SBIR-STTR Award

A process for the production and purification of non-infectious Human Immunodeficiency Virus-like particles from manunalian cells infected with recombinant vaccinia viruses may be developed. A procedure to produce milligram quantities of HIV-like particles will be designed using vaccinia recombinants that simultaneously express the env, gag, and pol genes of HIV. A systematic evaluation of several marmnalian cell lines and various culture conditions will be undertaken to maximize the yield of virus-like particles. In addition, processes will be developed for the characterization and purification of these particles consistent with their use as vaccine inimunogens in animal trials. The ability to produce conunercial quantities of non-infectious HIV particles, by these processes, will provide a safer alternative to the use of whole inactivated HIV as a vaccine for AIDS.Awardee's statement of the potential commercial applications of the research:If commercial quantities of non-infectious HIV particles can be produced using our vaccinia vector system, they will provide a safer alternative to the use of whole inactivated HIV as a vaccine for AIDS. In addition, vaccinia produced noninfectious HIV particles may provide an extremely safe and efficient antigen for boosting immune responses in individuals previously vaccinated with different HIV vaccines (e. g. recombinant vaccinia virus, subunit preparations) or as an immunogenic stimulant for therapeutic purposes in HIV infected individuals.National Institute of Allergy and Infectious Diseases (NIAID)

A process for the production and purification of non-infectious Human Immunodeficiency Virus-like particles produced after infection of mammalian cells with recombinant pox viruses, will be developed and scaled-up. A procedure to produce gram quantities of HIV-Iike particles will be designed using recombinants that simultaneously express the env, gag, and pol genes of HIV. HIV-Iike particles will be expressed in large-scale microcarrier cultures of Verocelis infected with the recombinant virus. Methods will be developed to process and purify the particles from large volumes of medium. Among the methods to be investigated and scaled-up are: (1) clarification by filtration and/or continuous flow centrifugation, and (2) purification by large-scale sucrose density gradient centrifugation and/or column chromatography. In addition, HPLC methodology and gpl2O ELISA will be developed to further characterize the particles. The immunogenicity of these non-infectious particles will be studied in conjunction with several adjuvant formulations in rodents. The ability to produce commercial quantities of non-infectious HIV particles by these processes will provide a completely safe immunotherapeutic for AIDS treatment as well as a safe alternative to the use of whole inactivated HIV as a vaccine for AIDS.Awardee's statement of the potential commercial applications of the research:If commercial quantities of non-infectious HIV particles can be produced using our vaccinia vector system they will provide a completely safe alternative to the use of whole inactivated HIV as a vaccine for AIDS. In addition, non-infectious HIV particles may provide an extremely safe and efficient antigen for boosting immune responses in individuals previous vaccinated with different HIV vaccines (e.g. recombinant vaccinia virus, subunit preparations) or as an immunogenetic stimulant for therapeutic purposes in HIV infected individuals.National Institute of Allergy and Infectious Diseases (NIAID)