Screening of Hemagglutinin(HA)Protein Vaccine for Influenza Viruses B/Yamagata Lineage
Objective:To screen the recombinant hemagglutinin(HA)protein with high expression level and hemagglutinin in-hibitory activity for B/Yamagata influenza virus.Methods:Based on the HA of B/Singapore/INFTT-16-0610/2016,the peptide sequences of GCN4pLL trimerization motif,GCN4pLL-cysteine,and GCN4pLL-the sequences from C-terminus of lamprey variable lymphocyte re-ceptor(VLR)-B antibodies were fused to C-terminus of HA ectodomain,to obtain HA mutant genes BY-T,BY-LLc,and BY-PLc.They were inserted into pFastBac1 separately and expressed using baculovirus expression vector system.After expression identification,Strep-Tactin affinity chromatography purification was performed to purify the mutant proteins followed by identification of the degree of oligomerization and hemagglutination activity of the purified mutant proteins.Anti-HA IgG titer and hemagglutination inhibition(HAI)antibody titer in sera of mice immunized with HA mutants were detected.Results:All HA mutants were effectively expressed.The BY-T mutant had the highest expression level,was easy to purify,had good purity,and existed in a polymeric form.The expression levels of BY-LLc and BY-PLc are secondary and exist in a highly polymerization form.The hemagglutination activities of the three HA mutant proteins were comparable,and all of them could effectively induce anti-HA IgG and HAI antibodies.Conclusion:The HA protein ectodomain mutant fused with oligomeric motif GCN4pLL at the C-terminus is easy to purify and has hemagglutination activity,which can induce anti-HA IgG and HAI antibodies in mice.The results provide a reference for the development strategy of recombinant protein vaccines against influenza virus B/Yamagata lineage.
B/YamagataHemagglutinin(HA)Recombinant protein vaccines
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