,这样使得可以从蛋白质的一级氨基酸序列预测出潜在的PTM。例如,N-x-S/T(第一位是天冬酰胺,第二位是非脯氨酸以外的任意氨基酸,第三位是丝氨酸或者苏氨酸)的序列模式预测出N-连接糖基化位点。引起PTM的氨基酸序列模式有可能来源于胚系基因序列,例如人胚系基因片段IGHV3-33天然地在FR3区域存在糖基化模式NST;也可能来源于体细胞高频突变。
可以通过氨基酸突变来破坏PTM的氨基酸序列模式,从而降低或者去除特定PTM的形成。根据抗体序列和PTM序列模式的不同,有不同的突变设计方法。一种方法是将“热点”氨基酸(如NS模式中的N或S)替换成物理化学性质相似的氨基酸(如把N突变为Q)。如果PTM序列模式来源于体细胞高频突变,而并不存在于胚系基因序列中,那么另一种方法可以是把该序列模式替换成对应的胚系基因序列。实际操作中,对同一个PTM序列模式可能采用多种突变设计方法。
表2列出了对抗体PR001476和PR002037的序列进行氨基酸突变得到的新的抗体分子序列。
表1抗B7H4抗体的胚系基因分析和PTM位点分析
表2抗原结合蛋白序列的突变位点设计
初始抗体 |
变体 |
可变区突变 |
重组抗体亚型 |
Fc突变 |
PR001476 |
PR002405 |
H:D54E;L:N92Q |
人IgG1 |
|
PR001476 |
PR002408 |
H:G55A;L:N92Q |
人IgG1 |
|
PR001476 |
PR002411 |
|
人IgG1 |
S239D,I332E |
PR001476 |
PR002418 |
H:G55A;L:N92Q |
人IgG1 |
S239D,I332E |
PR001476 |
PR003369 |
H:G55A,G101A;L:N92R |
人IgG1 |
S239D,I332E |
PR002037 |
PR002410 |
L:C87Y |
人IgG1 |
|
PR002037 |
PR002420 |
|
人IgG1 |
S239D,I332E |
PR002037 |
PR002421 |
L:C87Y |
人IgG1 |
S239D,I332E |
1.6.PR001476抗体亲和力成熟
通过定点突变的方法对PR001476这个分子进行改造以提高其结合B7H4的亲和力。此亲和力成熟的方法分为两轮。
第一轮,对PR001476这个分子的重链CDR3和轻链CDR3(按照Chothia CDR定义)的氨基酸逐点扫描,建立多个氨基酸位置的单点饱和突变库。对2饱和突变库进行筛选,信号超过野生型2倍的阳性分子被挑出来测序,并对这些阳性分子进一步鉴定,根据其对人B7H4的结合能力,选出若干个突变热点。
第二轮,将第一轮中饱和突变找到的热点进行随机组合,建立一个包含所有突变组合的库。然后对组合库进行筛选。阳性分子通过测序以及对人hB7H4的结合能力,选出若干个突变体。筛选出的突变体用对应的克隆号来表示,例如:PR001476-R1-25B3、PR001476-R1-26D7等等。
突变体构建到哺乳动物表达载体进行蛋白的表达纯化。然后运用FACS和FortebioOctet测试突变体与B7H4的结合能力。表2中的PR003369为PR001476衍生出的优选的突变体。
1.7.重组抗体制备和理化性质表征分析
1.7.1.抗体的表达和纯化
本实施例介绍了利用哺乳动物宿主细胞(例如,人胚肾细胞HEK293或中国仓鼠卵巢细胞CHO及其衍生细胞)、瞬时转染表达和亲和捕获分离等技术来制备抗体的一般方法。本方法适用于含有Fc区的目标抗体;目标抗体可以由一条或多条蛋白质多肽链组成;可以来源于一个或多个表达质粒。
将抗体多肽链的氨基酸序列通过密码子优化方法转换成核苷酸序列;合成编码的核苷酸序列并克隆到与宿主细胞兼容的表达载体上。将编码抗体多肽链的质粒按照特定比例同时转染哺乳动物宿主细胞,利用常规的重组蛋白表达和纯化技术,可以得到具有正确折叠和多肽链组装的重组抗体。具体地,将FreeStyleTM 293-F细胞(Thermo,#R79007)在FreeStyleTM F17 Expression Medium培养基(Thermo,#A1383504)中扩培。瞬时转染开始之前,调节细胞浓度至6–8×105细胞/mL,于37℃8%CO2摇床中培养24小时,细胞浓度在1.2×106细胞/mL。准备30mL培养的细胞。将编码抗体多肽链的质粒按照一定比例混合共计30μg质粒(质粒与细胞的比例为1μg:1mL)溶解于1.5mL Opti-MEM减血清培养基(Thermo,#31985088),并用0.22μm滤膜过滤除菌。再取1.5mL Opti-MEM溶入1mg/mL PEI(Polysciences,#23966-2)120μl,静置5分钟。把PEI缓慢加入质粒中,室温孵育10分钟,边摇晃培养瓶边缓慢滴入质粒PEI混合溶液,于37℃8%CO2摇床中培养5天。5天后观测细胞活率。收集培养物,以3300g转速离心10分钟后取上清;然后将上清高速离心去除杂质。用PBSpH7.4缓冲液平衡含有MabSelectTM(GE Healthcare,#71-5020-91)的重力柱(Bio-Rad,#7311550),2-5倍柱体积冲洗。将上清样品过柱;用5-10倍柱体积的PBS缓冲液冲洗柱子,再用pH3.5的0.1M甘氨酸洗脱目的蛋白,随后用pH 8.0的Tris-HCl调节至中性,最后用超滤管(Millipore,#UFC901024)浓缩换液至PBS缓冲液或者含有其他成分的缓冲液,得到纯化的重组抗体溶液。最后用NanoDrop(Thermo,NanoDropTM One)测定浓度,分装、存储备用。
1.7.2.利用SEC-HPLC分析蛋白纯度和聚体
本实施例使用分析型分子尺寸排阻层析色谱法(SEC)来分析蛋白样品的纯度和聚体形式。将分析型色谱柱TSKgel G3000SWxl(Tosoh Bioscience,#08541,5μm,7.8mm×30cm)连接到高压液相色谱仪HPLC(Agilent Technologies,Agilent 1260Infinity II),用PBS缓冲液室温下平衡至少1小时。适量蛋白样品(至少10μg)用0.22μm滤膜过滤后注射入系统,并设定HPLC程序:用PBS缓冲液将样品以1.0mL/分钟的流速流过色谱柱,最长时间为25分钟。HPLC将生成分析报告,报告出样品内不同分子尺寸组份的滞留时间。
1.7.3.利用HIC-HPLC分析蛋白纯度和疏水性
使用分析型疏水相互作用层析色谱法(HIC)来分析蛋白样品的纯度和疏水性。将分析型色谱柱TSKge1 Buty1-NPR(Tosoh Bioscience,14947,4.6mm×3.5cm)连接到高压液相色谱仪HPLC(Agilent Technologies,Agilent 1260Infinity II),用PBS缓冲液室温下平衡至少1小时。设定方法由16分钟内从100%流动相A(20mM组氨酸,1.8M硫酸铵,pH 6.0)至100%流动相B(20mM组氨酸,pH 6.0)的线性梯度,流速设定为0.7mL/分钟,蛋白样品浓度1mg/mL,进样体积20μl,检测波长280nm。采集后用ChemStation软件对色谱图进行积分并计算相关数据,生成分析报告,报告出样品内不同分子尺寸组份的滞留时间。
1.7.4.利用DSF测定蛋白分子的热稳定性
差示扫描荧光法(Differential Scanning Fluorimetry,DSF)是一种常用的高通量的测定蛋白质热稳定性的方法。它使用实时荧光定量PCR仪器通过监测与去折叠的蛋白分子结合的染料的荧光强度的变化,来反映蛋白质的变性的过程,从而反映出蛋白分子的热稳定性。本实施例利用DSF方法来测定蛋白分子热变性温度(Tm)。10μg蛋白加入96-孔PCR板(Thermo,#AB-0700/W),接着加入2μl 100×稀释的染料SYPROTM(Invitrogen,#2008138),然后加入缓冲液使得终体积为40μl每孔。将PCR板密封,放置于实时荧光定量PCR仪器(Bio-Rad CFX96 PCR System),先于25℃孵育5分钟,然后以0.2℃/0.2分钟的梯度逐渐从25℃升温至95℃,在测试结束时将温度降至25℃。使用FRET扫描模式并使用Bio-RadCFX Maestro软件进行数据分析并计算出样品的Tm。
1.8.制备抗B7H4全人重组抗体
将1.3-1.6中得到的抗B7H4的全人源IgG抗体和优化后抗体利用1.7.1所述方法进行制备并分析。表3和表4分别列出了小体积和大体积瞬时表达纯化的结果。另外,从现有文献中得到抗B7H4抗体序列(表5),在后续实验中作为对照。
表3抗B7H4抗体的表达
抗体 |
表达体系和体积 |
一步纯化后产量(mg/L) |
SEC-HPLC纯度(%) |
PR001476 |
HEK293-F(50mL) |
80.6 |
99.18 |
PR002037 |
HEK293-F(30mL) |
17.3 |
100 |
PR002038 |
HEK293-F(30mL) |
34.0 |
100 |
PR002408 |
HEK293-6E(40mL) |
37.0 |
99.03 |
PR002411 |
HEK293-6E(40mL) |
64.8 |
99.3 |
PR002418 |
HEK293-F(1.5mL) |
26.7 |
未测 |
PR003369 |
HEK293-F(30mL) |
6.6 |
100 |
PR002410 |
HEK293-6E(40mL) |
173.5 |
98.65 |
PR002420 |
HEK293-6E(40mL) |
5.7 |
99.16 |
PR002421 |
HEK293-F(1.5mL) |
73.3 |
未测 |
表4抗B7H4抗体的表达
表5对照抗体相关信息
1.9.抗B7H4抗体序列与编号
在本发明中,上述所列CDR的氨基酸序列均是按照Chothia定义规则所示出的。但是,本领域人员公知,在本领域中可以通过多种方法来定义抗体的CDR,例如基于序列可变性的Kabat定义规则(参见,Kabat等人,免疫学的蛋白质序列,第五版,美国国立卫生研究院,贝塞斯达,马里兰州(1991))和基于结构环区域位置的Chothia定义规则(参见JMolBiol 273:927-48,1997)。在本发明的技术方案中,还可以使用包含了Kabat定义和Chothia定义的Combined定义规则来确定可变结构域序列中的氨基酸残基。其中Combined定义规则即是将Kabat定义和Chothia定义的范围相结合,基于此取了一个更大的范围,详见表6。本领域技术人员应当理解的是,除非另有规定,否则术语给定抗体或其区(例如可变区)的“CDR”及“互补决定区”应了解为涵盖如通过本发明描述的上述已知方案中的任何一种界定的互补决定区。虽然本发明中请求保护的范围是基于Chothia定义规则所示出的序列,但是根据其他CDR的定义规则所对应的氨基酸序列也应当落在本发明的保护范围中。
表6本申请抗体CDR定义方法
|
Kabat |
Chothia |
Combined |
LCDR1 |
L24--L34 |
L24--L34 |
L24-L34 |
LCDR2 |
L50--L56 |
L50--L56 |
L50-L56 |
LCDR3 |
L89--L97 |
L89--L97 |
L89-L97 |
HCDR1 |
H31--H35 |
H26--H32 |
H26-H35 |
HCDR2 |
H50--H65 |
H52--H56 |
H50-H65 |
HCDR3 |
H95--H102 |
H95--H102 |
H95-H102 |
其中,Laa-Lbb可以指从抗体轻链的N端开始,第aa位(Chothia编码规则)至第bb位(Chothia编码规则)的氨基酸序列;Haa-Hbb可以指从抗体重链的N端开始,第aa位(Chothia编码规则)至第bb位(Chothia编码规则)的氨基酸序列。例如,L24-L34可以指从抗体轻链N端开始,按照Chothia编码规则的从第24位至第34位的氨基酸序列;H26-H35可以指从抗体重链N端开始,按照Chothia编码规则的从第26位至第35位的氨基酸序列。本领域技术人员应当知晓的是,在用Chothia编码CDR时,有些位置会有插入位点的情况(可参见http://bioinf.org.uk/abs/)。
表7列出了本发明的抗B7H4抗体以及对照抗体分子的序列所对应的CDR、可变区和轻重链的序列编号。PR003366是利用PR002410的可变区序列构建的单链可变区(scFv)同源二聚体分子(scFv-Fc结构)。
表7抗B7H4抗体的序列编号表
表8列出了本发明的抗B7H4抗体以及对照抗体分子的序列所对应框架区、Fv的序列编号。
表8抗B7H4抗体的框架区和Fv序列编号表
表9列出了本发明的抗B7H4抗体以及对照抗体分子的序列所对应的CDR序列。
表9抗B7H4抗体的CDR序列
实施例2.FACS检测抗B7H4抗体结合B7H4的能力
本实施例是为了研究抗人B7H4的H2L2单抗体外结合人/食蟹猴/小鼠B7H4的活性。采用过表达人B7H4的CHOK1细胞株(CHOK1/hu B7H4,和铂医药)、过表达食蟹猴B7H4的CHOK1细胞株(CHOK1/cyno B7H4,和铂医药)、过表达小鼠B7H4的CHOK1细胞株(CHOK1/m B7H4,和铂医药)和高表达人B7H4的细胞系SK-BR-3(
HTB-30)进行细胞水平上的抗体结合实验。简言之,消化CHOK1/hu B7H4细胞、CHOK1/cyno B7H4细胞、CHOK1/m B7H4细胞或SK-BR-3细胞,并用含2%BSA的PBS重悬。将细胞密度分别调整为1×10
6细胞/mL。以100μL细胞/孔接种于96孔V底板(Corning,#3894),随后加入100μL/孔,2倍于终浓度的3倍浓度梯度稀释的待测抗体。将细胞放置于4℃,避光孵育2小时。之后,加入100μL/孔预冷含2%BSA的PBS漂洗细胞两次,于500g、4℃下离心5分钟,弃上清。再加入100μL/孔荧光二抗(Alexa Fluor488-conjugated AffiniPure Goat Anti-Human IgG,FcγFragment Specific,Jackson,#109-545-098,1:500稀释),4℃,避光孵育1小时。用100μL/孔预冷含2%BSA的PBS洗涤细胞两次,于500g、4℃下离心5分钟,弃上清。最后,200μL/孔预冷含2%BSA的PBS重悬细胞,使用ACEA Novocyte3000流式细胞仪读取荧光发光信号值。
抗体结合细胞表面的人B7H4、食蟹猴B7H4和小鼠B7H4,以及结合肿瘤细胞SK-BR-3表面的B7H4的汇总如下(表10、表11、表12)。其中亲和力成熟变体PR003369与PR002418相比在肿瘤细胞的结合上有了明显的提高(表12)。
表10抗B7H4抗体(初始抗体)结合细胞表面的人B7H4、食蟹猴B7H4、小鼠B7H4以及结合肿瘤细胞SK-BR-3表面的B7H4
表11抗B7H4抗体(变体分子)结合细胞表面的食蟹猴B7H4、小鼠B7H4以及结合肿瘤细胞SK-BR-3表面的B7H4
表12 PR003369与PR002418与肿瘤细胞的结合
初始抗体结合细胞表面的人B7H4的结果见图1,结果显示,PR001476、PR002037有较好的人B7H4结合活性,PR002038结合活性较差;PR001476与PR002037及其PTM变体/DE突变抗体表达上清结合细胞表面的人B7H4的结果见图2,结果显示,PR002037、PR001476的PTM变体/DE突变没有明显影响抗体与人B7H4的结合活性。初始抗体结合细胞表面的食蟹猴B7H4的结果见图3,结果显示,PR002037、PR001476有较好的食蟹猴B7H4交叉结合活性,PR002038交叉结合活性较差;PR001476与PR002037及其PTM变体/DE突变抗体结合细胞表面的食蟹猴B7H4的结果见图4,结果显示,PR002037、PR001476的PTM变体/DE突变没有明显影响抗体与食蟹猴B7H4的交叉结合活性。初始抗体结合细胞表面的小鼠B7H4的结果见图5,结果显示,PR002037有较好的小鼠B7H4交叉结合活性,PR001476有较弱的小鼠B7H4交叉结合活性,PR002038无小鼠B7H4交叉结合活性;PR001476与PR002037及其PTM变体/DE突变抗体结合细胞表面的小鼠B7H4的结果见图6,结果显示,PR002037、PR001476的PTM变体/DE突变抗体与其母本相比保持了相似的小鼠B7H4交叉结合活性。初始抗体结合肿瘤细胞SK-BR-3表面的B7H4的结果见图7,结果显示,PR002037、PR001476与肿瘤细胞SK-BR-3表面的B7H4有较好的结合活性,PR002038结合活性较差;PR001476与PR002037及其PTM变体/DE突变抗体上清结合肿瘤细胞SK-BR-3表面的B7H4的结果见图8,结果显示,PR002037、PR001476的PTM变体/DE突变没有明显影响抗体与肿瘤细胞SK-BR-3表面的B7H4的结合活性;PR001476与PR002037及其PTM变体/DE突变抗体结合肿瘤细胞SK-BR-3表面的B7H4的结果见图9,结果显示,PR002037、PR001476的PTM变体/DE突变抗体保持了与肿瘤细胞SK-BR-3表面的B7H4的结合活性。其中PR001476的PTM变体/DE突变抗体与对照抗体1相比,有更强的结合活性(更低的EC50);PR002418与PR001476的亲和力成熟/DE突变抗体PR003369结合肿瘤细胞MDA-MB-468表面的B7H4的结果见图10,结果显示,亲和力成熟变体PR003369与PTM变体PR002418相比与肿瘤细胞MDA-MB-468表面的B7H4的结合有明显提高。
实施例3.ADCC活性检测
本实施例是为了研究抗人B7H4的H2L2单抗体外通过ADCC效应介导NK细胞杀伤靶细胞的活性。本实验采用人PBMC作为效应细胞,高表达B7H4的细胞系SK-BR-3、MDA-MB-468以及中表达B7H4的细胞系HCC-1954作为靶细胞。使用ACEA公司的RTCA仪器检测靶细胞的电导率反映杀伤效率。96孔板e-plate先用50μl完全培养基平衡。消化SK-BR-3、MDA-MB-468或HCC-1954细胞,重悬于含10%胎牛血清的RPM1640完全培养基中,稀释至4×105/mL,铺板50μl/孔于e-plate 96板中,即2×104/孔,37℃培养过夜。次日每孔加入新鲜50μl含2×105PBMC的培液,随后加入50μl 4×的浓度梯度稀释的抗体,抗体最高终浓度为10nM,每个抗体共8个浓度,设置两个重复。实时检测靶细胞的电导率,一般情况取4小时时间点的数据计算靶细胞杀伤效率=(1-样品/空白对照)×100%。
PR001476及其PTM变体/DE突变抗体对SK-BR-3肿瘤细胞的ADCC杀伤活性见图11,结果显示,Fc端的DE突变(PR002418)能明显增强抗体对SK-BR-3肿瘤细胞的ADCC杀伤活性;PR002037及其PTM变体/DE突变抗体对SK-BR-3肿瘤细胞的ADCC杀伤活性见图12,结果显示,Fc端的DE突变(PR002421)能明显增强抗体对SK-BR-3肿瘤细胞的ADCC杀伤活性。PR002418、PR002421与来自FivePrime的对照抗体对SK-BR-3肿瘤细胞的ADCC杀伤活性比较见图13(其中A为供体1,B为供体2数据),结果显示,PR002418与来自FivePrime对照抗体相比有相似的SK-BR-3肿瘤细胞的ADCC杀伤活性,PR002421杀伤活性稍弱。PR003369、PR002418、RP002421与来自FivePrime的对照抗体2对MDA-MB-468,HCC-1954肿瘤细胞的ADCC杀伤活性比较见图14(其中A为MDA-MB-468,B为HCC-1954数据),结果显示,PR003369、PR002418、RP002421对MDA-MB-468,HCC-1954肿瘤细胞都有ADCC杀伤活性,且杀伤活性与B7H4表达成正相关,即在高表达B7H4的MDA-MB-468细胞上杀伤活性更强,在中表达B7H4的HCC-1954细胞上杀伤活性较弱。亲和力成熟变体PR003369与PR002418,PR002421相比,进一步提高了ADCC杀伤活性。
实施例4.T细胞活化活性检测
为了检测抗B7H4抗体阻断T细胞免疫检查点从而活化T细胞的功能,本实验在HEK293T细胞上过表达全长B7H4以及scFv形式的抗人CD3抗体OKT3作为人工抗原递呈细胞(HEK293T/OS8/hB7H4,北京康源),使用人T细胞分离试剂盒(Miltenyi,#130-096-535)按说明书的方法分离T细胞,将人工抗原递呈细胞与T细胞共培养,检测抗B7H4抗体对T细胞活化的作用。具体地,将HEK293T-OS8-hB7H4按1×104/孔的密度铺板,培养过夜。分离人原代T细胞,按2×105/100μl/孔的密度加入HEK293T/OS8/hB7H4细胞中。随后加入100μL/孔,2×终浓度的5倍浓度梯度稀释的待测抗体,其中抗体最高终浓度为10nM,每个抗体共6个浓度,设置两个重复。培养3天后,收上清,ELISA方法检测IFN-γ的浓度。结果显示PR003369,PR002418,PR002421与对照抗体都可以促进T细胞活化,其中亲和力成熟变体PR003369比PTM变体PR002418以及对照抗体2在有更强的T细胞活化活性,其作用机制可能是阻断了B7H4与其在T细胞上的未知受体之间的作用。抗B7H4抗体阻断B7H4的免疫抑制信号从而活化T细胞的结果见图15(其中A为供体1,B为供体2)。
实施例5.抗体内化实验
使用Zenon pHrodo iFL IgG Labeling Reagents试剂盒(Invitrogen,#Z25611)检测抗体的内化。该试剂是一种带荧光染料的二抗,在中性pH时不发出荧光,当结合一抗并随抗体被内化到溶酶体,内化后在酸性pH环境下可自动发出明亮的荧光,可用FACS方法检测。具体方法如下:收集SK-BR-3细胞,离心弃上清后,用培养基重悬细胞,调整细胞浓度为3×106/mL。向96孔板内加入细胞悬液,50μl/孔,然后放入37℃恒温培养箱内孵育过夜。准备4×的待测抗体,最高浓度为40nM(4×),3倍稀释,共8个稀释度。准备4×的Zenon溶液,将25μl的待测抗体和25μl的4×Zenon标记溶液混合在一起,室温放置5分钟。然后将50μl标记过的抗体加入到含有细胞的96孔板内,37℃恒温培养箱内孵育24小时。消化细胞,在流式细胞仪上读取荧光值。结果表明,与对照抗体RP000014和其它抗体相比,PR003369具有最高的内化活性。抗B7H4抗体在SK-BR-3细胞上的内化结果见图16。
使用a-HFc-CL-MMAF试剂(Moradec,#AH-102-AF)检测抗体的内化。该试剂是一种带毒性基团MMAF的二抗,当结合一抗并随抗体被内化后在细胞内释放毒性基团杀死靶细胞,其机理类似于ADC的作用。具体方法如下:收集SK-BR-3细胞,离心弃上清后,用培养基重悬细胞,调整细胞浓度为1×105/mL。向96孔板内加入细胞悬液,50μl/孔,然后放入37℃恒温培养箱内孵育过夜。准备4×的待测抗体,最高浓度为40nM(4×),5倍稀释,共8个稀释度。准备4×的MMAF溶液(4μg/mL)。分别向含有细胞的96孔板内加入25μl的4×待测抗体和25μl的4×MMAF溶液,37℃恒温培养箱内孵育72小时。向实验孔内加入100μl的CTG溶液,用酶标仪读取CTG发光信号。图17结果表明,与对照抗体RP000014和其它抗体相比,PR003369具有最高的内化介导的细胞毒性活性。
实施例6.抗体药物偶联(ADC)
本实施例使用ADC偶联技术将一种毒性基团MMAE交联到抗B7H4抗体(PR003369抗体或者对照抗体1)上制备成ADC。产品的纯度参数如下,HPLC检测方法同1.7.2。
表13抗B7H4的H2L2抗体的ADC制备
为了研究抗体在MMAE基团交联后是否影响其对B7H4靶点的结合活性,采用高表达人B7H4的细胞系MDA-MB-468进行细胞水平上的抗体结合实验,方法同实施例2。图18结果显示MMAE基团交联基本没有影响抗体对肿瘤细胞表面的B7H4结合活性。
为了研究抗体在MMAE基团交联后是否影响其内化活性,使用Zenon pHrodo iFLIgG Labeling Reagents试剂盒(Invitrogen,#Z25611)检测抗体的内化。实验方法同实施列5。图19结果显示MMAE基团交联基本没有影响抗体在肿瘤细胞上的内化活性。
本实施例是为了研究抗体MMAE基团交联ADC的细胞杀伤活性。采用高表达B7H4的细胞系MDA-MB-468作为靶细胞,使用ACEA公司的RTCA仪器检测靶细胞的电导率反映杀伤效率。96孔板e-plate先用50μl完全培养基平衡。消化MDA-MB-468细胞,重悬于含10%胎牛血清的RPM1640完全培养基中,稀释至1×105/mL,铺板50μl/孔于e-plate 96板中,即5×103/孔,37℃培养过夜。次日每孔加入100μl 2×的浓度梯度稀释的抗体,抗体最高终浓度为50nM,每个抗体共8个浓度,设置两个重复。实时检测靶细胞的电导率,一般情况取96小时时间点的数据计算靶细胞杀伤效率=(1-样品/空白对照)×100%。图20结果显示亲和力成熟变体PR003369-ADC比对照抗体1-ADC有更高的肿瘤细胞杀伤效率。
实施例7.抗B7H4抗体与人B7H4重组蛋白的亲和力测定
7.1.用SPR方法测定亲和力
将10×HBS-EP+(GE Healthcare,#BR-1006-69)10倍稀释后,作为实验缓冲液。设置流速10μl/分钟,在芯片CM5(GE Healthcare,#BR-1005-30)的4个通道上偶联protein A:1)设置注入时间800s,将50mM NHS和200mM EDC以1:1体积比新鲜混合后注入4个通道;2)用pH4.5的醋酸钠(GE Healthcare,#BR-1003-50)将protein A稀释至20μg/mL,注入各通道800s;3)注入1M pH8.5乙醇胺800s,以封闭芯片表面剩余的活性羧基。封闭后继续用1×HBS-EP+缓冲液平衡仪器两小时,protein A最终偶联量约为2000RU。
在Biacore T200设置多循环动力学模式,每个循环包括抗体的捕获、分析物的结合以及芯片的再生。将抗体PR002418、PR002421、对照抗体1、对照抗体2均稀释至1μg/mL,以10μl/分钟的流速注入2、3、4通道30s,由预先偶联的Protein A捕获各抗体,捕获量约为160RU。将人B7-H4(Sino biological,#10738-H08H)依次按照0nM、1.5625nM、3.125nM、6.25nM、12.5nM、25nM、50nM的浓度梯度注入四个通道(对于对照抗体1,增加了一个最高浓度100nM),流速设置为30μl/分钟。对PR002418、PR002421、对照抗体2设置解离时间200s,对对照抗体1设置解离时间500s,注入时间均设为180s。最后以同样流速注入10mM甘氨酸-盐酸pH1.5(GE Healthcare,#BR-1003-54)30s,以再生芯片。
用Biacore T200分析软件2.0对实验结果进行分析,1通道作为参比通道扣除,分析模型选用1:1动力学拟合模型。结果见表14和图21的A-D,结果显示PR002421有最高的蛋白亲和力。
表14抗B7H4抗体结合人B7H4蛋白的亲和力(SPR方法)
抗体 |
抗原 |
抗体浓度(nM) |
ka(1/Ms) |
kd(1/s) |
KD(M) |
PR002418 |
人B7H4(his tag) |
50-1.5625 |
2.01E+05 |
4.75E-03 |
2.37E-08 |
PR002421 |
人B7H4(his tag) |
50-1.5625 |
3.50E+05 |
3.22E-04 |
9.21E-10 |
对照抗体2 |
人B7H4(his tag) |
50-1.5625 |
2.61E+05 |
1.16E-03 |
4.45E-09 |
对照抗体1 |
人B7H4(his tag) |
100-3.125 |
4.04E+04 |
1.89E-04 |
4.67E-09 |
7.2.用BLI方法测定亲和力
将10×动力学缓冲液(ForteBio,#18-1105)稀释至1×,用于亲和力测试以及抗原、抗体的稀释。通过生物膜干涉(BLI)技术,使用Octet Red 96e(Fortebio)分子相互作用分析仪来进行抗原抗体之间的结合动力学分析。
测定抗原和抗体的亲和力时,传感器转动速度为1000转/分钟。先将置于一列的AHC传感器(Fortebio,#18-5060)在测试缓冲液中平衡10分钟,然后用AHC传感器捕获B7-H4抗体,捕获高度0.7nm;AHC传感器在缓冲液中平衡120s后与2倍梯度稀释的人B7-H4(浓度为50-3.125nM以及0nM)结合180s,解离300s。最后将AHC传感器浸入10mM甘氨酸-盐酸pH 1.5溶液进行再生,以洗脱结合在传感器上的蛋白。
使用Octet Data Analysis软件(Fortebio,版本11.0)进行数据分析时,以0nM为参照孔,扣除参照信号(reference subtraction),选择“1:1Global fitting”方法进行数据拟合,计算出抗原与抗原结合蛋白结合的动力学参数,得到kon(1/Ms)值、kdis(1/s)值和KD(M)值(见表15),结果显示,亲和力成熟变体PR003369与PR002418相比在蛋白亲和力上有明显的提高。
表15抗B7H4抗体结合人B7H4蛋白的亲和力(BLI方法)
抗体 |
浓度(nM) |
KD(M) |
kon(1/Ms) |
kdis(1/s) |
Full R^2 |
PR002418 |
50-12.5 |
1.03E-08 |
2.75E+05 |
2.84E-03 |
0.9926 |
PR003369 |
50-12.5 |
1.39E-09 |
2.83E+05 |
3.94E-04 |
0.9981 |
实施例8.利用BLI方法测定抗B7H4抗体结合B7H4的表位竞争
使用ForteBio Octet Red96e平台对B7-H4抗体PR002418、PR002421、对照抗体1、对照抗体2进行表位竞争实验,实验缓冲液与上述相同。第一步,获取抗体的100%信号:用SA传感器(Fortebio,#18-5019)捕获B7-H4(Acro Biosystems,#B74-H82E2-200μg),捕获高度为0.25nm。将传感器在缓冲液中平衡120s后浸入稀释至100nM的各抗体中,时间240s,将抗体与B7-H4结合的最终信号记录为该抗体的100%信号。第二步,表位竞争实验:用SA传感器捕获B7-H4,捕获高度为0.25nm。将传感器浸入第一抗体中(浓度为100nM),时间240s,再将SA传感器浸入第一抗体和第二抗体的混合物中(两种抗体的终浓度均为100nM),时间240s,将传感器浸入抗体混合物后的信号差,记录为该抗体作为第二抗体的信号。抑制率通过下式计算,
抑制率(%)=(A-B)/A*100
A:某抗体的100%信号(从第一步中获得),B:该抗体作为第二抗体的信号(从第二步获得)。
若得到的抑制率大于85(%),则意味着两种抗体的表位完全重叠;若抑制率小于85(%),则意味着两种抗体结合的表位不完全重叠。
表16结果显示PR002418与PR002421与B7-H4的结合表位不同,且都与对照抗体1、对照抗体2的表位不同。其中PR002418结合一个独特的表位(第一表位),PR002421结合另一个表位(第二表位),对照抗体1,对照抗体2结合相同的表位(第三表位)。
表16抗体结合B7H4竞争性分析
实施例9.与B7家族其它成员的交叉反应性
将B7家族的蛋白(详见表17)分别用PBS稀释为1μg/mL,加至96孔板(Corning,#9018)中,每孔100μl,4℃下孵育过夜。弃去液体后用PBST缓冲液(pH 7.4,含0.05%tween-20)洗板3次,加入250μl 2%BSA封闭液,37℃条件下孵育1小时。弃去封闭液,并用PBST缓冲液(pH7.4,含0.05%吐温-20)洗板3次,将待测抗原结合蛋白,稀释成10nM和1nM 2个浓度,每个孔加入100μl,37℃孵育1小时,同型抗体作为对照。PBST缓冲液(pH7.4,含0.05%吐温-20)清洗3次后,加入稀释5000倍的羊抗人F(ab')2HRP二抗(Jackson ImmunoResearch,109-035-097),37℃条件下,避光孵育1小时。PBST缓冲液(pH7.4,含0.05%tween-20)清洗3次后,添加100μl/孔TMB(Biopanda,#TMB-S-003),室温避光放置约30分钟;每孔加入50μl/孔终止液(BBI life sciences,#E661006-0200)终止反应,置酶标仪(PerkinElemer,#Enspire)中测定450nm吸光度(OD450)值。图22说明本发明的抗体不与B7家族其它成员蛋白发生交叉反应。
表17本实施例所使用的B7家族蛋白的材料信息
B7家族其它成员 |
供应商 |
目录号 |
Human B7-1/CD80 Protein,Fc Tag(HPLC-verified) |
Acro |
B71-H5259 |
Human B7-2/CD86Protein,Fc Tag |
Acro |
CD6-H5257 |
Human B7-DC/PD-L2/CD273Protein,Recombinant(Fc Tag) |
Sino Biological |
H10292-H02H |
Human PD-L1/B7-H1 Protein,His Tag(HPLC verified) |
Acro |
PD1-H5229 |
CD275/ICOS ligand Protein,Human,Recombinant(Fc Tag) |
Sino Biological |
11559-H02H |
B7-H3/CD276Protein,Human,Recombinant(ECD,Fc Tag) |
Sino Biological |
11188-H02H |
Recombinant human B7H4 Fc Chimera |
R&D |
8870-B7-050 |
Recombinant human VISTA/B7H5/PD-1H Fc Chimera |
R&D |
7126-B7-050 |
B7-H6/NCR3LG1 Protein,Human,Recombinant(Fc Tag) |
Sino Biological |
16140-H02H |
Recombinant human B7H7/HHLA2 Fc Chimera |
R&D |
8084-B7-050 |
实施例10.血清稳定性分析
取30μl抗体稀释到270μl正常人血清中(血清浓度90%),抗体分成5份,分别放入37℃孵育0,1,4,7,14天,取出用液氮速冻后,保存于-80℃。流式方法检测抗体对SK-BR-3细胞上的B7H4的结合。
消化SK-BR-3或CHOK1/h B7H4细胞,并用含2%BSA的PBS重悬。将细胞密度分别调整为1×106细胞/mL。以100μL细胞/孔接种于96孔V底板(Corning,#3894),随后加入100μL/孔,2倍于终浓度的3倍浓度梯度稀释的待测抗体。将细胞放置于4℃,避光孵育2小时。之后,加入100μL/孔预冷含2%BSA的PBS漂洗细胞两次,于500g、4℃下离心5分钟,弃上清。再加入100μL/孔荧光二抗(Alexa Fluor 488-conjugated AffiniPure Goat Anti-Human IgG,FcγFragment Specific,Jackson,#109-545-098,1:500稀释),4℃,避光孵育60分钟。用100μL/孔预冷含2%BSA的PBS洗涤细胞两次,于500g、4℃下离心5分钟,弃上清。最后,200μL/孔预冷含2%BSA的PBS重悬细胞,使用ACEA Novocyte3000流式细胞仪读取荧光发光信号值。图23结果显示抗B7H4抗体PR002418、PR002037、PR003369以及对照抗体1、对照抗体2在37℃人血清中14天具有很好的稳定性。
实施例11.在C57BL/6小鼠体内的药代动力学
选取体重18~22克的雌性Nu/Nu小鼠6只,按20mg/kg的剂量通过尾静脉注射给与药物;一组3只于给药前以及给药后15分钟、24小时(1天)、第4天、和第10天采集全血,另一组3只于只于给药前以及给药后5小时、第2天、第7天、和第14天采集全血。将全血静置30分钟使其凝固,随后在4℃下以2,000rpm离心5分钟并将分离的血清样品在-80℃下冻存直至分析。本实施例采用ELISA方法来定量测定小鼠血清中的药物浓度。ELISAFc端总体检测方法,通过包被于96孔板的山羊抗人Fc多克隆抗体来捕获小鼠血清中的含有人Fc的融合蛋白,然后加入HRP标记的山羊抗人Fc第二抗体来检测。使用Phoenix WinNonlin软件8.2版,选用非房室模型(NCA)对血药浓度数据进行分析以评价其药代动力学。
表18为PR002418、PR002421和对照抗体2(PR002962)的药代动力学参数。图24的结果表明,在Fc端总体检测方法下,从前14天的数据计算,PR002418在小鼠体内的半衰期约为6.56天,PR002421在小鼠体内的半衰期约为6.64天,对照抗体2在小鼠体内的半衰期约为5.90天。该结果表明,PR002418和PR002421在小鼠体内的半衰期略长于对照抗体2。
表18抗B7H4抗体的药代动力学参数
PK参数 |
PR002421 |
PR002418 |
PR002962 |
T1/2(hr) |
159.3 |
157.5 |
141.5 |
Vd(mL/kg) |
93.3 |
89.3 |
93.3 |
AUCall(μg/mL hr) |
36,710 |
37,786 |
33,913 |
Cl(mL/hr/kg) |
0.42 |
0.41 |
0.48 |
C0(μg/mL) |
437.9 |
387.6 |
440.4 |
实施例12.抗B7H4抗体的抗肿瘤药效
12.1.BALB/c nude小鼠MDA-MB-468肿瘤模型
细胞接种当天每只BALB/c nude小鼠皮下接种1×107MDA-MB-468肿瘤细胞,细胞重悬在PBS与Matrigel(1:1)混合液中(0.1mL/只),皮下接种。当小鼠平均瘤体积在135mm3进行分组,25只小鼠被分为5组,给药周期为每周2次,共进行了12次给药,腹腔给药。开始给药后,每周称量体重及瘤体积两次,瘤体积计算方式为:肿瘤体积(mm3)=0.5×肿瘤长径×肿瘤短径2。给药后第39天结束实验,所有小鼠进行安乐处理。
BALB/c nude小鼠MDA-MB-468肿瘤模型的体内抗瘤效果参见图25,具体地,溶媒对照组小鼠在给药后第39天平均肿瘤体积为1054mm3。测试药PR002418(5mg/kg)治疗组在给药后第39天平均肿瘤体积为606mm3,相较溶媒对照组有显著性差异(p值为0.015),肿瘤抑制率TGI(%)为42.45%。测试药PR002418(15mg/kg)治疗组在给药后第39天平均肿瘤体积为532mm3,相较溶媒对照组有显著性差异(p值为0.007),肿瘤抑制率TGI(%)为49.47%。测试药PR002421(5mg/kg)治疗组在给药后第39天平均肿瘤体积为665mm3,相较溶媒对照组无显著性差异(p值为0.07),肿瘤抑制率TGI(%)为36.86%。测试药PR002421(15mg/kg)治疗组在给药后第39天平均肿瘤体积为335mm3,相较溶媒对照组有显著性差异(p值为0.018),肿瘤抑制率TGI(%)为68.23%。
12.2.NSG小鼠重建人PBMC免疫系统的MDA-MB-468肿瘤模型
细胞接种当天每只NCG小鼠皮下接种5×106MDA-MB-468肿瘤细胞,细胞重悬在PBS与Matrigel(1:1)混合液中(0.1mL/只),皮下接种。当小鼠平均瘤体积在126mm3进行分组,30只小鼠被分为5组,每只小鼠静脉接种5×106人的PBMC,细胞重悬在200μl PBS中。次日开始给药,给药周期为一周两次,共进行了8次给药,腹腔给药。开始给药后,每周称量体重及瘤体积两次,瘤体积计算方式为:肿瘤体积(mm3)=0.5×肿瘤长径×肿瘤短径2。给药后第36天结束实验观察,随后所有小鼠进行安乐死处理。
NSG小鼠重建人PBMC免疫系统的MDA-MB-468肿瘤模型的体内抗瘤效果参见图26,具体地,溶媒对照组小鼠在给药后第36天平均肿瘤体积为942mm3。测试药PR002418(15mg/kg)治疗组在给药后第36天平均肿瘤体积为585mm3,相较溶媒对照组无显著性差异(p值为0.073),肿瘤抑制率TGI(%)为37.91%。测试药PR002421(15mg/kg)治疗组在给药后第36天平均肿瘤体积为670mm3,相较溶媒对照组无显著性差异(p值为0.200),肿瘤抑制率TGI(%)为28.87%。测试药PR003369(15mg/kg)治疗组在给药后第36天平均肿瘤体积为354mm3,相较溶媒对照组有显著性差异(p值为0.008),肿瘤抑制率TGI(%)为62.38%。测试药对照抗体2(15mg/kg)治疗组在给药后第36天平均肿瘤体积为533mm3,相较溶媒对照组有显著性差异(p值为0.028),肿瘤抑制率TGI(%)为43.41%。
实施例13.免疫组化染色(IHC)
病理组织芯片购买自桂林泛谱生物技术有限公司。包括BRC1021乳腺癌组织芯片、EMC1021子宫内膜癌组织芯片、OVC1021卵巢癌组织芯片、MNO1021正常组织芯片。石蜡切片为4um厚度,带上阳性对照组织。脱蜡,水洗;抗原修复:PH6(柠檬酸),125°加热5分钟,闷10分钟,后在室温冷却30分钟;水洗后用0.3%过氧化氢5分钟,然后用TBST冲洗3次5分钟;Dako封闭液直接使用,在室温内用孵育盒封闭20分钟;甩去封闭液,上一抗,抗体稀释液为Dako直接使用,在室温内用孵育盒孵育60分钟,Rabbit IgG替代对照;TBST洗三次,每次五分钟;二抗,Anti-Rabbit(EnVision+System-HRP Labelled Polymer),在室温内用孵育盒孵育30分钟;TBST洗三次,每次五分钟;DAB显色,0.85mL蒸馏水,按照试剂ABC顺序加入试剂各50μl,在室温内用孵育盒孵育5分钟;蒸馏水清洗,用苏木素复染。显微镜下观察、封片、读片。
图27结果显示,B7H4在正常组织的肾上腺、肾皮质、膀胱、乳腺、输卵管、食道、输尿管和子宫内膜有轻微的表达,在其它组织中未见表达(图27,A)。相反,在乳腺癌、卵巢癌和子宫内膜瘤上高度表达(图27,B)。比如在卵巢癌中,IHC评分在2-4分的占102个总样品的67.65%,在子宫内膜瘤中,IHC评分在2-4分的占98个总样品的62.24%(图27,C)。
实施例14.B7H4×CD3双特异性抗体的结构和设计
所选的抗B7H4抗体和抗CD3抗体用于制备双特异性抗体。这种B7H4×CD3双特异性抗体的制备双抗可以同时结合两个靶点,其中一端可以识别肿瘤细胞表面特异表达的B7H4,而另一端可以结合T细胞上的CD3分子。当B7H4×CD3双抗分子结合到肿瘤细胞表面后,可以招募并激活肿瘤细胞附近的T细胞,从而杀死肿瘤细胞。
图28的A与B为“1+1”Fab-Fc-scFv非对称结构分子;C与D为具有“1+1”Fab-Fc-crossFab非对称结构的B7H4×CD3双抗分子。对于“1+1”非对称结构分子,结构(1)和(2)涉及三条蛋白链,其分别包含相应抗B7H4抗体的重链及轻链,以及上述抗CD3抗体的scFv多肽链(见图28,A和B)。对于“1+1”非对称结构分子,结构(3)和(4)涉及四条蛋白链,其分别包含相应抗B7H4抗体的重链及轻链,以及上述抗CD3抗体体的重链及轻链(见图28,C和D)。
E与F、G与H、I与J为具有“2+1”非对称结构的B7H4×CD3双抗分子。对于“2+1”非对称结构分子,结构(5)和(6)涉及四条蛋白链,其分别包含相应抗B7H4抗体的重链及轻链,以及上述抗CD3抗体的重链及轻链(见图28,E和F)。对于“2+1”非对称结构分子,结构(7)(8)和(9)(10)涉及三条蛋白链,其分别包含相应抗B7H4抗体的重链及轻链,以及上述抗B7H4和CD3抗体的scFv的多肽链(见图28,G、H、I和J)。
为了将具有错配的重链(例如抗CD3抗体的两条重链错配)的副产物形成最小化,使用了突变的异源二聚体Fc区,其携带“knob-hole”突变和改造的二硫键,如WO2009080251和WO2009080252中所述。B7H4×CD3双特异性抗体具有IgG1的Fc,并在Fc的CH3上携带L234A、L235A及P329G(根据EU索引编号)突变。通过同时共转染三个或四个不同的哺乳动物表达载体产生每一个双特异性抗体,分别编码:1)相应的抗B7H4抗体的重链,其在Fc区携带“Hole”突变以产生异源二聚体抗体,Fc的CH3携带L234A、L235A和P329G突变。2)相应的抗CD3抗体的重链,其在Fc区携带“knob”突变以产生异源二聚体抗体,Fc的CH3携带L234A、L235A、P329G突变。3)相应的抗CD3抗体的轻链。4)相应的抗B7H4抗体的轻链。人IgG1 Fc区的“knob”突变由以下组成:T366W,“Hole”突变由以下组成:T366S、L368A、Y407V。此外,可以包括“knob”Fc区的S354C和“Hole”Y349C形成一对二硫键以增加稳定性和异源二聚体抗体产量。
表19、表20和表21列出了本实施例所构建的B7H4×CD3双抗分子,表中的结构编号对应于图28。表22为连接肽的序列。表23列出了CD3单抗分子所对应的序列号。B7H4单抗分子来源于表7。表24列出了B7H4×CD3双抗分子所对应的序列号。表25列出了双抗分子的第一和第二抗原结合结构域相应的CDR序列的序列编号。
表19具有“1+1”Fab-Fc-scFv非对称结构的B7H4×CD3双抗分子
表20具有“1+1”Fab-Fc-crossFab非对称结构的B7H4×CD3双抗分子
表21具有“2+1”非对称结构的B7H4×CD3双抗分子
表22连接肽序列
连接肽名字 |
长度 |
序列 |
|
GS_5 |
5 |
GGGGS |
SEQ ID NO:133 |
GS_15 |
15 |
GGGQSGQGGSGGGGS |
SEQ ID NO:134 |
GS_20 |
20 |
GGGGSGGGGSGGGGSGGGGS |
SEQ ID NO:135 |
表23本实施例的CD3抗体的序列编号表
抗体编号 |
轻链 |
重链 |
VL |
VH |
LCDR1 |
LCDR2 |
LCDR3 |
HCDR1 |
HCDR2 |
HCDR3 |
PR000627 |
|
131 |
86 |
76 |
46 |
53 |
63 |
8 |
20 |
34 |
PR001924 |
111 |
97 |
86 |
78 |
46 |
53 |
63 |
10 |
20 |
34 |
PR001848 |
110 |
96 |
86 |
78 |
46 |
53 |
63 |
10 |
20 |
34 |
PR003886 |
110 |
107 |
86 |
84 |
46 |
53 |
63 |
10 |
20 |
34 |
表24本实施例的B7H4×CD3双抗分子的序列编号表
表25 B7H4×CD3双抗分子的抗原结合结构域的CDR的序列编号表
表26 B7H4×CD3双抗分子蛋白的表达
实施例15.FACS检测B7H4×CD3双抗结合B7H4的能力
消化SK-BR-3细胞。使用人T细胞分离试剂盒(Miltenyi,#130-096-535)按说明书的方法分离T细胞。用含2%BSA的PBS重悬。将细胞密度分别调整为1×106细胞/mL。以100μL细胞/孔接种于96孔V底板(Corning,#3894),随后加入100μL/孔,2倍于终浓度的3倍浓度梯度稀释的待测抗体。将细胞放置于4℃,避光孵育2小时。之后,加入100μL/孔预冷含2%BSA的PBS漂洗细胞两次,于500g、4℃下离心5分钟,弃上清。再加入100μL/孔荧光二抗(AlexaFluor 488-conjugated AffiniPure Goat Anti-Human IgG,FcγFragment Specific,Jackson,#109-545-098,1:500稀释),4℃,避光孵育1小时。用100μL/孔预冷含2%BSA的PBS洗涤细胞两次,于500g、4℃下离心5分钟,弃上清。最后,200μL/孔预冷含2%BSA的PBS重悬细胞,使用ACEA Novocyte3000流式细胞仪读取荧光发光信号值。
“1+1”非对称结构的B7H4×CD3双抗分子结合SK-BR-3细胞的结果见图29,A-C。结果显示,双抗分子与肿瘤细胞SK-BR-3表面的B7H4有较好的结合活性。图29A为抗B7H4端为Fab结构,抗CD3端为ScFv多肽链的双抗分子的结合活性,其中抗B7H4端来源于PR001476及其PTM突变体抗体的ScFv多肽链的双抗分子PR002849、PR002850和PR002851比抗B7H4端来源于PR002037PTM突变体抗体的ScFv多肽链的双抗分子PR002852有更强的结合活性;图29B为抗B7H4端为Fab结构,抗CD3端为Cross Fab结构的双抗分子的结合活性,其中抗B7H4端来源于PR001476及其PTM突变体抗体的双抗分子PR002855、PR002856和PR002857比抗B7H4端来源于PR002037PTM突变体抗体的双抗分子PR002858有更强的结合活性;图29C为抗B7H4端为ScFv多肽链,抗CD3端为Fab结构的双抗分子的结合活性,由于PR003733与PR003899在抗B7H4端为相同的ScFv多肽链,其结合活性一致。“2+1”非对称结构的B7H4×CD3双抗分子结合SK-BR-3细胞的结果见图29,D。结果显示,2价的抗B7H4端也有很高的结合活性。“1+1”非对称结构的B7H4×CD3双抗分子结合人T细胞的结果见图29,E-G;“2+1”非对称结构的B7H4×CD3双抗分子结合人T细胞的结果见图29,H。结果显示,抗CD3端为Fab结构或ScFv多肽链都可以结合在T细胞表面,而弱抗CD3端的双抗分子PR003899在FACS水平无法看到结合。
实施例16.T细胞杀伤实验
本实验采用人原代T细胞作为效应细胞,高表达B7H4的细胞系SK-BR-3,或MDA-MB-468,B7H4中表达的细胞系HCC-1954或B7H4阴性的细胞系MDA-MB-231作为靶细胞。使用ACEA公司的RTCA仪器检测靶细胞的电导率反映杀伤效率。96孔板e-plate先用50μl完全培养基平衡。消化靶细胞,重悬于含10%胎牛血清的RPM1640完全培养基中,稀释至4×105/mL,铺板50μl/孔于e-plate 96板中,即2×104/孔、37℃培养过夜。次日使用美天妮T细胞分离试剂盒(Miltenyi,#130-096-535)按说明书的方法分离原代T细胞。每孔加入新鲜50μl含2×105T细胞的培液,随后加入50μl 4×的浓度梯度稀释的抗体,抗体最高终浓度为10nM,每个抗体共8个浓度,设置两个重复。实时检测靶细胞的电导率,一般情况取24小时时间点的数据计算靶细胞杀伤效率=(1-样品/空白对照)×100%。24小时收上清,ELISA方法检测IFN-γ的浓度。ELISA检测方法参照IFN gamma Human Uncoated ELISA Kit(Thermo,#88-7316-77)试剂盒操作说明。
其中,“1+1”非对称结构的B7H4×CD3双抗分子激活T细胞并杀伤靶细胞SK-BR-3的结果见图30,A-K。图30的A、B为抗B7H4端为Fab结构,抗CD3端为ScFv多肽链的双抗分子的杀伤活性和细胞因子IFN-γ的产生,其中抗B7H4端来源于PR002037PTM突变体抗体的ScFv多肽链的双抗分子PR002852有最强的杀伤活性和细胞因子IFN-γ产生。图30的C、D为抗B7H4端为Fab结构,抗CD3端为Cross Fab结构的双抗分子的杀伤活性和细胞因子IFN-γ的产生,其中抗B7H4端来源于PR002037PTM突变体抗体的双抗分子PR002852有最强的杀伤活性和细胞因子IFN-γ产生。图30的E、F为抗B7H4端为ScFv多肽链,抗CD3端为Fab结构的双抗分子的杀伤活性和细胞因子IFN-γ的产生,具有强CD3端的双抗分子PR003733与弱的CD3端双抗分子PR003899相比,其杀伤活性和细胞因子IFN-γ的产生均高于后者。图30的G-L为比较PR003733与PR003899在其它肿瘤细胞MDA-MB-468、HCC-1954和MDA-MB-231上的杀伤活性和细胞因子分泌,结果显示PR003733在MDA-MB-468、HCC-1954细胞的体外杀伤以及细胞因子IFN-γ的产生上均高于PR003899;MDA-MB-231为不表达B7H4的阴性对照细胞,两个抗体均没有任何作用。“2+1”非对称结构的B7H4×CD3双抗分子激活T细胞并杀伤靶细胞SK-BR-3的结果见图30,M-N。“2+1”非对称结构的双抗分子均可以杀伤SK-BR-3细胞并产生细胞因子IFN-γ,且抗CD3端为Cross Fab结构的双抗分子PR003001、PR003008比抗CD3端为ScFv多肽链的双抗分子PR002987、PR002994有更强的杀伤活性和细胞因子IFN-γ产生。
实施例17.NSG小鼠重建人PBMC免疫系统的肿瘤模型
细胞接种当天每只NCG小鼠皮下接种5×106MDA-MB-468肿瘤细胞,细胞重悬在PBS与Matrigel(1:1)混合液中(0.1mL/只),皮下接种。当小鼠平均瘤体积在126mm3进行分组,18只小鼠被分为3组,每只小鼠静脉接种5×106人的PBMC,细胞重悬在200μl PBS中。次日开始给药,给药周期为一周一次,共进行了3次给药,静脉给药。开始给药后,每周称量体重及瘤体积两次,瘤体积计算方式为:肿瘤体积(mm3)=0.5×肿瘤长径×肿瘤短径2。给药后第36天结束实验观察,随后所有小鼠进行安乐处理。
溶媒对照组小鼠在给药后第36天平均肿瘤体积为942mm3。测试药PR003733(2mg/kg)治疗组在给药后第36天平均肿瘤体积为590mm3,相较溶媒对照组有显著性差异(p值为0.048),肿瘤抑制率TGI(%)为37.31%。测试药PR003899(2mg/kg)治疗组在给药后第36天平均肿瘤体积为0mm3,肿瘤完全消退,相较溶媒对照组有显著性差异(p值为0.0001),肿瘤抑制率TGI(%)为100%(参见图31,A)。
在HCC-1954模型中,细胞接种当天每只NCG小鼠皮下接种5×106HCC-1954肿瘤细胞,细胞重悬在PBS与Matrigel(1:1)混合液中(0.1mL/只),皮下接种。当小鼠平均瘤体积在102mm3进行分组,15只小鼠被分为3组,每只小鼠静脉接种3×106人的PBMC,细胞重悬在200μl PBS中。次日开始给药,给药周期为一周一次,共进行了2次给药,静脉给药。开始给药后,每周称量体重及瘤体积两次,瘤体积计算方式为:肿瘤体积(mm3)=0.5×肿瘤长径×肿瘤短径2。给药后第16天结束实验观察,随后所有小鼠进行安乐处理。
溶媒对照组小鼠在给药后第16天平均肿瘤体积为622mm3。测试药PR003733(0.5mg/kg)治疗组在给药后第16天平均肿瘤体积为450mm3,相较溶媒对照组无显著性差异(p值为0.1),肿瘤抑制率TGI(%)为27.64%。测试药PR003899(0.5mg/kg)治疗组在给药后第16天平均肿瘤体积为322mm3,相较溶媒对照组有显著性差异(p值为0.0028),肿瘤抑制率TGI(%)为48.27%(参见图31,B)。
两个肿瘤模型的药效实验均显示,PR003899的药效要好于PR003733。
SEQUENCE LISTING
<110> 和铂医药(上海)有限责任公司
<120> 抗B7H4抗体及其双抗和应用
<130> P22111092CP
<150> CN202010618159.5
<151> 2020-06-30
<160> 135
<170> PatentIn version 3.5
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Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
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Ser Val Lys Val Ser Cys Lys Ala Ser
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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
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Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
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Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
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Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
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Ser Met Arg Val Ser Cys Lys Ala Ser
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Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
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Gly Tyr Thr Phe Thr Ser Tyr
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Gly Phe Thr Phe Asn Thr Tyr
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Gly Phe Thr Phe Arg Ser Phe
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Gly Phe Thr Phe Ser Thr Tyr
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Glu Asp Thr Phe Ser Ser Tyr
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Gly Gly Ser Ile Lys Ser Gly Ser Tyr
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Trp Ile Gly Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
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Gly Asp Ile
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Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val
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Gly Arg Ile
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Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
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Ala Val Ile
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Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
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Ser Arg Ile
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Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
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Gly Gly Thr
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Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile
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Gly Asn Ile
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Tyr Pro Gly Gly Gly Tyr
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Arg Ser Lys Tyr Asn Asn Tyr Ala
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Ser Tyr Asp Gly Ser Asn
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Ala Pro Ile Phe Gly Thr
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Ser Tyr Glu Gly Ser Asn
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Ser Tyr Asp Ala Ser Asn
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Tyr Tyr Ser Gly Ser
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Thr Asn Tyr Asn Glu Lys Phe Lys Gly Arg Val Thr Ile Thr Arg Asp
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Thr Ser Thr Ser Thr Ala Tyr Leu Glu Leu Ser Ser Leu Arg Ser Glu
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Asp Thr Ala Val Tyr Tyr Cys Ala Arg
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Thr Tyr Tyr Ala Asp Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp
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Asp Ser Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu
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Asp Thr Ala Val Tyr Tyr Cys Val Arg
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Glu Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp
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Asn Ser Lys Asp Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Gly
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Asp Thr Ala Val Tyr Tyr Cys Ala Lys
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Thr Tyr Tyr Ala Asp Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp
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Asp Ser Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu
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Asp Thr Ala Val Tyr Tyr Cys Val Arg
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Thr Asn Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp
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Lys Ser Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu
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Asp Thr Ala Val Tyr Tyr Cys Ala Arg
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Thr Tyr Tyr Asn Pro Ser Leu Arg Ser Arg Val Thr Ile Ser Val Asp
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Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala
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Asp Thr Ala Val Tyr Tyr Cys Ala Arg
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Thr Tyr Tyr Ala Asp Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp
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Asp Ser Lys Asn Thr Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu
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Asp Thr Ala Val Tyr Tyr Cys Thr Arg
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Leu Ala Gly Ser Ser Tyr Arg Gly Ala Met Asp Ser
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His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr
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Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr
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Gly Gly Pro Tyr Phe Asp Tyr
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Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro
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Gly Gly Ala Leu Arg Trp Tyr Phe Ala Tyr
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Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
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Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
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Asp Arg Val Thr Ile Thr Cys
20
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Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
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Thr Val Thr Leu Thr Cys
20
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Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
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Glu Arg Ala Thr Leu Ser Cys
20
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Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
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Glu Arg Ala Thr Leu Ser Cys
20
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Asp Ile Gln Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
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Glu Arg Ala Thr Leu Ser Cys
20
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Lys Ala Ser Gln Gly Phe Asn Lys Tyr Val Ala
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Arg Ser Ser Thr Gly Ala Val Thr Thr Ser Asn Tyr Ala Asn
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Arg Ala Ser Gln Ser Ile Ser Ser Asn Leu Gly
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Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala
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Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr
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Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly Leu Ile Gly
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<223> 合成的
<400> 51
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr
1 5 10 15
<210> 52
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 52
Tyr Thr Ser Thr Leu Gln Pro
1 5
<210> 53
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 53
Gly Thr Asn Lys Arg Ala Pro
1 5
<210> 54
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 54
Gly Ala Ser Thr Arg Ala Thr
1 5
<210> 55
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 55
Gly Asp Tyr Ile Arg Ala Thr
1 5
<210> 56
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 56
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Arg Asp Tyr Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys
20 25 30
<210> 57
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 57
Trp Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala
1 5 10 15
Leu Thr Leu Leu Gly Ala Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys
20 25 30
<210> 58
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 58
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys
20 25 30
<210> 59
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 59
Gly Ile Pro Ala Arg Val Ser Gly Gly Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Cys Cys
20 25 30
<210> 60
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 60
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Ile Ala Thr Tyr Tyr Cys
20 25 30
<210> 61
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 61
Gly Ile Pro Ala Arg Val Ser Gly Gly Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys
20 25 30
<210> 62
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 62
Leu Gln Tyr Gly Asp Leu Leu Tyr Ala
1 5
<210> 63
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 63
Ala Leu Trp Tyr Ser Asn Leu Trp Val
1 5
<210> 64
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 64
Gln Gln Tyr Asn Ser Trp Pro Pro Leu Thr
1 5 10
<210> 65
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 65
Gln Gln Tyr Lys Asn Trp Pro Phe Thr
1 5
<210> 66
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 66
Gln Gln Tyr Val Asp Leu Pro Ile Thr
1 5
<210> 67
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 67
Gln Gln Tyr Gln Ser Trp Pro Pro Leu Thr
1 5 10
<210> 68
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 68
Gln Gln Tyr His Ser Phe Pro Phe Thr
1 5
<210> 69
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 69
Gln Gln Tyr Arg Ser Trp Pro Pro Leu Thr
1 5 10
<210> 70
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 70
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
1 5 10
<210> 71
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 71
Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
1 5 10
<210> 72
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 72
Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
1 5 10
<210> 73
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 73
Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys
1 5 10
<210> 74
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 74
Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
1 5 10
<210> 75
<211> 121
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 75
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Asp Ile Tyr Pro Gly Gly Gly Tyr Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Gly Ser Ser Tyr Arg Gly Ala Met Asp Ser Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 76
<211> 125
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 76
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val
35 40 45
Gly Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 77
<211> 119
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 77
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 78
<211> 125
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 78
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 79
<211> 116
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 79
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Met Arg Val Ser Cys Lys Ala Ser Glu Asp Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Thr Ala Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 80
<211> 119
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 80
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Glu Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 81
<211> 119
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 81
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Ala Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 82
<211> 120
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 82
Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Lys Ser Gly
20 25 30
Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
35 40 45
Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser
50 55 60
Leu Arg Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe
65 70 75 80
Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 83
<211> 119
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 83
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Ala Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Ala Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 84
<211> 125
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 84
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val
35 40 45
Gly Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Thr Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 85
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 85
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Gly Phe Asn Lys Tyr
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Thr Leu Gln Pro Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Gly Asp Leu Leu Tyr
85 90 95
Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 86
<211> 109
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 86
Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu Leu Gly Ala
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 87
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 87
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 88
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 88
Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Val Ser Gly
50 55 60
Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Cys Cys Gln Gln Tyr Lys Asn Trp Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 89
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 89
Asp Ile Gln Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Asp Tyr Ile Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Val Asp Leu Pro Ile
85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 90
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 90
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gln Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 91
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 91
Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Val Ser Gly
50 55 60
Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Asn Trp Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 92
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 92
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr His Ser Phe Pro Phe
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 93
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 93
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Arg Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 94
<211> 451
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 94
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Ile Gly Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Asp Ile Tyr Pro Gly Gly Gly Tyr Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Thr Ser Thr Ala Tyr
65 70 75 80
Leu Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Gly Ser Ser Tyr Arg Gly Ala Met Asp Ser Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 95
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 95
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 96
<211> 455
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 96
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
225 230 235 240
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
245 250 255
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
260 265 270
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
305 310 315 320
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
325 330 335
Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
340 345 350
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
355 360 365
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
370 375 380
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
385 390 395 400
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
420 425 430
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
435 440 445
Leu Ser Leu Ser Pro Gly Lys
450 455
<210> 97
<211> 228
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 97
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys
225
<210> 98
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 98
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Met Arg Val Ser Cys Lys Ala Ser Glu Asp Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Thr Ala Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 99
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 99
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Glu Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 100
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 100
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Ala Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 101
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 101
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 102
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 102
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Ala Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 103
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 103
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Met Arg Val Ser Cys Lys Ala Ser Glu Asp Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Thr Ala Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Asp Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 104
<211> 450
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 104
Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Lys Ser Gly
20 25 30
Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
35 40 45
Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser
50 55 60
Leu Arg Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe
65 70 75 80
Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 105
<211> 450
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 105
Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Lys Ser Gly
20 25 30
Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
35 40 45
Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser
50 55 60
Leu Arg Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe
65 70 75 80
Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 106
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 106
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Ala Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Ala Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 107
<211> 455
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 107
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val
35 40 45
Gly Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Thr Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
225 230 235 240
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
245 250 255
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
260 265 270
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
305 310 315 320
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
325 330 335
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
340 345 350
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
355 360 365
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
370 375 380
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
385 390 395 400
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
420 425 430
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
435 440 445
Leu Ser Leu Ser Pro Gly Lys
450 455
<210> 108
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 108
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Gly Phe Asn Lys Tyr
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Thr Leu Gln Pro Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Gly Asp Leu Leu Tyr
85 90 95
Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 109
<211> 215
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 109
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 110
<211> 215
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 110
Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu Leu Gly Ala
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro
100 105 110
Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu
115 120 125
Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro
130 135 140
Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala
145 150 155 160
Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala
165 170 175
Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg
180 185 190
Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr
195 200 205
Val Ala Pro Thr Glu Cys Ser
210 215
<210> 111
<211> 442
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 111
Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu Leu Gly Ala
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro
100 105 110
Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu
115 120 125
Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro
130 135 140
Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala
145 150 155 160
Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala
165 170 175
Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg
180 185 190
Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr
195 200 205
Val Ala Pro Thr Glu Cys Ser Asp Lys Thr His Thr Cys Pro Pro Cys
210 215 220
Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
225 230 235 240
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
245 250 255
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
260 265 270
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
275 280 285
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
290 295 300
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
305 310 315 320
Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
325 330 335
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
340 345 350
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
355 360 365
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
370 375 380
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
385 390 395 400
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
405 410 415
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
420 425 430
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440
<210> 112
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 112
Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Val Ser Gly
50 55 60
Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Cys Cys Gln Gln Tyr Lys Asn Trp Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 113
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 113
Asp Ile Gln Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Asp Tyr Ile Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Val Asp Leu Pro Ile
85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 114
<211> 215
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 114
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gln Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 115
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 115
Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Val Ser Gly
50 55 60
Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Asn Trp Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 116
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 116
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr His Ser Phe Pro Phe
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 117
<211> 215
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 117
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Asn
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Arg Ser Trp Pro Pro
85 90 95
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 118
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 118
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser
355 360 365
Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 119
<211> 483
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 119
Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu Leu Gly Ala
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gln Ser Gly Gln Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
115 120 125
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu
130 135 140
Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala Met Asn Trp
145 150 155 160
Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val Gly Arg Ile Arg
165 170 175
Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp
180 185 190
Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr Leu Tyr Leu Gln
195 200 205
Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg
210 215 220
His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr Trp Gly
225 230 235 240
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Glu Pro Lys Ser Ser
245 250 255
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
260 265 270
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
275 280 285
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
290 295 300
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
305 310 315 320
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
325 330 335
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
340 345 350
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile
355 360 365
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
370 375 380
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
385 390 395 400
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
405 410 415
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
420 425 430
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
435 440 445
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
450 455 460
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
465 470 475 480
Pro Gly Lys
<210> 120
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 120
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Glu Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser
355 360 365
Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 121
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 121
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Ala Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser
355 360 365
Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 122
<211> 446
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 122
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Met Arg Val Ser Cys Lys Ala Ser Glu Asp Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Thr Ala Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro
340 345 350
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 123
<211> 442
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 123
Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu Leu Gly Ala
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro
100 105 110
Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu
115 120 125
Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro
130 135 140
Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala
145 150 155 160
Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala
165 170 175
Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg
180 185 190
Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr
195 200 205
Val Ala Pro Thr Glu Cys Ser Asp Lys Thr His Thr Cys Pro Pro Cys
210 215 220
Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
225 230 235 240
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
245 250 255
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
260 265 270
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
275 280 285
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
290 295 300
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
305 310 315 320
Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
325 330 335
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu
340 345 350
Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr
355 360 365
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
370 375 380
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
385 390 395 400
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
405 410 415
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
420 425 430
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440
<210> 124
<211> 669
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 124
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Gly Gly
210 215 220
Gly Gly Ser Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser
225 230 235 240
Pro Gly Gly Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val
245 250 255
Thr Thr Ser Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala
260 265 270
Pro Arg Gly Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro
275 280 285
Ala Arg Phe Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu
290 295 300
Leu Gly Ala Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp
305 310 315 320
Tyr Ser Asn Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
325 330 335
Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser
340 345 350
Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp
355 360 365
Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro
370 375 380
Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn
385 390 395 400
Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys
405 410 415
Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val
420 425 430
Glu Lys Thr Val Ala Pro Thr Glu Cys Ser Asp Lys Thr His Thr Cys
435 440 445
Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu
450 455 460
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
465 470 475 480
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
485 490 495
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
500 505 510
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
515 520 525
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
530 535 540
Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys
545 550 555 560
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys
565 570 575
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys
580 585 590
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
595 600 605
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
610 615 620
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
625 630 635 640
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
645 650 655
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
660 665
<210> 125
<211> 668
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 125
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Gly Ser Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr
450 455 460
Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly
465 470 475 480
Ala Val Thr Thr Ser Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly
485 490 495
Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp
500 505 510
Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu
515 520 525
Thr Leu Leu Gly Ala Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala
530 535 540
Leu Trp Tyr Ser Asn Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr
545 550 555 560
Val Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro
565 570 575
Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile
580 585 590
Ser Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser
595 600 605
Ser Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser
610 615 620
Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln
625 630 635 640
Trp Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser
645 650 655
Thr Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser
660 665
<210> 126
<211> 710
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 126
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Gly Gly
210 215 220
Gly Gly Ser Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser
225 230 235 240
Pro Gly Gly Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val
245 250 255
Thr Thr Ser Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala
260 265 270
Pro Arg Gly Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro
275 280 285
Ala Arg Phe Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu
290 295 300
Leu Gly Ala Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp
305 310 315 320
Tyr Ser Asn Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
325 330 335
Gly Gly Gly Gln Ser Gly Gln Gly Gly Ser Gly Gly Gly Gly Ser Glu
340 345 350
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser
355 360 365
Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala
370 375 380
Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val Gly
385 390 395 400
Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser
405 410 415
Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr Leu
420 425 430
Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr
435 440 445
Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala
450 455 460
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Glu Pro
465 470 475 480
Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
485 490 495
Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
500 505 510
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
515 520 525
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
530 535 540
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
545 550 555 560
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
565 570 575
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly
580 585 590
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
595 600 605
Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn
610 615 620
Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
625 630 635 640
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
645 650 655
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
660 665 670
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
675 680 685
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
690 695 700
Ser Leu Ser Pro Gly Lys
705 710
<210> 127
<211> 702
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 127
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Phe
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asp Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Gly Gly Leu Arg Trp Tyr Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Gly Ser Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr
450 455 460
Val Ser Pro Gly Gly Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly
465 470 475 480
Ala Val Thr Thr Ser Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly
485 490 495
Gln Ala Pro Arg Gly Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp
500 505 510
Thr Pro Ala Arg Phe Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu
515 520 525
Thr Leu Leu Gly Ala Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala
530 535 540
Leu Trp Tyr Ser Asn Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr
545 550 555 560
Val Leu Gly Gly Gly Gln Ser Gly Gln Gly Gly Ser Gly Gly Gly Gly
565 570 575
Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly
580 585 590
Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr
595 600 605
Tyr Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp
610 615 620
Val Gly Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala
625 630 635 640
Asp Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser
645 650 655
Thr Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val
660 665 670
Tyr Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp
675 680 685
Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
690 695 700
<210> 128
<211> 455
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 128
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
225 230 235 240
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
245 250 255
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
260 265 270
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
305 310 315 320
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
325 330 335
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
340 345 350
Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys
355 360 365
Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
370 375 380
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
385 390 395 400
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
420 425 430
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
435 440 445
Leu Ser Leu Ser Pro Gly Lys
450 455
<210> 129
<211> 477
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 129
Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Val Ser Gly
50 55 60
Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Asn Trp Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
115 120 125
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser Ser
130 135 140
Met Arg Val Ser Cys Lys Ala Ser Glu Asp Thr Phe Ser Ser Tyr Ala
145 150 155 160
Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
165 170 175
Gly Thr Ala Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe Gln
180 185 190
Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met
195 200 205
Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Gly Gly Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
225 230 235 240
Val Ser Ser Ala Ser Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys
245 250 255
Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu
260 265 270
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
275 280 285
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
290 295 300
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
305 310 315 320
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
325 330 335
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
340 345 350
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
355 360 365
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser
370 375 380
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys
385 390 395 400
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
405 410 415
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
420 425 430
Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
435 440 445
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
450 455 460
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
465 470 475
<210> 130
<211> 455
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 130
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val
35 40 45
Gly Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp
50 55 60
Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Thr Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe
100 105 110
Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
225 230 235 240
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
245 250 255
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
260 265 270
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
305 310 315 320
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
325 330 335
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
340 345 350
Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys
355 360 365
Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
370 375 380
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
385 390 395 400
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
420 425 430
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
435 440 445
Leu Ser Leu Ser Pro Gly Lys
450 455
<210> 131
<211> 483
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 131
Gln Ala Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly
35 40 45
Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Trp Thr Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Leu Gly Asp Lys Ala Ala Leu Thr Leu Leu Gly Ala
65 70 75 80
Gln Pro Glu Asp Glu Ala Glu Tyr Phe Cys Ala Leu Trp Tyr Ser Asn
85 90 95
Leu Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gln Ser Gly Gln Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu
115 120 125
Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Lys Leu
130 135 140
Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Thr Tyr Ala Met Asn Trp
145 150 155 160
Val Arg Gln Ala Ser Gly Lys Gly Leu Glu Trp Val Gly Arg Ile Arg
165 170 175
Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp Ser Val Lys Asp
180 185 190
Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Thr Leu Tyr Leu Gln
195 200 205
Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg
210 215 220
His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe Ala Tyr Trp Gly
225 230 235 240
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Glu Pro Lys Ser Ser
245 250 255
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
260 265 270
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
275 280 285
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
290 295 300
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
305 310 315 320
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
325 330 335
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
340 345 350
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
355 360 365
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
370 375 380
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
385 390 395 400
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
405 410 415
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
420 425 430
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
435 440 445
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
450 455 460
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
465 470 475 480
Pro Gly Lys
<210> 132
<211> 480
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成的
<400> 132
Glu Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Val Ser Gly
50 55 60
Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Lys Asn Trp Pro Phe
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
115 120 125
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser Ser
130 135 140
Met Arg Val Ser Cys Lys Ala Ser Glu Asp Thr Phe Ser Ser Tyr Ala
145 150 155 160
Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
165 170 175
Gly Thr Ala Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe Gln
180 185 190
Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met
195 200 205
Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Gly Gly Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
225 230 235 240
Val Ser Ser Gly Gly Gly Ala Ser Glu Pro Lys Ser Cys Asp Lys Thr
245 250 255
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
260 265 270
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
275 280 285
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
290 295 300
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
305 310 315 320
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
325 330 335
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
340 345 350
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
355 360 365
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
370 375 380
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
385 390 395 400
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
405 410 415
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
420 425 430
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
435 440 445
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
450 455 460
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
465 470 475 480
<210> 133
<211> 5
<212> PRT
<213> Artificial Sequence
<220>
<223> GS_5
<400> 133
Gly Gly Gly Gly Ser
1 5
<210> 134
<211> 15
<212> PRT
<213> Artificial Sequence
<220>
<223> GS_15
<400> 134
Gly Gly Gly Gln Ser Gly Gln Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 135
<211> 20
<212> PRT
<213> Artificial Sequence
<220>
<223> GS_20
<400> 135
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1 5 10 15
Gly Gly Gly Ser
20