Separation of Tissue and Serum Acid Phosphatase lsoenzymes by Ion-Exchange Column Chromatography

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Abstract
I describe a simple, rapid ion-exchange column-chroma-tographic technique for separating the acid phosphatase (EC 3.1.3.2) isoenzymes in human serum and tissue. Ex¬tracts of platelets, spleen, liver, erythrocytes, and prostate were used to determine optimum conditions for separating these isoenzymes. Samples layered on mini-columns of DEAE-Sephadex A-50 were eluted stepwise with sodium chloride (100, 200, and 300 mmol/liter, buffered with tris(hydroxymethyl)aminomethane). Activity in column effluents was measured with p-nitrophenol phosphate as substrate, and their isoenzyme content was assessed by electrophoresis on polyacrylamide gel. Comparison of activity patterns so derived for various tissues revealed prostatic tissue to be a rich source of acid phosphatase isoenzyme 2 activity. Evaluation of sera from six patients with prostatic cancer revealed isoenzyme patterns with prominent amounts of isoenzyme 2 (3.8 to 27.6 U/liter). Sera from 10 healthy laboratory technicians contained isoenzyme 2 in the range of 0.3-0.5 U/liter. Samples from two patients with abnormally high activity owing to non¬prostatic conditions (Gaucher's disease and carcinoma of lung) exhibited less than 2 U of isoenzyme 2 per liter and acid phosphatase isoenzymes 3-5 that were 50- to 100-fold the normal range. Quantification of isoenzyme 2 by DEAE-Sephadex column chromatography as de¬scribed appears to provide a more sensitive and specific approach to diagnosis of prostatic cancer.

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