Validation of an HIV-1 inactivation protocol that is compatible with intracellular drug analysis by mass spectrometry.
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.
van Kampen
Jeroen J A
JJ
Department of Neurology, Laboratory of Neuro-Oncology, Erasmus MC, Rotterdam, The Netherlands.
Verschuren
Esther J
EJ
Burgers
Peter C
PC
Luider
Theo M
TM
de Groot
Ronald
R
Osterhaus
Albert D M E
AD
Gruters
Rob A
RA
D019380:Anti-HIV Agents / Q000032:analysis* / Q000494:pharmacology; D002457:Cell Extracts / Q000737:chemistry; D002478:Cells, Cultured / Q000821:virology; D000431:Ethanol / Q000737:chemistry; D015497:HIV-1 / Q000187:drug effects* / Q000302:isolation & purification; D006801:Humans; D000432:Methanol / Q000737:chemistry; D019438:Ritonavir / Q000032:analysis* / Q000494:pharmacology; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D038301:Virus Inactivation
D019380:Anti-HIV Agents; D002457:Cell Extracts; D000431:Ethanol; D019438:Ritonavir; D000432:Methanol