An investigation into the lability of the bioactivity of human growth hormone using the ESTA bioassay.
We compared the bioactivity attributable to human growth hormone (hGH) in serum samples, determined at the time of their collection, with that after storage for 2-18 months at -20 degrees C. The samples were obtained from volunteers and patients who underwent provocative tests of hGH secretion, and the bioactivity was determined in the ESTA bioassay, which is based upon the use of Nb2 cells. We report that, in some subjects, the bioactivity of samples collected at the response peaks deteriorated on storage for as little as 2 months. The decrease in hGH bioactivity was systematic in that it consistently declined so as to approach the values initially determined by an immunoassay (Hybritech IRMA). This differential lability was a characteristic of the peak samples, and was not observed for either samples collected before and after the peaks of hGH secretion or for purified preparations of hGH which were subjected to a range of freeze/thaw and storage regimens. We suggest that this unusual lability is indicative of transient shifts in the spectrum of the variants of hGH which are present in the circulation following stimulation by provocative agents. This study emphasises the need to minimise the risk of introducing storage artefacts in investigations into the responses of hGH to provocation.
Dattani
M T
MT
Endocrine Unit, University College London Medical School, UK.
Ealey
P A
PA
Pringle
P J
PJ
Hindmarsh
P C
PC
Brook
C G
CG
Marshall
N J
NJ
D000328:Adult; D000818:Animals; D001681:Biological Assay / Q000379:methods; D001682:Biological Availability; D002648:Child; D004355:Drug Stability; D005260:Female; D005615:Freezing; D019382:Human Growth Hormone / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D006801:Humans; D015592:Immunoradiometric Assay; D008223:Lymphoma; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D013048:Specimen Handling; D013997:Time Factors; D014407:Tumor Cells, Cultured
D011994:Recombinant Proteins; D019382:Human Growth Hormone