Microwave-assisted preparation of affinity medium

Abstract

Microwave assistance was used for preparing polyethylene glycol (PEG)–Cibacron blue 3GA and Sepharose CL-4B–Cibacron blue 3GA affinity materials. The former was used as the affinity macroligand in a PEG–dextran aqueous two-phase system for purification of alcohol dehydrogenase and EcoRI. The Sepharose CL-4B–Cibacron blue 3GA was used for affinity chromatography of the above two enzymes. It was found that microwave assistance could reduce the time of PEG–dye preparation to 5 min (from 7 h). Similarly, Sepharose CL-4B–Cibacron blue 3GA preparation time could be reduced to 21 min (from 3.5 h). The performances of affinity macroligand PEG–dye and the affinity medium Sepharose–dye prepared by conventional methods and with microwave assistance were similar during purification of these enzymes.