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Validation of an automated system for aliquoting of HIV-1 Env-pseudotyped virus stocks

: Schultz, Anke; Germann, Anja; Fuss, Martina; Sarzotti-Kelsoe, Marcella; Ozaki, Daniel; Montefiori, David; Zimmermann, Heiko; Briesen, Hagen von

Fulltext urn:nbn:de:0011-n-4842722 (3.9 MByte PDF)
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Created on: 13.2.2018

PLoS one. Online journal 13 (2018), No.1, Art. e0190669, 20 pp.
ISSN: 1932-6203
Journal Article, Electronic Publication
Fraunhofer IBMT ()

The standardized assessments of HIV-specific immune responses are of main interest in the preclinical and clinical stage of HIV-1 vaccine development. In this regard, HIV-1 Envpseudotyped viruses play a central role for the evaluation of neutralizing antibody profiles and are produced according to Good Clinical Laboratory Practice- (GCLP-) compliant manual and automated procedures. To further improve and complete the automated production cycle an automated system for aliquoting HIV-1 pseudovirus stocks has been implemented. The automation platform consists of a modified Tecan-based system including a robot platform for handling racks containing 48 cryovials, a Decapper, a tubing pump and a safety device consisting of ultrasound sensors for online liquid level detection of each individual cryovial. With the aim to aliquot the HIV-1 pseudoviruses in an automated manner under GCLP-compliant conditions a validation plan was developed where the acceptance criteria Ðaccuracy, precision as well as the specificity and robustnessÐwere defined and summarized. By passing the validation experiments described in this article the automated system for aliquoting has been successfully validated. This allows the standardized and operator independent distribution of small-scale and bulk amounts of HIV-1 pseudovirus stocks with a precise and reproducible outcome to support upcoming clinical vaccine trials.