Use of triazine dyes in the affinity chromatographic purification of alkaline phosphatase from calf intestine.

Dye-ligand chromatography was examined as a method for the purification of alkaline phosphatase (EC.3.1.3.1). Forty six dye-Matrex Gels were assessed for their ability to bind alkaline phosphatase. Most dye adsorbents bound significant quantities of the enzyme. Three dye columns were examined in more detail for their selectivity using gradients of potassium chloride to desorb enzyme protein. Purification of alkaline phosphatase using Cibacron blue 3GA-Sepharose 6B chromatography was enhanced by using affinity elution. The best purifications (290-fold) were obtained using pulsed elution with the substrate alpha-naphthyl phosphate although the inhibitor, inorganic phosphate, was also useful (128-fold purification).