Preparation of Polyclonal Antibody Against Recombinant CP of Potato virus A and Its Application to DAS-ELISA Detection

doi:10.7668/hbnxb.2017.01.007

Abstract

Abstract: The purpose was to prepare polyclonal antibody against recombinant CP of Potato virus A,and apply it to DAS-ELISA.Taking pET22b(+) as the initial vector, CP gene prokaryotic expression vector of Potato virus A (pET22b-ACP) was constructed,the recombinant strain BL21 (pET22b-ACP) was induced with IPTG and specific recombinant CP of 30 kDa was obtained.Antiserum with the titer of 1:512k was prepared by using high-purity recombinant CP as antigen to immune rabbits.Using PVA recombinant CP as antigen,visual inspection and OD value measurement results indicated that reactions were respectively positive in indirect ELISA assay with the purified antibody (IgG) against the recombinant CP,in direct ELISA assay with the alkaline phosphatase-conjugated IgG (IgG-AP),and in DAS-ELISA determination with IgG against the recombinant CP and its AP-conjugated IgG.When PVA positive standard substance was used as antigen,reactions were also positive respectively in the three kinds of ELISA detections mentioned above.The polyclonal antibody against PVA recombinant CP and its enzyme-labeled antibody could meet the requirements of DAS-ELISA detection of Potato virus A.

Key words: Potato virus A, Recombinant CP, Polyclonal antibody, Enzyme-conjugated antibody, DAS-ELISA detection

CLC Number:

SONG Zhicheng, FEI Xinmin, YANG Yu, QIAO Lixian, WANG Jingshan, LI Guangcun, GUO Baotai. Preparation of Polyclonal Antibody Against Recombinant CP of Potato virus A and Its Application to DAS-ELISA Detection[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(1): 41-46. doi: 10.7668/hbnxb.2017.01.007.

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