The Estimation of Erythropoietin (Epo): Principles, Problems and Progress

  • P. Mary Cotes
Part of the NATO ASI Series book series (volume 8)


Assay methods and estimates of the concentration of erythropoietin in normal human serum, when determined by different methods, are reviewed. The heterogeneity of estimates is discussed in relation to the principles upon which valid assays are based. Similarities between preparations of erythropoietin, standard and test samples compared, may give rise to problems and these are discussed. Advances in assay technology are considered.


Normal Human Serum Friend Virus Fetal Mouse Liver Rabbit Bone Marrow Bone Marrow Adherent Cell 
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  1. 1.
    Goldwasser E and Kung Ck-H (1971). Purification of erythropoietin. Proc Natl Acad Sci USA 68: 697–698.PubMedCrossRefGoogle Scholar
  2. 2.
    Miyake T, Kung CK-H and Goldwasser E (1977). Purification of human erythropoietin. J Biol Chem 252: 5558–5564.PubMedGoogle Scholar
  3. 3.
    Yanagawa S, Hirade K, Ohnoda H, Sasaki R, Chiba H, Ueda M and Goto M (1984). Isolation of human erythropoietin with monoclonal antibodies. J Biol Chem 259: 2707–2710.PubMedGoogle Scholar
  4. 4.
    Krystal G, Pankratz HRC, Farber NM and Smart JE (1986). Purification of human erythropoietin to homogeneity by a rapid five-step procedure. Blood 67: 71–79.PubMedGoogle Scholar
  5. 5.
    Browne JK, Cohen AM, Egrie JC, Lai PH, Lin FK, Strickland T, Watson E and Stebbing N (1986). Erythropoietin:gene cloning, protein structure and biological properties. Cold Spring Harbor Symp Quant Biol (in press).Google Scholar
  6. 6.
    Cotes PM (1986). Estimation of erythropoietin and its sec- retion in normal subjects. In: Fisher JW (ed) Kidney Hormones Vol III Academic Press, London pp. 535–558.Google Scholar
  7. 7.
    Sawatzki G(1987),Isolation of anti-erythropoietin by auto- mated Immuno-affinity FPLC using recombinant EPO. NATO ASI series. This volume.Google Scholar
  8. 8.
    McDonald JD, Lin F-K and Goldwasser E(1986).Cloning, sequ- encing, and evolutionary analysis of the mouse erythropoietin gene. Mol Cell Biol 6: 842–848.Google Scholar
  9. 9.
    Sue JM and Sytowski AJ (1983). Site specific antibodies to human erythropoietin directed towards the NH2-terminal region. Proc Natl Acad Sci USA 80: 3651–3655.PubMedCrossRefGoogle Scholar
  10. 10.
    Annable L, Cotes PM and Mussett MV(1972).The second international reference preparation of erythropoietin, human urinary, for bioassay. Bull Wld Hlth Org 47: 99–112.Google Scholar
  11. 11.
    Brandan NC, Cotes PM and Espada J (1981). In vitro assay of erythropoietin in fetal mouse liver cultures. I. Comparison of radioactive tracers and evidence of assay specificity. Br J Haematol 47: 461–468.Google Scholar
  12. 12.
    Weiss TL and Goldwasser E (1981). The biological properties of endotoxin-free human erythropoietin. Biochem J 198: 17–21.PubMedGoogle Scholar
  13. 13.
    Lappin TRJ, Elder E, Shannon JS, Taylor T and Bridges JM (1987). The effect of heat-treatment on the estimation of bioactive erythropoietin in serum using the mouse spleen cell assay. NATO ASI Series. This volume.Google Scholar
  14. 14.
    Berson SA and Yallow RS (1973). Radioimmunoassay 2.1.General. In: Berson SA and Yalow RS (eds) Methods in Investiagtive and Diagnostic Endocrinology. Vol 2A North-Holland, Amsterdam, pp. 84–120.Google Scholar
  15. 15.
    Egrie JC and Lane J (1987). Development of a radioimmunoassay for erythropoietin using recombinant erythropoietin-derived reagents. NATO ASI Series. This volume.Google Scholar
  16. 16.
    Brandon NC, Cotes PM and Espada (1981). In vitro assay of erythropoietin in fetal mouse liver cultures. II Effects of human transferrin bound iron andof serum on the stimulation of incorporation of H-thymidine into DNA. Br J Haematol 47: 469–478.CrossRefGoogle Scholar
  17. 17.
    Garcia JF and Schooley JC (1965). Dose-response relationships for erythropoietin given in serum or in saline. Proc Soc Exp Biol Med 120: 614–616.PubMedGoogle Scholar
  18. 18.
    Bozzini CE, Alippi RM and Barvelo AC (1987). Enhanced erythropoiesis induced by hypoxia in hypertransfused, posthypoxic, post-anemic or testosterone-treated polycythemic rodents. NATO ASI Series. This volume.Google Scholar
  19. 19.
    Lin FK (1987). The molecular biology of erythropoietin.NATO ASI Series. This volume.Google Scholar
  20. 20.
    Krantz SB, Sawyer ST, Kourny MJ and Bondurant MC (1987). Use of purified erythropoietin responsive cells produced by the anemia strain of Friend virus to study the action of erythropoietin (Ep). NATO ASI Series. This volume.Google Scholar
  21. 21.
    Bonanou-Tzedaki SA, Setchenska MS and Arnstein HRV (1986). Stimulation of adenylate cyclase activity of rabbit bone marrow immature erythroblasts by erythropoietin and haemin. Eur J Biochem 155: 363–370.PubMedCrossRefGoogle Scholar
  22. 22.
    Reid CDL, Prouse PJ, Baptista LC, Gumpel JM and Chanarin I (1984). The mechanism of anemia in rheumatoid arthritis: effects of bone marrow adherent cells and of serum on in vitro erythropoiesis. Br J Haematol 58: 607–615.PubMedCrossRefGoogle Scholar
  23. 23.
    Farrant J, Clark JC, Lee H, Knight SC and O’Brian J (1980) Conditions for measuring DNA synthesis in PHA stimulated human lymphocytes in 20u1 hanging drops with various cell concentrations and conditions of culture. J Immunol Meth 33: 301–312.CrossRefGoogle Scholar
  24. 24.
    Clemons GK (1986). Comparison of radioimmunoassay and bioassay of erythropoietin. In: Zanjani ED, Tavassoli M and Ascensao JL (eds) Humoral and cellular regulation of erythropoiesis. Spectrum, New York, In press.Google Scholar
  25. 25.
    Cotes PM, Canning CE and Gaines Das RE (1983). Modification of a radioimmunoassay for human serum erythropoietin to provide increased sensitivity and investigate nonspecific serum responses. In: Hunter WM and Corrie JET (eds) Immunoassays in Clinical Chemistry. Chruchill Livingstone, Edinburgh pp. 106–112 and 124–127.Google Scholar
  26. 26.
    Cotes PM (1982). Immunoreactive erythropoietin in serum. I. Evidence for the validity of the assay method and the physiological relevance of estimates. Brit J Haematol 50: 427–438.CrossRefGoogle Scholar
  27. 27.
    Egrie JC, Cotes PM, Lane J,Gaines Das RE and Tam RC(1985). Development of valid radioimmunoassays (RIA) for human erythropoietin (Epo) using recombinant Epo as tracer and immunogen. Blood 66:Suppl 1, 149a.Google Scholar
  28. 28.
    Cotes PM, Gaines Das RE and Tam RC (1987). The use of 125 recombinant DNA derived humanerythropoietin ( R-HuEpo) as a replacement for I human urinary erythropoietin in a radioimmunoassay for human Epo. NATO ASI Series, this volume.Google Scholar
  29. 29.
    Chappel SC, Ulloa-Aguirre A and Coutafaris C (1983). Biosynthesis and secretion of follicle stimulating hormone. Endocrine Rev 4: 179–211.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1987

Authors and Affiliations

  • P. Mary Cotes
    • 1
  1. 1.Section of HaematologyClinical Research CentreHarrow Middx.UK

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