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-like particles will be designed using recombinants that simultaneously express the env, gag, and pol genes of HIV. HIV-like particles will be expressed in large-scale microcarrier cultures of Vero cells 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 gp120 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 the treatment of AIDS as well as a safe alternative to the use of whole inactivated HIV as a vaccine for AIDS.
