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猪瘟病毒囊膜糖蛋白 E rns的表位分析及其单克隆抗体轻链可变区cDNA 的测序
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张富强， 李志华， 张念祖
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（云南省热带亚热带动物病毒病重点实验室， 云南 昆明650224）
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摘要：
应用抗CSFV Alfort Tübingen毒株E rns结构蛋白的b422、24/16单克隆抗体，筛选了噬菌体展示的12肽随机肽库，进行了E rns结构蛋白表位分析。研究发现，b422、24/16单克隆抗体识别的表（拟）位基序分别为DKNR(Q)G、 A（T）CxYxKN，定位于E rns结构蛋白的384～386及322～323位、380～386位氨基酸区域。二者识别的表（拟）位基序存在共有序列KN，针对的是E rns蛋白中的相似抗原区，但其拟位的侧翼序列及ELISA、免疫印迹分析结果均存在差异。自杂交瘤细胞中提取总RNA，对单克隆抗体轻链可变区cDNA进行了测序。结果证实，b422、24/16单克隆抗体虽然来源于同一批次融合的杂交瘤细胞系，识别相似抗原区，但属于不同的单克隆抗体。 关键词：
猪瘟病毒； 单克隆抗体； 表位； 可变区； 测序
中图分类号：S 855.3：Q 523.8 文献标识码：
A 文章编号：1000-6419(2004)08-0009-05 |
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Mapping of epitope and determination of cDNA sequences oflight chain variable region of MAbs against envelope glycoprotein E rns of classical swine fever virus
ZHANG Fuqiang, LI Zhihua, ZHANG Nianzu
(Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory, Kunming 650224, China) |
Abstract: Analysis of E rns epitope by screening a 12mer random peptide phage display library was reported using the monoclonal antibodies (MAbs) b422 and 24/16,which raisedagainst CSFV strain alfort T bingen and reacted with E rns protein. The epitope or mimotope motifs DKNR(Q)G,A(T)CxYxKN, recognized by MAbs b422 and 24/16 respectively, had been identified and located around amino acid (aa) position 384-386/322-323 and 380-386 of E rns protein. MAbs b422 and 24/16 shared partial binding sequence KN and recognized a imilar antigenic domain but showed distinct diversities of flank sequence and reactivities by ELISA and Western blot analysis. Sequencing both cDNA of light chain variable region of two MAbs from total RNA of the hybridoma cells were carried out. The results showed MAb b422 and MAb 24/16 are different monoclonal antibodies though both MAbs derived from the same fusion cells and recognized the similar antigenic domain.
Key words: Classical swine fever virus; monoclonal antibody; epitope; variable region; sequencing |
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