Preparation and Potential Application of High-titer Polyclonal Antibody Against Capsid Protein of Porcine Circovirus Type Ⅱ

doi:10.13560/j.cnki.biotech.bull.1985.2019-1209

Abstract

Abstract: Capsid protein（Cap）is the major antigen protein of porcine circovirus（PCV）and plays critical roles in viral infection and elicitation of host immune responses. To prepare a high-titer polyclonal antibody against PCV2b Cap,a PCV2b Cap truncated protein without nuclear localization signal peptide was expressed by Escherichia coli prokaryotic expression system,and soluble recombinant protein was acquired. White rabbits were then immunized with the purified soluble recombinant protein to generate antiserum and anti-Cap IgG was further purified by caprylic acid-ammonium sulfate precipitation. ELISA analysis showed that the titer of the purified anti-Cap antibody was 1∶10⁷,indicating that the antibody was really in high-titer. The antibody was then applied to infect PCV2b-infected PK-15 cells or mouse tissues as well as clinical samples,and then they were detected by immunoblotting,indirect immunofluorescence,or immunohistochemistry. The results showed that the antibody specifically bound with target antigens. In summary,the prepared high-tier anti-Cap polyclonal antibody in this study may provide a candidate tool for the diagnosis and research of PCV2b-assciated diseases.

Key words: porcine circovirus type 2, capsid protein, prokaryotic expression, polyclonal antibody, virus detection

ZHANG Wen, LIU Zhao-zhen, LI Jun-shuo, SHANG Ying-li. Preparation and Potential Application of High-titer Polyclonal Antibody Against Capsid Protein of Porcine Circovirus Type Ⅱ[J]. Biotechnology Bulletin, 2020, 36(7): 72-79.

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