μMACS™ VitalVirus HIV Isolation Kit
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 load analysis, 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, for example, CD44. These host cell proteins are incorporated either actively or passively when the virus buds from the cell membrane. Many of the cellular proteins present in the HIV envelope retain their biological function, suggesting that they could play a role in viral pathogenesis.1 In addition, the presence of certain host cell type-specific antigens in the viral envelope serve as markers of the cellular origin of the virus particle.3 The μMACS™ VitalVirus HIV Isolation Kit enables the rapid and efficient magnetic isolation of infectious HIV-1 virions from culture-derived HIV-1, human plasma or serum, and other bodily fluids, for example, cerebral spinal fluid, breast milk, or cervical lavage. 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-1. Efficient labeling. 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 like - 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. 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. To isolate virus originating specifically from lymphoid cells (T cell-derived HIV) the CD26 surface marker can be used; while CD36 would be the marker of choice to isolate virus of myeloid origin (macrophage-derived HIV-1). The special protocol "Isolation of infectious HIV-1", available at www.miltenyibiotec.com, describes the indirect method of magnetic virus isolation for viral compartmentalization studies. Enrichment of HIV-infected cells In order to study the impact of HIV infection on normal cell function, it is necessary to isolate a pure population of HIV-infected cells. Cells infected with HIV express HIV-transmembrane proteins such as gp415 which provide a target on the cell surface for immunomagnetic cell sorting. Monoclonal antibodies specific for gp41 can be used to label a cell population that contains HIV-infected cells. The cells can then be labeled with indirect MACS® MicroBeads and the HIV-infected cell population enriched using a suitable cell separation column.6 The enriched cell fraction can be studied in downstream applications such as: - Isolation of HIV-infected cells for gene expression studies - Study of HIV-infected cells isolated from bystander effects Columns μ Columns (included in kit) - for virion isolation Cell Separation Columns - for isolation of virus-infected cells 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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