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

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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.

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