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Affinity capture of a biotinylated retrovirus on macroporous monolithic adsorbents: Towards a rapid single‐step purification process
Author(s) -
Williams Sharon L.,
Eccleston Mark E.,
Slater Nigel K.H.
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20382
Subject(s) - biotinylation , streptavidin , adsorption , glycidyl methacrylate , chromatography , monolith , chemistry , tenax , acrylamide , methacrylate , nuclear chemistry , biotin , polymer , biochemistry , organic chemistry , polymerization , monomer , catalysis
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 × 10 5 cfu/ml of adsorbent were demonstrated (cf. Fractogel streptavidin, at 3.9 × 10 5 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.