Optimised concentration and purification of retroviruses using membrane chromatography

D. J. McNally; David Darling; F. Farzaneh; P. R. Levison; N. K. H. Slater

doi:10.1016/j.chroma.2014.03.023

Optimised concentration and purification of retroviruses using membrane chromatography

D. J. McNally^*, David Darling, F. Farzaneh, P. R. Levison, N. K. H. Slater

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

35 Citations (Scopus)

Abstract

The ability of an anion exchange membrane to purify a gamma-retrovirus was assessed and optimised with respect to different loading and wash buffers. Recoveries of infectious virus greater than 50% were consistently obtained, while specific titre was increased up to one thousand fold when compared to the material loaded. Specific proteins removed and retained by this optimised process were identified by mass spectrometry. It was possible to successfully bind and elute the equivalent of 1.27 x 10(8) Ifu/ml of ion exchange membrane. This could then be highly concentrated, with infectious virus concentrated to a maximum of 420-fold compared to the load.

Original language	English
Pages (from-to)	24-32
Number of pages	9
Journal	Journal of Chromatography A
Volume	1340
DOIs	https://doi.org/10.1016/j.chroma.2014.03.023
Publication status	Published - 2 May 2014

Keywords

Chromatography
Gene therapy
Virus
Membrane adsorption
Retrovirus
GENE-THERAPY VECTORS
AFFINITY-CHROMATOGRAPHY
LENTIVIRAL VECTORS
EFFICIENT CONCENTRATION
MONOLITHIC ADSORBENTS
HOST PROTEINS
PARTICLES
CAPTURE
DNA

Access to Document

10.1016/j.chroma.2014.03.023

Cite this

@article{f53b5c5e9c21459f8d0435860f182cd8,

title = "Optimised concentration and purification of retroviruses using membrane chromatography",

abstract = "The ability of an anion exchange membrane to purify a gamma-retrovirus was assessed and optimised with respect to different loading and wash buffers. Recoveries of infectious virus greater than 50% were consistently obtained, while specific titre was increased up to one thousand fold when compared to the material loaded. Specific proteins removed and retained by this optimised process were identified by mass spectrometry. It was possible to successfully bind and elute the equivalent of 1.27 x 10(8) Ifu/ml of ion exchange membrane. This could then be highly concentrated, with infectious virus concentrated to a maximum of 420-fold compared to the load.",

keywords = "Chromatography, Gene therapy, Virus, Membrane adsorption, Retrovirus, GENE-THERAPY VECTORS, AFFINITY-CHROMATOGRAPHY, LENTIVIRAL VECTORS, EFFICIENT CONCENTRATION, MONOLITHIC ADSORBENTS, HOST PROTEINS, PARTICLES, CAPTURE, DNA",

author = "McNally, {D. J.} and David Darling and F. Farzaneh and Levison, {P. R.} and Slater, {N. K. H.}",

year = "2014",

month = may,

day = "2",

doi = "10.1016/j.chroma.2014.03.023",

language = "English",

volume = "1340",

pages = "24--32",

journal = "Journal of Chromatography A",

issn = "0021-9673",

publisher = "Elsevier",

}

TY - JOUR

T1 - Optimised concentration and purification of retroviruses using membrane chromatography

AU - McNally, D. J.

AU - Darling, David

AU - Farzaneh, F.

AU - Levison, P. R.

AU - Slater, N. K. H.

PY - 2014/5/2

Y1 - 2014/5/2

N2 - The ability of an anion exchange membrane to purify a gamma-retrovirus was assessed and optimised with respect to different loading and wash buffers. Recoveries of infectious virus greater than 50% were consistently obtained, while specific titre was increased up to one thousand fold when compared to the material loaded. Specific proteins removed and retained by this optimised process were identified by mass spectrometry. It was possible to successfully bind and elute the equivalent of 1.27 x 10(8) Ifu/ml of ion exchange membrane. This could then be highly concentrated, with infectious virus concentrated to a maximum of 420-fold compared to the load.

AB - The ability of an anion exchange membrane to purify a gamma-retrovirus was assessed and optimised with respect to different loading and wash buffers. Recoveries of infectious virus greater than 50% were consistently obtained, while specific titre was increased up to one thousand fold when compared to the material loaded. Specific proteins removed and retained by this optimised process were identified by mass spectrometry. It was possible to successfully bind and elute the equivalent of 1.27 x 10(8) Ifu/ml of ion exchange membrane. This could then be highly concentrated, with infectious virus concentrated to a maximum of 420-fold compared to the load.

KW - Chromatography

KW - Gene therapy

KW - Virus

KW - Membrane adsorption

KW - Retrovirus

KW - GENE-THERAPY VECTORS

KW - AFFINITY-CHROMATOGRAPHY

KW - LENTIVIRAL VECTORS

KW - EFFICIENT CONCENTRATION

KW - MONOLITHIC ADSORBENTS

KW - HOST PROTEINS

KW - PARTICLES

KW - CAPTURE

KW - DNA

U2 - 10.1016/j.chroma.2014.03.023

DO - 10.1016/j.chroma.2014.03.023

M3 - Article

SN - 0021-9673

VL - 1340

SP - 24

EP - 32

JO - Journal of Chromatography A

JF - Journal of Chromatography A

ER -

Optimised concentration and purification of retroviruses using membrane chromatography

Abstract

Keywords

Access to Document

Fingerprint

Cite this