Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10537098 | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics | 2015 | 7 Pages |
Abstract
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 20Â 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 20Â 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 22/20Â 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.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Gerard Such-SanmartÃn, Nicolai Bache, Jaume Bosch, Ricardo Gutiérrez-Gallego, Jordi Segura, Ole N. Jensen,