Alkaline Phosphatase Purification: A Comparison of Prep-PAGE with Cyclic Processes

Publisher Website
Google Scholar
Abstract
Enzyme purification has been investigated in a preparative-scale polyacrylamide gel electrophoresis system, the Buchler Poly-Prep 200. Experimental results are presented for the optimization of this process using human placental alkaline phosphatase as a typical enzyme. The variables considered are electric field strength, feed concentration, buffer ionic strength, and buffer pH. Enzymes may also be purified by adsorption/desorption onto an ion exchange resin such as DEAE Sepharose in either of two cyclic processes-parametric pumping or cycling zone adsorption. Comparison of the three processes indicates that polyacrylamide gel electrophoresis has the highest purification factor and the greatest enzyme activity recovered, but also the lowest rate of production.