Profiling of HIV Proteins in Cerebrospinal Fluid

HIV-1 proteins are rarely identified during mass spectrometry-based proteomic profiling studies of body fluids from HIV-1-infected people even when elaborated fractionation schema and highly sensitive instruments are used. Genotyping of HIV-1 isolated from body fluids does not provide exact information about �characteristics of circulating proteins and is a limiting factor in expanding an important segment of our knowledge about the course of infection. Therefore, we propose that in vitro amplification of freshly �isolated virus followed by sucrose cushion purification will yield sufficient amounts of viral proteins for mass spectrometric characterization. This chapter provides protocols for virus propagation using CD4+ T cell line or human macrophages, virus purification, and preparation of samples for two-dimensional electrophoresis and mass spectrometry analyses.