Human prostatic acid phosphatase in seminal plasma: purifications, simple preparation of antibody and development of a new immunoenzymatic assay.
Prostatic acid phosphatase (PAP) was purified from human seminal plasma by precipitation with ammonium sulfate and by serial chromatographies with concanavalin A-Sepharose, Sephadex G-100, and carboxymethyl-cellulose. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis, both with and without sodium dodecyl sulfate. The purified enzyme also gave a single precipitin line upon immunoelectrophoresis, when reacted with rabbit antiserum to seminal plasma. A simple method was developed for the preparation of monospecific antibody to PAP. In addition, a new immunoenzymatic assay system for PAP is reported, utilizing filter paper discs as a solid-phase to which monospecific antibody to PAP was covalently coupled. By this method, serum PAP in a range of 1 to 7,000 ng/ml could be accurately measured within 4 hr.
D000135:Acid Phosphatase / Q000276:immunology / Q000302:isolation & purification*; D000596:Amino Acids / Q000032:analysis; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007106:Immune Sera; D007122:Immunoelectrophoresis; D007124:Immunoenzyme Techniques; D008297:Male; D011467:Prostate / Q000201:enzymology; D012661:Semen / Q000201:enzymology*
D000596:Amino Acids; D007106:Immune Sera; D000135:Acid Phosphatase