猪腺病毒3型Hexon蛋白多克隆抗体制备与免疫活性鉴定

doi:10.16431/j.cnki.1671-7236.2015.01.001

Abstract

Abstract: This study was aimed to prepare polyclonal antibodies against porcine adenovirus serotype 3 (PADV-3) Hexon protein.The recombinant prokaryotic expression plasmid pET28a-Hexon was constructed and transformed into BL21 (DE3), 1 mmol/L concentration of IPTG was used to induce the target gene expression which was then analyzed by SDS-PAGE and Western blotting.The expressed proteins were emulsified with freund's adjuvant, the mixture was used to immunize the rabbit to prepare the polyclonal antibodies against PADV-3 Hexon protein.The immunocompetence and titers of the antibody were identified by immunoperoxidase monolayer assay (IPMA).The results showed that prokaryotic expressed Hexon protein existed as inclusion bodies, with a molecular weight of 105 ku, eukaryotic expressed Hexon protein was only observed in the cytoplasm of HEK293 cells.PADV-3 Hexon-specific polyclonal antibody showed perfect activity which recognized the PADV-3 propagated in PK-15 cells and the recombinant Hexon protein expressed in HEK293 cells specifically, it was IPMA antibody positive when diluted in 1:1 600 ratio.The prepared PADV-3 Hexon polyclonal antibodies showed wonderful activity and specificity.

Key words: porcine adenovirus serotype 3; Hexon gene; polyclonal antibody

CLC Number:

S852.4

WANG Zhen-nan, TANG Qing-hai, YAN Ai, LIU Yun-fu. Preparation and Immuno-activity Identification of Polyclonal Antibodies against Porcine Adenovirus Serotype 3 Hexon Protein[J]. , 2015, 42(1): 1-8.

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Preparation and Immuno-activity Identification of Polyclonal Antibodies against Porcine Adenovirus Serotype 3 Hexon Protein

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