Monoclonal antibodies to beta subunit of choriogonadotropin: development and use in a sandwich ELISA pregnancy test.
Balb/c mice were immunized with beta subunit isolated and purified from crude human chorionic gonadotropin preparations. Spleen cells from the higher titered mouse were fused with Sp 2/0 myeloma cells. Four specific secreting hybridomas were obtained. Specificity, affinity, and suitability of secreted antibodies for use in enzyme immunoassays were studied. Ascites of the selected hybridoma was raised; the antibody was purified by protein A-affinity chromatography and coupled to horseradish peroxidase. This conjugate was employed in a simultaneous sandwich enzyme immunoassay on microtiter plates sensitized with goat polyclonal antibody to measure the hormone. The test has a sensitivity of 10 mlU/ml either on urine, serum, or plasma samples when read in a microplate reader. The results can also be evaluated by the naked eye, with a sensitivity of 20 mlU/ml. No cross reactivity was detected with other human gonadotropins.
Gariglio
R
R
Research and Development Department, Wiener Laboratories, Rosario, Argentina.
Toplikar
E
E
Carlomagno
A
A
Rojkin
F
F
Knecher
L
L
Capriotti
G
G
Lorenzo
L
L
D000818:Animals; D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin / Q000032:analysis / Q000276:immunology*; D018997:Chorionic Gonadotropin, beta Subunit, Human; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology*; D011247:Pregnancy; D011258:Pregnancy Tests / Q000379:methods* / Q000706:statistics & numerical data; D011863:Radioimmunoassay; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D018997:Chorionic Gonadotropin, beta Subunit, Human; D010446:Peptide Fragments