The Preparation Process And Safety Evaluation Research Of H1/H5 Subtype Influenza Recombinant Adenovirus Vaccine

Objective: Established for H1, multi-epitope H5 subtype influenza virus adenovirus purification process of the candidate vaccine pac Ad5-H5HA-EHA-H1HA1 new and to evaluate its immunogenicity and acute toxicity.Methods: Using Source 30 Q as chromatography media and Sepharose 4FF as molecular sieve chromatography, purified the recombinant adenovirus pac Ad5-H5HA-EHA-H1HA1 new and pac Ad5-H5 HA respectively with two-step method purified, and identified the purified adenovirus in this research.Finally, evaluated the recombinant adenovirus’ s acute toxicity by mice,which meet the requirements of the recombinant adenovirus immunogenicity assessment.Results: Using Source 30 Q as chromatography media and Sepharose 4FF as molecular sieve chromatography, purified the recombinant adenovirus pac Ad5-H5HA-EHA-H1HA1 new and pac Ad5-H5 HA respectively with two-step method purified,the overall recovery rate is 41%. PCR identification results show that the target gene: H5 HA and H1HA1 are not lost. Compared with the unpurified recombinant adenovirus, as the immunogenicity assay results showed,after immunized mice,purified recombinant adenovirus pac Ad5-H5-EHA-H1HA1 new and pac Ad5-H5 HA could induce higher levels responses of cellular immunity and humoral immune. Acute toxicity test results show that purified recombinant adenovirus vaccine candidates in mice had no toxic effect.Conclusion: Purified recombinant adenovirus pac Ad5-H5HA-EHA-H1HA1 new has improved immunogenicity and immune mice, no acute toxicity,and better security.