Monoclonal antibody capture fluorometric enzyme linked immunosorbent assayfor detection of porcine growth hormone in plasma

A specific and sensitive fluorometric enzyme-linked immunosorbent assay (EL ISA) was developed to measure growth hormone in swine plasma. Homogeneous p ig growth hormone (pGH) was purified from pituitary gland, and used to rais e monoclonal (mAb) and polyclonal antibodies. The mAb showing the highest r eactivity was used as solid-phase antibody to capture pGH, while the rabbit biotin-labelled antibodies against pGH, followed by beta-galactosidase-avi din-conjugate, were used for detection of mAb-bound pGH. The calibration gr aph was linear ever the range 0.05-25 ng ml(-1). Plasma did not interfere w ith the ability to measure pGH acid the assay proved suitable for measuring pGH in plasma without any prior treatment of samples. Cross-reactivity and recovery tests confirmed the specificity of the method. The intra- and int er-assay coefficients of variation ranged between 7.2% and 9.7% and 12.1% a nd 18.7%, respectively. The analytical specificity of the method was valida ted by provocative tests for GH using the GH-releasing hexapeptide GHRP-6 i n prepubertal gilts, The ELISA developed is sensitive, reproducible and sui table for measuring GH in pig plasma, and offers an alternative to radioimm unoassay. (C)1999 Elsevier Science B.V. All rights reserved.