Characterization of Polyclonal Antibodies Against PRRSV Nonstructural Protein 1α(Nsp1α)Generated by Immunization with a Synthetic Peptide
The synthetic peptide antibodies of PRRSV non-structural protein Nsp1α have not been reported yet.In this study,the synthetic peptide of PRRSV Nsp1α was used to prepare polyclonal antibodies for further analysis.First,the antigenic peptide sequence of PRRSV non-structural protein Nsp1α was predicted by using bioinformatics technology.Then,the antigenic peptide was synthesized according to this sequence and coupled with the KLH carrier protein.The coupled antigenic peptide was emulsified with Freund's adjuvant to immunize rabbits for preparation of polyclonal antibodies.Finally,the polyclonal antibodies were analyzed in ELISA,Western blot and indirect immunofluorescence.The results show that the ELISA titer of the polyclonal antibodies exceeded 105 and the specific Nsp1α band was visualized in Western blotting,including samples from transient transfection and infection with different virus strains.Additionally,the polyclonal antibodies confirmed virus-infected and uninfected samples in indirect immunofluorescence.Therefore,the synthetic peptide polyclonal antibodies against PRRSV Nsp1α could be used in the detection of PRRSV Nsp1α expression,thus providing an effective tool for further research on the function of PRRSV Nsp1α.
Porcine reproductive and respiratory syndrome virussynthetic peptidepolyclonal antibodynonstructural protein
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