Immunogenicity of hemagglutinin of H7 N9 expressed in yeast Pichia pastoris

VernacularTitle:H7 N9流感病毒血凝素HA在毕赤酵母中的表达及免疫原性分析
Author: Sha WANG ; Shaohong CHANG ; Bo LIU ; Xin GONG ; Wei XU ; Jun WU
Publication Type:Journal Article
Keywords: H7N9 influenza virus; hemagglutinins; Pichia; hemagglutination-inhibition
From: Military Medical Sciences 2015;(6):448-452
CountryChina
Language:Chinese
Abstract: Objective To express the hemagglutinin of H7N9 in Pichia pastoris and analyze its immunogenicity. Methods The HA [1 -525 amino acids(aa)] of H7N9 [A/Hangzhou/1/2013(H7N9)] lacking the C-terminal transmembrane anchor-coding sequence was amplified by PCR and cloned into the expression vector pPICZαA.The plasmid pPICZ-HA7-S-was transformed into P.pastoris and the HA1-525 was detected by ELISA.Then the HA1-525 was precipitated by PEG20000.After immunization with the HA1-525 , the anti-HA7 antibody in mouse serum was detected by ELISA and hemagglutinin inhibition ( HI) test.Results The HA1-525 was expressed in P.pastoris after being induced with methanol. Western blotting confirmed that there were specific dispersion bands and the molecular weight of HA1-525 decreased to 58 × 103 after being digested by endo H.Anti-HA7 antibody was found in serum of HA1-525 immunized mice and the hemaggluti-nation-inhibition titers reached 1∶700 after the third doses.Conclusion This study shows the HA1-525 expressed in P.pastoris can induce the neutralizing antibody in mice.