An automated HIV-1 env-pseudotyped virus production for global HIV vaccine trials

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• dc.contributor.author Meyerhans, Andreasca
• dc.contributor.author Schultz, Ankeca
• dc.contributor.author Koch, Stefanieca
• dc.contributor.author Fuss, Martinaca
• dc.contributor.author Mazzotta, Angela S.ca
• dc.contributor.author Sarzotti-Kelsoe, Marcellaca
• dc.contributor.author Ozaki, Daniel A.ca
• dc.contributor.author Montefiori, David Cca
• dc.contributor.author Briesen, Hagen vonca
• dc.contributor.author Zimmermann, Heikoca
• dc.date.accessioned 2015-05-13T08:08:03Z
• dc.date.available 2015-05-13T08:08:03Z
• dc.date.issued 2012ca
• dc.description.abstract Background: Infections with HIV still represent a major human health problem worldwide and a vaccine is the only long-term option to fight efficiently against this virus. Standardized assessments of HIV-specific immune responses in vaccine trials are essential for prioritizing vaccine candidates in preclinical and clinical stages of development. With respect to neutralizing antibodies, assays with HIV-1 Env-pseudotyped viruses are a high priority. To cover the increasing demands of HIV pseudoviruses, a complete cell culture and transfection automation system has been developed. Methodology/Principal Findings: The automation system for HIV pseudovirus production comprises a modified Tecan-based Cellerity system. It covers an area of 5×3 meters and includes a robot platform, a cell counting machine, a CO2 incubator for cell cultivation and a media refrigerator. The processes for cell handling, transfection and pseudovirus production have been implemented according to manual standard operating procedures and are controlled and scheduled autonomously by the system. The system is housed in a biosafety level II cabinet that guarantees protection of personnel, environment and the product. HIV pseudovirus stocks in a scale from 140 ml to 1000 ml have been produced on the automated system. Parallel manual production of HIV pseudoviruses and comparisons (bridging assays) confirmed that the automated produced pseudoviruses were of equivalent quality as those produced manually. In addition, the automated method was fully validated according to Good Clinical Laboratory Practice (GCLP) guidelines, including the validation parameters accuracy, precision, robustness and specificity. Conclusions: An automated HIV pseudovirus production system has been successfully established. It allows the high quality production of HIV pseudoviruses under GCLP conditions. In its present form, the installed module enables the production of 1000 ml of virus-containing cell culture supernatant per week. Thus, this novel automation facilitates standardized large-scale productions of HIV pseudoviruses for ongoing and upcoming HIV vaccine trials.en
• dc.description.sponsorship This article derived from a collaboration of the Fraunhofer Institute for Biomedical Engineering, the ‘‘Global HIV Vaccine Research Cryorepository Consortium’’ (principal investigator: HVB) and the ‘‘Comprehensive Antibody Vaccine Immune Monitoring Consortium’’ (principal investigator: DM) within the‘‘Collaboration for AIDS Vaccine Discovery’’, funded by the Bill and Melinda Gates Foundation (#OPP38580_01). Automation of cell culture and HIV pseudovirus production was established in exclusive cooperation with TECAN funded as a strategic investment by the Fraunhofer Society, Germany.en
• dc.format.mimetype application/pdfca
• dc.identifier.citation Schultz A, Koch S, Fuss M, Mazzotta AS, Sarzotti-Kelsoe M, Ozaki DA et al. An automated HIV-1 env-pseudotyped virus production for global HIV vaccine trials. PLoS ONE. 2012;7(12):e51715. DOI: 10.1371/journal.pone.0051715ca
• dc.identifier.doi http://dx.doi.org/10.1371/journal.pone.0051715
• dc.identifier.issn 1932-6203ca
• dc.identifier.uri http://hdl.handle.net/10230/23563
• dc.language.iso engca
• dc.publisher Public Library of Science (PLoS)ca
• dc.relation.ispartof PLoS ONE. 2012;7(12):e51715
• dc.rights © 2012 Schultz et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are crediteden
• dc.rights.accessRights info:eu-repo/semantics/openAccessca
• dc.subject.other VIH (Virus) -- Transmissióca
• dc.title An automated HIV-1 env-pseudotyped virus production for global HIV vaccine trialsen
• dc.type info:eu-repo/semantics/articleca
• dc.type.version info:eu-repo/semantics/publishedVersionca