Detection and differentiation of 22 kDa and 20 kDa Growth Hormone proteoforms in human plasma by LC-MS/MS

Human growth hormone (GH) is suspected to be widely and illegally used in sport to improve athletes' performance. For the detection of GH abuse, blood samples are screened for abnormal ratios between the 22 and 20 kDa GH proteoforms that demonstrate the administration of the synthetic hormone....

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Bibliographic Details
Published in:Biochimica et biophysica acta 2015-04, Vol.1854 (4), p.284-290
Main Authors: Such-Sanmartín, Gerard, Bache, Nicolai, Bosch, Jaume, Gutiérrez-Gallego, Ricardo, Segura, Jordi, Jensen, Ole N
Format: Article
Language:English
Subjects:
Adult
Blood Chemical Analysis - methods
Chromatography, Liquid - methods
Exercise - physiology
Female
Human Growth Hormone - blood
Humans
Immunoassay - methods
Male
Molecular Weight
Protein Isoforms - blood
Proteolysis
Sensitivity and Specificity
Tandem Mass Spectrometry - methods
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Summary:Human growth hormone (GH) is suspected to be widely and illegally used in sport to improve athletes' performance. For the detection of GH abuse, blood samples are screened for abnormal ratios between the 22 and 20 kDa GH proteoforms that demonstrate the administration of the synthetic hormone. Current detection methods are based on classical immunoassays as they provide sufficient sensitivity for the detection of GH proteoforms. These antibody based methods, however, suffer from unclear selectivity and potential cross-reactivity towards similar proteins. For unambiguous GH detection, we report a Mass Spectrometry ImmunoAssay (MSIA) that first enriches GH from plasma with an antibody of relatively low specificity, and subsequently quantifies the 22 and 20 kDa proteoforms by Selected Reaction Monitoring (SRM) LC-MS/MS analysis. This method proved superior to an antibody-free strategy based on GH purification by protein precipitation. Using GH-MSIA we successfully quantified the 22/20 kDa GH ratio in post-exercise capillary plasma extracted from two individuals. This GH-MSIA is applicable to anti-doping and GH-related disease analysis.
ISSN:0006-3002
1570-9639
1878-2434
DOI:10.1016/j.bbapap.2014.12.022