Preparation and efficacy evaluation of polyclonal antibodies against serum albumin of five species
Objective To prepare polyclonal antibodies against the serum albumin of human,cattle,sheep,pig and horse,and evaluate their efficacy in the identification of human serum albumin(HSA).Methods The specific polypeptides of human,cattle,sheep,pig and horse serum albumin were prepared by bioinformatics and polypeptide synthesis method,which were coupled with keyhole limpet hemocyanin(KLH)to prepare the peptide antigen after the purity was identified by high performance liquid chromatography(HPLC).Male New Zealand white rabbits were immunized with polypeptide antigens of five species subcutaneously,with 2 for each kind of antigen.The antiserum was then obtained and purified by Protein A affinity chromatography to prepare the polyclonal antibody.The titers and specificity of the polyclonal antibodies were deter-mined by ELISA and Western blot respectively,and the prepared five species of serum albumin were used to identify HSA products.Results The synthetic peptides of human,cattle,sheep,pig and horse serum albumin had a purity of over 91％,and the corresponding polyclonal antibodies all had the titer of 1∶160 000,which showed specific binding with the correspond-ing antigens and effectively identified the HSA products.Conclusion The polyclonal antibodies of human cattle,sheep,pig and horse serum albumin prepared in this study have good specificity and the preparation process is simple and rapid,suit-able for the mass production,which lays a foundation of the development of HSA rapid identification kit.
Human serum albumin(HSA)PolypeptidePolyclonal antibodyImmuno-double-diffusion method
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