ELISA for Determination of Human Growth Hormone: Recognition of Helix 4 Epitopes.
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.
Moura
Juliana F.
JF
DeLacerda
Luiz
L
Sandrini
Romolo
R
Borba
Fernanda M.
FM
Castelo
Denise N.
DN
Sade
Elis R.
ER
Sella
Sandra
S
Minozzo
João C.
JC
Callefe
Luis G.
LG
Figueiredo
Bonald C.
BC