针对猪瘟病毒E2蛋白的嵌合猪源化单克隆抗体的表达及抗病毒活性鉴定

发布时间：2018-04-12 21:48

本文选题：猪瘟病毒 + 单克隆抗体　；参考：《生物工程学报》2017年08期

【摘要】：猪瘟(Classical swine fever,CSF)是严重危害养猪业的一种烈性传染病,常造成巨大的经济损失,是世界动物卫生组织要求必须申报的动物疫病之一。猪瘟的病原是猪瘟病毒(Classical swine fever virus,CSFV),CSFV的结构蛋白由衣壳蛋白(C)和囊膜糖蛋白(E~(rns)、E1、E2)构成。E2蛋白是CSFV主要的保护性抗原,可以诱导机体产生中和抗体,从而抵抗CSFV的感染。此前,本团队制备了一株针对CSFV E2蛋白的鼠源单克隆抗体HQ06。文中将HQ06抗体重链和轻链可变区基因与猪源恒定区基因嵌合后克隆至真核表达载体,利用中国仓鼠卵巢(CHO)细胞制备一株针对CSFV E2蛋白的嵌合猪源化单克隆抗体c HQ06。应用ELISA、Western blotting试验证实了c HQ06与CSFV E2蛋白具有良好的反应性;中和试验结果表明c HQ06可以中和CSFV。综上所述,本研究应用CHO细胞稳定表达了具有良好反应性和中和活性的针对CSFV E2蛋白的嵌合猪源化单克隆抗体c HQ06,为研究CSFV E2蛋白结构、功能以及开发新型的CSFV诊断和治疗制剂奠定基础。
[Abstract]:Classical swine epidemic (CSFs) is a severe infectious disease in swine industry, which often causes huge economic losses and is one of the animal diseases that must be declared by the World Organization of Animal Health (OIE).The structural protein of classical swine fever virus (CSFV) of CSFV is composed of capsid protein (C) and envelope glycoprotein E1E2). E2 protein is the main protective antigen of CSFV, which can induce the production of neutralizing antibody and resist the infection of CSFV.Earlier, the team prepared a murine monoclonal antibody against CSFV E2, HQ06.In this paper, the heavy chain and light chain variable region genes of HQ06 antibody were chimeric with porcine constant region gene and cloned into eukaryotic expression vector. A chimeric porcine monoclonal antibody c HQ06against CSFV E2 protein was prepared by using Chinese hamster ovary Cho cells.The reaction of c HQ06 to CSFV E2 protein was confirmed by Elisa Western blotting test, and the neutralization test showed that c HQ06 could neutralize CSFV E2 protein.To sum up, CHO cells were used to stably express the chimeric porcine monoclonal antibody c HQ06against CSFV E2 protein with good reactivity and neutralization activity, which was used to study the structure of CSFV E2 protein.Function and development of new CSFV diagnostic and therapeutic agents lay the foundation.
【作者单位】：天津农学院动物科学与动物医学学院;中国农业科学院哈尔滨兽医研究所;
【基金】：国家自然科学基金(No.31672537)资助~~
【分类号】：S852.4

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