Microwave-assisted preparation of affinity medium

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 7h). Similarly, Sepharose CL-4B-Cibacron blue 3GA preparation time could be reduced to 21 min (from 3.5h). 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.