A NEW SCALABLE PURIFICATION PROCEDURE FOR PROSTATIC SECRETORY PROTEIN(PSP94) FROM HUMAN SEMINAL PLASMA

A simple three-step procedure for the purification of native prostate secretory protein (PSP94) from human seminal plasma is described. The steps are ammonium sulfate fractionation followed by a Macro-Prep S su pport (cation) flowthrough process and the final Macro-Prep high Q sup port (anion exchange) chromatography using two step-gradient elution. The benefits of this procedure lie in its simplicity, speed, and relat ively inexpensive materials, thus providing advantages over the previo usly reported schemes. The purity of the product as judged by single b and on SDS-polyacrylaminde gel electrophoresis was equivalent to that attained by analytical HPLC anion exchange procedure. Yields were in t he range of 18-25 mg highly pure PSP94 per 50 ml of seminal plasma. Th e desalted, lyophilized, purified PSP94 sample was characterized by SD S-PAGE, Western blot, and N-terminal sequencing. All parameters tested confirm its identity. Preliminary data show that this procedure is su itable for a large-scale production. The direct quantitation of PSP94 by SDS-PAGE and densitometric image analysis at various purification s teps for evaluating the recovery of PSP94 is described. The results ob tained show that this is an efficient strategy for preparation of high ly purified native PSP94. (C) 1996 Academic Press, Inc.