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Problems with Growth Hormone Doping in Sports: Isoform Methods

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Book cover Hormone Use and Abuse by Athletes

Part of the book series: Endocrine Updates ((ENDO,volume 29))

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Abstract

Human growth hormone (GH), a 191 amino acid proteohormone, is abused in sports. Although there were only few positive test result reported from accredited antidoping laboratories until today, there is enough evidence to assume its wider abuse for performance enhancement: Former athletes have confessed to have abused GH, and ampoules containing recombinant GH have been found in the possession of active athletes or trainers by police and customs officials. For a long time, the detection of GH abuse by testing laboratories was not possible. The main reason why it was difficult to develop an appropriate test was the identity in amino acid sequence between pituitary and recombinant GH, making a distinction by means of physicochemical characterization impossible. Over the last decade, two approaches have been followed to detect abuse of GH. One approach, based on characterization of changes in concentrations of proteins which are produced in response to GH (the “marker approach”), is described elsewhere in this book. Here, we are describing an alternative strategy, called the “differential immunoassay” or “isoform approach.” Pituitary GH consists of a spectrum of molecular isoforms, whereas recombinant hGH preparations are made of the 22 kD isoform only. After injection, secretion of the pituitary-derived isoforms is blunted by negative feedback, and the 22 kD isoform becomes predominant in circulation. Specific immunoassays employing monoclonal antibodies with a preference for the one or the other isoform can be used to detect these changes. Two sets of specific assays have been validated for this purpose and are currently implemented in antidoping laboratories. Future directions for developing the isoform approach include the specific measurement of the 20 kD isoform, which is reduced after application of 22 kD GH, and also the identification of GH isoforms by methods other than immunoassays, including plasmon surface resonance, two dimensional gel electrophoresis, and mass spectrometry.

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References

  1. Baumann G. Growth hormone heterogeneity: Genes, isohormones, variants, and binding ­proteins. Endocr Rev. 1991;12:424–49

    Article  CAS  PubMed  Google Scholar 

  2. Hirt H, Kimelman J, Birnbaum MJ, Chen EY, Seeburg PH, Eberhardt NL, Barta A. The human growth hormone gene locus: Structure, evolution, and allelic variations. DNA. 1987;6:59–70

    Article  CAS  PubMed  Google Scholar 

  3. Igout A, Van Beeumen J, Frankenne F, Scippo ML, Devreese B, Hennen G. Purification and biochemical characterization of recombinant human placental growth hormone produced in Escherichia coli. Biochem J. 1993;295:719–24

    CAS  PubMed  Google Scholar 

  4. Cooke NE, Ray J, Watson MA, Estes PA, Kuo BA, Liebhaber SA. Human growth hormone gene and the highly homologous growth hormone variant gene display different splicing patterns. J Clin Invest. 1988;82:270–5

    Article  CAS  PubMed  Google Scholar 

  5. Wallace JD, Cuneo RC, Bidlingmaier M, Lundberg PA, Carlsson L, Boguszewski CL, Hay J, Healy ML, Napoli R, Dall R, Rosen T, Strasburger CJ. The response of molecular isoforms of growth hormone to acute exercise in trained adult males. J Clin Endocrinol Metab. 2001;86:200–6

    Article  CAS  PubMed  Google Scholar 

  6. Nindl BC. Exercise modulation of growth hormone isoforms: Current knowledge and future directions for the exercise endocrinologist. Br J Sports Med. 2007;41:346–8; discussion 348

    Google Scholar 

  7. Baumann G. Growth hormone heterogeneity in human pituitary and plasma. Horm Res. 1999;51:2–6

    Article  CAS  PubMed  Google Scholar 

  8. Popii V, Baumann G. Laboratory measurement of growth hormone. Clin Chim Acta. 2004; 350:1–16

    Article  CAS  PubMed  Google Scholar 

  9. Baumann G, Stolar MW, Buchanan TA. The metabolic clearance, distribution, and degradation of dimeric and monomeric growth hormone (GH): Implications for the pattern of circulating GH forms. Endocrinology. 1986;119:1497–501

    Article  CAS  PubMed  Google Scholar 

  10. Baumann G, Amburn K, Shaw MA. The circulating growth hormone (GH)-binding protein complex: A major constituent of plasma GH in man. Endocrinology. 1988;122:976–84

    Article  CAS  PubMed  Google Scholar 

  11. Kratzsch J, Selisko T, Birkenmeier G. Identification of transformed alpha 2-macroglobulin as a growth hormone-binding protein in human blood. J Clin Endocrinol Metab. 1995;80: 585–90

    Article  CAS  PubMed  Google Scholar 

  12. Bidlingmaier M, Freda PU. Measurement of human growth hormone by immunoassays: Current status, unsolved problems and clinical consequences. Growth Horm IGF Res. 2010; 20:19–25

    Google Scholar 

  13. Pokrajac A, Wark G, Ellis AR, Wear J, Wieringa GE, Trainer PJ. Variation in GH and IGF-I assays limits the applicability of international consensus criteria to local practice. Clin Endocrinol (Oxf). 2007;67:65–70

    Article  CAS  Google Scholar 

  14. Wieringa GE, Trainer PJ. Commentary: Harmonizing growth hormone measurements: Learning lessons for the future. J Clin Endocrinol Metab. 2007;92:2874–5

    Article  CAS  PubMed  Google Scholar 

  15. Boguszewski CL, Hynsjo L, Johannsson G, Bengtsson BA, Carlsson LM. 22-kD growth hormone exclusion assay: A new approach to measurement of non-22-kD growth hormone isoforms in human blood. Eur J Endocrinol. 1996;135:573–82

    Article  CAS  PubMed  Google Scholar 

  16. Wada M, Uchida H, Ikeda M, Tsunekawa B, Naito N, Banba S, Tanaka E, Hashimoto Y, Honjo M. The 20-kilodalton (kDa) human growth hormone (hGH) differs from the 22-kDa hGH in the complex formation with cell surface hGH receptor and hGH-binding protein circulating in human plasma. Mol Endocrinol. 1998;12:146–56

    Article  CAS  PubMed  Google Scholar 

  17. Keller A, Wu Z, Kratzsch J, Keller E, Blum WF, Kniess A, Preiss R, Teichert J, Strasburger CJ, Bidlingmaier M. Pharmacokinetics and pharmacodynamics of GH: Dependence on route and dosage of administration. Eur J Endocrinol. 2007;156:647–53

    Article  CAS  PubMed  Google Scholar 

  18. Ishikawa M, Yokoya S, Tachibana K, Hasegawa Y, Yasuda T, Tokuhiro E, Hashimoto Y, Tanaka T. Serum levels of 20-kilodalton human growth hormone (GH) are parallel those of 22-kilodalton human GH in normal and short children. J Clin Endocrinol Metab. 1999;84:98–104

    Article  CAS  PubMed  Google Scholar 

  19. Leung KC, Howe C, Gui LY, Trout G, Veldhuis JD, Ho KK. Physiological and pharmacological regulation of 20-kDa growth hormone. Am J Physiol Endocrinol Metab. 2002;283:E836–43

    CAS  PubMed  Google Scholar 

  20. Tsushima T, Katoh Y, Miyachi Y, Chihara K, Teramoto A, Irie M, Hashimoto Y. Serum ­concentration of 20K human growth hormone (20K hGH) measured by a specific enzyme-linked immunosorbent assay. Study Group of 20K hGH. J Clin Endocrinol Metab. 1999;84:317–22

    Article  CAS  PubMed  Google Scholar 

  21. Sinha YN, Jacobsen BP. Human growth hormone (hGH)-(44-191), a reportedly diabetogenic fragment of hGH, circulates in human blood: Measurement by radioimmunoassay. J Clin Endocrinol Metab. 1994;78:1411–8

    Article  CAS  PubMed  Google Scholar 

  22. Wu Z, Bidlingmaier M, Friess SC, Kirk SE, Buchinger P, Schiessl B, Strasburger CJ. A new nonisotopic, highly sensitive assay for the measurement of human placental growth hormone: Development and clinical implications. J Clin Endocrinol Metab. 2003;88:804–11

    Article  CAS  PubMed  Google Scholar 

  23. Celniker AC, Chen AB, Wert RM, Jr., Sherman BM. Variability in the quantitation of circulating growth hormone using commercial immunoassays. J Clin Endocrinol Metab. 1989;68:469–76

    Article  CAS  PubMed  Google Scholar 

  24. Bidlingmaier M, Suhr J, Ernst A, Wu Z, Keller A, Strasburger CJ, Bergmann A. High-sensitivity chemiluminescence immunoassays for detection of growth hormone doping in sports. Clin Chem. 2009;55:445–53

    Article  CAS  PubMed  Google Scholar 

  25. Hayakawa M, Shimazaki Y, Tsushima T, Kato Y, Takano K, Chihara K, Shimatsu A, Irie M. Metabolic effects of 20-kilodalton human growth hormone (20K-hGH) for adults with growth hormone deficiency: Results of an exploratory uncontrolled multicenter clinical trial of 20K-hGH. J Clin Endocrinol Metab. 2004;89:1562–71

    Article  CAS  PubMed  Google Scholar 

  26. Bidlingmaier M, Strasburger CJ. Technology insight: Detecting growth hormone abuse in athletes. Nat Clin Pract Endocrinol Metab. 2007;3:769–77

    Article  CAS  PubMed  Google Scholar 

  27. Bidlingmaier M, Wu Z, Strasburger CJ. Test method: GH. Baillieres Best Pract Res Clin Endocrinol Metab. 2000;14:99–109

    Article  CAS  PubMed  Google Scholar 

  28. Wu Z, Bidlingmaier M, Dall R, Strasburger CJ. Detection of doping with human growth hormone. Lancet. 1999;353:895

    Article  CAS  PubMed  Google Scholar 

  29. Wallace JD, Cuneo RC, Bidlingmaier M, Lundberg PA, Carlsson L, Boguszewski CL, Hay J, Boroujerdi M, Cittadini A, Dall R, Rosen T, Strasburger CJ. Changes in non-22-kilodalton (kDa) isoforms of growth hormone (GH) after administration of 22-kDa recombinant human GH in trained adult males. J Clin Endocrinol Metab. 2001;86:1731–7

    Article  CAS  PubMed  Google Scholar 

  30. Saugy M, Robinson N, Saudan C, Baume N, Avois L, Mangin P. Human growth hormone doping in sport. Br J Sports Med. 2006;40 Suppl 1:i35–9

    Article  PubMed  Google Scholar 

  31. Gutierrez-Gallego R, Bosch J, Such-Sanmartin G, Segura J. Surface plasmon resonance immuno assays – A perspective. Growth Horm IGF Res. 2009;19:388–98

    Article  CAS  PubMed  Google Scholar 

  32. Kohler M, Puschel K, Sakharov D, Tonevitskiy A, Schanzer W, Thevis M. Detection of recombinant growth hormone in human plasma by a 2-D PAGE method. Electrophoresis. 2008;29:4495–502

    Article  CAS  PubMed  Google Scholar 

  33. Kohler M, Thomas A, Puschel K, Schanzer W, Thevis M. Identification of human pituitary growth hormone variants by mass spectrometry. J Proteome Res. 2009;8:1071–6

    Article  CAS  PubMed  Google Scholar 

  34. Segura J, Gutierrez-Gallego R, Ventura R, Pascual JA, Bosch J, Such-Sanmartin G, Nikolovski Z, Pinyot A, Pichini S. Growth hormone in sport: Beyond Beijing 2008. Ther Drug Monit. 2009;31:3–13

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Endocrine Laboratory, Medizinische Klinik – Innenstadt, Ludwig-Maximilians-University, Munich, Germany

    Martin Bidlingmaier

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  1. Martin Bidlingmaier
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  2. Zida Wu
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  3. Christian J. Strasburger
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Correspondence to Martin Bidlingmaier .

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Editors and Affiliations

  1. , Division of Endocrinology, Diabetology a, University of Torino, Torino, 10126, Italy

    Ezio Ghigo

  2. , Division of Endocrinology, Diabetology a, University of Torino, Torino, 10126, Italy

    Fabio Lanfranco

  3. , Division of Clinical Endocrinology, Charité - Universitätsmedizin, Berlin, 10117, Germany

    Christian J. Strasburger

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Bidlingmaier, M., Wu, Z., Strasburger, C.J. (2011). Problems with Growth Hormone Doping in Sports: Isoform Methods. In: Ghigo, E., Lanfranco, F., Strasburger, C. (eds) Hormone Use and Abuse by Athletes. Endocrine Updates, vol 29. Springer, Boston, MA. https://doi.org/10.1007/978-1-4419-7014-5_14

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  • DOI: https://doi.org/10.1007/978-1-4419-7014-5_14

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  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4419-7013-8

  • Online ISBN: 978-1-4419-7014-5

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