Purification and Identification of Polyclonal Antibodies against SHN3

Objective: To purify polyclonal antibodies against SHN3 by affinity chromatography with immunologic specificity peptide fragment. Method: SHN3 immunologic specificity gene( 90bp) was divided 4 equal length parts( 48bp). And subcloned into prokaryotic expression vector Pmal- C2. These recombinant vectors were transformed into E. coliBL21( DE3). Recombinant peptide expression were induced by IPTG. Western blotting method was used to screen the best immunologic specificity peptide fragment. The polyclonal antibodies was purified by affinity chromatography and checked by Western blotting. Result: The immunologic specificity peptide fragment part- 3 of SHN3 was screened successfully,and the SHN3 polyclonal antibodies were purified with this peptide by affinity chromatography. The affinity chromatography of SHN3 polyclonal antibodies was improved significantly. Conclusion: Affinity chromatograph for SHN3 polyclonal antibodies purification was successfully established and immunologic specificity was significantly improved. The method laid a foundation for subsequent SHN3 research and application.