Molecular template formation on serum albumin under a high-pressure condition

A method of the formation of a molecular template on bovine serum albumin ( BSA) under a high-pressure condition has been developed. The BSA-adsorbed a minopropylsilica gel (APS-BSA) was suspended with cholestenone (4-cholesten -3-one) as a template molecule in aqueous ethanol, and the suspension was l eft standing under 200-800 MPa for 4-12 hrs. The solid residue was thorough ly washed with THF and ethyl acetate to extract cholestenone. The "template d"-APS-BSA gel selectively adsorbed cholestenone as the substrate in the su spended solution, including cholestenone. Also, the gel was used as a liqui d-chromatographic packing material to examine its substrate- or structure-s electivity for some steroid compounds. A large-capacity factor for choleste none was observed when an aqueous methanol solution was used as a mobile ph ase. This shows that the templated-APS-BSA gel has an appreciable molecular -recognition ability, and can be used as a selective adsorbent for a chroma tographic separation. The selectivity has persisted for several months afte r intermittent applications. Furthermore, the templated-APS-BSA gel was ana lyzed using an IAsys plus, bio-molecule interaction analysis system by a re sonant-mirror detector, to observe the binding amounts of the substrate. On ly cholestenone showed an extremely higher value of the binding efficiency with the gel compared to any other steroid compounds tested. It has thus be en shown that high pressure can be a parameter for molecular template forma tion on BSA, which means that a packing material with a highly selective mo lecular recognition site could be conveniently produced.