Purification of monoclonal antibodies raised against prostate-specific acid phosphatase for use in vivo in radioimaging of prostatic cancer.
In developing diagnostic reagents for the radioimaging of prostatic cancer, methods were optimized for the purification of two mouse IgG1 monoclonal antibodies raised against prostate-specific acid phosphatase and produced in cell culture. Two different two-step methods were selected. One method consisted of two successive ion exchange chromatographic steps on Mono S and Mono Q; in the other method, Mono S chromatography was followed by hydrophobic interaction (Alkyl Superose) chromatography. In both cases, fast protein liquid chromatography (FPLC) instrumentation was used. The antibodies were purified from cell culture media containing fetal calf serum (1-5%). Highly pure (greater than 95%) IgG1 antibodies, free of contaminating serum-derived proteins or column materials, were obtained in good yield (greater than 90% recovery). The purified antibodies completely retained their immunological reactivity towards prostate-specific acid phosphatase and were sterile and pyrogen-free. Since the monoclonal antibodies produced were intended for applications in vivo, an essential feature of the methods selected was the availability of in situ cleaning procedures for sterilization of the gel materials and for the inactivation of viruses and pyrogens in the gels. The methods developed could be readily scaled up for preparative purposes.
Hakalahti
L
L
Biocenter, University of Oulu, Finland.
Vihko
P
P
D000135:Acid Phosphatase / Q000276:immunology*; D000180:Acrylic Resins; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D014408:Biomarkers, Tumor / Q000276:immunology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D008297:Male; D051379:Mice; D011467:Prostate / Q000201:enzymology* / Q000276:immunology; D011471:Prostatic Neoplasms / Q000000981:diagnostic imaging* / Q000201:enzymology; D011877:Radionuclide Imaging
D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D007074:Immunoglobulin G; C023049:sephacryl S 200; D000135:Acid Phosphatase