Radioimmunoassay of growth hormone-releasing hormone (GHRH) with a polyclonal antibody against synthetic GHRH(1–29)-Gly 4-Cys-NH 2: Method and clinical studies

Abstract

A radioimmunoassay (RIA) for growth hormone-releasing hormone (GHRH) using a polyclonal antibody against synthetic GHRH(1–29)-Gly 4-Cys-NH 2 has been developed. The antiserum (RBM105) showed full cross-reactivity with GHRH-(1–44)NH 2, GHRH-(1–40)OH, GHRH-(1–37)OH and GHRH-(3–44)NH 2, and probably recognized the region of Ala 4 to Lys 12 of GHRH. Since the sensitivity of the GHRH RIA was 1.5 pg/tube, the lowest detectable plasma level was 5 ng/l when an extract of 0.3 ml of plasma per tube was used. On gelfiltration chromatography, the GHRH immunoreactivity of normal plasma was eluted in the same position as synthetic GHRH. The plasma GHRH concentration in healthy subjects was 20.5 ± 6.5 ng/l (mean ± SD), and in patients with hypothalamic disorders was 17.4 ± 2.0 ng/l. In contrast, the plasma GHRH level in hemodialysis-dependent, chronic renal failure (CRF-HD) patients (38.7 ± 13.1 ng/l) was significantly higher than normal. The acromegalic patients were 24.3 ±11.9 ng/l, except for one patient with ectopic GHRH syndrome (990 ng/l): his plasma GHRH level reached 7,100 ng/l during operation, and then decreased logarithmically to 70 ng/l after 6 h. Somatostatin at concentrations of 10 and 1,000 nmol/l significantly suppressed (GHRH release) from primary culture cells of the GHRH-producing tumor from 17.3 ± 0.92 ng/2× 10 5 cells to f9.98 ± 3.61 and 4.32 ± 1.01 ng/2× 10 5( cells, respectively after 48 h. These data indicate that this GHRH RIA is useful for determining the plasma GHRH concentration in normal and diseased states and also for in vitro studies of GHRH release.