Preparation And Detection Of Polyclonal Antibody Of Porcine β-defensin-2

Antimicrobial peptides (AMPs) are a class of proteins secreted by animals against pathogenic microorganisms, which played impotmant role in innate immunity. Porcine β-defensin-2(PBD-2) is an important antimicrobial peptide. It is mainly secreted by epithelial tissue, and widely exists in the skin, tongue, respiratory tract, spleen, thymus, gastrointestinal tract, liver.et al.it had broad antimicrobial activity. Because of their specific antibacterial mechanism, which made bacteria hard to resisit it, AMPs are a popular research area recently.At present there are not monoclonal antibody or polyclonal antibody of β-defensin-2in the market.In order to identify β-defensin-2 in pigs This study aimed to produce polyclonal antibody against porcine β-defensin-2.In this study, the BL-pET-pBD2 was inducted and purified. The purity of target protein is above 90%, which was used to immunize rabbits, after four times injection, the antiserum was collected.Precipitated by saturated ammonium sulfate salt and purified by DEAE cellulose column chromatography, Serum was purified. The titer of antibodies was 1:12800 by indirect ELISA determination.Western blowing indicated that the specific detection of polyclonal antibody was sensitive, which was the base for further study.