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Affinity capture of a biotinylated retrovirus on macroporous monolithic adsorbents: Towards a rapid single‐step purification process

Affinity capture of a biotinylated retrovirus on macroporous monolithic adsorbents: Towards a... A streptavidin derivitised macroporous monolith was developed to enable single‐step capture of chemically biotinylated Moloney Murine Leukaemia Virus (MoMuLV) from crude, unclarified cell culture supernatant. Monoliths were prepared by aqueous cryopolymerisation of acrylamide with N,N″‐methylene‐bis (acrylamide) and glycidyl methacrylate (Arvidsson et al. (2003) J Chrom A 986:275–290). Streptavidin was immobilised to the epoxy functionalised monoliths. Particulate‐containing cell culture supernatant was passed through the monolith without preclarification of the feedstock and adsorption capacities of 2 × 105 cfu/ml of adsorbent were demonstrated (cf. Fractogel streptavidin, at 3.9 × 105 cfu/ml of adsorbent). The specific titre of the recovered fraction was increased by 425‐fold; however, recoveries of less than 8% were achieved. Adsorption of nonbiotinylated MoMuLV on the streptavidin‐coated monolith was not observed. © 2005 Wiley Periodicals, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology and Bioengineering Wiley

Affinity capture of a biotinylated retrovirus on macroporous monolithic adsorbents: Towards a rapid single‐step purification process

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References (10)

Publisher
Wiley
Copyright
Copyright © 2005 Wiley Periodicals, Inc., A Wiley Company
ISSN
0006-3592
eISSN
1097-0290
DOI
10.1002/bit.20382
pmid
15643627
Publisher site
See Article on Publisher Site

Abstract

A streptavidin derivitised macroporous monolith was developed to enable single‐step capture of chemically biotinylated Moloney Murine Leukaemia Virus (MoMuLV) from crude, unclarified cell culture supernatant. Monoliths were prepared by aqueous cryopolymerisation of acrylamide with N,N″‐methylene‐bis (acrylamide) and glycidyl methacrylate (Arvidsson et al. (2003) J Chrom A 986:275–290). Streptavidin was immobilised to the epoxy functionalised monoliths. Particulate‐containing cell culture supernatant was passed through the monolith without preclarification of the feedstock and adsorption capacities of 2 × 105 cfu/ml of adsorbent were demonstrated (cf. Fractogel streptavidin, at 3.9 × 105 cfu/ml of adsorbent). The specific titre of the recovered fraction was increased by 425‐fold; however, recoveries of less than 8% were achieved. Adsorption of nonbiotinylated MoMuLV on the streptavidin‐coated monolith was not observed. © 2005 Wiley Periodicals, Inc.

Journal

Biotechnology and BioengineeringWiley

Published: Mar 30, 2005

Keywords: affinity chromatography; monolith; Moloney Murine leukaemia virus (MoMuLV); streptavidin; biotin

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