Advertisement

Analytical and Bioanalytical Chemistry

, Volume 388, Issue 7, pp 1521–1529 | Cite as

Procedures for monitoring recombinant erythropoietin and analogues in doping control

  • Jordi Segura
  • José A. Pascual
  • Ricardo Gutiérrez-Gallego
Review

Abstract

The present report summarizes the main analytical strategies developed to identify the presence of recombinant erythropoietin (EPO) administered as a doping agent. Indirect evidence is based on the analysis of blood parameters (haemoglobin, haematocrit, reticulocytes, macrocytes, etc.) and serum markers (concentration of EPO and serum transferrin receptors, etc.). The problem of intertechnique comparison for reliable results evaluation is emphasized, especially for serum markers. Charge differences between isoforms of recombinant EPO and native urinary EPO are the grounds for the isoelectric focusing–double blotting–chemiluminescence detection method presently approved for doping control. Works addressing its advantages and limitations are presented and commented on. The chemical bases of the differential detection are highlighted and some future approaches for detection are also presented. The appearance and detectability of EPO analogues and mimetics susceptible for abuse are also addressed.

Keywords

Erythropoietin Detection Blood Urine Analogues Mimetics 

Notes

Acknowledgements

The authors are grateful for financing received from the Spanish (Ministerio de Educación y Ciencia; Consejo Superior de Deportes) and Catalan (Departament d’Universitats, Recerca i Societat de l’informació; Consell Catalá de l’Esport) authorities for supporting the preparation of the present review. Special thanks are addressed also to WADA for supporting several research projects mentioned in this review. The scientific contribution from other staff members of the Pharmacology Research Unit at IMIM is also gratefully acknowledged.

References

  1. 1.
    Faulds D, Sorkin EM (1989) Drugs 38:863–899Google Scholar
  2. 2.
    Lappin TR, Rich IN (1996) Clin Lab Haematol 18:137–145CrossRefGoogle Scholar
  3. 3.
    Lacombe C, Mayeux P (1998) Haematologica 83:724–732Google Scholar
  4. 4.
    Maiese K, Li F, Chong ZZ (2005) JAMA 293:90–95CrossRefGoogle Scholar
  5. 5.
    Badia R, de la Torre R, Segura J (1992) Biol Clin Hematol 14:177–184Google Scholar
  6. 6.
    Pascual JA, Belalcazar V, de Bolos C, Gutiérrez R, Llop E, Segura J (2004) Ther Drug Monit 26:175–179CrossRefGoogle Scholar
  7. 7.
    Peltre G, Thormann W (2003) Evaluation report of the EPO test. Internal report to WADAGoogle Scholar
  8. 8.
    Macmillan D, Bill RM, Sage KA, Fern D, Flitsch SL (2001) Chem Biol 8:133–145CrossRefGoogle Scholar
  9. 9.
    Elliott S, Lorenzini T, Asher S, Aoki K, Brankow D, Buck L, Busse L, Chang D, Fuller J, Grant J, Hernday N, Hokum M, Hu S, Knudten A, Levin N, Komorowski R, Martin F, Navarro R, Osslund T, Rogers G, Rogers N, Trail G, Egrie J (2003) Nat Biotechnol 21:414–421CrossRefGoogle Scholar
  10. 10.
    Kochendoerfer GG, Chen SY, Mao F, Cressman S, Traviglia S, Shao H, Hunter CL, Low DW, Cagle EN, Carnevali M, Gueriguian V, Keogh PJ, Porter H, Stratton SM, Wiedeke MC, Wilken J, Tang J, Levy JJ, Miranda LP, Crnogorac MM, Kalbag S, Botti P, Schindler-Horvat J, Savatski L, Adamson JW, Kung A, Kent SB, Bradburne JA (2003) Science 299:884–887CrossRefGoogle Scholar
  11. 11.
    Macdougall IC (1998) Nephrol Dial Transplant 13(Suppl 2):23–27CrossRefGoogle Scholar
  12. 12.
    Barbone FP, Johnson DL, Farrell FX, Collins A, Middleton SA, McMahon FJ, Tullai J, Jolliffe LK (1999) Nephrol Dial Transplant 14(Suppl 2):80–84CrossRefGoogle Scholar
  13. 13.
    Goldberg J, Jin Q, Ambroise Y, Satoh S, Desharnais J, Capps K, Boger DL (2002) J Am Chem Soc 124:544–555CrossRefGoogle Scholar
  14. 14.
    Qureshi SA, Kim RM, Konteatis Z, Biazzo DE, Motamedi H, Rodrigues R, Boice JA, Calaycay JR, Bednarek MA, Griffin P, Gao YD, Chapman K, Mark DF (1999) Proc Natl Acad Sci USA 96:12156–12161CrossRefGoogle Scholar
  15. 15.
    Audran M, Gareau R, Matecki S, Durand F, Chenard C, Sicart MT, Marion B, Bressolle F (1999) Med Sci Sports Exerc 31:639–645CrossRefGoogle Scholar
  16. 16.
    Breymann C (2000) Baillieres Best Pract Res Clin Endocrinol Metab 14:135–145CrossRefGoogle Scholar
  17. 17.
    Vergouwen PC, Collee T, Marx JJ (1999) Int J Sports Med 20:538–541CrossRefGoogle Scholar
  18. 18.
    Schmidt W, Biermann B, Winchenbach P, Lison S, Boning D (2000) Int J Sports Med 21:133–138CrossRefGoogle Scholar
  19. 19.
    Schumacher YO, Grathwohl D, Barturen JM, Wollenweber M, Heinrich L, Schmid A, Huber G, Keul J (2000) Int J Sports Med 21:380–385CrossRefGoogle Scholar
  20. 20.
    Parisotto R, Gore CJ, Hahn AG, Ashenden MJ, Olds TS, Martin DT, Pyne DB, Gawthorn K, Brugnara C (2000) Int J Sports Med 21:471–479CrossRefGoogle Scholar
  21. 21.
    Magnani M, Corsi D, Bianchi M, Paiardini M, Galluzzi L, Gargiullo E, Parisi A, Pigozzi F (2001) Blood Cells Mol Dis 27:559–571CrossRefGoogle Scholar
  22. 22.
    Parisotto R, Gore CJ, Emslie KR, Ashenden MJ, Brugnara C, Howe C, Martin DT, Trout GJ, Hahn AG (2000) Haematologica 85:564–572Google Scholar
  23. 23.
    Parisotto R, Wu M, Ashenden MJ, Emslie KR, Gore CJ, Howe C, Kazlauskas R, Sharpe K, Trout GJ, Xie M (2001) Haematologica 86:128–137Google Scholar
  24. 24.
    Gore CJ, Parisotto R, Ashenden MJ, Stray-Gundersen J, Sharpe K, Hopkins W, Emslie KR, Howe C, Trout GJ, Kazlauskas R, Hahn AG (2003) Haematologica 88:333–344Google Scholar
  25. 25.
    Sharpe K, Ashenden MJ, Schumacher YO (2006) Haematologica 91:356–363Google Scholar
  26. 26.
    Lippi G, Guidi G (2000) Clin Chem Lab Med 38:13–19CrossRefGoogle Scholar
  27. 27.
    Ashenden MJ, Sharpe K, Damsgaard R, Jarvis L (2004) Am J Clin Pathol 121:816–825CrossRefGoogle Scholar
  28. 28.
    Bechensteen AG, Lappin TR, Marsden J, Muggleston D, Cotes PM (1993) Br J Haematol 83:663–664Google Scholar
  29. 29.
    Lindstedt G, Lundberg PA (1998) Scand J Clin Lab Invest 58:441–458CrossRefGoogle Scholar
  30. 30.
    Marsden JT, Sherwood RA, Peters TJ (1999) Ann Clin Biochem 36(Pt 3):380–387Google Scholar
  31. 31.
    Benson EW, Hardy R, Chaffin C, Robinson CA, Konrad RJ (2000) J Clin Lab Anal 14:271–273CrossRefGoogle Scholar
  32. 32.
    Owen WE, Roberts WL (2004) Clin Chim Acta 340:213–217CrossRefGoogle Scholar
  33. 33.
    Abellan R, Ventura R, Pichini S, Remacha AF, Pascual JA, Pacifici R, Di Giovannandrea R, Zuccaro P, Segura J (2004) J Pharm Biomed Anal 35:1169–1177CrossRefGoogle Scholar
  34. 34.
    Kuiper-Kramer EP, Huisman CM, van Raan J, van Eijk HG (1996) Eur J Clin Chem Clin Biochem 34:645–649Google Scholar
  35. 35.
    Suominen P, Punnonen K, Rajamaki A, Irjala K (1997) Clin Chem 43:1641–1646Google Scholar
  36. 36.
    Yeung GS, Kjarsgaard JC, Zlotkin SH (1998) Eur J Clin Nutr 52:801–804CrossRefGoogle Scholar
  37. 37.
    Akesson A, Bjellerup P, Vahter M (1999) Scand J Clin Lab Invest 59:77–81CrossRefGoogle Scholar
  38. 38.
    Suominen P, Punnonen K, Rajamaki A, Majuri R, Hanninen V, Irjala K (1999) Clin Chem 45:1302–1305Google Scholar
  39. 39.
    Cotton F, Thiry P, Boeynaems J (2000) Clin Biochem 33:263–267CrossRefGoogle Scholar
  40. 40.
    Wians FH Jr, Urban JE, Kroft SH, Keffer JH (2001) Clin Chim Acta 303:75–81CrossRefGoogle Scholar
  41. 41.
    Paritpokee N, Bhokaisawan N, Wiwanitkit V, Boonchalermvichian C (2001) Asian Pac J Allergy Immunol 19:207–211Google Scholar
  42. 42.
    Thomas C, Thomas L (2002) Clin Chem 48:1066–1076Google Scholar
  43. 43.
    Abellan R, Ventura R, Pichini S, Sarda MP, Remacha AF, Pascual JA, Palmi I, Bacosi A, Pacifici R, Zuccaro P, Segura J (2004) J Immunol Methods 295:89–99CrossRefGoogle Scholar
  44. 44.
    Lasne F, de Ceaurriz J (2000) Nature 405:635CrossRefGoogle Scholar
  45. 45.
    Wide L, Bengtsson C, Berglund B, Ekblom B (1995) Med Sci Sports Exerc 27:1569–1576Google Scholar
  46. 46.
    Lasne F, Martin L, Crepin N, de Ceaurriz J (2002) Anal Biochem 311:119–126CrossRefGoogle Scholar
  47. 47.
    Lasne F (2001) J Immunol Methods 253:125–131CrossRefGoogle Scholar
  48. 48.
    Egrie JC, Browne JK (2001) Nephrol Dial Transplant 16(Suppl 3):3–13Google Scholar
  49. 49.
    Catlin DH, Breidbach A, Elliott S, Glaspy J (2002) Clin Chem 48:2057–2059Google Scholar
  50. 50.
    Pascual JA, Lasne F, Howe C, Saugy M, Hemmersbach P, Belalcazar V, Martin L, Grainger J, Pfister Y, Mangin P, Nissen-Lie G, Skibeli V (2002) Recent Adv Doping Anal 10:135144Google Scholar
  51. 51.
    Breidbach A, Catlin DH, Green GA, Tregub I, Truong H, Gorzek J (2003) Clin Chem 49:901–907CrossRefGoogle Scholar
  52. 52.
    Catlin D, Howe C, Lasne F, Nissen-Lie G, Pascual J, Saugy M (2004) WADA technical document—TD2004EPO. WADA. http://www.wada-ama.org/rtecontent/document/td2004epo_en.pdf
  53. 53.
    Belalcazar V, Gutiérrez Gallego R, Llop E, Segura J, Pascual JA (2006) Electrophoresis 27:4387–4395CrossRefGoogle Scholar
  54. 54.
    Lasne F, Martin L, de Ceaurriz J (2005) Recent Adv Doping Anal 13:297–304Google Scholar
  55. 55.
    Belalcazar V, Pascual JA, Abanades S, Farré M, de la Torre R (2005) Recent Adv Doping Anal 13:427–431Google Scholar
  56. 56.
    Nelson K, Trout GJ, Howe C, Kazlauskas R (2004) Recent Adv Doping Anal 12:309–316Google Scholar
  57. 57.
    Abellan R, Ventura R, Remacha AF, Rodríguez FA, Pascual JA, Segura J (2007) J Sport Sci (in press)Google Scholar
  58. 58.
    Berglund B, Aulin KP, Wide L (2003) Scand J Med Sci Sports 13:124–127CrossRefGoogle Scholar
  59. 59.
    World Anti-Doping Agency (2005) Clarification about the EPO detection method. http://www.wada-ama.org/rtecontent/document/EPO_QA.pdf
  60. 60.
    Khan A, Grinyer J, Truong ST, Breen EJ, Packer NH (2005) Clin Chim Acta 358:119–130CrossRefGoogle Scholar
  61. 61.
    Rabin OP, Lasne F, Pascual JA, Saugy M, Delbeke FJ, Van Eenoo P (2006) Clin Chim Acta 373:186–187CrossRefGoogle Scholar
  62. 62.
    Beullens M, Delanghe JR, Bollen M (2006) Blood 107:4711–4713CrossRefGoogle Scholar
  63. 63.
    Khan A, Packer N (2006) Clin Chim Acta 373:188CrossRefGoogle Scholar
  64. 64.
    Beullens M, Delanghe JR, Bollen M (2006) Blood 108:1179–1780Google Scholar
  65. 65.
    Lasne F (2006) Blood 108:1778–1779CrossRefGoogle Scholar
  66. 66.
    Catlin D, Green G, Sekera M, Scott P, Starcevic B (2006) Blood 108:1778–1780CrossRefGoogle Scholar
  67. 67.
    Takeuchi M, Takasaki S, Miyazaki H, Kato T, Hoshi S, Kochibe N, Kobata A (1988) J Biol Chem 263:3657–3663Google Scholar
  68. 68.
    Skibeli V, Nissen-Lie G, Torjesen P (2001) Blood 98:3626–3634CrossRefGoogle Scholar
  69. 69.
    Nagano M, Stubiger G, Marchetti M, Gmeiner G, Allmaier G, Reichel C (2005) Electrophoresis 26:1633–1645CrossRefGoogle Scholar
  70. 70.
    Hokke CH, Bergwerff AA, van Dedem GW, van Oostrum J, Kamerling JP, Vliegenthart JF (1990) FEBS Lett 275:9–14CrossRefGoogle Scholar
  71. 71.
    Llop E, Gutiérrez Gallego R, Belalcazar V, Gerwig GJ, Kamerling JP, Segura J, Pascual JA (2004) Recent Adv Doping Anal 12:297–307Google Scholar
  72. 72.
    Nimtz M, Martin W, Wray V, Kloppel KD, Augustin J, Conradt HS (1993) Eur J Biochem 213:39–56CrossRefGoogle Scholar
  73. 73.
    Tsuda E, Goto M, Murakami A, Akai K, Ueda M, Kawanishi G, Takahashi N, Sasaki R, Chiba H, Ishihara H (1988) Biochemistry 27:5646–5654CrossRefGoogle Scholar
  74. 74.
    Van Rooijen JJ, Kamerling JP, Vliegenthart JF (1998) Eur J Biochem 256:471–487CrossRefGoogle Scholar
  75. 75.
    Kawasaki N, Haishima Y, Ohta M, Itoh S, Hyuga M, Hyuga S, Hayakawa T (2001) Glycobiology 11:1043–1049CrossRefGoogle Scholar
  76. 76.
    Neususs C, Demelbauer U, Pelzing M (2005) Electrophoresis 26:1442–1450CrossRefGoogle Scholar
  77. 77.
    Balaguer E, Neususs C (2006) Anal Chem 78:5384–5393CrossRefGoogle Scholar
  78. 78.
    Yu B, Cong H, Liu H, Li Y, Liu F (2005) J Sep Sci 28:2390–2400CrossRefGoogle Scholar
  79. 79.
    Jelkmann W (2007) Eur J Haematol 78:183–205CrossRefGoogle Scholar
  80. 80.
    Schellekens H, Ryff JC (2002) Trends Pharmacol Sci 23:119–121CrossRefGoogle Scholar
  81. 81.
    Schellekens H (2004) Nat Biotechnol 22:1357–1359CrossRefGoogle Scholar
  82. 82.
    Schellekens H (2004) Trends Biotechnol 22:406–410CrossRefGoogle Scholar
  83. 83.
    Macdougall IC (2005) Curr Hematol Rep 4:436–440Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Jordi Segura
    • 1
    • 2
  • José A. Pascual
    • 1
    • 2
  • Ricardo Gutiérrez-Gallego
    • 1
    • 2
  1. 1.Pharmacology Research Unit, Municipal Institute for Medical ResearchIMIM-Hospital del MarBarcelonaSpain
  2. 2.Department of Experimental and Health SciencesCEXS, Pompeu Fabra UniversityBarcelonaSpain

Personalised recommendations