Development of an in-house radioimmunoassay for human growth hormone.
A simple and sensitive double-antibody radioimmunoassay for human growth hormone (HGH) was developed, optimised and validated. The anti-hGH sera raised in 2 rabbits were highly specific with low cross-reactions of 0.19% and 0.3% with human placental lactogen and 0.21% and 0.13% with human prolactin. The mean sensitivity of the assay determined from 28 assays was found to be 0.4 +/- 0.2 mIU/L. Mean recovery of added exogenous hGH was 98.8 +/- 6.8%. Linearity studies of samples diluted at 1:2, 1:4 and 1:8 gave values of 101.3 +/- 5.3%, 109.6 +/- 13.4% and 97.3 +/- 13% respectively of those expected. The reproducibility of the assay was good; within assay coefficient of variation for serum samples with GH concentrations of 2.7, 13.6 and 28.2 mU/l ranged from 5.1 to 8.3% while the inter-assay precision varied from 4.9 to 10.3%. The in-house assay showed good correlation (r = 0.96, p less than 0.001) with a commercial HGH RIA kit (Dainabot, Japan). A reference normal adult fasting GH level of less than 7 mIU/l was established from 95 samples assayed by this method.
Wan Nazaimoon
W M
WM
Radiochemistry Division, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur.
Satgunasingam
N
N
Khalid
B
B
D000818:Animals; D013006:Growth Hormone / Q000032:analysis*; D006801:Humans; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D011933:Reagent Kits, Diagnostic
D011933:Reagent Kits, Diagnostic; D013006:Growth Hormone