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  • Viral Transfer Technology
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Production and concentration of pseudotyped HIV-1-based gene transfer vectors

Gene Therapy volume 7, pages 910–913 (2000)Cite this article

Abstract

Strategies to generate highly concentrated HIV-1 vector pseudotypes involving different envelope (Env) proteins including the vesicular stomatitis virus (VSV) G glycoprotein, the Moloney murine leukemia virus (MLV) 4070A amphotropic Env and the rabies G glycoprotein were established. Virus stocks were prepared by transient transfection using standard cell culture media or serum-free media. Such stocks were concentrated 50- to 300-fold by ultracentrifugation or by ultrafiltration using Centricon Plus-80 units yielding titers of up to 109transducing units per milliliter. There was no loss in titer with any of the pseudotypes tested. Thus, like lentiviral vectors pseudotyped with VSV-G, HIV-1-based vectors pseudotyped with the MLV 4070A amphotropic Env and the rabies G glycoprotein resist inactivation during concentration. This opens up the possibility to generate highly concentrated HIV-1 vector stocks carrying alternative Env proteins on a large scale.

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Acknowledgements

I am grateful to Yasuhiro Takeuchi for providing plasmid ALF. I thank Simon Tang and Hideki Mochizuki for their help during the early phase of this work.

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  1. Developmental and Metabolic Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD

    J Reiser

  2. Louisiana State University Gene Therapy Program, Louisiana State University School of Medicine, New Orleans, LA, USA

    J Reiser

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Reiser, J. Production and concentration of pseudotyped HIV-1-based gene transfer vectors. Gene Ther 7, 910–913 (2000). https://doi.org/10.1038/sj.gt.3301188

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  • DOI: https://doi.org/10.1038/sj.gt.3301188

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