Research On The Production Process Of The HIV-1 Recombinant Vaccines

The safe and effective human immunodeficiency virus type 1(HIV-1)vaccine is expected to have the greatest positive impact on ultimate elimination of the AIDS pandemic.Despite decades of efforts,effective vaccine against HIV-1 still remains elusive.In the past years,only RV144 Thai trial reduced HIV-1 acquisition by 31%,but cannot delay disease progression in vaccinated individuals.On the basis of the "prime-boost" vaccination strategy of RV144 experiment,we introduced a virus-like particle(VLP)protein capable of forming a natural trimer as a new vaccine and established the mature DNA,MVA and VLP vaccine production process to apply in the DNA prime-MVA/VLP boost immunization strategy to strengthen immunity.In this study,the immunogen design of vaccine is based on the major epidemic strain B/C recombinant subtype which has a high incidence of AIDS in China.Among them,the recombinant DNA vaccine contains HIV-1 B/C subtype Gag/Pol and Env gene,and we explored bacteria growth conditions and depicted the growth curve by orthogonal experiment,and then scaled up to 5L fermenter to explore the production process and determined the best harvest time of bacteria,and then gained large quantities of purified DNA vaccine plasmid.CEF cells infected with recombinant MVA vaccine can secrete the HIV-1 B/C subtype VLP,and we established the purification method of amplifying a large number of the recombinant virus,repeating freezing and thawing and sonication to release viruses from CEF cells,and using sucrose ultracentrifugation to collect viruses.As a byproduct of the recombinant MVA vaccine,the recombinant VLP vaccine was secreted into the supernatant of CEF cells infected with recombinant MVA vaccine,and the supernatant was collected to produce the recombinant VLP vaccine by concentrating and ultracentrifugation.The DNA prime-MVA/VLP boost immunization strategy is hopping to be used in BALB/c mice experiments.