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Chemistry and Assays of Erythropoietin

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Erythropoietin

Abstract

The current therapeutic use of erythropoietin (Epo) represents the culmination of a difficult struggle to isolate, purify and characterise the hormone. Epo is the principal regulatory hormone of the circulating red cell mass [1,2] and is normally present in minute concentrations in serum at 15–30 milliunits/ml, or approximately 10−11 M [3–5]. The study of Epo has been hampered for many years by its extreme scarcity and the lack of a convenient assay.

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References

  1. Goldwasser E (1975) Erythropoietin and the differentiation of red blood cells. Fed Proc 34: 2285–2292

    PubMed  CAS  Google Scholar 

  2. Graber SE, Krantz SB (1978) Erythropoietin and the control of red cell production. Annu Rev Med 29: 51–66

    Article  PubMed  CAS  Google Scholar 

  3. Koeffler HP, Goldwasser E (1981) Erythropoietin radioimmunoassay in evaluating patients with polycythemia. Ann Intern Med 94: 44–47

    PubMed  CAS  Google Scholar 

  4. Garcia JF, Ebbe SN, Hollander L, Cutting HO, Miller ME, Cronkite EP (1982) Radioimmunoassay of erythropoietin: circulating levels in normal and polycythemic human beings. J Lab Clin Med 99: 624–635

    PubMed  CAS  Google Scholar 

  5. Cotes PM (1982) Immunoreactive erythropoietin in serum. 1. Evidence for the validity of the assay method and the physiological relevance of estimates. Br J Haematol 50: 427–438

    Article  PubMed  CAS  Google Scholar 

  6. Davis JM, Arakawa T, Strickland TW, Yphantis DA (1987) Characterization of recombinant human erythropoietin produced in Chinese hamster ovary cells. Biochemistry 26: 2633–2638

    Article  PubMed  CAS  Google Scholar 

  7. Mok M, Spivak JL (1982) Protease activity in human urine erythropoietin preparations. Exp Hematol 10: 300–306

    PubMed  CAS  Google Scholar 

  8. Lowy PH (1970) Preparation and chemistry of erythropoietin. In: Gordon AS (ed) Regulation of hematopoiesis. Appleton-Century-Crofts, East Norwalk, pp 395–412

    Google Scholar 

  9. Espada J, Brandan NC, Dorado M (1973) Effect of chemical and enzymatic agents on the biological activity of erythropoietin. Acta Physiol Latinoam 23: 193–201

    CAS  Google Scholar 

  10. Lee-Huang S (1980) A new preparative method for isolation of human erythropoietin with hydrophobic interaction chromatography. Blood 56: 620–624

    PubMed  CAS  Google Scholar 

  11. Wang FF, Kung CK-H, Goldwasser E (1985) Some chemical properties of human erythropoietin. Endocrinology 116: 2286–2292

    Article  PubMed  CAS  Google Scholar 

  12. Miyake T, Kung CK-H, Goldwasser E (1977) Purification of human erythropoietin. J Biol Chem 252: 5558–5564

    PubMed  CAS  Google Scholar 

  13. Lin F-K, Suggs S, Lin C-H, Browne JK, Smalling R, Egrie JC, Chen KK, Fox GM, Martin F, Stabinsky Z, Badrawi SM, Lai P-H, Goldwasser E (1985) Cloning and expression of the human erythropoietin gene. Proc Natl Acad Sci USA 82: 7580–7584

    Article  PubMed  CAS  Google Scholar 

  14. Jacobs K, Shoemaker C, Rudersdorf R, Neill SD, Kaufmann RJ, Mufson A, Seehra J, Jones SS, Hewick R, Fritsch EF, Kawakita M, Shimizu T, Miyake T (1985) Isolation and characterization of genomic and cDNA clones of human erythropoietin. Nature 313: 806–810

    Article  PubMed  CAS  Google Scholar 

  15. Lai P-H, Everett R, Wang F-F, Arakawa T, Goldwasser E (1986) Structural characterization of human erythropoietin. J Biol Chem 261: 3116–3121

    PubMed  CAS  Google Scholar 

  16. Fyhrquist F, Rosenlöf K, Grönhagen-Riska C, Hortling L, Tikkanen I (1984) Is renin substrate an erythropoietin precursor? Nature 308: 649–652

    Article  PubMed  CAS  Google Scholar 

  17. Powell JS, Berker KL, Lebo RV, Adamson JW (1986) Human erythropoietin gene: high level expression in stably transfected mammalian cells and chromosome localization. Proc Natl Acad Sci USA 83: 6465–6469

    Article  PubMed  CAS  Google Scholar 

  18. Watkins PC, Eddy R, Hoffman N, Stranislovitis P, Beck AK, Galli J, Vellucci V, Gusella JF, Shows TB (1986) Regional assignment of the erythropoietin gene to human chromosome region 7pter–q22. Cytogenet Cell Genet 42: 214–218

    Article  PubMed  CAS  Google Scholar 

  19. Recny MA, Scoble HA, Kim Y (1987) Structural characterization of natural human urinary and recombinant DNA — derived erythropoietin. J Biol Chem 262: 17156–17163

    PubMed  CAS  Google Scholar 

  20. Wang FF, Kung CK-H, Goldwasser E (1985) Some chemical properties of human erythropoietin. Endocrinology 116: 2286–2292

    Article  PubMed  CAS  Google Scholar 

  21. McDonald JD, Lin F-K, Goldwasser E (1986) Cloning, sequencing and evolutionary analysis of the mouse erythropoietin gene. Mol Cell Biol 6: 842–848

    PubMed  CAS  Google Scholar 

  22. Chou PY, Fasman GD (1978) Empirical predictions of protein conformation. Annu Rev Biochem 47: 251–276

    Article  PubMed  CAS  Google Scholar 

  23. Goldwasser E, McDonald J, Beru N (1987) The molecular biology of erythropoietin and the expression of its gene. In: Rich IN (ed) Molecular and cellular aspects of erythropoietin and erythropoiesis. Springer, Berlin Heidelberg New York pp 11–21 (NATO ASI series, vol H8)

    Chapter  Google Scholar 

  24. Neuberger A, Gottschalk A, Marshall RD, Spiro RG (1972) Carbohydrate-peptide linkages in glycoproteins and methods for their elucidation. In: Gottschalk A (ed) The glycoproteins: their composition, structure and function. Elsevier/North Holland, Amsterdam, pp 450–490

    Google Scholar 

  25. Dordal MS, Wang FF, Goldwasser E (1985) The role of carbohydrate in erythropoietin action. Endocrinology 116: 2293–2299

    Article  PubMed  CAS  Google Scholar 

  26. Egrie JC, Strickland TW, Lane J, Aoki K, Cohen AM, Smalling R, Trail G, Lin FK, Browne JK, Hinds DK (1986) Characterization and biological effects of recombinant human erythropoietin. Immunobiol ogy 172: 213–224

    Article  CAS  Google Scholar 

  27. Takeuchi M, Takasaki S, Inoue N, Kobata A (1988) Sensitive method for carbohydrate composition analysis of glycoproteins by high-performance liquid chromatography. J Chromatogr 400: 207–213

    Article  Google Scholar 

  28. Goldwasser E, Kung CK-H, Eliason JF (1974) On the mechanism of erythropoietin-induced differentiation. 13. The role of sialic acid in erythropoietin action. J Biol Chem 249: 4202–4206

    PubMed  CAS  Google Scholar 

  29. Lowy PH, Keighley G, Borsook H (1960) Inactivation of erythropoietin by neuraminidase and by mild substitution reactions. Nature 185: 102–103

    Article  PubMed  CAS  Google Scholar 

  30. Briggs DW, Fisher JW, George WJ (1974) Hepatic clearance of intact and desialylated erythropoietin. Am J Physiol 227: 1385–1388

    PubMed  CAS  Google Scholar 

  31. Schooley JC (1985) Neuraminidase increases DNA synthesis of spleen cells induced by native and asialylated erythropoietin. Exp Hematol 13: 994–998

    PubMed  CAS  Google Scholar 

  32. Sasaki H, Bothner B, Dell A, Fukuda M (1987) Carbohydrate structure of erythropoietin expressed in Chinese hamster ovary cells by a human erythropoietin cDNA. J Biol Chem 262: 12059–12076

    PubMed  CAS  Google Scholar 

  33. Goto M, Akai K, Murakami A, Hashimoto C, Tsuda E, Ueda M, Kawanishi G, Takahashi N, Ishimoto A, Chiba H, Sasaki R (1988) Production of recombinant human erythropoietin in mammalian cells: host cell dependency of the biological activity of the cloned glycoprotein. Biotechnology 6: 67–71

    Article  CAS  Google Scholar 

  34. Takeuchi M, Takasaki S, Miyazaki H, Kato T, Hoshi S, Kochibe N, Kobata A (1988) Comparative study of the asparagine — linked sugar chains of human erythropoietins purified from urine and the culture medium of recombinant Chinese hamster ovary cells. J Biol Chem 263: 3657–3663

    PubMed  CAS  Google Scholar 

  35. Morell AG, Irvine RA, Sternlieb I, Scheinberg IH, Ashwell G (1968) Physical and chemical studies on ceruloplasmin in vivo. J Biol Chem 243: 155–159

    PubMed  CAS  Google Scholar 

  36. Sherwood JB, Goldwasser E (1979) A radioimmunoassay for erythropoietin. Blood 54: 885–893

    PubMed  CAS  Google Scholar 

  37. Garcia JF, Sherwood JB, Goldwasser E (1979) Radioimmunoassay of erythropoietin. Blood Cells 5: 405–419

    PubMed  CAS  Google Scholar 

  38. Fried W, Plzak LF, Jacobson LO, Goldwasser E (1957) Studies on erythropoiesis. III. Factors controlling erythropoietin production. Proc Soc Exp Biol Med 94: 237–241

    PubMed  CAS  Google Scholar 

  39. Jacobson LO, Goldwasser E, Plzak LF, Fried W (1957) Studies on erythropoiesis. IV. Reticulocyte response of hypophysectomized and polycythemic rodents to erythropoietin. Proc Soc Exp Biol Med 94: 243–249

    PubMed  CAS  Google Scholar 

  40. Cotes PM, Bangham DR (1961) Bioassay of erythropoietin in mice made polycythaemic by exposure to air at reduced pressure. Nature 191: 1065–1067

    Article  PubMed  CAS  Google Scholar 

  41. Goldwasser E, Eliason JF, Sikkema D (1975) An assay for erythropoietin in vitro at the milliunit level. Endocrinology 97: 315–323

    Article  PubMed  CAS  Google Scholar 

  42. Dunn CDR, Jarvis JH, Greenman JM (1975) A quantitative bioassay for erythropoietin using mouse fetal liver cells. Exp Hematol 3: 65–78

    PubMed  CAS  Google Scholar 

  43. Bessler H, Notti I, Djaldetti M (1980) Quantitative determination of human plasma erythropoietin using embryonic mouse liver erythroblasts. Acta Haematol 63: 204–210

    Article  PubMed  CAS  Google Scholar 

  44. Krystal G (1983) A simple microassay for erythropoietin based on 3H-thymidine incorporation into spleen cells from phenylhydrazine — treated mice. Exp Hematol 11: 649–660

    PubMed  CAS  Google Scholar 

  45. Sherwood JB (1984) The chemistry and physiology of erythropoietin. Academic, New York pp 161–211, Vitamins and hormones, vol 41

    Google Scholar 

  46. Lappin TRJ, Elder GE, McKibbin SH, McNamee PT, McGeown MG, Bridges JM (1985) The effect of transferrin saturation on the estimation of erythropoietin by the mouse spleen cell microassay. Exp Hematol 13: 1007–1013

    PubMed  CAS  Google Scholar 

  47. Goldwasser E (1973) Erythropoietin. Methods in investigative and diagnostic endocrinology, p III, nonpituitary hormones. Elsevier, Amsterdam, p 1097

    Google Scholar 

  48. Hunter WM, Greenwod FC (1962) Preparation of iodine — 131I-labelled human growth hormone of high specific activity. Nature 194: 495–496

    Article  PubMed  CAS  Google Scholar 

  49. Tejedor G, Ballestra JPG (1982) Iodination of biological samples without loss of functional activity. Anal Biochem 127: 143–149

    Article  PubMed  CAS  Google Scholar 

  50. Fraker PJ, Speck Jr JC (1978) Protein and cell membrane iodinations with a sparingly soluble chloroamide, l,3,4,6-tetrachloro-3a,6a-diphenylglycoluril. Biochem Biophys Res Commun 80: 849–857

    Article  PubMed  CAS  Google Scholar 

  51. Rege AB, Brookins J, Fisher JW (1982) A radioimmunoassay for erythropoietin: serum levels in normal human subjects and patients with hemopoietic disorders. J Lab Clin Med 100: 829–843

    PubMed  CAS  Google Scholar 

  52. Garcia JF, Clemons GK (1983) The radioimmunoassay of erythropoietin. In: Lawrence JH, Winchell S (eds) Recent advances in nuclear medicine, vol 6. Grune and Stratton, New York, pp 19–40.

    Google Scholar 

  53. Sawyer ST, Krantz SB, Goldwasser E (1987) Binding and Receptor — mediated endocytosis of erythropoietin in Friend Virus — infected erythroid cells. J Biol Chem 262: 5554–5562

    PubMed  CAS  Google Scholar 

  54. Mizoguchi H, Ohta K, Suzuki T, Murakami A, Ueda M, Sasaki R, Chiba H (1987) Basic conditions for radioimmunoassay of erythropoietin, and plasma levels of erythropoietin in normal subjects and anaemic patients. Acta Haematol Jpn 50: 15–24

    CAS  Google Scholar 

  55. Urabe A, Saito T, Fukamachi H, Kubota M, Takaku F (1987) Serum erythropoietin titers in the anemia of chronic renal failure and other hematological states. Int J Cell Cloning 5: 202–208

    Article  PubMed  CAS  Google Scholar 

  56. Egrie JC, Lane J (1987) Development of a radioimmunoassay for erythropoietin using recombinant erythropietin — derived reagents. In: Rich IN (ed) Molecular and cellular aspects of erythropoietin and erythropoiesis. Springer, Berlin Heidelberg New York pp 395–407 (NATO ASI series, vol H8)

    Chapter  Google Scholar 

  57. Lappin TRJ, Elder GE, Taylor T, McMullin MF, Bridges JM (1988) Comparison of the mouse spleen cell assay and a radioimmunoassay for the measurement of serum erythropoietin. Br J Haemato 170: 117–120

    Article  Google Scholar 

  58. Cotes PM (1987) The estimation of erythropoietin (Epo): principles, problems and progress. In: Rich IN (ed) Molecular and cellular aspects of erythropoietin and erythropoiesis. Springer, Berlin Heidelberg New York pp 377–387 (NATO ASI series, vol H8)

    Chapter  Google Scholar 

  59. Erslev AJ, Wilson J, Caro J (1987) Erythropoietin titers in anemic nonuremic patients. J Lab Clin Med 109: 429–433

    PubMed  CAS  Google Scholar 

  60. Dunn CDR, Lange RD (1980) Erythropoietin titers in normal human serum. An appraisal of assay techniques. Exp Hematol 8: 231–235

    PubMed  CAS  Google Scholar 

  61. de Klerk G, Vet RJ, Rosengarten PC, Goudsmit R (1980) Comparison of the hemagglutination inhibition assay kit for erythropoietin (ESF) with the fetal mouse liver cell bioassy in vitro. Blood 55: 955–959

    PubMed  Google Scholar 

  62. Cotes PM, Doré CJ, Liu Yin JA, Lewis SM, Messinezy M, Pearson TC, Reid C (1986) Determination of serum immunoreactive erythropoietin in the investigation of erythrocytosis. N Engl J Med 315: 283–287

    Article  PubMed  CAS  Google Scholar 

  63. Caro J, Brown S, Miller O, Murray T, Erslev AJ (1979) Erythropoietin levels in uremic nephric and anephric patients. J Lab Clin Med 93: 449–458

    PubMed  CAS  Google Scholar 

  64. Zaroulis CG, Hoffman BJ, Kourides IA (1981) Serum concentrations of erythropoietin measured by radioimmunoassay in hematologic disorders and chronic renal failure. Am J Hematol 11: 85–92

    Article  PubMed  CAS  Google Scholar 

  65. Fisher JW, Lertora JL, Lindholm DD, Tornyos K, Moriyama Y (1973) Erythropoietin production and inhibitors in serum in the anemia of uremia. Proc Clin Dialysis Transplant Forum 3: 22–23

    CAS  Google Scholar 

  66. McGonigle RJS, Husserl F, Wallin JD, Fisher JW (1984) Hemodialysis and continuous ambulatory peritoneal dialysis effects on erythropoiesis in renal failure. Kidney Int 25: 430–436

    Article  PubMed  CAS  Google Scholar 

  67. Sherwood JB, Carmichael LD, Goldwasser E (1988) The heterogeneity of circulating human serum erythropoietin. Endocrinology 122: 1472–1477

    Article  PubMed  CAS  Google Scholar 

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Authors
  1. T. R. J. Lappin
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  2. A. P. Maxwell
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Editors and Affiliations

  1. Physiologisches Institut, Medizinische Universität zu Lübeck, Ratzeburger Allee 160, 2400, Lübeck 1, Federal Republic of Germany

    Wolfgang Jelkmann  & Andreas J. Gross  & 

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© 1989 Springer-Verlag Berlin Heidelberg

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Lappin, T.R.J., Maxwell, A.P. (1989). Chemistry and Assays of Erythropoietin. In: Jelkmann, W., Gross, A.J. (eds) Erythropoietin. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-83745-6_2

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  • DOI: https://doi.org/10.1007/978-3-642-83745-6_2

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-83747-0

  • Online ISBN: 978-3-642-83745-6

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