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| HIV Isolation and Propagation |
| 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 propagation of HIV isolates in PBMC or hematopoietic cell lines. |
| Benefits |
Easy - Isolate intact HIV-1 virions from plasma, serum, or other bodily fluids without extensive sample processing
Fast - Isolate HIV-1 in less than 1 hour
Gentle - Obtain viable and highly infectious HIV-1 for downstream studies
Versatile - Start with plasma, serum or bodily fluid such as cerebral spinal fluid, cervical lavage or breast milk
Background. The envelope of human immunodeficiency virus 1 (HIV-1) contains not only virus-encoded proteins, but also host cell proteins5-7. It has been shown 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. The μMACS VitalVirus HIV Isolation Kit contains MicroBeads coupled to a monoclonal antibody specific against CD44H isoform for magnetic labeling and further isolation of HIV virions (see figure 1).
Downstream applications. 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 applications such as
- Development of primary isolates
- Inhibitor screening in the presence of neutralizing antibodies or HIV serum inhibitors
- Drug resistance phenotyping
- Enrichment and detection of HIV-1 from various sample types such as cerebral spinal fluid, cervical lavage or breast milk
- Direct RT-PCR and genotyping 2.
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 Kit4. The special protocol Isolation of infectious HIV-1 describes the indirect method of magnetic virus isolation for viral compartmentalization studies.
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-11. |
| HIV Infectivity Enhancement Reagent |
The HIV Infectivity Enhancement Reagent simplifies and accelerates HIV cell culture procedures. Simply incubate the reagent with the viral supernatant 30 min before addition to the target cells.
Efficient - Up to 100-fold more target cells are infected.
High integration - Up to 100-fold more proviral copies are inserted into the host cell genome.
Fast - Reduce culture times to viral harvest by 50%.
High yield - Up to 100-fold more virus is present in the cell culture supernatant. |
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| Figure 1 |
| MACS® MicroBeads for HIV-1 isolation bind specifically to the host cell protein CD44, which is incorporated in the virus envelope4. |
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| Figure 2 |
| HIV-1 isolation procedure |
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| Favorites / Prices |
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| Details |
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| Products |
| μMACS VitalVirus HIV Isolation Starting Kit |
- for 20 isolations
Components
- 1 μMACS VitalVirus HIV Isolation Kit
- 1 μMACS Separator
- 1 MACS MultiStand
Download datasheet
130-092-833
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| μMACS VitalVirus HIV Isolation Kit |
- for 20 isolations
Components
- 1 mL CD44 MicroBeads
- 4 mL Equilibration Buffer for protein applications
- 30 mL Virus Wash Buffer
- 6 mL Virus Lysis Buffer
- 20 μ Columns
Download datasheet
130-092-805
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| MultiMACS VitalVirus HIV Isolation Kit (12×8) |
- for 96 isolations
Components
- 5×1 mL CD44 MicroBeads
- 5×4 mL Equilibration Buffer for protein applications
- 5×30 mL Virus Wash Buffer
- 5×6 mL Virus Lysis Buffer
- 12 Multi-8 Columns
- 1 MultiColumn Frame
- 1 Microtiter Plate, U-bottom
- 1 Deep Well Block, 2.5 mL
Download datasheet
130-092-806
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| MultiMACS VitalVirus HIV Isolation Kit (4×96) |
- for 384 isolations
Components
- 4× reagents of (12×8) MultiMACS VitalVirus HIV Isolation Kit
- 4 Multi-96 Columns
- 4 Deep Well Blocks, 2.5 mL
- 4 Microtiter Plates, U-bottom
Download datasheet
130-092-807
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| HIV Infectivity Enhancement Reagent |
- 20 HIV infections
Components
- 1 mL HIV Infectivity Enhancement Reagent
Download datasheet
130-095-093
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| μMACS Streptavidin Starting Kit |
- for 20 isolations
Components
- 1 μMACS Streptavidin Kit
- 1 μMACS Separator
- 1 MACS MultiStand
130-091-287
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| μMACS Streptavidin Kit |
- for 20 isolations
Components
- 2 mL μMACS Streptavidin MicroBeads
- 4 mL Equilibration Buffer for nucleic acids applications
- 4 mL Equilibration Buffer for protein applications
- 20 μ Columns
Download datasheet
130-074-101
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| References |
| 1. Prado et al. (2010) J. Virol. 84(1): 492-502 |
| 2. Piantadosi et al. (2009) J. Virol. 83: 7805-7814 |
| 3. Terry et al. (2009) Virology 388: 294-304 |
| 4. Lupo and Butera (2004) MACS&more 8(1): 16–19. |
| 5. Lawn et al. (2000) J. Virol. 74: 139–145. |
| 6. Tremblay et al. (1998) Immunol. Today 19: 346–351. |
| 7. Ott (1997) Rev. Med. Virol. 7: 167–180. |
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