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pp 1-19 | Cite as

High-Grade Purification of Third-Generation HIV-Based Lentiviral Vectors by Anion Exchange Chromatography for Experimental Gene and Stem Cell Therapy Applications

  • Hazal Banu Olgun
  • Hale M. Tasyurek
  • Ahter D. Sanlioglu
  • Salih Sanlioglu
Protocol
Part of the Methods in Molecular Biology book series

Abstract

Lentiviral vectors (LVs) have been increasingly used in clinical gene therapy applications particularly due to their efficient gene transfer ability, lack of interference from preexisting viral immunity, and long-term gene expression they provide. Purity of LVs is essential in in vivo applications, for a high therapeutic benefit with minimum toxicity. Accordingly, laboratory scale production of LVs frequently involves transient cotransfection of 293T cells with packaging and transfer plasmids in the presence of CaPO4. After clearance of the cellular debris by low-speed centrifugation and filtration, lentivectors are usually concentrated by high-speed ultracentrifugation in sucrose cushion. Concentrated viral samples are then purified by anion exchange chromatography (AEX) after benzonase treatment to remove the residual cellular DNA. Here, we describe an improved practical method for LV purification using AEX, useful for experimental studies concerning gene and stem cell therapy.

Keywords

Anion exchange chromatography Gene and cell therapy Lentivirus 

Notes

Acknowledgments

This study is supported by grants from Akdeniz University Scientific Research Administration Division (TYL-2015-1027) and the Scientific and Technological Research Council of Turkey (TUBITAK-112S114).

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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Hazal Banu Olgun
    • 1
  • Hale M. Tasyurek
    • 1
  • Ahter D. Sanlioglu
    • 1
  • Salih Sanlioglu
    • 1
  1. 1.Human Gene and Cell Therapy Center of Akdeniz University Hospitals and ClinicsAntalyaTurkey

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