Effect of Fetal Bovine Serum on Foamy and Lentiviral Vector Production

Powers, John M.; Trobridge, Grant D.

doi:10.1089/hgtb.2013.097

Cited by 1 publication

(1 citation statement)

References 18 publications

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“…Of note, distinct batches of sera from the same supplier may yield different LV production capacity, as previously described. 24 Therefore, we advise testing serum batches before committing on large-scale LV production. We measured the infectious titer, physical particles, and infectivity of LV-containing supernatant from independent productions.…”

Section: Resultsmentioning

confidence: 99%

Laboratory-Scale Lentiviral Vector Production and Purification for Enhanced Ex Vivo and In Vivo Genetic Engineering

Soldi

Sergi

Unali

et al. 2020

Molecular Therapy - Methods & Clinical Development

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Lentiviral vectors (LVs) are increasingly employed in gene and cell therapy. Standard laboratory production of LVs is not easily scalable, and research-grade LVs often contain contaminants that can interfere with downstream applications. Moreover, purified LV production pipelines have been developed mainly for costly, large-scale, clinical-grade settings. Therefore, a standardized and cost-effective process is still needed to obtain efficient, reproducible, and properly executed experimental studies and preclinical development of ex vivo and in vivo gene therapies, as high infectivity and limited adverse reactions are important factors potentially influencing experimental outcomes also in preclinical settings. We describe here an optimized laboratory-scale workflow whereby an LV-containing supernatant is purified and concentrated by sequential chromatographic steps, obtaining biologically active LVs with an infectious titer and specific activity in the order of 10 9 transducing unit (TU)/mL and 5 × 10 4 TU/ng of HIV Gag p24, respectively. The purification workflow removes >99% of the starting plasmid, DNA, and protein impurities, resulting in higher gene transfer and editing efficiency in severe combined immunodeficiency (SCID)-repopulating hematopoietic stem and progenitor cells (HSPCs) ex vivo , as well as reduced activation of inflammatory responses ex vivo and in vivo as compared to TU-matched, laboratory-grade vectors. Our results highlight the value of accessible purified LV production for experimental studies and preclinical testing.

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Section: Resultsmentioning

confidence: 99%