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Journal of Virology, October 2005, p. 13190-13194, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.13190-13194.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Conjugation of Lentivirus to Paramagnetic Particles via Nonviral Proteins Allows Efficient Concentration and Infection of Primary Acute Myeloid Leukemia Cells

Lucas Chan,¹ Darren Nesbeth,¹ Taylor MacKey,¹ Joanna Galea-Lauri,^1, Joop Gäken,¹ Francisco Martin,^2, Mary Collins,² Ghulam Mufti,¹ Farzin Farzaneh,^1* and David Darling¹

King's College London, Department of Haematological and Molecular Medicine, The Rayne Institute, 123 Coldharbour Lane, London SE5 9NU, United Kingdom,¹ Department of Immunology and Molecular Pathology, Windeyer Institute of Medical Sciences, University College London, London, United Kingdom²

Received 11 July 2005/ Accepted 18 July 2005

Nonviral producer cell proteins incorporated into retroviral vector surfaces profoundly influence infectivity and in vivo half-life. We report the purification and concentration of lentiviral vectors using these surface proteins as an efficient gene transduction strategy. Biotinylation of these proteins and streptavidin paramagnetic particle concentration enhances titer 400- to 2,500-fold (to 10⁹ CFU/ml for vesicular stomatitis virus G protein and 5 x 10⁸ for amphotropic murine leukemia virus envelope). This method also uses newly introduced membrane proteins (B7.1 and LNGFR) directed to lentiviral surfaces, allowing up to 17,000-fold concentrations. Particle conjugation of lentivirus allows facile manipulation in vitro, resulting in the transduction of 48 to 94% of human acute myeloid leukemia blasts.

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* Corresponding author. Mailing address: King's College London, Department of Haematological and Molecular Medicine, The Rayne Institute, 123 Coldharbour Lane, London SE5 9NU, United Kingdom. Phone: 44 20 7848 5901. Fax: 44 20 7733 3877. E-mail: farzin.farzaneh{at}kcl.ac.uk.

Present address: R&D Office, Institute of Child Health, Great Ormond Street Hospital, 30 Guilford Street, London, United Kingdom.

Present address: IPB Lopez Neyra, Parque tecnológico de la Salud, Avda del conocimiento S/N, Granada 18100, Spain.

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Journal of Virology, October 2005, p. 13190-13194, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.13190-13194.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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