Functional characterization of monoclonal antibodies specific to growth hormone receptor

Upon engagement with appropriate ligands, receptors can be activated to initiate various metabolic and morphological changes in living cells. An attempt was made in this study to generate monoclonal antibodies (mAb) specific to recombinant rat growth hormone receptor (GHR) and subsequently to investigate their ability to act as biologically active ligands. Three mAbs, designated 1A9, 1H2 and 2C3, were produced and all were highly reactive with GHR in an enzyme-linked immunosorbent assay. In contrast to 1H2, 1A9 and 2C3 competed with radioactive growth hormone (GH) tracer for the binding to GHR in a radioreceptor assay, suggesting that the GH-binding sites of GHR were identical, or very close to its epitopes recognized by 1A9 and 2C3. The molecular interaction evaluated by the BIAcore technology further demonstrated the separate GHR epitopes for 1A9 and 2C3. 2C3 apparently targeted the precise GH-binding sites of GHR, while the antigenic determinants for 1A9 were not at the site, but adjacent to it. Functional analysis showed that 2C3 promoted the growth of hypophysectomized rats, whereas others failed to do so. Therefore, findings from the present study suggest that these mAbs recognize distinct GHR epitopes and are useful for investigating the structure-function relationship of GHR. Furthermore, 2C3 may prove important as a biologically active agonist for better understanding of the process of GHR activation relevant to growth. (C) 1997 Elsevier Science Ltd.