Mapping of Epitope and Determination of cDNA Sequences of Variable Region of MAbs Against E2 Protein of Classical Swine Fever Virus

ZHANG Fu-qiang; LI Zhi-hua; ZHANG Nian-zu

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December 2004

ZHANG Fu-qiang, LI Zhi-hua and ZHANG Nian-zu. Mapping of Epitope and Determination of cDNA Sequences of Variable Region of MAbs Against E2 Protein of Classical Swine Fever Virus[J]. Virologica Sinica, 2004, 19(6): 591-597.

Citation: ZHANG Fu-qiang, LI Zhi-hua, ZHANG Nian-zu. Mapping of Epitope and Determination of cDNA Sequences of Variable Region of MAbs Against E2 Protein of Classical Swine Fever Virus .VIROLOGICA SINICA, 2004, 19(6) : 591-597.

猪瘟病毒E2蛋白表位分析及其单克隆抗体可变区cDNA测序

1.
云南省热带亚热带动物病毒病重点实验室，云南昆明

通讯作者： 张富强*, ;

摘要

猪瘟病毒（CSFV）囊膜表面结构糖蛋白E2 (gp55)是诱导机体产生中和抗体及激发保护性免疫应答的主要抗原蛋白。E2和Erns与细胞表面受体的相互作用介导病毒对细胞的感染过程。本文采用抗CSFV（Alfort Tübingen 毒株）E2结构蛋白的单克隆抗体c2410和a18，淘选噬菌体展示的12肽随机肽库，对CSFV E2蛋白表位进行分析。研究发现：单克隆抗体c2410和a18识别同一线性表位，定位于E2蛋白的832-837位氨基酸（SPTTLR），但二者在ELISA和免疫印迹分析中对不同表（拟）位的反应性存在差异。自杂交瘤细胞中提取总RNA，对单克隆抗体轻链和重链可变区cDNA进行序列分析。结果表明：c2410和a18虽然来源于同一批次融合的杂交瘤细胞系，识别同一表位，但仍属于不同的单克隆抗体。
- 猪瘟病毒
- , 单克隆抗体
- , 表位
- , 可变区
- , 测序

Mapping of Epitope and Determination of cDNA Sequences of Variable Region of MAbs Against E2 Protein of Classical Swine Fever Virus

1.
Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory, Kunming 650224, China

Corresponding author: ZHANG Fu-qiang,

Abstract

Structural and envelope glycoprotein E2 (gp55) of Classical swine fever virus (CSFV) is the most antigenic protein being responsible for eliciting neutralizing antibodies and conferring protective immunity. Infection of cells with CSFV is mediated by the interaction of E2 and Erns with the cell surface receptor. In this paper we report the analysis of E2 epitope by screening a 12-mer random peptide phage display library using the monoclonal antibodies (MAbs), c2410 and a18, raised against CSFV strain alfort Tübingen and reacted with the E2 structural protein. MAbs, c2410 and a18 recognized the same linear epitope, located at aa832- aa 837 (SPTTLR) of E2 protein, but showed some different reactivities with the mimotopes by ELISA and Western blot analysis. Sequencing both cDNA of light chain and heavy chain variable regions of the two MAbs from total RNA of the hybridoma cells was carried out. The results showed that MAb c2410 and MAb a18 are different monoclonal antibodies, though both MAbs derived from the same fusion and recognized the same epitope.
- CSFV
- , Monoclonal antibody
- , Epitope
- , Variable region
- , Sequencing

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Mapping of Epitope and Determination of cDNA Sequences of Variable Region of MAbs Against E2 Protein of Classical Swine Fever Virus

Corresponding author: ZHANG Fu-qiang,

1. Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory, Kunming 650224, China

Abstract: Structural and envelope glycoprotein E2 (gp55) of Classical swine fever virus (CSFV) is the most antigenic protein being responsible for eliciting neutralizing antibodies and conferring protective immunity. Infection of cells with CSFV is mediated by the interaction of E2 and Erns with the cell surface receptor. In this paper we report the analysis of E2 epitope by screening a 12-mer random peptide phage display library using the monoclonal antibodies (MAbs), c2410 and a18, raised against CSFV strain alfort Tübingen and reacted with the E2 structural protein. MAbs, c2410 and a18 recognized the same linear epitope, located at aa832- aa 837 (SPTTLR) of E2 protein, but showed some different reactivities with the mimotopes by ELISA and Western blot analysis. Sequencing both cDNA of light chain and heavy chain variable regions of the two MAbs from total RNA of the hybridoma cells was carried out. The results showed that MAb c2410 and MAb a18 are different monoclonal antibodies, though both MAbs derived from the same fusion and recognized the same epitope.