Abstract
We recently described the generation of mouse monoclonal antibodies directed against hCG using a synthetic peptide immunogen: the carboxy-terminal peptide (CTP) of β-hCG. The CTP, residues 109–145, is a portion unique to hCG and is not present in other glycoprotein hormones. These monoclonal antibodies react with β-hCG-CTP and whole hCG but do not react with hLH. In the present study, a two-site sandwich ELISA specific for hCG has been developed using these CTP-induced monoclonal antibodies. Two monoclonal antibodies were paired: β-hCG-CTP-a6 was immobilized to give a hCG-specific solid phase, and a biotinylated monoclonal antibody against β-hCG was used as the indicator antibody. This second antibody was introduced during the hCG-capture step because of its agonist effect on hCG-capture by β-hCG-CTP-a6 solid phase. This ELISA system can detect 2.2 IU/1 hCG in a serum-free medium and 10.4 IU/1 in a medium containing 20% human serum. We did not observe any cross-reactivity with hLH up to concentrations of 5000 IU/1. The hCG-ELISA correlated well ( r = 0.99) with a control RIA in an assay of 78 human sera from healthy males, pregnant or non-pregnant females and from patients with choriocarcinoma, hydatidiform mole, or teratocarcinoma.
Published Version
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