HIV Research
HIV Isolation and Cell Culture The VitalVirus HIV Isolation Kits have been developed to isolate viable and infectious HIV-1 from clinical samples. Once isolated, the HIV Infectivity Enhancement Reagent enables the efficient and rapid expansion of HIV isolates in PBMC or hematopoeitic cell lines. VitalVirus HIV Isolation Kits Research with intact HIV-1 virions from plasma, serum, or other bodily fluids without extensive sample processing Rapid magnetic virus isolation with μMACS™ VitalVirus HIV Isolation Kit Viable and infectious HIV-1 for downstream studies such as viral genotyping, viral phenotyping, neutralization studies or development of primary isolates. The envelope of human immunodeficiency virus 1 (HIV-1) contains not only virus-encoded proteins, but also host cell proteins1, 2, 3. It has been determined that CD44 is the most effective host cell marker for the general labeling and capture of HIV-1 from patient samples and culture-derived virus, independent of the origin of the virus (lymphoid or myeloid cells).4 Accordingly, the μMACS VitalVirus HIV Isolation Kit contains CD44 MicroBeads for magnetic labeling of HIV virions (see figure 1). These MicroBeads are coupled to a monoclonal antibody that recognises the CD44H isoform present on hematopoietic cells. Gentle magnetic isolation. Once the virions are magnetically labeled with CD44 MicroBeads, MACS® Column Technology enables a very gentle isolation to yield viable and infectious HIV-1 for downstream research applications such as Enrichment of live and infectious HIV-1 (see figure 2) for downstream studies, including neutralization studies, and the development of primary isolates. Enrichment of infectious HIV-1 in the presence of neutralizing antibodies or other uncharacterized HIV serum inhibitors. Enrichment of infectious HIV-1 for drug resistance phenotyping Enrichment and detection of HIV-1 from normal, dilute, and/or difficult samples, for example, cerebral spinal fluid, cervical lavage, or breast milk. Enrichment of HIV-1 for direct RT-PCR and genotyping 5 Optimized PBMC stimulation protocol. The use of the 3x3 Stimulation Protocol for peripheral blood mononuclear cells (PBMCs) in combination with the µMACS™ VitalVirus HIV Isolation Kit greatly increases the success rate of the isolation of infectious HIV-16. Alternative markers for indirect HIV-1 isolation. It is also possible to isolate virus via other host cell surface molecules that are incorporated into the HIV-1 membrane using biotinylated monoclonal antibodies and the μMACS Streptavidin Kit.4 The special protocol "Isolation of infectious HIV-1", describes the indirect method of magnetic virus isolation for viral compartmentalization studies. HIV Infectivity Enhancement Reagent The HIV Infectivity Enhancement Reagent is a unique tool to simplify and accelerate HIV cell culture procedures. A 30 minute coincubation of the viral supernatant and reagent before addition to the target cells is all that is required for the following benefits: More infected target cells. Up to 100-fold more target cells are infected in the presence of the reagent. Increase in integration events. Up to 100-fold more proviral copies are inserted into the host cell genome. Accelerated kinetics of infection. Reduce culture times to viral harvest by 50%. Increased viral yields. Up to 100-fold more virus is present in the cell culture supernatant. The HIV Infectivity Enhancement Reagent uses the presence of host proteins in both the virus envelope and the target cells to tether HIV on the surface of target lymphocytes and macrophages 7. Further information Isolation of infectious HIV-1 [PDF; 105,6 KB] MACS&more 8-1 (2004) [PDF; 1,9 MB] VitalVirus HIV isolation flyer [PDF; 118,5 KB] Isolation of infectious HIV using stimulated PBMCs [PDF; 162,3 KB]		Figure 1 MACS® MicroBeads for HIV-1 isolation bind specifically to the host cell protein CD44, which is incorporated in the virus envelope4. Figure 2 HIV-1 isolation procedure

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