中国畜牧兽医2016，Vol.43Issue(7)：1659-1666,8.DOI:10.16431/j.cnki.1671-7236.2016.07.001
犬细小病毒NY株VP2蛋白多克隆抗体的制备与鉴定
Preparation and Identification of Canine Parvovirus NY Strain VP2 Protein Polyclonal Antibody
摘要
Abstract
This study was aimed to prepare canine parvovirus (CPV)VP2 protein polyclonal anti-body.The recombinant expression vector pET28a-CPV-VP2 was constructed and transfromed into E.coli BL21 (DE3),the expression of recombinant proteins was induced by IPTG from which the fusion protein was identified by SDS-PAGE.The target protein was purified and emulsify with ad-j uvant,the prepared immunogen was inoculated into rabbit by subcutaneous inj ections to prepare of VP2 protein specific polyclonal antibody.The immuno-activity,titers,neutralization titers of the prepared polyclonal antibody were determined by immunoperoxidase monolayer assay (IPMA).The results showed that the expressed recombinant protein VP2 (rVP2)existed in the form of inclusion body with a molecular weight of 72 ku.The prepared polyclonal antibody titer was 1 600 dilution,the virus titer was 107 TCID50/mL,the neutralizing titer was 1∶2 884.The antibodies showed specific reaction with CPV.In conclusion,rVP2 specific polyclonal antibody showed wonderful immunocompetence,specificity and neutralizing activity,providing foundation for the development of genetic vaccine and clinical therapeutic method.关键词
犬细小病毒/VP2 基因/多克隆抗体/免疫活性Key words
caine parvirus/VP 2 gene/polyclonal antibody/immunocompetence分类
农业科技引用本文复制引用
毛倩倩,崔尚金,周灵,唐青海,卜宾,唐存多,焦铸锦,姚伦广,阚云超,杨建伟..犬细小病毒NY株VP2蛋白多克隆抗体的制备与鉴定[J].中国畜牧兽医,2016,43(7):1659-1666,8.基金项目
河南省重点科技攻关项目(142102110101) （142102110101）
河南省高等学校重点科研项目(16A230023) （16A230023）
南阳师范学院引进人才专项项目(70640) （70640）
