Lentiviral Vector Purification Using Nanofiber Ion-Exchange Chromatography

Jelena Ruscic; Christopher Perry; Tarit Mukhopadhyay; Yasu Takeuchi; Daniel G. Bracewell

Molecular Therapy: Methods & Clinical Development (Dec 2019)

Lentiviral Vector Purification Using Nanofiber Ion-Exchange Chromatography

Jelena Ruscic,
Christopher Perry,
Tarit Mukhopadhyay,
Yasu Takeuchi,
Daniel G. Bracewell

Affiliations

Jelena Ruscic: Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK
Christopher Perry: Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK; Division of Infection and Immunology, University College London, The Rayne Building, 5 University Street, London WC1E 6EJ, UK; Advanced Therapies Division, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK
Tarit Mukhopadhyay: Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK
Yasu Takeuchi: Division of Infection and Immunology, University College London, The Rayne Building, 5 University Street, London WC1E 6EJ, UK; Advanced Therapies Division, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK
Daniel G. Bracewell: Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK; Corresponding author: Daniel G. Bracewell, Department of Biochemical Engineering, University College London, Bernard Katz Building, Gower Street, London WC1E 6BT, UK.

Journal volume & issue: Vol. 15
pp. 52 – 62

Abstract

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Lentiviral vectors (LVs) are used in cell and gene therapies due to their ability to transduce both dividing and non-dividing cells while carrying a relatively large genetic payload and providing long-term gene expression via gene integration. Current cultivation methods produce titers of 105–107 transduction unit (TU)/mL; thus, it is necessary to concentrate LVs as well as remove process- and product-related impurities. In this work, we used a packaging cell line WinPac-RD-HV for LV production to simplify upstream processing. A direct capture method based on ion-exchange chromatography and cellulose nanofibers for LV concentration and purification was developed. This novel scalable stationary phase provides a high surface area that is accessible to LV and, therefore, has potential for high-capacity operation compared to traditional bead-based supports. We were able to concentrate LVs 100-fold while achieving a two-log removal of host cell protein and maintaining up to a 90% yield of functional vector.

Published in Molecular Therapy: Methods & Clinical Development

ISSN: 2329-0501 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Cytology
Website: http://www.cell.com/molecular-therapy-family/methods/latest-content

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