Abstract

A process of column chromatography of human erythrocyte acetylcholinesterase, using a cationic exchange resin, was investigated as a possible method for the primary partial purification of the enzyme on a large scale. The investigation was based, first; on a detailed study of the enzymatic stability to variation in hydrogen-ion and salt concentrations for periods of time extending to forty eight hours and, second; on an investigation of the degree of adsorption by the resin of the enzyme at various pH values within the pH range of enzyme stability. A method for the analysis of the enzyme activity of large numbers of samples, without the sacrifice of accuracy, was devised and described in which enzyme is buffered at the optimum pH for maximum activity. Chromatography of human erythrocyte hemolysate on columns of Amberlite IRC-50 at pH 7.0 was found to yield two separate fractions of acetylcholinesterase with a purification of 20 to 30 fold of the larger fraction. A 3 to 4 fold purification was obtained on partially purified cholinesterase extracts with two fractions also being obtained. This purification process was found to be readily reproducible and applicable on a large scale with a minimum of material, equipment, and labor.

Degree

College and Department

Physical and Mathematical Sciences; Chemistry and Biochemistry

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