title: Purification of HIV-1 virions by subtilisin digestion or CD45 immunoaffinity depletion for biochemical studies.
abstract: The presence of cellular proteins outside and inside retroviruses can indicate the roles they play in viral biology. However, experiments examining retroviruses can be complicated by the contamination of even highly purified virion preparations with nonviral particles (either microvesicles or exosomes). Two useful methods have been developed that can remove contaminating particles from virus stocks to produce highly pure virus preparations. One approach, the subtilisin digestion procedure, enzymatically removes the proteins outside the virions. While this method is well suited for the analysis of the interior proteins in the virions, it removes the extracellular domains of the integral membrane proteins on the virion. To preserve the proteins on the exterior of the virion for biochemical studies, a CD45 immunoaffinity depletion procedure that removes vesicles by capture with antibody-linked microbeads is employed. These methods allow for the isolation of highly purified virion preparations that are suitable for a wide variety of experiments, including the biochemical characterization of cellular proteins both on and in HIV virions, examination of virion/cell interactions, and imaging of virions.
authors.affiliation: AIDS Vaccine Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, MD, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Ott
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D008863:Microspheres; D020860:Subtilisin / Q000378:metabolism*; D014771:Virion / Q000302:isolation & purification*
chemical_list: D017493:Leukocyte Common Antigens; D020860:Subtilisin