犬细小病毒NY株VP2蛋白多克隆抗体的制备与鉴定

doi:10.16431/j.cnki.1671-7236.2016.07.001

Abstract

Abstract:

This study was aimed to prepare canine parvovirus (CPV) VP2 protein polyclonal antibody.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 adjuvant,the prepared immunogen was inoculated into rabbit by subcutaneous injections 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.

Key words: caine parvirus; VP2 gene; polyclonal antibody; immunocompetence

CLC Number:

S852.65⁺5

MAO Qian-qian, ZHOU Ling, TANG Qing-hai, BU Bin, TANG Cun-duo, JIAO Zhu-jin, YAO Lun-guang, KAN Yun-chao, YANG Jian-wei, CUI Shang-jin. Preparation and Identification of Canine Parvovirus NY Strain VP2 Protein Polyclonal Antibody[J]. , 2016, 43(7): 1659-1666.

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Preparation and Identification of Canine Parvovirus NY Strain VP2 Protein Polyclonal Antibody

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