Validation of an HIV-1 inactivation protocol that is compatible with intracellular drug analysis by mass spectrometry |
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Authors: | van Kampen Jeroen J A Verschuren Esther J Burgers Peter C Luider Theo M de Groot Ronald Osterhaus Albert D M E Gruters Rob A |
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Affiliation: | Department of Neurology, Laboratory of Neuro-Oncology, Erasmus MC, Rotterdam, The Netherlands. |
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Abstract: | Mass spectrometry is a powerful tool for studying the intracellular pharmacokinetics of antiretroviral drugs. However, the biohazard of HIV-1 calls for a safety protocol for such analyses. To this end, we extracted HIV-1 producing cells with methanol or ethanol at 4 degrees C. After extraction, no viral infectivity was detected, as shown by a reduction in infectious titers of more than 6log. In addition, this protocol is compatible with the quantitative analysis of antiretroviral drugs in cell extracts using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. Thus, using this protocol, infectious HIV-1 is inactivated and antiretroviral drugs are extracted from cells in a single step. |
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