ELISA for Determination of Human Growth Hormone: Recognition of Helix 4 Epitopes

• Published in: Journal of biomedicine & biotechnology Vol. 2004; no. 3; pp. 143 - 149
• Main Authors: Moura, Juliana F., DeLacerda, Luiz, Sandrini, Romolo, Borba, Fernanda M., Castelo, Denise N., Sade, Elis R., Sella, Sandra, Minozzo, João C., Callefe, Luis G., Figueiredo, Bonald C.
• Format: Journal Article
• Language: English
• Published: United States Hindawi Publishing Corporation 2004
• ISSN: 1110-7243; 1110-7251
• DOI: 10.1155/S1110724304308090

Human growth hormone (hGH) signal transduction initiates with a receptor dimerization in which one molecule binds to the receptor through sites 1 and 2. A sandwich enzyme-linked immunosorbent assay was developed for quantifying hGH molecules that present helix 4 from binding site 1. For this, horse anti-rhGH antibodies were eluted by an immunoaffinity column constituted by sepharose-rhGH. These antibodies were purified through a second column with synthetic peptide correspondent to hGH helix 4, immobilized to sepharose, and used as capture antibodies. Those that did not recognize synthetic peptide were used as a marker antibody. The working range was of 1.95 to 31.25 ng/mL of hGH. The intra-assay coefficient of variation (CV) was between 4.53% and 6.33%, while the interassay CV was between 6.00% and 8.27%. The recovery range was between 96.0% to 103.8%. There was no cross-reactivity with human prolactin. These features show that our assay is an efficient method for the determination of hGH.