Tracking growth hormone abuse in sport: A comparison of distinct isoform-based assays

• Published in: Analytica chimica acta Vol. 733; pp. 56 - 63
• Main Authors: Bosch, J., Ueki, M., Such-Sanmartín, G., Segura, J., Gutiérrez-Gallego, R.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 06.07.2012; Elsevier
• Subjects: Analytical chemistry; antibodies; Antibodies, Monoclonal - analysis; Antibodies, Monoclonal - immunology; binding properties; Chemistry; Doping; Doping in Sports; Exact sciences and technology; Growth hormone; human growth; Human Growth Hormone - analysis; Human Growth Hormone - immunology; Humans; Immunoassay; Immunoassay - methods; immunoassays; Isoform; Male; males; Marker proteins; Miscellaneous; monitoring; Protein Isoforms - analysis; Protein Isoforms - immunology; Recombinant Proteins - analysis; Recombinant Proteins - immunology; Single-Blind Method; somatotropin; sports; Substance Abuse Detection - methods; Surface Plasmon Resonance; Immunoassay; rhGH; LIA; Doping; hGH; phGH; WADA; mAb; Marker proteins; SPR; ILMA; WOO; Isoform; NIBSC; Growth hormone; Human; Hormone; Chemical analysis; Antibody; Use; Time; Concentration; Marker; Capture; Protein; Immunological method; Variant; Signal; Surface plasmon resonance; Application; Comparative study
• ISSN: 0003-2670; 1873-4324; 1873-4324
• DOI: 10.1016/j.aca.2012.04.028

[Display omitted] ► Detection of recombinant human growth hormone abuse in sport. ► Comparison between different immunoassays employed. ► Dissection of the antibody specificity of all antibodies by SPR. ► Final validation with a dedicated human clinical trial. Detecting recombinant human growth hormone (rhGH) abuse in sport remains one of the major challenges in doping control. We have compared two different approaches to detect the hGH (human growth hormone) abuse. The first measures the concentrations of the 22kDa hGH isoform (rec assay) and pituitary derived isoforms (pit assay) and a ratio rec/pit is obtained. The second measures the concentrations of 22 and 20kDa hGH isoforms and also a ratio 22/20kDa is derived. Using a single set (nine healthy male subjects, 7 days, 0.026mg/kg/day of rhGH, 2 week wash out period) both approaches were compared. To quantify the agreement between the immunoassays, B.A. (Bland–Altman) analysis and P.r. (Pearson correlation) were used. To fully understand the assay readings, all relevant antibodies were characterised by surface plasmon resonance (SPR). In either approach the ratio numerator produces similar results and the denominator determines both signal-amplitude and time-frame of possible application. The rec vs pit approach displays a higher distinctive capacity to detect hGH abuse but the complex binding properties of the capture antibodies make it very difficult to evaluate the precise contributions of the individual hGH variants to the assay result. In the 22 vs 20 approach, the 20kDa hGH concentration measures determine its applicability. Both approaches are based on a different principle, should be preferably applied within 24h after rhGH administration, and are perfectly comparable given the results obtained. The reduced time frame of application indicates that their principle application should be preferably in an out-of-competition setting.