传染性法氏囊病毒VP2蛋白抗原表位多肽P22的免疫原性及生物学功能鉴定  

Preparation and Identification of the Epitope Peptide P22 from VP2 of Infectious Bursal Disease Virus

作　　者：宋幸辉[1,2] 王睿[1,2] 王选年[2,3] 鲍登克[2] 杨苏珍[2] 卢清侠[2] 王寅彪[2] 赵东[2] 张改平[2] 

机构地区：[1]河南农业大学,郑州450002 [2]河南省农业科学院农业部动物免疫学重点开放实验室,郑州450002 [3]新乡学院生物技术研究所,新乡453003

出　　处：《畜牧兽医学报》2011年第5期692-697,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30871885)

摘　　要：为鉴定IBDV VP2蛋白线性B细胞抗原表位肽P22的免疫原性及生物学功能,作者采用固相多肽合成方法合成多肽P22,经纯化后用SMCC法分别与牛血清白蛋白(BSA)和细胞穿膜肽(PNT)进行偶联,得到免疫抗原P22-BSA和检测抗原P22-PNT;用P22-BSA免疫BALB/c小鼠3只,经3次免疫后得到多抗血清3份。应用ELISA检测了抗P22抗体的特异性,并通过抗P22多肽抗体作为特异性抗体检测了P22多肽结合CEF细胞的特性。结果经鉴定3份多抗血清均与P22多肽特异性反应,效价均达到1∶105,且能特异性检测P22多肽与CEF细胞的特异性结合。本试验成功制备了P22免疫原,获得了效价高、特异性较好的鼠源抗P22多抗血清,并证明P22多肽与CEF细胞能够特异性结合,为进一步研究多肽疫苗的设计或试纸条的研制及表位单抗的制备提供了新的思路和基础。The aim of this study was to identify the immunogenicity and the cell binding ability of the linear B cell epitope peptide P22 from VP2 of IBDV.The peptide P22 was synthesized by a solid-phase method.The purified peptide was conjugated with bovine serum albumin（BSA） and cell penetrating peptide（PNT） using SMCC method to prepare the immunogen P22-BSA and the detecting antigen P22-PNT.After immunizing BALB/c mice with P22-BSA,polyclonal antibodies were obtained and identified with ELISA and immunohistochemical assay.Results showed that the titers of the 3 polyclonal antibodies of P22 all reached to 1：105,and the anti-P22 polyclonal antibodies could effectively bind the P22 peptide previously incubated with CEF cells.In this study,the immunogen P22,and the anti-P22 polyclonal antibody were successfully prepared for further study of the infection mechanism of IBDV,and lays the foundation for the design of peptide vaccine and the development of new drugs.

关 键 词：IBDV VP2蛋白 P22多肽 抗原表位 免疫 

分 类 号：S852.659.4[农业科学—基础兽医学]