Production and Characterization of Specific Monoclonal Antibodies of the Human Thyroid Stimulating Hormone

Abstract

Spleen cells from BALB/c mice immunized with human thyroid stimulating hormone (β-subunit) were fused with mouse myeloma cells (P3/X63-Ag8) and five hybridomas secreting monoclonal antibodies (MAbs) were obtained. These hybridomas specifically recognize (hTSH) and do not cross-react with the other human glycoprotein hormones such as: luteinizing hormone (LH), follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hcG). The MAbs were of the IgG₁ subclass and ascitic fluid from these hybridomas was purified by affinity chromatography on Protein A-sepharose CL-4B column to isolate the IgG₁ active fraction. The affinity constant of these MAbs ranged from 3.2 × 10¹⁰ to 1.5 × 10¹¹ M^-1.