猫杯状病毒VP1蛋白的单克隆抗体制备及抗原表位鉴定  

Preparation of Monoclonal Antibodies against Feline Calicivirus VP1 Protein and Identification of Antigenic Epitopes

作　　者：张泽宇 董宁宁 谈晓梅 李传锋 朱杰[2] 刘光清[2] 张伟[1] 孟春春 ZHANG Zeyu;DONG Ningning;TAN Xiaomei;LI Chuanfeng;ZHU Jie;LIU Guangqing;ZHANG Wei;MENG Chunchun(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830000,China;Team for Companion Animal Biosafety and Prevention Technology,Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China)

机构地区：[1]新疆农业大学动物医学院,乌鲁木齐830000 [2]中国农业科学院上海兽医研究所伴侣动物生物安全与防控技术团队,上海200241

出　　处：《畜牧兽医学报》2024年第12期5784-5791,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：上海市自然科学基金(23ZR1477100);中央级公益性科研院所基本科研业务费专项资金项目(2023JB05)。

摘　　要：本研究旨在制备并鉴定针对猫杯状病毒(feline calicivirus,FCV)VP1蛋白的单克隆抗体(monoclonal antibodies,mAbs)。本研究使用SH14株FCV全病毒作为免疫原,通过腹腔注射法免疫BALB/c小鼠,制备了针对VP1蛋白的mAb。通过ELISA筛选,成功获得特异性单克隆抗体3A3C1株。对3A3C1株mAb进行了全面的生物学特性鉴定,3A3C1株mAb确认为IgG_(1)型,轻链为κ链。Western blot和间接免疫荧光检测的结果显示,3A3C1株mAb能特异性地识别并结合FCV SH14株、F9株和GZ22。此外,本研究还深入探究了3A3C1株mAb的线性抗原表位。通过分段表达VP1蛋白并进行详细的Western blot分析,最终确定3A3C1株mAb的线性抗原表位位于VP1蛋白的^(426)PSRLTPAGDYAITSG^(440)区域。本研究制备的3A3C1株mAb不仅是理解FCV免疫学特性的重要工具,也对发展FCV诊断方法和疫苗策略具有潜在应用价值。The purpose of this study was to prepare and identify monoclonal antibodies(mAbs)against the VP1 protein of feline calicivirus(FCV)strain SH14.In this study,the whole virus of the SH14 strain of FCV was used as an immunogen to immunize BALB/c mice through intraperitoneal injection,and mAbs against the VP1 protein were prepared.Through ELISA screening,a specific monoclonal antibody clone,3A3C1,was successfully obtained.A comprehensive biological characterization of the 3A3C1 clone mAb was conducted,confirming it as an IgG_(1)type with aκlight chain.The results of Western blot and immunofluorescence assay(IFA)showed that the 3A3C1 clone mAb could specifically recognize and bind to FCV strains SH14,F9,and GZ22.In addition,this study further explored the linear antigenic epitope of the 3A3C1 clone mAb.By segmentally expressing the VP1 protein and conducting detailed Western blot analysis,the linear antigenic epitope of the 3A3C1 clone mAb was finally determined to be located in the^(426)PSRLTPAGDYAITSG^(440)region of the VP1 protein.The 3A3C1 clone mAb prepared in this study is not only an important tool for understanding the immunological characteristics of FCV but also has potential applications in developing FCV diagnostic methods and vaccine strategies.

关 键 词：猫杯状病毒 单克隆抗体 抗原表位 衣壳蛋白 

分 类 号：S852.659.6[农业科学—基础兽医学]