A rapid spin through oil results in higher cell-associated concentrations of antiretrovirals compared with conventional cell washing

Bioanalysis. 2015;7(12):1447-55. doi: 10.4155/bio.15.70.

Abstract

Background: Determination of cell-associated antiretroviral drug concentrations is necessary for research into reservoirs of HIV. Variation exists in cell-associated drug concentrations among research groups. One cause for this may be washing cells during processing. We explored spinning cells through oil to minimize this variability.

Methods & results: Raltegravir, atazanavir, darunavir, efavirenz, lopinavir and ritonavir concentrations were assessed in CEM.ss T cells washed with HBSS and oil-spun cells. Oil-spun cells had significantly higher concentrations for all drugs compared with samples washed with HBSS.

Conclusion: The decline in cell-associated drug concentrations with saline washes compared with a single spin through oil shows the utility of a spin through oil. Oil centrifugation results in high cell-associated drug concentrations, and can be done in a fast, efficient manner.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Anti-Retroviral Agents / analysis
  • Anti-Retroviral Agents / chemistry*
  • Cell Line
  • Chromatography, High Pressure Liquid
  • HIV Protease Inhibitors / analysis
  • HIV Protease Inhibitors / chemistry
  • Humans
  • Isotonic Solutions / chemistry
  • Isotope Labeling
  • Mass Spectrometry
  • Oils / chemistry*

Substances

  • Anti-Retroviral Agents
  • HIV Protease Inhibitors
  • Hanks Balanced Salt Solution
  • Isotonic Solutions
  • Oils