Use of monoclonal antibodies to pregnanediol-3 alpha-glucuronide for the development of a solid phase chemiluminescence immunoassay.
Monoclonal antibodies to pregnanediol-3 alpha-glucuronide were produced by hybridomas between P3-X63-Ag8 variants and spleen cell of mice immunized with a bovine serum albumin conjugate of the homologous hapten. The ascites fluid collected from mice inoculated with the cloned hybridoma cells contained antibodies with high specificity and affinity to pregnanediol-3 alpha-glucuronide. A sensitive solid-phase chemiluminescence immunoassay for urinary pregnanediol-3 alpha-glucuronide was established utilizing these antibodies. The assay was validated in terms of specificity, accuracy, sensitivity and precision. When urine samples were assayed for pregnanediol-3 alpha-glucuronide, the results obtained by the solid-phase chemiluminescence immunoassay method and the conventional gas liquid chromatographic method agreed well (n = 30, r = 0.96). The method may be of value for monitoring luteal function since it is fast, sensitive and does not require the use of radioisotopes or purification of the biological sample. Monoclonal antibody preparations facilitate rigorous standardization of the assay.
D000818:Animals; D000911:Antibodies, Monoclonal; D002849:Chromatography, Gas; D005260:Female; D005965:Glucuronates / Q000276:immunology / Q000652:urine; D006801:Humans; D006822:Hybrid Cells / Q000276:immunology; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000378:metabolism; D008163:Luminescent Measurements; D008297:Male; D008598:Menstruation; D051379:Mice; D009101:Multiple Myeloma / Q000378:metabolism; D009374:Neoplasms, Experimental / Q000378:metabolism; D011276:Pregnanediol / Q000031:analogs & derivatives* / Q000276:immunology / Q000652:urine; D013696:Temperature; D013997:Time Factors
D000911:Antibodies, Monoclonal; D005965:Glucuronates; D007074:Immunoglobulin G; C022270:pregnanediol-3 alpha-glucuronide; D011276:Pregnanediol