Serum growth hormone-binding protein (GHBP) in domestic animals as measured by ELISA2

Abstract

This research was conducted to develop and characterize a competitive ELISA for bovine serum growth hormone-binding protein (GHBP) using recombinant bovine GHBP and a polyclonal rabbit antiserum. In addition to bovine, however, the assay was found to measure some activity in equine, chicken, porcine, ovine, and human sera. The reference standard curve had an effective range of 3 to 200 ng/mL. Recovery of increasing amounts of GHBP added to ovine serum was 103% but seemed to overestimate the amount of GHBP at low concentrations (intercept = 2.5 ng/mL). Recovery from bovine and porcine serum was near ideal but seems to be overestimated at concentrations higher than 50 ng/microL. Within and between assay coefficients of variation were 12.1 and 18.9%, respectively, for a sheep serum pool. Neither exogenous GH (20 ng/mL) nor prolactin (100 ng/mL) interfered with the measurement of GHBP in serum. The GHBP activity measured in increasing doses of serum from ovine, porcine, and bovine inhibited the assay in a parallel manner. This observation suggests that the GHBP antiserum contains antibodies that are directed toward epitopes of GHBP, which are common among these species. Serum GHBP concentrations were similar among samples from a line of miniature Brahman and normal stature Brahman and Angus cattle. In mature ewes, there were no differences in serum GHBP among three different breed types. An increase (P < .0001) in serum GHBP was observed in pigs between 1 and 6 mo of age but no sex effect was observed.(ABSTRACT TRUNCATED AT 250 WORDS)