CN113234148A - 全人源的新冠IgK单链抗体及其应用 - Google Patents

全人源的新冠IgK单链抗体及其应用 Download PDF

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CN113234148A
CN113234148A CN202110547125.6A CN202110547125A CN113234148A CN 113234148 A CN113234148 A CN 113234148A CN 202110547125 A CN202110547125 A CN 202110547125A CN 113234148 A CN113234148 A CN 113234148A
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方涛
程艳兵
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Wuhan Feisha Genomics Medicine Co ltd
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Abstract

本发明涉及生物医药技术领域，具体公开了全人源的新冠IgK单链抗体及其应用。本发明通过BCR全长测序，通过分析不同康复者共有而在正常人群中没有的BCR序列，筛选出一类新冠特异性的IgK BCR抗体全长。同时基于抗体全长序列设计特定抗体特异性引物，进行扩增富集后并进行单克隆的制备与后续的克隆筛选测序，获得了BCR序列全长，该序列进行表达后可获得具有中和活性的病毒特异性抗体。通过获得的全人源抗体可以寻找一种能与新冠特异性结合的单链抗体，这类特异识别新冠的单链抗体本身或者其可变区序列，经过直接表达或经基因工程改造成其它的抗体形式，可以作为抗新冠药物开发、疫苗生产、检测标志物开发等新冠相关应用中。

Description

全人源的新冠IgK单链抗体及其应用
技术领域
本发明涉及生物医药技术领域，具体涉及全人源的新冠IgK单链抗体及其应用。
背景技术
新型冠状病毒是一种具有包膜的、不分节段的正链单股RNA病毒，颗粒呈圆形或椭圆形，直径约80～120nm，属于网巢病毒目冠状病毒科Betacoronavirus。病毒粒子被宿主细胞提供的脂质双层所包裹，其中含有核酸及核衣壳蛋白，有三种主要蛋白：包膜蛋白(E蛋白)、膜蛋白(M蛋白)和刺突蛋白(S蛋白)。此病毒每组基因组长度约三万个核苷酸左右，基因序列显示SARS-CoV-2属于乙型冠状病毒属谱系β(Betacoronavirus Lineageβ,Sarbecovirus)进化树中分支较长的一种病毒，与中华菊头蝠中发现的冠状病毒相似，例如MERS-CoV或SARS-CoV。对病毒的生物遗传学分析显示，同属人类冠状病毒的SARS病毒分离株AY274119相比MERS病毒分离株KC164505、JX869059等在亲缘关系上距离SARS-CoV-2病毒更近。
2019年末，该病爆发并向全球扩散。病毒潜伏期平均大约3-7天，最长不超过14天。大多数患者的表现以下呼吸道症状为主，常见临床表现包括发热、四肢乏力、干咳等症状，其他表现包含鼻塞、流鼻涕、头痛、咽痛、咳血，咳痰、或腹泻等。有部分患者仅表现为低热、轻微乏力等，无肺炎表现。还有部分患者无任何临床表现。病毒感染严重后，会引发多种并发症包含急性呼吸窘迫综合征(ARDS)、脓毒症休克、全身炎症反应综合征、难以纠正的代谢性酸中毒、急性心肌损伤，和出凝血功能障碍等。
针对这场危及全人类的群体大流行，揭示人体对新冠侵染后的免疫过程，获取新冠病毒的B细胞受体对于新冠特异性药物的筛选、病毒疫苗的生产研发及其重要。B细胞受体(B cell receptor，BCR)是B细胞抗原识别决定性表面分子，其本质是一种膜表面免疫球蛋白(membrane immunoglobulin,mIg)。BCR具有抗原结合特异性，每个个体的BCR多样性高达5×10^13，构成容量巨大的BCR库，赋予个体识别各种抗原、产生特异性抗体的巨大潜能。
BCR的结构包括重链和轻链。BCR的重链(heavy chain，H)由65～100种可变区(VH)、2种多变区(DH)、6种结合区(JH)和恒定区(CH)四部分基因片段组成；轻链(lightchain，L)由可变区、结合区和恒定区三部分基因片段组成。发育过程中的B细胞在重组酶(RAG1、RAG2)作用下，形成了多样性高达1～2×10^11的BCR。同时，由其形成互补决定区(complementarities determining region，CDR)：CDR1、CDR2和CDR3区氨基酸序列的多样性，特别是编码CDR3的基因，由于其位于轻链V、J或重链V、D、J片段的连接处，可以通过V(D)J的重排和(或)两个基因片段的连接间丢失或插入数个核苷酸，进一步增加BCR的多样性，而形成具有功能的BCR编码基因(B细胞克隆)。
BCR测序是通过高通量测序技术检测靶向扩增后的BCR重链和轻链，全面解析BCR基因重排碱基序列，以及各序列的丰度的测序技术。BCR测序常用于评价各种免疫相关疾病和遗传性突变引起的某个物种所有B细胞或特定B细胞激活介导的细胞免疫反应中BCR基因重排碱基序列，以及各序列的丰度，用于研究不同B细胞克隆的转录情况和相互间关系，从而揭示更深层次的B细胞功能特异性，继而解释体液免疫应答耐受以及高频突变在B细胞应答识别抗原异常等相关生命现象。传统的BCR测序都是采用使用Illumina的测序仪进行2×300bp或2×150bp的双端测序方法来对BCR进行测序，该方法的测序准确率高，但对于部分长度超过600bp的BCR序列，该方法仅能获得两端的序列，会存在中间部分关键可变区序列缺失的问题。
PacBio公司正式推出Sequel II新一代测序系统后，依赖于PacBio SMRT测序技术独有的CCS测序模式，能够通过滚环测序提升reads准确度，同时结合聚合酶试剂的优化，酶读长大幅提升，在获得高精准度HiFi reads的同时能够保证10kb以上的插入片段读长，将解决原有的Illumina等二代测序平台下无法完整覆盖整个片段区域的问题。通过进行HiFi测序，可以获得准确性达到99.5％以上的长读长序列。
目前，针对新冠病毒侵染人体后引起的BCR免疫反应，常规的方法是采用二代测序平台对抗体的可变区进行测序后筛选新冠相关抗体，但是受限于测序的读长，仅能够获得可变区的部分序列，无法进行准确的抗体类型识别与编码翻译识别，而基于三代测序平台PacBio的新冠病毒BCR全长扩增测序可轻松读取病毒侵染后的BCR全长序列，突破了二代测序读长较短的局限性，提高了针对新冠病毒特异性BCR抗体的分辨能力，极大的提高了新冠特异性BCR抗体鉴定的分辨率与准确率。特别是在PacBio公司正式推出Sequel II新一代测序系统后，依赖于PacBio SMRT测序技术独有的HiFi测序模式，能够通过滚环测序提升reads准确度，目前采用HiFi技术进行BCR抗体全长测序的准确率可以达到99.5％以上，通过进行HiFi测序，可以获得极高准确性的新冠BCR抗体全长序列，用于新冠相关BCR抗体序列筛选、基于全长抗体序列的表达翻译与后续新冠中和反应评估等。
BCR一般包括与膜结合的免疫球蛋白分子和Ig-α/Ig-β信号转导组件，通过二硫键连接着免疫球蛋白分子和信号转导组件。BCR包括以下两部分：1.某一亚型(IgD、IgM、IgK、IgG或者IgE)的膜结合免疫球蛋白(mIg)。除了C端疏水的膜结合区和胞内区，这些膜结合免疫球蛋白和分泌型的免疫球蛋白单体是相同的，有两条重链(IgHs)和两条轻链(IgLs)；2.信号转导组件：Ig-α/Ig-β的异二聚体(CD79)，由二硫键连接。两个亚基都是跨膜蛋白，在胞内区都有免疫受体酪氨酸的活化基序(ITAM)。
免疫球蛋白有两个轻链，称为lambda型或kappa型，还有两条重链，它们的结构中有特定的变量，以适应体内的结合和精确的功能。轻链之所以“轻”，是因为它们由大约220个氨基酸组成，而重链通常由500多个组成。完整的免疫球蛋白在人体系统中承担着两个不同的角色；免疫球蛋白可作为细胞表面的抗原受体，或在细胞液中自由循环，以识别和消除抗原化合物或病原体。免疫球蛋白的功能决定与其发生和分化过程有关。免疫球蛋白是由作为抗体的白细胞。一种抗体分子是由一种称为B淋巴细胞(B细胞)的白细胞产生的。每个产生免疫球蛋白的B细胞只被编码来表达和产生一种单一形式的免疫球蛋白轻链。卡帕链在2号染色体上有一个由免疫球蛋白编码的特定位点kappa基因座(IgK)和另一种基因lambda链，在22号染色体上有一个由免疫球蛋白lambda位点(IgL)编码的位点。血清免疫球蛋白轻链中，kappa链与lambda链的比率约为2:1。
目前为止，未能通过筛选方式获得新冠特异性的IgK BCR抗体全长。
发明内容
本发明目的在于提供一种全人源的新冠IgK单链抗体，这类特异识别新冠的单链抗体本身或者其可变区序列，经过直接表达或经基因工程改造成其它的抗体形式后，可以作为抗新冠药物开发、疫苗生产、检测标志物开发等进行新冠相关应用。
本发明提供一种全人源的新冠IgK单链抗体，包括如SEQ ID No：37-SEQ ID No：72中至少一种所示的氨基酸序列。
本发明还提供，编码上述新冠IgK单链抗体的基因序列，优选地，包括如SEQ IDNo：1-SEQ ID No：36中至少一种与上述氨基酸对应的核苷酸序列。
本发明还提供，包含上述新冠IgK单链抗体中所述基因序列的文库。
本发明还提供，上述文库的制备方法，对不同新冠康复者共有而在正常人群中没有的BCR序列进行分析并筛选出新冠特异性的IgK BCR抗体序列。
进一步地，所述分析筛选过程为：对新冠康复者与正常人群进行BCR全长测序，将测序的原始数据进行HiFi的一致性校正后，获得了质量值在Q20以上的BCR全长一致性序列，经过与BCR数据库中的抗体恒定区序列进行比对后，获得了每个样本测序数据中的不同类别的BCR抗体序列。
本发明还提供，包含上述新冠IgK单链抗体中所述基因序列的表达载体。
本发明还提供，包含上述新冠IgK单链抗体中所述基因序列的宿主细胞。
本发明还提供上述新冠IgK单链抗体在制备新冠病毒治疗药物、药物载体及检测标记物中的应用。
相比现有技术，本发明的有益效果为：
1.本发明通过使用BCR全长扩增子测序的方法对新冠康复者与健康人群进行了BCR全长扩增建库测序，获得了质量值在Q20以上的BCR全长扩增子序列，获得的BCR序列包括完整的启动子至终止密码子的的区域，所获得的序列为全人源，无需进一步进行整合即可进行后续的表达验证。
2.本发明通过PacBio HiFi测序获得高质量的BCR序列，经与数据库进行比对后，根据人的BCR序列在恒定区的保守性，对获得的BCR全长序列进行转录方向校正与分类，构建新冠康复者与建库人群不同类别的BCR抗体全长数据库；传统的二代BCR测序技术仅进行部分可变区的测序，无法准确进行抗体的聚类筛选。
3.本发明针对获取的不同类型的BCR序列，基于不同类别抗体恒定区BCR序列的一致性，通过定位位于恒定区上的终止密码子位置，将获得的抗体直接翻译为蛋白质氨基酸序列，在蛋白质水平上对抗体进行比较；传统的二代BCR测序方法不涉及BCR序列的恒定区，无法进行准确的翻译，仅能在DNA序列的水平上进行比较。
4.本发明通过对新冠康复者与正常人群的不同类别的抗体蛋白质氨基酸序列进行比较，找到新冠康复者共有而在正常人群中没有的BCR抗体，这些抗体为新冠特异性抗体；其抗体DNA序列为全人源的新冠特异性抗体全长序列，在经过直接表达或经基因工程改造成其它的抗体形式后，可以作为抗新冠药物开发、疫苗生产、检测标志物开发等进行新冠相关应用中；传统的新冠相关应用均是基于可变区的部分序列进行基因工程改造后再进行表达，由于其序列并非全人源，且必须经过改造，可能引入某些未知的安全问题。
5.本发明通过对获取的新冠特异性抗体序列设计特异性引物，该引物扩增区域包括启动子至终止密码子的全部区域，使用该引物在新冠康复者的cDNA作为模板进行PCR扩增后，将扩增产物转移至大肠杆菌表达载体中，通过构建单克隆后进行一代测序，基于扩增片段大小筛选获取的目标序列进行测序，在测序数据中找到了单一的新冠特异性的抗体DNA序列，获取的DNA序列为单一的纯的全人源特定抗体DNA序列，无需进行人工合成即可直接作为抗新冠药物开发、疫苗生产、检测标志物开发的原始DNA反应物。
附图说明
为了更清楚地说明本发明实施例的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，应当理解，以下附图仅示出了本发明的某些实施例，因此不应被看作是对范围的限定，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他相关的附图。
图1是本发明新冠相关特异性抗体筛选与获取原理示意图。
图2是本发明特异性引物扩增富集新冠相关BCR序列电泳图示意图。
图3是本发明单克隆制备培养示意图。
图4是本发明单克隆扩增筛选电泳图。
图5是本发明单克隆一代测序峰图。
具体实施方式
以下实例用于说明本发明，但不限制本发明的范围。在不背离本发明精神和实质的前提下，对本发明的方法、步骤或条件所作的修改或替换，均属于本发明的范围。
如图1所示，本发明提供的全人源的新冠单链抗体获取流程如下：
1.通过进行新冠康复者与健康人群的BCR全长扩增子建库测序，获取质量值Q20以上的BCR全长序列，分别构建新冠康复者与健康人群的BCR抗体库；
2..在获得高质量的BCR抗体序列后，与抗体数据库进行比对，构建BCR抗体恒定区序列确定抗体类别，由于同类型的BCR抗体相似性非常高，且进行翻译时翻译的终止密码子位置也是一致，基于终止密码子位置将BCR抗体序列翻译为蛋白质氨基酸序列；
3.通过比较新冠康复者共有，而健康人群没有的BCR蛋白质氨基酸序列，找到新冠特异性相关的不同类别的BCR序列；
4.基于找到的新冠特异性序列设计特异引物，进行扩增特异性富集新冠相关的抗体序列；
5.将抗体序列转移至感受态细胞中，制备单克隆，挑选单克隆进行扩增后，选择特定片段大小的扩增产物进行一代测序，基于一代测序的结果筛选获得的全人源的新冠相关的BCR序列。
在本发明中，获得了新冠特异性IgK序列36条，获得的新冠特异性IgK序列如SEQID No：1-SEQ ID No：36所示，经翻译后的氨基酸序列如SEQ ID No：37-SEQ ID No：72所示。所有的序列均包含VDJ区域，在5’端包括起始密码子ATG，在3’端包括终止密码子TGA，该抗体序列为全人源序列，可安全的应用于后续疫苗生产与抗体药物研发等应用中。
根据本发明的新冠特异性IgK中特异性核苷酸或氨基酸序列，可在体外人工合成与此相同的抗体轻重链基因的核苷酸序列或编码相同氨基酸的核苷酸序列，从而获得相同的抗体基因或用于相关基因的改造，而获得IgK抗体或相关蛋白。
在本发明中，提出的包含上述新冠IgK单链抗体中所述基因序列的文库，所述文库为：通过对新冠康复者与健康人群的BCR全长扩增子建库测序，获取质量值Q20以上的BCR全长序列，分别构建新冠康复者与健康人群的BCR抗体库；经与数据库进行比对后，根据人的BCR序列在恒定区的保守性，对获得的BCR全长序列进行转录方向校正与分类，构建新冠康复者与建库人群不同类别的BCR抗体全长数据库。
在本发明中，提出包含所述基因序列的表达载体，根据本领域的常识，载体可以为原核细胞表达载体、真核细胞表达载体或昆虫细胞表达载体。
在本发明中，提出包含所述表达载体的宿主细胞，根据本领域的常识，所述的宿主细胞可以为原核表达细胞、真核表达细胞或昆虫细胞，所述原核表达细胞优选大肠杆菌。
实施例一：新冠康复者与正常人群BCR全长扩增测序
新冠康复者与正常人群的BCR全长扩增测序参照前期申请的专利方法(公开号CN111662970A)进行，主要流程如下：
1.全血样本Total RNA的提取
取新鲜全血样本1mL，使用TriZol LS进行全血样本Total RNA提取，提取后使用Nanidrop 2000C测定RNA样本的浓度与纯度，使用Agilent 2100测定样本的完整性，达到合格标准的样本(总量>1μg，完整性RIN值>7)的样本进行后续实验。
2.总RNA中cDNA第一链的合成
实验操作流程如下：
1)Oligo dT逆转录引物与poly(A)结合，如表1所示。
表1：
Figure BDA0003073959360000091
轻弹混匀，瞬时离心，70℃孵育5min后立即置于冰上。
2)逆转录合成cDNA第一链，配制表2中的反应。
表2：
Figure BDA0003073959360000092
Figure BDA0003073959360000101
轻弹混匀，瞬时离心，42℃孵育75min。反应完成后立即置于冰上，加入1μL的BCRTemplate Switching Oligo，轻弹混匀，瞬时离心，42℃孵育15min。
3.BCR cDNA的全长扩增
BCR cDNA的全长扩增包括两轮半巢式扩增反应，第一轮扩增进行BCR序列的初步富集，第二轮扩增时采用内部的巢式引物，进一步提高扩增的特异性，使扩增条带单一。
1)BCR cDNA全长的第一轮PCR扩增
取一新的0.2mL PCR管，加入表3中的试剂。
表3：
Figure BDA0003073959360000102
充分混匀，瞬时离心，置于PCR仪上进行PCR反应：98℃2min；98℃20s、65℃15s、72℃45s，18cycles；72℃5min。
2)BCR cDNA全长的第二轮PCR扩增
取一新的0.2mL PCR管，加入表4中的试剂。
表4：
Figure BDA0003073959360000103
Figure BDA0003073959360000111
10组混合引物均需要单独进行扩增，可以提高扩增反应的稳定性。
充分混匀，瞬时离心，置于PCR仪上进行PCR反应：98℃2min；98℃20s、65℃15s、72℃30s，20cycles；72℃5min。
反应结束后按照AMPure磁珠的使用说明对扩增产物进行磁珠纯化，最后使用10μL洗脱缓冲液洗脱，取1μL纯化产物，使用无核酸酶水稀释5倍后进行Qubit定量。
4.BCR全长扩增子片段混样
根据Qubit定量结果，将同一样本的不同扩增产物进行等量混样，要求混样后的总量在1μg以上，用于后续建库。
5.文库构建
1)末端修复
取1μg的全基因组扩增子样本，进行末端修复反应体系的配制，配制表5中的反应。
表5：
Figure BDA0003073959360000112
Figure BDA0003073959360000121
充分混匀，瞬时离心，20℃孵育30min。
反应完成后，按照AMPure磁珠的使用说明进行1X磁珠纯化，去除反应时加入的酶与Buffer，最后使用14μL洗脱缓冲液洗脱，获得片段末端加A的粘性末端。
2)连接带barcode的测序接头
通过末端修复与加A后，在加入具有与A末端匹配的带barcode的测序接头，在连接酶的作用下即可实现接头的连接。反应体系如表6。
表6：
Figure BDA0003073959360000122
混匀，瞬时离心，20℃孵育60min，反应结束后65℃孵育10min，置于冰上。进行外切酶消化，反应体系如表7。
表7：
Figure BDA0003073959360000123
混匀，瞬时离心，37℃孵育60min，置于冰上。按照AMPure磁珠的使用说明进行磁珠纯化，最后使用20μL洗脱缓冲液洗脱，获得的适用于PacBio测序平台的哑铃型的环状文库。
3)文库质检与上机测序
取1μL文库进行Qubit定量，获得文库浓度；取1μL文库进行安捷伦2100的片段大小分析，将全长扩增文库在PacBio Sequel II测序平台上进行混合测序，每个样本获得了约60G的测序数据。
实施例二：新冠康复者与正常人群BCR免疫组库分析，获得新冠特异性IgK单链抗体
将测序的原始数据进行HiFi的一致性校正后，获得了质量值在Q20以上的BCR全长一致性序列，经过与BCR数据库中的抗体恒定区序列进行比对后，获得了每个样本测序数据中的不同类别的BCR抗体序列，同时基于恒定区的终止密码子位置，将DNA序列翻译为蛋白质多肽序列。通过比较新冠康复者共有的，而正常人群中没有的BCR多肽序列，获得了新冠特异性相关的抗体全长。本次获得的新冠特异性IgK序列36条，所有的序列均包含VDJ区域，在5’端包括起始密码子ATG，在3’端包括终止密码子TGA，该抗体序列为全人源序列，可安全的应用于后续疫苗生产与抗体药物研发等应用中。获得的新冠特异性IgK序列如SEQ IDNo：1-SEQ ID No：36所示，经翻译后的氨基酸序列如SEQ ID No：37-SEQ ID No：72所示。
实施例三：基于以新冠康复者样本设计特异性引物，进行单克隆BCR抗体库的构建
基于抗体库筛选的IgK抗体序列，在起始密码子的与终止密码子的外测设计特异性引物，通过PCR扩增特异性富集目标抗体序列。由于抗体存在多重重组的特性，获取的扩增产物是一类与引物匹配的抗体序列集合，无法直接进行一代测序与后续应用，需要将抗体序列进行克隆实验，获取单个抗体序列的单克隆，并挑选单克隆进行测序，验证所获得的单克隆序列为目标抗体。具体流程如下：
1.目标抗体IgK抗体序列引物设计
基于测序筛选的目标序列抗体设计引物，该引物扩增的目标片段包括起始密码子与终止密码子区域，覆盖整个抗体表达区域的全长，包含实施例二公开抗体的所有序列，具体设计的引物序列如表8所示。
表8：IgK抗体序列引物
Figure BDA0003073959360000141
其中，Primer_ID_01～02为R端通用引物，该引物在恒定区内，所有IgK抗体序列均可与该引物进行匹配，使用时混合在一起作为R端通用引物使用；Primer_ID_03～19为F端引物，该引物序列基于筛选的新冠相关特异性抗体设计，与R端引物组合在一起为一对引物，用于目标抗体序列的扩增。
2.扩增富集获取新冠相关特异性抗体
本步骤使用的扩增模板为新冠康复者样本的单链cDNA样本。取一新的0.2mL PCR管，加入如表9所示试剂。
表9：
Figure BDA0003073959360000151
充分混匀，瞬时离心，置于PCR仪上进行PCR反应：98℃2min；98℃20s、65℃15s、72℃120s，35cycles；72℃5min。
反应结束后进行电泳检测，电泳结果示意图如图2所示。切胶获取片段大小在1.5～2k的目标特异性片段。
3.目标片段的单克隆制备
本步骤使用表10中的试剂。
表10：
Figure BDA0003073959360000152
Figure BDA0003073959360000161
LB固体培养基配制：取本品8克，溶解于250ml蒸馏水中，121℃高压灭菌15分钟，至不烫手时加入250μL氨苄青霉素，混匀倒平板备用(每个平板15mL左右)。
LB液体培养基配制：取本品1克，溶解于40ml蒸馏水中，121℃高压灭菌15分钟，分装至2mL无菌离心管中备用。
配制好培养基后进行载体连接反应，配制如表11中的反应体系。
表11：
Figure BDA0003073959360000162
轻弹管底混匀，低速瞬时离心收集所有液体于离心管底，室温25℃反应5min。反应结束后，将离心管置于冰上。
将Fast-T1感受态细胞从-70℃拿出，迅速置于冰上融化，取20μL感受态细胞加入目的载体连接反应产物，轻弹管壁混匀(避免用枪吸打)，冰上静置30min。
42℃水浴热激30s后，迅速置于冰上静置2min，切勿摇动离心管。
向离心管中加入200μL LB液体培养基(不含抗生素)，混匀后置于37℃，200rpm摇床中复苏5min。
复苏后颠倒混匀直接取200μL，涂布在含氨苄青霉素的LB固体培养基平板上，将平板正置于37℃培养箱10min，待菌液被完全吸收后，倒置平板，过夜培养，单克隆制备培养结果示意图如图3所示。
4.单克隆筛选与测序鉴定
在平板培养基上挑选单克隆菌落至10μL ddH2O中混匀作为模板；使用2×RapidTaq Master Mix(Vazyme#P222)进行扩增，反应体系如表12所示。
表12：
Figure BDA0003073959360000171
放置在PCR仪上进行如表13所示的扩增反应程序。
表13：
Figure BDA0003073959360000172
将扩增获得的扩增产物进行凝胶电泳检测，获得图4所示的电泳图，挑选扩增条带亮度高、扩增条带单一且片段大小在1.5～5k的扩增产物进行双端单克隆一代测序鉴定，测序峰图如图5所示，根据一代测序的序列与目标抗体序列进行一致性比对，选择序列完全一致的单克隆进一步进行扩增放大，即获得了高纯度的单一的新冠相关抗体特异性序列，该序列可以直接转移至假病毒体系中，进行后续的抗体效价验证，也可以直接转移至抗体表达体系中进行表达。
序列表
<110> 武汉菲沙基因组医学有限公司
<120> 全人源的新冠IgK单链抗体及其应用
<160> 91
<170> SIPOSequenceListing 1.0
<210> 1
<211> 714
<212> DNA
<213> IgK
<400> 1
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagcttct tttcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgtatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtata ttactgtcag cagtatggta gctcacctcc ctcgatcacc 360
ttcggccaag ggacacggct ggagattaaa cgaactgtgg ctgcaccatc tgtcttcatc 420
ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat 480
aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt 540
aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag cctcagcagc 600
accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc 660
catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg ttag 714
<210> 2
<211> 720
<212> DNA
<213> IgK
<400> 2
atgaggctcc ctgctcagct cctggggctg ctaatgctct ggatacctgg atccagtgca 60
gatattgtga tgacccagac tccactctct ctgtccgtca cccctggaca gccggcctcc 120
atctcctgca agtctagtca gagcctcctg catagtgatg gaaagaccta tttgtattgg 180
tacctgcaga agccaggcca gcctccacag ctcctgatct atgaagtttc caaccggttc 240
tctggagtgc cagataggtt cagtggcagc gggtcaggga cagatttcac actgaaaatc 300
agccgggtgg aggctgagga tgttggggtt tattactgca tgcaaagtat acagcttccg 360
ctcactttcg gcggagggac caaggtggag atcaaacgaa ctgtggctgc accatctgtc 420
ttcatcttcc cgccatctga tgagcagttg aaatctggaa ctgcctctgt tgtgtgcctg 480
ctgaataact tctatcccag agaggccaaa gtacagtgga aggtggataa cgccctccaa 540
tcgggtaact cccaggagag tgtcacagag caggacagca aggacagcac ctacagcctc 600
agcagcaccc tgacgctgag caaagcagac tacgagaaac acaaagtcta cgcctgcgaa 660
gtcacccatc agggcctgag ctcgcccgtc acaaagagct tcaacagggg agagtgttag 720
<210> 3
<211> 720
<212> DNA
<213> IgK
<400> 3
atgaggctcc ctgctcagct cctggggctg ctaatgctct ggatccctgg atccagtgcg 60
gatattgtga tgacccagac tccactctct ctgtccgtca cccctggaca gccggcctcc 120
atctcctgca agtctagtca gagcctcctg catagtgatg gaaagaccta tttgtattgg 180
tacctgcaga agccaggcca gtctccacag ctcctaatct atgaagtttc cagccggttc 240
tctggagtgc cagataggtt cagtggcagc gggtcaggga cagatttcac actgaaaatc 300
agccgggtgg aggctgagga tgttggggtt tattactgca tgcaagctat acaccttccg 360
ttcacttttg gccaggggac caagctggag atcaaacgaa ctgtggctgc accatctgtc 420
ttcatcttcc cgccatctga tgagcagttg aaatctggaa ctgcctctgt tgtgtgcctg 480
ctgaataact tctatcccag agaggccaaa gtacagtgga aggtggataa cgccctccaa 540
tcgggtaact cccaggagag tgtcacagag caggacagca aggacagcac ctacagcctc 600
agcagcaccc tgacgctgag caaagcagac tacgagaaac acaaagtcta cgcctgcgaa 660
gtcacccatc agggcctgag ctcgcccgtc acaaagagct tcaacagggg agagtgttag 720
<210> 4
<211> 720
<212> DNA
<213> IgK
<400> 4
atgaggctcc ctgctcagct cctggggctg ctaatgctct gggtctctgg atccagtggg 60
gatattgtga tgactcagtc tccactctcc ctgcccgtca cccctggaga gccggcctcc 120
atctcctgca ggtctagtca gagcctcctg catggtaatg gatacaacta tttggattgg 180
tacctgcaga agccagggca gtctccacag ctcctgatct atttgggttc taatcgggcc 240
tccggggtcc ctgacaggtt cagtggcagt gcatcaggca cagattttac actgaaaatc 300
agcagagtgg aggctgagga tgttggggtt tattactgca tgcaggctct ccaaagtccg 360
atcaccttcg gccaagggac acgactggag attaaacgaa ctgtggctgc accatctgtc 420
ttcatcttcc cgccatctga tgagcagttg aaatctggaa ctgcctctgt tgtgtgcctg 480
ctgaataact tctatcccag agaggccaaa gtacagtgga aggtggataa cgccctccaa 540
tcgggtaact cccaggagag tgtcacagag caggacagca aggacagcac ctacagcctc 600
agcagcaccc tgacgctgag caaagcagac tacgagaaac acaaagtcta cgcctgcgaa 660
gtcacccatc agggcctgag ctcgcccgtc acaaagagct tcaacagggg agagtgttag 720
<210> 5
<211> 720
<212> DNA
<213> IgK
<400> 5
atgaggctcc ctgctcagct cctggggctg ctaatgctct gggtcccagg atccagtggg 60
gatgttgtga tgactcagtc tccactctcc ctgcccgtca cccttggaca gccggcctcc 120
atctcctgca gggctagtca aagcctcgga cagagtgatg gatccaccta cttgacttgg 180
tttcagcaga ggccaggcca atctccaagg cgcctaatct ataaggtttc ttaccgggac 240
tctggggtcc cagacagatt cagcggcagt gggtcaggca ctgatttcac actgaaaatc 300
agcagggtgg aggctgagga tgttgggact tattactgca tgcaaggtat acattggccc 360
tggacgctcg gccaagggac caaggtggaa atcaaacgaa ctgtggctgc accatctgtc 420
ttcatcttcc cgccatctga tgagcagttg aaatctggaa ctgcctctgt tgtgtgcctg 480
ctgaataact tctatcccag agaggccaaa gtacagtgga aggtggataa cgccctccaa 540
tcgggtaact cccaggagag tgtcacagag caggacagca aggacagcac ctacagcctc 600
agcagcaccc tgacgctgag caaagcagac tacgagaaac acaaagtcta cgcctgcgaa 660
gtcacccatc agggcctgag ctcgcccgtc acaaagagct tcaacagggg agagtgttag 720
<210> 6
<211> 723
<212> DNA
<213> IgK
<400> 6
atgaggctcc ctgctcagct cctggggctg ctaatgctct gggtccctgg atccagtgag 60
gatgttgtga tgacccagac tccactctcc ctgcccgtca cccctggaga gccggcctcc 120
atctcctgca ggtctagtca gagcctcttc gatagtgctg atggaaacac ctatttggac 180
tggtacctgc agaagccagg gcagtctcca caactcctga tctatatgac ttcctatcgg 240
gcctctggag tcccagacag gttcagtggc agtgggtcag gcactgattt cacactgaaa 300
atcagcaggg tggaggctga ggatgttgga gtttattact gcatgcaacg tatagagttt 360
ccttacactt ttggccaggg gaccaagctg gagatcaaac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 7
<211> 723
<212> DNA
<213> IgK
<400> 7
atgaggctcc ctgctcagct cctggggctg ctaatgctct gggtccctgg atccagtgag 60
gatattgtga tgacccagac tccactctcc ctgcccgtca cccctggaga gccggcctcc 120
atctcctgca ggtctagtca gagcctcttg gatagtgatg atggaaacac ctatttggac 180
tggtacctgc agaagccagg gcagtctcca cagctcctga tctatacgct ttcctatcgg 240
gcctctggag tcccagacag gttcagtggc agtgggtcag gcactgattt cacactgaaa 300
atcagcaggg tggaggctga ggatgttgga gtttattact gcatgcaacg tatagagttt 360
ccgtacactt ttggccaggg gaccaagctg gagatcaaac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 8
<211> 723
<212> DNA
<213> IgK
<400> 8
atgaggctcc ctgctcagct cctggggctg ctaatgctct gggtccctgg atccagtgag 60
gatattgtga tgacccagac tccactctcc ctgcccgtca cccctggaga gccggcctcc 120
atctcctgca ggtctagtca gagcctcttg gatagtgatg atggaaacac ctatttggac 180
tggtacctgc agaagccagg gcagtctcca cagctcctga tctatacgct ttcctatcgg 240
gcctctggag tcccagacag gttcagtggc agtgggtcag gcactgattt cacactgaaa 300
atcagcaggg tggaggctga ggatgttgga gtttattact gcatgcaacg tatagagttt 360
ccgctcactt ttggccaggg gaccaagctg gagatcaaac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 9
<211> 723
<212> DNA
<213> IgK
<400> 9
atggtgttgc agacccaggt cttcatttct ctgttgctct ggatctctgg tgcctgcggg 60
gccatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 120
atcaactgca agtccagcca gactctttta tacagctcca acaacaggga ctacttaggt 180
tggtaccagc agaaatcagg acagcctcca cgtctcctca tttcctgggc atctacccgg 240
gaatccgggg tccctgaccg attcattggc agcgggtctg ggacagattt cactctcacc 300
atcagcagcc tgcaggctga agatgtggca atttattact gtcagcaata ctataatgct 360
ccaatcactt tcggcggagg gaccaaggtg gaaatcagac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gctacaacag gggagagtgt 720
tag 723
<210> 10
<211> 723
<212> DNA
<213> IgK
<400> 10
atggtgttgc agacccaggt cttcatttct ctgttgctct ggatctctgg tgcctacggg 60
gacatcgtga tgagccagtc tccagactcc ctgactgtgt ctctgggcga gagggccacc 120
atcaactgca agtccagcca gagtgttttc tataactcca accataagaa ctacttagct 180
tggtaccagc agaaacctgg acagcctcct aaggtgctca tttactgggc atctaccagg 240
gagtccgggg tccctgaccg gttcagtggc agcgggtctg ggacagagtt cactctcacc 300
atcagcagcc tgcaggctga agatgtggca gtttattact gtcaacaata ttttaataag 360
cccatcactt tcggccctgg gaccaaagtg gacatcaaac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 11
<211> 723
<212> DNA
<213> IgK
<400> 11
atggtgttgc agacccaggt cttcatttct ctgttgctct ggatctctgg tgcctacggg 60
gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 120
atcaactgca agtccagcca gagtatttta tacagctcca acaataagaa ttacttagct 180
tggttccagc agaaaccagg acagcctcct aaactgctca tttactgggc atctacccgg 240
gaatccgggg tccctgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 300
atcagcagcc tgcaggctga agatgtggca gtttattact gtcagcaata ttatcgtatt 360
ccttggacgt tcggccaagg gaccaaggtg gaaatcagac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 12
<211> 723
<212> DNA
<213> IgK
<400> 12
atggtgttgc agacccaggt cttcatttct ctgttgctct ggatctctgg tgcctacggg 60
gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 120
atcaactgca agtccagcca gagtgtttta tacagctcca acaataagaa ctacttagct 180
tggtaccagc agaaaccagg acagcctcct aagctgctca tttactgggc atctacccgg 240
gaatccgggg tccctgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 300
atcagcagcc tgcaggctga agatgtggca gtttattact gtcagcaata ttatagtact 360
ccattcactt tcggccctgg gaccaaagtg gatatcaaac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 13
<211> 723
<212> DNA
<213> IgK
<400> 13
atggtgttgc agacccaggt cttcatttct ctgttgctct ggatctctgg tgtctacggg 60
gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 120
atcaactgca agtccagcca gagtgtttta tacagctcca acaataagaa ctacttagat 180
tggtaccagc agagaccagg acagcctcct aaactgctca tttactgggc atctacccgg 240
gaatccgggg tccctggccg attcagtggc agcgggtctg ggacagattt cactctcacc 300
atcagcagcc tgcaggctga agatgcggca gtttattact gtcaccaata ttatagtagt 360
ccgttcacct ttggccaggg gaccaagctg gagatcaaac gaactgtggc tgcaccatct 420
gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 480
ctgctgaata acttctatcc cagagaggcc aaagtacagt ggaaggtgga taacgccctc 540
caatcgggta actcccagga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 600
ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 660
gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 720
tag 723
<210> 14
<211> 732
<212> DNA
<213> IgK
<400> 14
atggtgttgc agacccaggt cttcatttct ctgttgctct ggacctctgt gaccacaggt 60
gccttcgggg acatcgtgat gacccagtct ccagactccc tggctgtgtc tctgggcgag 120
agggccacca tcagctgcaa gtccagccag cgtgttttat acagttccaa caataagtat 180
tacttggctt ggtaccagca gaaaccaggt cagtctccta agttgctgat ttattgggca 240
tctacccgag aatccggggt ccctgatcga ttcagcggca gcgggtctgg gacagatttc 300
actctcacca tcagcagcct gcagtctgaa gatgtggcag tttattactg ccaacaatat 360
tatagtactc cttggacgtt cggccggggg accaaggtgg aaatcaagcg aactgtggct 420
gcaccatctg tcttcatctt cccgccatct gatgagcagt tgaaatctgg aactgcctct 480
gttgtgtgcc tgctgaataa cttctatccc agagaggcca aagtacagtg gaaggtggat 540
aacgccctcc aatcgggtaa ctcccaggag agtgtcacag agcaggacag caaggacagc 600
acctacagcc tcagcagcac cctgacgctg agcaaagcag actacgagaa acacaaagtc 660
tacgcctgcg aagtcaccca tcagggcctg agctcgcccg tcacaaagag cttcaacagg 720
ggagagtgtt ag 732
<210> 15
<211> 726
<212> DNA
<213> IgK
<400> 15
atggtgttgc agacccaggt cttcatttct ctgttgctct ggatctctgg tgccttcggg 60
gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gacggccacc 120
atcaactgca agtccagcca gagtgttttc tacagctcca acaataagaa ctacttagct 180
tggtaccagc agaaaccagg acagcctcct aagctgctcg tttactgggc atctacccgg 240
gaatccgggg tccctgaccg attcagtggc agcgggtctg ggacagattt cactctcacc 300
atcagcagcc tgcaggctga agatgtggct atttattact gtcagcaata ttatggtgct 360
cctccaagga cgttcggcca agggaccaag gtggaaatca aacgaactgt ggctgcacca 420
tctgtcttca tcttcccgcc atctgatgag cagttgaaat ctggaactgc ctctgttgtg 480
tgcctgctga ataacttcta tcccagagag gccaaagtac agtggaaggt ggataacgcc 540
ctccaatcgg gtaactccca ggagagtgtc acagagcagg acagcaagga cagcacctac 600
agcctcagca gcaccctgac gctgagcaaa gcagactacg agaaacacaa agtctacgcc 660
tgcgaagtca cccatcaggg cctgagctcg cccgtcacaa agagcttcaa caggggagag 720
tgttag 726
<210> 16
<211> 726
<212> DNA
<213> IgK
<400> 16
atgaggctcc ctgctcagct cctggggctg ctaatgctct gggtctctgg atccagtggg 60
gatattgtga tgactcagtc tccactctcc ctgcccgtca cccctggaga gccggcctcc 120
atctcctgca ggtctagtca gagcctcctg catagtaatg gatacaacta tttggattgg 180
tacctgcaga agccagggca gtctccacag ctcctgatct atttgggttc taatcgggcc 240
tccggggtcc ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc 300
agcagagtgg aggctgagga tgttggggtt tattactgca tgcaagctct acaaactcct 360
ccggagtaca cttttggcca ggggaccaag ctggagatca aacgaactgt ggctgcacca 420
tctgtcttca tcttcccgcc atctgatgag cagttgaaat ctggaactgc ctctgttgtg 480
tgcctgctga ataacttcta tcccagagag gccaaagtac agtggaaggt ggataacgcc 540
ctccaatcgg gtaactccca ggagagtgtc acagagcagg acagcaagga cagcacctac 600
agcctcagca gcaccctgac gctgagcaaa gcagactacg agaaacacaa agtctacgcc 660
tgcgaagtca cccatcaggg cctgagctcg cccgtcacaa agagcttcaa caggggagag 720
tgttag 726
<210> 17
<211> 708
<212> DNA
<213> IgK
<400> 17
atggaagccc cagctcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct 180
ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 240
aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 300
gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccatt cactttcggc 360
cctgggacca aagtggatat caaacgaact gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttag 708
<210> 18
<211> 708
<212> DNA
<213> IgK
<400> 18
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtattagc agcaactact tagcctggta ccagctgaaa 180
cctggccagg gtcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag aagcctggtg 300
cctgaagatt ttgcactgta ttactgtcag cagtatggta gctcgccgaa gacgttcggc 360
caggggacca aggtagaaat caaacgaact gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttag 708
<210> 19
<211> 708
<212> DNA
<213> IgK
<400> 19
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcacctca gacttttggc 360
caggggacca agctggagat caaacgaact gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttag 708
<210> 20
<211> 708
<212> DNA
<213> IgK
<400> 20
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca acagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtcta ttactgtcag cagtatggta gctcacctct cactttcggc 360
ccagggacca aagtggatat caaacgaact gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttag 708
<210> 21
<211> 708
<212> DNA
<213> IgK
<400> 21
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttatc agcacctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccact cactttcggc 360
cctgggacca aagtggatat caaacgaact gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttag 708
<210> 22
<211> 708
<212> DNA
<213> IgK
<400> 22
atggaagccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccactgga 60
gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcaagttag cctggtacca gcagaaacct 180
ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 240
aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 300
gaagattttg cagtttatta ctgtcagcag tataataact ggcctccgat caccttcggc 360
caagggacac gactggatat taaacgaact gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgttag 708
<210> 23
<211> 711
<212> DNA
<213> IgK
<400> 23
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcacctcc gtggacgttc 360
ggccaaggga ccaaggtgga aatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 24
<211> 711
<212> DNA
<213> IgK
<400> 24
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccccc gtacactttt 360
ggccagggga ccaagctgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 25
<211> 711
<212> DNA
<213> IgK
<400> 25
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcttcgaa gctcactttc 360
ggcggaggga ccaaggtgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 26
<211> 711
<212> DNA
<213> IgK
<400> 26
atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga 60
gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 120
ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 180
cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 240
gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag 300
cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcacctcc gctcactttc 360
ggcggaggga ccaaggtgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 27
<211> 711
<212> DNA
<213> IgK
<400> 27
atggacatga gagtcctcgc tcagctcctg gggctcctgc tgctctgttt cccaggtgcc 60
agatgtgaca tccagatgac ccagtctcca tcctcactgt ctgcatctgt aggagacaga 120
gtcaccatca cttgtcgggc gagtcagggc attagcaagt atttactctg gtttcaacag 180
aaaccaggaa aagcccctaa gtccctaatc tatggtgcgt cctttttgca aagtggggtc 240
ccatcgaggt tcagcggcac tggctctggc acagatttca ctctcaccat cagccgccta 300
cagcctgaag attttgccac ttattactgt cagcaatatg atacttatcc tcccactttc 360
ggcggaggga ccagggtgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 28
<211> 711
<212> DNA
<213> IgK
<400> 28
atggacatga gggtccccgc tcagctcctg gggctcctgc tgctctggct cccaggtgcc 60
agatgtgaca tccagatgac ccagtctcct tccaccctgt ctgcatctgt aggagacaga 120
gtcatcatca cttgccgggc cagccagaat attaataggt gggtggcctg gtatcaacag 180
aaaccaggga aagccccaaa cctcctcatc tatcaggcat ctactttaga aggtggggtc 240
ccatcaaggt tcagcggcag tggatctggg acagaattta ctctcaccat cagcagcctg 300
cagcctggtg attttgcaac ttattactgc catcagtatc atagttattc gtacgctttt 360
ggccagggga ccaagctgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 29
<211> 711
<212> DNA
<213> IgK
<400> 29
atggacatga gggtccccgc tcagctcctg gggctcctgc tgctctggct cccaggtgcc 60
aaatgtgaca tccagatgac ccagtctcct tccaccctgt ctgcatctgt aggagacaga 120
gtcaccatca cttgccgggc cagtcagagt attagtagct ggttggcctg gtatcagcag 180
aaaccaggga aagcccctaa gctcctgatc tataaggcgt ctagtttaga aagtggggtc 240
ccatcaaggt tcagcggcag tggatctggg acagaattca ctctcaccat cagcagcctg 300
cagcctgatg attttgcaac ttattactgc caacagtata atagttatcc gtacactttt 360
ggccagggga ccaagctgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 30
<211> 711
<212> DNA
<213> IgK
<400> 30
atggacatga gggtccccgc tcagctcctg gggctcctgc tgctctggct cccaggtgcc 60
aaatgtgaca tccagatgac ccagtctcct tccaccctgt ctgcatctgt aggagacaga 120
gtcaccatca cttgccgggc cagtcagagt attagtagct ggttggcctg gtatcagcag 180
aaaccaggga aagcccctaa gctcctgatc tataaggcat ctagtttaga aagtggggtc 240
ccatcaaggt tcagcggcag tggatctggg acagaattca ctctcaccat cagcagcctg 300
cagcctgatg attttgcaac ttattactgc caacagtata atagttatcc tatcactttc 360
ggcggaggga ccaaggtgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 31
<211> 711
<212> DNA
<213> IgK
<400> 31
atggacatga gartcctcgc tcagctcctg gggctcctgc tgctctgttt cccaggtgcc 60
agatgtgaca tccagatgac ccagtctcca tcctcactgt ctgcatctgt aggagacaga 120
gtcaccatca cttgtcgggc gagtgagggc attagcaatt ttgtggcctg gtttcagcag 180
gaaccaggga aagcccctaa gtccctgatc tatggtgcat ccagtttgca aagtggggtc 240
ccatcacgat tcagcggcag tggatctggg acagatttca ctctcgccat cactaacctg 300
cagcctgaag attttggtac ttatttctgc caacagtata aaagtaaccc tcgcactttt 360
ggcccgggga ccagagtgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 32
<211> 711
<212> DNA
<213> IgK
<400> 32
atggacatga gggtccccgc tcagctcctg gggctcctgc tactctggct ccgaggtgcc 60
agatgtgaca tccagatgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120
gtcaccatca cttgccgggc aagtcagagc attagcagct atttaaattg gtatcagcag 180
aaaccaggga aagcccctaa gctcctgatc tatgctgcat ccagtttgca aagtggggtc 240
ccatcaaggt tcagtggcag tggatctggg acagatttca ctctcaccat cagcagtctg 300
caacctgaag attttgcaac ttactactgt caacagagtt acagtacccc gctcactttc 360
ggcggaggga ccaaggtgga gatcaaacga actgtggctg caccatctgt cttcatcttc 420
ccgccatctg atgagcagtt gaaatctgga actgcctctg ttgtgtgcct gctgaataac 480
ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540
tcccaggaga gtgtcacaga gcaggacagc aaggacagca cctacagcct cagcagcacc 600
ctgacgctga gcaaagcaga ctacgagaaa cacaaagtct acgcctgcga agtcacccat 660
cagggcctga gctcgcccgt cacaaagagc ttcaacaggg gagagtgtta g 711
<210> 33
<211> 762
<212> DNA
<213> IgK
<400> 33
atgcgatggc tgtacggggg aagagctgct cagttaggac ccagagggaa ccatggaaac 60
ccccagcgca gcttctcttc ctcctgctac tctggctccc agatacccac cggagaaatt 120
gtgttgacgc agtctccagg caccctgtct ttgtctccag gggaaagagc caccctctcc 180
tgcagggcca gtcagagtgt tagcagcagc tacttagcct ggtaccagca gaaacctggc 240
caggctccca ggctcctcat ctatggtgca tccagcaggg ccactggcat cccagacagg 300
ttcagtggca gtgggtctgg gacagacttc actctcacca tcagcagact ggagcccgaa 360
gattttgcag tgtattactg tcagcagtat ggtagctcag cctacacttt tggccagggg 420
accaagctgg agatcaaacg aactgtggct gcaccatctg tcttcatctt cccgccatct 480
gatgagcagt tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc 540
agagaggcca aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag 600
agtgtcacag agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg 660
agcaaagcag actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg 720
agctcgcccg tcacaaagag cttcaacagg ggagagtgtt ag 762
<210> 34
<211> 699
<212> DNA
<213> IgK
<400> 34
atggacatga gggtccccgc tcagctcctg gggctcctgc tgctctggct cccaggtgcc 60
aaatgtgaca tccagatgac ccagtctcct tccaccctgt ctgcatctgt aggagacaga 120
gtcaccatca cttgccgggc cagtcagagt attagtagct ggttggcctg gtatcagcag 180
aaaccaggga aagcccctaa gctcctgatc tataaggcat ctagtttaga aagtggggtc 240
ccatcaaggt tcagcggcag tggatctggg acagaattca ctctcaccat cagcagcctg 300
cagcctgatg attttgcaac ttattactgc caacagtata agacgttcgg ccaagggacc 360
aaggtggaaa tcaaacgaac tgtggctgca ccatctgtct tcatcttccc gccatctgat 420
gagcagttga aatctggaac tgcctctgtt gtgtgcctgc tgaataactt ctatcccaga 480
gaggccaaag tacagtggaa ggtggataac gccctccaat cgggtaactc ccaggagagt 540
gtcacagagc aggacagcaa ggacagcacc tacagcctca gcagcaccct gacgctgagc 600
aaagcagact acgagaaaca caaagtctac gcctgcgaag tcacccatca gggcctgagc 660
tcgcccgtca caaagagctt caacagggga gagtgttag 699
<210> 35
<211> 717
<212> DNA
<213> IgK
<400> 35
atggacatga gggtccccgc tcagctcctg gggctcctgc tactctggct ccgaggtgcc 60
agatgtgaca tccagatgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120
gtcaccatca cttgccgggc aagtcagagc attagcagct atttaaattg gtatcagcag 180
aaaccaggga aagcccctaa gctcctgatc tatgctgcat ccagtttgca aagtggggtc 240
ccatcaaggt tcagtggcag tggatctggg acagatttca ctctcaccat cagcagtctg 300
caacctgaag attttgcaac ttactactgt caacagagtt acagtacccc tccggagaac 360
acttttggcc aggggaccaa gctggagatc aaacgaactg tggctgcacc atctgtcttc 420
atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 480
aataacttct atcccagaga ggccaaagta cagtggaagg tggataacgc cctccaatcg 540
ggtaactccc aggagagtgt cacagagcag gacagcaagg acagcaccta cagcctcagc 600
agcaccctga cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc 660
acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgttag 717
<210> 36
<211> 702
<212> DNA
<213> IgK
<400> 36
atgagggtcc ccgctcagct cctggggctc ctgctgctct ggctcccagg tgccagatgt 60
gacatccagt tgacccagtc tccatccttc ctgtctgcat ctgtaggaga cagagtcacc 120
atcacttgcc gggccagtca gggcattagc agttatttag cctggtatca gcaaaaacca 180
gggaaagccc ctaagctcct gatctatgct gcatccactt tgcaaagtgg ggtcccatca 240
aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 300
gaagattttg caacttatta ctgtcaacag cttaatagtt tgtacacttt tggccagggg 360
accaagctgg agatcaaacg aactgtggct gcaccatctg tcttcatctt cccgccatct 420
gatgagcagt tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc 480
agagaggcca aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag 540
agtgtcacag agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg 600
agcaaagcag actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg 660
agctcgcccg tcacaaagag cttcaacagg ggagagtgtt ag 702
<210> 37
<211> 237
<212> PRT
<213> IgK
<400> 37
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Phe Phe Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Val Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Pro Ser Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu
115 120 125
Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
130 135 140
Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
145 150 155 160
Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala
165 170 175
Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys
180 185 190
Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp
195 200 205
Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu
210 215 220
Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 38
<211> 239
<212> PRT
<213> IgK
<400> 38
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Ile Pro
1 5 10 15
Gly Ser Ser Ala Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser
20 25 30
Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser
35 40 45
Leu Leu His Ser Asp Gly Lys Thr Tyr Leu Tyr Trp Tyr Leu Gln Lys
50 55 60
Pro Gly Gln Pro Pro Gln Leu Leu Ile Tyr Glu Val Ser Asn Arg Phe
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
Cys Met Gln Ser Ile Gln Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys
115 120 125
Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
130 135 140
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
145 150 155 160
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
165 170 175
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
180 185 190
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
195 200 205
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
210 215 220
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 39
<211> 239
<212> PRT
<213> IgK
<400> 39
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Ile Pro
1 5 10 15
Gly Ser Ser Ala Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser
20 25 30
Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser
35 40 45
Leu Leu His Ser Asp Gly Lys Thr Tyr Leu Tyr Trp Tyr Leu Gln Lys
50 55 60
Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Glu Val Ser Ser Arg Phe
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
Cys Met Gln Ala Ile His Leu Pro Phe Thr Phe Gly Gln Gly Thr Lys
115 120 125
Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
130 135 140
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
145 150 155 160
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
165 170 175
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
180 185 190
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
195 200 205
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
210 215 220
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 40
<211> 239
<212> PRT
<213> IgK
<400> 40
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Ser
1 5 10 15
Gly Ser Ser Gly Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro
20 25 30
Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Leu Leu His Gly Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys
50 55 60
Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Ala Ser Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
Cys Met Gln Ala Leu Gln Ser Pro Ile Thr Phe Gly Gln Gly Thr Arg
115 120 125
Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
130 135 140
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
145 150 155 160
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
165 170 175
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
180 185 190
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
195 200 205
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
210 215 220
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 41
<211> 239
<212> PRT
<213> IgK
<400> 41
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Pro
1 5 10 15
Gly Ser Ser Gly Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro
20 25 30
Val Thr Leu Gly Gln Pro Ala Ser Ile Ser Cys Arg Ala Ser Gln Ser
35 40 45
Leu Gly Gln Ser Asp Gly Ser Thr Tyr Leu Thr Trp Phe Gln Gln Arg
50 55 60
Pro Gly Gln Ser Pro Arg Arg Leu Ile Tyr Lys Val Ser Tyr Arg Asp
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Thr Tyr Tyr
100 105 110
Cys Met Gln Gly Ile His Trp Pro Trp Thr Leu Gly Gln Gly Thr Lys
115 120 125
Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
130 135 140
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
145 150 155 160
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
165 170 175
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
180 185 190
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
195 200 205
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
210 215 220
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 42
<211> 240
<212> PRT
<213> IgK
<400> 42
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Pro
1 5 10 15
Gly Ser Ser Glu Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro
20 25 30
Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Leu Phe Asp Ser Ala Asp Gly Asn Thr Tyr Leu Asp Trp Tyr Leu Gln
50 55 60
Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Met Thr Ser Tyr Arg
65 70 75 80
Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
100 105 110
Tyr Cys Met Gln Arg Ile Glu Phe Pro Tyr Thr Phe Gly Gln Gly Thr
115 120 125
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 43
<211> 240
<212> PRT
<213> IgK
<400> 43
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Pro
1 5 10 15
Gly Ser Ser Glu Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro
20 25 30
Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Leu Leu Asp Ser Asp Asp Gly Asn Thr Tyr Leu Asp Trp Tyr Leu Gln
50 55 60
Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Thr Leu Ser Tyr Arg
65 70 75 80
Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
100 105 110
Tyr Cys Met Gln Arg Ile Glu Phe Pro Tyr Thr Phe Gly Gln Gly Thr
115 120 125
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 44
<211> 240
<212> PRT
<213> IgK
<400> 44
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Pro
1 5 10 15
Gly Ser Ser Glu Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro
20 25 30
Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Leu Leu Asp Ser Asp Asp Gly Asn Thr Tyr Leu Asp Trp Tyr Leu Gln
50 55 60
Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Thr Leu Ser Tyr Arg
65 70 75 80
Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
100 105 110
Tyr Cys Met Gln Arg Ile Glu Phe Pro Leu Thr Phe Gly Gln Gly Thr
115 120 125
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 45
<211> 240
<212> PRT
<213> IgK
<400> 45
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser
1 5 10 15
Gly Ala Cys Gly Ala Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Thr
35 40 45
Leu Leu Tyr Ser Ser Asn Asn Arg Asp Tyr Leu Gly Trp Tyr Gln Gln
50 55 60
Lys Ser Gly Gln Pro Pro Arg Leu Leu Ile Ser Trp Ala Ser Thr Arg
65 70 75 80
Glu Ser Gly Val Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Ile Tyr
100 105 110
Tyr Cys Gln Gln Tyr Tyr Asn Ala Pro Ile Thr Phe Gly Gly Gly Thr
115 120 125
Lys Val Glu Ile Arg Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Tyr Asn Arg Gly Glu Cys
225 230 235 240
<210> 46
<211> 240
<212> PRT
<213> IgK
<400> 46
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser
1 5 10 15
Gly Ala Tyr Gly Asp Ile Val Met Ser Gln Ser Pro Asp Ser Leu Thr
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser
35 40 45
Val Phe Tyr Asn Ser Asn His Lys Asn Tyr Leu Ala Trp Tyr Gln Gln
50 55 60
Lys Pro Gly Gln Pro Pro Lys Val Leu Ile Tyr Trp Ala Ser Thr Arg
65 70 75 80
Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu
85 90 95
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr
100 105 110
Tyr Cys Gln Gln Tyr Phe Asn Lys Pro Ile Thr Phe Gly Pro Gly Thr
115 120 125
Lys Val Asp Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 47
<211> 240
<212> PRT
<213> IgK
<400> 47
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser
1 5 10 15
Gly Ala Tyr Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser
35 40 45
Ile Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Phe Gln Gln
50 55 60
Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg
65 70 75 80
Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr
100 105 110
Tyr Cys Gln Gln Tyr Tyr Arg Ile Pro Trp Thr Phe Gly Gln Gly Thr
115 120 125
Lys Val Glu Ile Arg Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 48
<211> 240
<212> PRT
<213> IgK
<400> 48
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser
1 5 10 15
Gly Ala Tyr Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser
35 40 45
Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln
50 55 60
Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg
65 70 75 80
Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr
100 105 110
Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Phe Thr Phe Gly Pro Gly Thr
115 120 125
Lys Val Asp Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 49
<211> 240
<212> PRT
<213> IgK
<400> 49
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser
1 5 10 15
Gly Val Tyr Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser
35 40 45
Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Asp Trp Tyr Gln Gln
50 55 60
Arg Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg
65 70 75 80
Glu Ser Gly Val Pro Gly Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Ala Ala Val Tyr
100 105 110
Tyr Cys His Gln Tyr Tyr Ser Ser Pro Phe Thr Phe Gly Gln Gly Thr
115 120 125
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
130 135 140
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
145 150 155 160
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
180 185 190
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
195 200 205
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235 240
<210> 50
<211> 243
<212> PRT
<213> IgK
<400> 50
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Thr Ser
1 5 10 15
Val Thr Thr Gly Ala Phe Gly Asp Ile Val Met Thr Gln Ser Pro Asp
20 25 30
Ser Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile Ser Cys Lys Ser
35 40 45
Ser Gln Arg Val Leu Tyr Ser Ser Asn Asn Lys Tyr Tyr Leu Ala Trp
50 55 60
Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala
65 70 75 80
Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser
85 90 95
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Val
100 105 110
Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Trp Thr Phe Gly
115 120 125
Arg Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val
130 135 140
Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser
145 150 155 160
Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln
165 170 175
Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val
180 185 190
Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu
195 200 205
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu
210 215 220
Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg
225 230 235 240
Gly Glu Cys
<210> 51
<211> 241
<212> PRT
<213> IgK
<400> 51
Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser
1 5 10 15
Gly Ala Phe Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Thr Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser
35 40 45
Val Phe Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln
50 55 60
Lys Pro Gly Gln Pro Pro Lys Leu Leu Val Tyr Trp Ala Ser Thr Arg
65 70 75 80
Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
85 90 95
Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Ile Tyr
100 105 110
Tyr Cys Gln Gln Tyr Tyr Gly Ala Pro Pro Arg Thr Phe Gly Gln Gly
115 120 125
Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
130 135 140
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
145 150 155 160
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
165 170 175
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
180 185 190
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
195 200 205
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
210 215 220
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
225 230 235 240
Cys
<210> 52
<211> 241
<212> PRT
<213> IgK
<400> 52
Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Met Leu Trp Val Ser
1 5 10 15
Gly Ser Ser Gly Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro
20 25 30
Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys
50 55 60
Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
Cys Met Gln Ala Leu Gln Thr Pro Pro Glu Tyr Thr Phe Gly Gln Gly
115 120 125
Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
130 135 140
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
145 150 155 160
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
165 170 175
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
180 185 190
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
195 200 205
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
210 215 220
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
225 230 235 240
Cys
<210> 53
<211> 235
<212> PRT
<213> IgK
<400> 53
Met Glu Ala Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
50 55 60
Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
100 105 110
Asn Trp Pro Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 54
<211> 235
<212> PRT
<213> IgK
<400> 54
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Ile Ser Ser Asn Tyr Leu Ala Trp Tyr Gln Leu Lys Pro Gly Gln Gly
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Arg Ser Leu Val Pro Glu Asp Phe Ala Leu Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Lys Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 55
<211> 235
<212> PRT
<213> IgK
<400> 55
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Gln Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 56
<211> 235
<212> PRT
<213> IgK
<400> 56
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Leu Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 57
<211> 235
<212> PRT
<213> IgK
<400> 57
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ile Ser Thr Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Leu Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 58
<211> 235
<212> PRT
<213> IgK
<400> 58
Met Glu Ala Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Lys Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
50 55 60
Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn
100 105 110
Asn Trp Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Asp Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 59
<211> 236
<212> PRT
<213> IgK
<400> 59
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 60
<211> 236
<212> PRT
<213> IgK
<400> 60
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 61
<211> 236
<212> PRT
<213> IgK
<400> 61
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Ser Lys Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 62
<211> 236
<212> PRT
<213> IgK
<400> 62
Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Asp Thr Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
50 55 60
Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro
65 70 75 80
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
85 90 95
Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr
100 105 110
Gly Ser Ser Pro Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 63
<211> 236
<212> PRT
<213> IgK
<400> 63
Met Asp Met Arg Val Leu Ala Gln Leu Leu Gly Leu Leu Leu Leu Cys
1 5 10 15
Phe Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Gly Ile Ser Lys Tyr Leu Leu Trp Phe Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Ser Leu Ile Tyr Gly Ala Ser Phe Leu Gln Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Thr Gly Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Arg Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
Tyr Asp Thr Tyr Pro Pro Thr Phe Gly Gly Gly Thr Arg Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 64
<211> 236
<212> PRT
<213> IgK
<400> 64
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Thr
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Ile Ile Thr Cys Arg Ala Ser
35 40 45
Gln Asn Ile Asn Arg Trp Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Asn Leu Leu Ile Tyr Gln Ala Ser Thr Leu Glu Gly Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Gly Asp Phe Ala Thr Tyr Tyr Cys His Gln
100 105 110
Tyr His Ser Tyr Ser Tyr Ala Phe Gly Gln Gly Thr Lys Leu Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 65
<211> 236
<212> PRT
<213> IgK
<400> 65
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Pro Gly Ala Lys Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Thr
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Ser Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
Tyr Asn Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 66
<211> 236
<212> PRT
<213> IgK
<400> 66
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Pro Gly Ala Lys Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Thr
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Ser Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
Tyr Asn Ser Tyr Pro Ile Thr Phe Gly Gly Gly Thr Lys Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 67
<211> 235
<212> PRT
<213> IgK
<400> 67
Met Asp Met Arg Leu Ala Gln Leu Leu Gly Leu Leu Leu Leu Cys Phe
1 5 10 15
Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu
20 25 30
Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu
35 40 45
Gly Ile Ser Asn Phe Val Ala Trp Phe Gln Gln Glu Pro Gly Lys Ala
50 55 60
Pro Lys Ser Leu Ile Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro
65 70 75 80
Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ala Ile
85 90 95
Thr Asn Leu Gln Pro Glu Asp Phe Gly Thr Tyr Phe Cys Gln Gln Tyr
100 105 110
Lys Ser Asn Pro Arg Thr Phe Gly Pro Gly Thr Arg Val Glu Ile Lys
115 120 125
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
130 135 140
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
145 150 155 160
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
165 170 175
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
180 185 190
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
195 200 205
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
210 215 220
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 68
<211> 236
<212> PRT
<213> IgK
<400> 68
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Ser Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
Ser Tyr Ser Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 69
<211> 253
<212> PRT
<213> IgK
<400> 69
Met Arg Trp Leu Tyr Gly Gly Arg Ala Ala Gln Leu Gly Pro Arg Gly
1 5 10 15
Asn His Gly Asn Pro Gln Arg Ser Phe Ser Ser Ser Cys Tyr Ser Gly
20 25 30
Ser Gln Ile Pro Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr
35 40 45
Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser
50 55 60
Gln Ser Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly
65 70 75 80
Gln Ala Pro Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly
85 90 95
Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
100 105 110
Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln
115 120 125
Gln Tyr Gly Ser Ser Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
130 135 140
Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
145 150 155 160
Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
165 170 175
Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala
180 185 190
Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys
195 200 205
Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp
210 215 220
Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu
225 230 235 240
Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
245 250
<210> 70
<211> 232
<212> PRT
<213> IgK
<400> 70
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Pro Gly Ala Lys Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Thr
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Ser Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
Tyr Lys Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val
115 120 125
Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys
130 135 140
Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg
145 150 155 160
Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
165 170 175
Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser
180 185 190
Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys
195 200 205
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
210 215 220
Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 71
<211> 238
<212> PRT
<213> IgK
<400> 71
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
20 25 30
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
35 40 45
Gln Ser Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys
50 55 60
Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val
65 70 75 80
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
85 90 95
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
100 105 110
Ser Tyr Ser Thr Pro Pro Glu Asn Thr Phe Gly Gln Gly Thr Lys Leu
115 120 125
Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
130 135 140
Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu
145 150 155 160
Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175
Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser
180 185 190
Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala
195 200 205
Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
210 215 220
Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 72
<211> 233
<212> PRT
<213> IgK
<400> 72
Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Leu Pro
1 5 10 15
Gly Ala Arg Cys Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly
35 40 45
Ile Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
50 55 60
Lys Leu Leu Ile Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn
100 105 110
Ser Leu Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr
115 120 125
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
130 135 140
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
145 150 155 160
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
165 170 175
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
180 185 190
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
195 200 205
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
210 215 220
Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 73
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
ggtgaaagat gagctggagg 20
<210> 74
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
tcaggctgga actgagga 18
<210> 75
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
tgctcagtta ggacccagag 20
<210> 76
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
atgcatggat gtacggggag 20
<210> 77
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
gtcaggacac agcatggaca 20
<210> 78
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
tgctcagtta ggacccagac 20
<210> 79
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
tcctcagttc accttctcac a 21
<210> 80
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
cctcagttca ccttctcaca atg 23
<210> 81
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
tcctcagttc accttctcac c 21
<210> 82
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
tcagttcacc ttctcaccat ga 22
<210> 83
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
ttctcagttc atcttctcac c 21
<210> 84
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
tcagttcatc ttctcaccat ga 22
<210> 85
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
agctacaaca ggcaggcag 19
<210> 86
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 86
ggggagctac aacaggca 18
<210> 87
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 87
aggtcagggc caaggttatg 20
<210> 88
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 88
ggactgacta ccacctgcag 20
<210> 89
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 89
ttccgatcta atgcgatggc 20
<210> 90
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 90
agacgtgtgc tcttccgatc t 21
<210> 91
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 91
gagctgctca gttaggacc 19

Claims (9)

1.一种全人源的新冠IgK单链抗体，其特征在于，包括如SEQ ID No：37-SEQ ID No：72中至少一种所示的氨基酸序列。
2.编码权利要求1所述的新冠IgK单链抗体的基因序列。
3.根据权利要求2所述的新冠IgK单链抗体，其特征在于，包括如SEQ ID No：1-SEQ IDNo：36中至少一种与所述氨基酸对应的核苷酸序列。
4.包含权利要求2所述新冠IgK单链抗体中所述基因序列的文库。
5.权利要求4所述文库的制备方法，其特征在于，对不同新冠康复者共有而在正常人群中没有的BCR序列进行分析并筛选出新冠特异性的IgK BCR抗体序列。
6.根据权权利要求5所述的制备方法，其特征在于，所述分析筛选过程为：对新冠康复者与正常人群进行BCR全长测序，将测序的原始数据进行HiFi的一致性校正后，获得了质量值在Q20以上的BCR全长一致性序列，经过与BCR数据库中的抗体恒定区序列进行比对后，获得了每个样本测序数据中的不同类别的BCR抗体序列。
7.包含权利要求3所述新冠IgK单链抗体中所述基因序列的表达载体。
8.包含权利要求3所述新冠IgK单链抗体中所述基因序列的宿主细胞。
9.权利要求1所述的新冠IgK单链抗体在制备新冠病毒治疗药物、药物载体及检测标记物中的应用。
CN202110547125.6A 2021-05-19 2021-05-19 全人源的新冠IgK单链抗体及其应用 Pending CN113234148A (zh)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11732030B2 (en) 2020-04-02 2023-08-22 Regeneron Pharmaceuticals, Inc. Anti-SARS-CoV-2-spike glycoprotein antibodies and antigen-binding fragments
US11999777B2 (en) 2020-06-03 2024-06-04 Regeneron Pharmaceuticals, Inc. Methods for treating or preventing SARS-CoV-2 infections and COVID-19 with anti-SARS-CoV-2 spike glycoprotein antibodies

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111732655A (zh) * 2020-07-01 2020-10-02 中国人民解放军军事科学院军事医学研究院 靶向RBD的高中和活性抗SARS-CoV-2全人源单克隆抗体及应用
CN111778218A (zh) * 2020-06-04 2020-10-16 山东宽和正生物医药有限公司 噬菌体展示抗体库及基于其淘筛获得的针对新冠病毒SARS-CoV-2的单克隆抗体

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111778218A (zh) * 2020-06-04 2020-10-16 山东宽和正生物医药有限公司 噬菌体展示抗体库及基于其淘筛获得的针对新冠病毒SARS-CoV-2的单克隆抗体
CN111732655A (zh) * 2020-07-01 2020-10-02 中国人民解放军军事科学院军事医学研究院 靶向RBD的高中和活性抗SARS-CoV-2全人源单克隆抗体及应用

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11732030B2 (en) 2020-04-02 2023-08-22 Regeneron Pharmaceuticals, Inc. Anti-SARS-CoV-2-spike glycoprotein antibodies and antigen-binding fragments
US11999777B2 (en) 2020-06-03 2024-06-04 Regeneron Pharmaceuticals, Inc. Methods for treating or preventing SARS-CoV-2 infections and COVID-19 with anti-SARS-CoV-2 spike glycoprotein antibodies

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