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22-kD growth hormone exclusion assay: a new approach to measurement of non-22-kD growth hormone isoforms in human blood.

Artikel i vetenskaplig tidskrift
Författare C L Boguszewski
L Hynsjö
Gudmundur Johannsson
B A Bengtsson
L M Carlsson
Publicerad i European journal of endocrinology
Volym 135
Nummer/häfte 5
Sidor 573-82
ISSN 0804-4643
Publiceringsår 1996
Publicerad vid Institutionen för invärtesmedicin
Sidor 573-82
Språk en
Länkar www.ncbi.nlm.nih.gov/entrez/query.f...
Ämnesord Adolescent, Adult, Animals, Antibodies, Monoclonal, Antibody Specificity, Evaluation Studies as Topic, Female, Human Growth Hormone, blood, chemistry, deficiency, Humans, Isomerism, Male, Middle Aged, Molecular Weight, Rats, Recombinant Proteins, Sensitivity and Specificity
Ämneskategorier Endokrinologi, Medicinsk laboratorievetenskap

Sammanfattning

Human growth hormone (GH) exists in a variety of isoforms. In the pituitary, the most abundant isoform is 22-kD GH (22 K GH), while other isoforms (non-22 K GH) are present in variable amounts. In human plasma, the GH heterogeneity contributes to the wide variability in GH levels measured by different immunoassays. The physiological role of the non-22 K GH isoforms is poorly understood, but they may represent a spectrum of agonists or antagonists of the GH receptor. It is possible that increased amounts of non-22 K GH isoforms in the circulation contribute to the growth failure observed in some short children and may be involved in the pathophysiology of acromegaly and other unrelated diseases. Currently, there is no method available to evaluate the ratio of non-22 K GH isoforms to total GH in large sets of serum samples. In this report, a novel assay procedure is described in which monomeric and dimeric isoforms of 22 K GH are removed from serum and non-22 K GH isoforms are quantitated. The 22 K GH exclusion assay (22 K GHEA) was established as a screening method to identify conditions in which the ratio of non-22 K GH isoforms to total GH in human blood is altered. A 22 K GH-specific monoclonal antibody (MCB) is used for binding to 22 K GH in serum. Magnetic beads coated with rat anti-mouse immunoglobulin G and a magnetic device are used to remove the 22K GH-MCB complexes from serum. The non-22 K GH isoforms are measured by a polyclonal antibody-based immunoradiometric assay (GH-IRMA). The assay procedure was optimized systematically by statistical experimental designs. In serum spiked with monomeric or dimeric 22 K GH, the 22 K GH extraction was efficient at GH levels up to 100 microg/l (range 96.3-100%). The intra- and interassay precision for non-22K GH levels of 3.9 microg/l were 2.6% and 8.7%, respectively, while for levels of 0.6 microg/l, which were very close to the detection limits of the assay, the coefficients were 17.0% and 21.6%, respectively. The percentage of non-22 K GH isoforms determined in serum samples from three different groups of subjects showed clearly distinctive values. The 22 K GHEA is a new method for evaluation of non-22 K GH isoforms in human blood under different physiological and pathophysiological conditions.

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