Skip to main content

Direct Methods for Distinction Between Endogenous and Exogenous Erythropoietin

  • Chapter
  • First Online:
Book cover Hormone Use and Abuse by Athletes

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

  • 1040 Accesses

Abstract

Since the commercialization of the first recombinant human erythropoietin (rhEPO) product (epoetin-a) in 1989 as a treatment for acute anemia, rhEPO detection hasrepresented a continuous challenge for the anti doping fight. Indeed, it appeared rapidly that this ergogenic hormone would be abused by athletes looking for an arepsicial performance enhancer. Hemoglobin is one of the principal modulators of aerobic power [1, 2] and, consequently, of performance in endurance sports [3]. By stimulating the red blood cells production, EPO is known to raise hemoglobin concentration in a dose-dependant and predictable way.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Kanstrup IL, Ekblom B. Blood volume and hemoglobin concentration as determinants of maximal aerobic power. Med Sci Sports Exerc. 1984;16(3):256–262.

    CAS  PubMed  Google Scholar 

  2. Gledhill N, Warburton D, Jamnik V. Haemoglobin, blood volume, cardiac function, and aerobic power. Can J Appl Physiol. 1999;24(1):54–65.

    CAS  PubMed  Google Scholar 

  3. Craig NP, Norton KI, Bourdon PC, et al. Aerobic and anaerobic indices contributing to track endurance cycling performance. Eur J Appl Physiol Occup Physiol. 1993;67(2):150–158.

    Article  CAS  PubMed  Google Scholar 

  4. Wang MD, Yang M, Huzel N, Butler M. Erythropoietin production from CHO cells grown by continuous culture in a fluidized-bed bioreactor. Biotechnol Bioeng. 2002;77(2):194–203.

    Article  CAS  PubMed  Google Scholar 

  5. Choi D, Kim M, Park J. Erythropoietin: physico- and biochemical analysis. J Chromatogr B Biomed Appl. 1996;687(1):189–199.

    Article  CAS  PubMed  Google Scholar 

  6. Wide L, Bengtsson C, Berglund B, Ekblom B. Detection in blood and urine of recombinant erythropoietin administered to healthy men. Med Sci Sports Exerc. 1995;27(11):1569–1576.

    CAS  PubMed  Google Scholar 

  7. Lasne F, de Ceaurriz J. Recombinant erythropoietin in urine. Nature. 2000;405(6787):635.

    Article  CAS  PubMed  Google Scholar 

  8. Lasne F, Martin L, Crepin N, de Ceaurriz J. Detection of isoelectric profiles of erythropoietin in urine: differentiation of natural and administered recombinant hormones. Anal Biochem. 2002;311(2):119–126.

    Article  CAS  PubMed  Google Scholar 

  9. Lasne F. Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures. J Immunol Methods. 2003;276(1–2):223–226.

    Article  CAS  PubMed  Google Scholar 

  10. Wide L, Bengtsson C. Molecular charge heterogeneity of human serum erythropoietin. Br J Haematol. 1990;76(1):121–127.

    Article  CAS  PubMed  Google Scholar 

  11. Catlin DH, Breidbach A, Elliott S, Glaspy J. Comparison of the isoelectric focusing patterns of darbepoetin alfa, recombinant human erythropoietin, and endogenous erythropoietin from human urine. Clin Chem. 2002;48(11):2057–2059.

    CAS  PubMed  Google Scholar 

  12. Breidbach A, Catlin DH, Green GA, Tregub I, Truong H, Gorzek J. Detection of recombinant human erythropoietin in urine by isoelectric focusing. Clin Chem. 2003;49(6 Pt 1):901–907.

    Article  CAS  PubMed  Google Scholar 

  13. WADA technical document TD2007EPO. 2007.

    Google Scholar 

  14. Lasne F, Martin L, Martin JA, de CJ. Isoelectric profiles of human erythropoietin are different in serum and urine. Int J Biol Macromol. 2007;41(3):354–357.

    CAS  Google Scholar 

  15. Lönnberg M, Drevin M, Carlsson J. Ultra-sensitive immunochromatographic assay for quantitative determination of erythropoietin. J Immunol Methods. 2008;339(2):236–244.

    Article  PubMed  Google Scholar 

  16. Franke WW, Heid H. Pitfalls, errors and risks of false-positive results in urinary EPO drug tests. Clin Chim Acta. 2006;373(1–2):189–190.

    Article  CAS  PubMed  Google Scholar 

  17. Kahn A, Baker M. Non-specific binding of monoclonal human erythropoietin antibody AE7A5 to Escherichia coli and Saccharomyces cerevisiae proteins. Clin Chim Acta. 2006;379:173–175.

    Article  Google Scholar 

  18. Beullens M, Delanghe JR, Bollen M. False-positive detection of recombinant human erythropoietin in urine following strenuous physical exercise. Blood. 2006;107(12):4711–4713.

    Article  CAS  PubMed  Google Scholar 

  19. Khan A, Grinyer J, Truong ST, Breen EJ, Packer NH. New urinary EPO drug testing method using two-dimensional gel electrophoresis. Clin Chim Acta. 2005;358(1–2):119–130.

    Article  CAS  PubMed  Google Scholar 

  20. Rabin OP, Lasne F, Pascual JA, Saugy M, Delbeke FJ, Van EP. New urinary EPO drug testing method using two-dimensional gel electrophoresis. Clin Chim Acta. 2006;373(1–2):186–187.

    Article  CAS  PubMed  Google Scholar 

  21. Reichel C. Idenepsication of zinc-alpha-2-glycoprotein binding to clone AE7A5 antihuman EPO antibody by means of nano-HPLC and high-resolution high-mass accuracy ESI-MS/MS. J Mass Spectrom. 2008;43(7):916–923.

    Article  CAS  PubMed  Google Scholar 

  22. Peltre G, Thormann W. Evaluation Report of the Urine EPO Test. Bern: Council of the World Anti-Doping Agency (WADA); 2003.

    Google Scholar 

  23. Lamon S, Robinson N, Sottas PE, et al. Possible origins of undetectable EPO in urine samples. Clin Chim Acta. 2007;385(1–2):61–66.

    Article  CAS  PubMed  Google Scholar 

  24. Macdougall IC, Ashenden M. Current and upcoming erythropoiesis-stimulating agents, iron products, and other novel anemia medications. Adv Chronic Kidney Dis. 2009;16(2):117–130.

    Article  PubMed  Google Scholar 

  25. Barbone FP, Johnson DL, Farrell FX, et al. New epoetin molecules and novel therapeutic approaches. Nephrol Dial Transplant. 1999;14(Suppl 2):80–84.

    Article  CAS  PubMed  Google Scholar 

  26. Lamon S, Robinson N, Mangin P, Saugy M. Detection window of Darbepoetin-alpha following one single subcutaneous injection. Clin Chim Acta. 2007;379(1–2):145–149.

    Article  CAS  PubMed  Google Scholar 

  27. Macdougall IC. CERA (continuous erythropoietin receptor activator): a new erythropoiesis-stimulating agent for the treatment of anemia. Curr Hematol Rep. 2005;4(6):436–440.

    CAS  PubMed  Google Scholar 

  28. Wattendorf U, Merkle HP. PEGylation as a tool for the biomedical engineering of surface modified microparticles. J Pharm Sci. 2008;97(11):4655–4669.

    Article  CAS  PubMed  Google Scholar 

  29. Macdougall IC. Recent advances in erythropoietic agents in renal anemia. Semin Nephrol. 2006;26(4):313–318.

    Article  PubMed  Google Scholar 

  30. Ashenden M, Varlet-Marie E, Lasne F, Audran M. The effects of microdose recombinant human erythropoietin regimens in athletes. Haematologica. 2006;91(8):1143–1144.

    CAS  PubMed  Google Scholar 

  31. Caldini A, Moneti G, Fanelli A, et al. Epoetin alpha, epoetin beta and darbepoetin alfa: two-dimensional gel electrophoresis isoforms characterization and mass spectrometry analysis. Proteomics. 2003;3(6):937–941.

    Article  CAS  PubMed  Google Scholar 

  32. Kohler M, Ayotte C, Desharnais P, et al. Discrimination of recombinant and endogenous urinary erythropoietin by calculating relative mobility values from SDS gels. Int J Sports Med. 2008;29(1):1–6.

    Article  CAS  PubMed  Google Scholar 

  33. Reichel C, Kulovics R, Jordan V, Watzinger M, Geisendorfer T. SDS-PAGE of recombinant and endogenous erythropoietins: benefits and limitations of the method for application in doping control. Drug Test Anal. 2009;1:43–50.

    Article  CAS  PubMed  Google Scholar 

  34. Stubiger G, Marchetti M, Nagano M, Reichel C, Gmeiner G, Allmaier G. Characterisation of intact recombinant human erythropoietins applied in doping by means of planar gel electrophoretic techniques and matrix-assisted laser desorption/ionisation linear time-of-flight mass spectrometry. Rapid Commun Mass Spectrom. 2005;19(5):728–742.

    Article  PubMed  Google Scholar 

  35. Guan F, Uboh CE, Soma LR, et al. Differentiation and idenepsication of recombinant human erythropoietin and darbepoetin Alfa in equine plasma by LC-MS/MS for doping control. Anal Chem. 2008;80(10):3811–3817.

    Article  CAS  PubMed  Google Scholar 

  36. Guan F, Uboh CE, Soma LR, Birksz E, Chen J. Idenepsication of darbepoetin alfa in human plasma by liquid chromatography coupled to mass spectrometry for doping control. Int J Sports Med. 2009;30(2):80–86.

    Article  CAS  PubMed  Google Scholar 

  37. Groleau PE, Desharnais P, Cote L, Ayotte C. Low LC-MS/MS detection of glycopeptides released from pmol levels of recombinant erythropoietin using nanoflow HPLC-chip electrospray ionization. J Mass Spectrom. 2008;43(7):924–935.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Martial Saugy .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this chapter

Cite this chapter

Lamon, S., Robinson, N., Saugy, M. (2011). Direct Methods for Distinction Between Endogenous and Exogenous Erythropoietin. 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_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-4419-7014-5_17

  • Published:

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4419-7013-8

  • Online ISBN: 978-1-4419-7014-5

  • eBook Packages: MedicineMedicine (R0)

Publish with us

Policies and ethics