Skip to main content
SpringerLink
Log in

Analysis of erythropoietin by capillary electrophoresis

  • Chapter
Handbook of Capillary Electrophoresis Applications

Abstract

Human erythropoietin (EPO) is a glycoprotein hormone responsible for regulating the growth and maturation of erythroid progenitor cells [1]. The site of biosynthesis is in the liver in the fetus and switches to the kidney in adults [2–4]. It is maintained in circulation at about 10 pM under normal physiological conditions, and may be excreted in urine [5]. EPO was first purified in small quantities from the urine of aplatic anemia patients [5]. The first functional recombinant human erythropoietin was cloned, expressed [6] and made available as one of the most celebrated therapeutic drugs in recent years. Recombinant EPO (r-EPO) and natural EPO purified from the urine showed no significant difference in their functional activity in vivo or their carbohydrate structures [7, 8].

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

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 259.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 329.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 329.99
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

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Goldwasser, E.; Kung, C.K.H. (1968) Chemistry and purification of erythropoietin: progress in the purification of erythropoietin. Ann. N Y. Acad. Sci. 149, 49–53.

    Article  CAS  Google Scholar 

  2. Jacobson, L.O.; Goldwasser, E.; Fried, W.; Plzak, L. (1957) Role of kidney in erythropoiesis. Nature 179, 633–634.

    Article  CAS  Google Scholar 

  3. Fried, W. (1972) The liver as a source of extrarenal erythropoietin production. Blood 40, 671–677.

    CAS  Google Scholar 

  4. Zanjani, E.D.; Ascensao, J.L.; McGlave, P.B.; Banisadre, M.; Ash, R.C. (1981) Studies on the liver to kidney switch of erythropoietin production. J. Clin. Invest. 67, 1183–1188.

    Article  CAS  Google Scholar 

  5. Miyake, T.; Kung, C.K.H.; Goldwasser, E. (1977) Purification of human erythropoietin. J. Biol. Chem. 252, 5558–5564.

    CAS  Google Scholar 

  6. Lin, F.K.; Suggs, J.; Lin, C.H.; Browne, J.; Smalling, R.; Egrie, J.C.; Chen, K.K.; Fox, G.M.; Martin, F.; Stabinsky, Z.; Badrawi, S.M.; Lai, P.S.; Goldwasser, E. (1985) Cloning and expression of human erythropoietin gene. Proc. Natl Acad. Sci. USA 82, 7580–7584.

    Article  CAS  Google Scholar 

  7. Sasaki, H.; Brothner, 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.

    CAS  Google Scholar 

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

    CAS  Google Scholar 

  9. Goldwasser, E.; Kung, C.K.H.; Ellason, J. (1974) On the mechanism of erythropoietin-induced differentiation. J. Biol. Chem. 249, 4202–4206.

    CAS  Google Scholar 

  10. Kobata, A. (1992) Structures and functions of the sugar chains of glycoproteins. Eur. J. Biochem. 209, 485–501.

    Article  Google Scholar 

  11. Takeuchi, M.; Inoue, N.; Strickland, T.W.; Kubota, M.; Wada, M.; Shimizu, R.; Hoshi, S.; Kozutsumi, H.; Takasaki, S.; Kobata, A. (1989) Relationship between sugar chain structure and biological activity of recombinant human erythropoietin produced in Chinese hamster ovary cells. Proc. Natl Acad. Sci. USA 86, 7819–7822.

    Article  CAS  Google Scholar 

  12. Rice, K.G.; Takahashi, N.; Namiki, Y.; Tran, A.D.; Lisi, P.J.; Lee, Y.C. (1992) Quantitative mapping of the N-Minked sialyloligosaccharides of recombinant erythropoietin: combination of direct high-performance anion-exchange chromatography and 2-aminopyridine derivatization. Anal. Biochem. 206, 278–287.

    Article  CAS  Google Scholar 

  13. Watson, E.; Bhide, A.; Van Halbeck, H. (1994) Structural determination of intact major sialylated oligosaccharide chains of recombinant human erythropoietin expressed in Chinese hamster ovary cells. Glycobiology 4, 227–237.

    Article  CAS  Google Scholar 

  14. Tsuda, E.; Goto, M.; Murakami, A.; Akai, K.; Ueda, M.; Kawanishi, G.; Takahashi, N.; Sasaki, R.; Chiba, H.; Ishihara, H.; Mori, M.; Tejima, S.; Endo, S.; Arata, Y. (1988) Comparative structural study of the N-linked oligosaccharides of urinary and recombinant erythropoietins. Biochemistry 27, 5646–5654.

    Article  CAS  Google Scholar 

  15. Nimitz, M.; Martin, W.; Wray, V.; Kloppel, K.D.; Augustin, J.; Conradt, H.S. (1993) Structures of sialylated oligosaccharides of human erythropoietin expressed in recombinant BHK-21 cells. Eur. J. Biochem. 213, 39–56.

    Article  Google Scholar 

  16. Ohashi, H.; Miyata, M.; Ishi, Y.; Takeuchi, M.; Takasago, A.; Suzuki, T.; Sudo, T. (1989) Purification and characterization of human recombinant erythropoietin expressed in human cervix carcinoma HeLa cells. Trends Anim. Cell Culture Technol. 1, 115–120.

    Google Scholar 

  17. Tran, A.D.; Huynh, C.T.; Park, S.; Ryall, R.R.; Lisi, P.J.; Lane, P.A. (1991) Separation of carbohydrate-mediated microheterogeneity of recombinant human erythropoietin by free solution capillary electrophoresis. J. Chromatogr. 542, 459–471.

    Article  CAS  Google Scholar 

  18. McCormick, R.M. (1988) Capillary zone electrophoretic separation of peptides and proteins using low pH buffers in modified silica capillaries. Anal. Chem. 60, 2322–2328.

    Article  CAS  Google Scholar 

  19. Chen, F.-T.A.; Kelly, L.; Palmieri, R.; Biehler, R.; Schwartz, H. (1992) Use of high ionic strength buffers for the separation of proteins and peptides with capillary electrophoresis. J. Liq. Chromatogr. 15, 114–119.

    Google Scholar 

  20. Watson, E.; Yao, F. (1993) Capillary electrophoretic separation of human recombinant erythropoietin (r-HuEPO) glycoforms. Anal Biochem. 210, 389–393.

    Article  CAS  Google Scholar 

  21. Benedek, K.; Thiede, S.J. (1994) High-performance capillary electrophoresis of proteins using sodium dodecylsulfate-poly(ethylene oxide). J. Chromatogr. 676, 209–217.

    Article  CAS  Google Scholar 

  22. Narhi, L.O.; Arakawa, T.; Aoki, K.H.; Elmore, R.; Rohde, M.F.; Boone, T.; Strickland, T.W. (1991) The effect of carbohydrate on the structure and stability of erythropoietin. J. Biol. Chem. 266, 23022–23026.

    CAS  Google Scholar 

  23. Rush, R.S.; Derby, P.L.; Strickland, T.W.; Rohde, M.F. (1993) Peptide mapping and evaluation of glycopeptide microheterogeneity derived from endoproteinase dogestion of erythropoietin by affinity high-performance capillary electrophoresis. Anal Chem. 65, 1834–1842.

    Article  CAS  Google Scholar 

  24. Sasaki, H.; Ochi, N.; Dell, A.; Fukuda, M. (1988) Site-specific glycosylation of human recombinant erythropoietin expressed in Chinese hamster ovary cells. Biochemistry 27, 8618–8626.

    Article  CAS  Google Scholar 

  25. Boss, H.J.; Rohde, M.F.; Rush, R.S. (1995) Multiple sequenctial fraction collection of peptides and glycoproteins by high-performance capillary electrophoresis. Anal. Biochem. 230, 123–129.

    Article  CAS  Google Scholar 

  26. Hermentin, P.; Doenges, R.; Witzel, R.; Hokke, C.H.; Vliegenthart, J.P.G.; Kamerling, J.P.; Conradt, H.S.; Nimtz, M.; Brazel, D. (1994) A strategy for mapping N-glycans by high-performance capillary electrophoresis. Anal. Biochem. 221, 29–41.

    Article  CAS  Google Scholar 

  27. Evangelista, R.A.; Liu, M.-S.; Chen, F.-T.A. (1995) Characterization of 9-aminopyrene-1,4,6-trisulfonate derivatized sugars by capillary electrophoresis with laser-induced fluorescence detection. Anal Chem. 67, 2239–2245.

    Article  CAS  Google Scholar 

  28. Evangelista, R.A.; Chen, F.-T.A.: unpublished results. We thank Dr Fu-Kuen Lin of Amgen for providing the r-EPO sample.

    Google Scholar 

Download references

Authors
  1. F.-T. A. Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. National Institute of Health Sciences, Ministry of Health & Welfare, 1-18-1, Kamiyoga, Setagaya, Tokyo, Japan

    Hideharu Shintani (Section Director) (Section Director)

  2. Slovak Technical University, Bratislava, Slovakia

    J. Polonský (Professor of Analytical Chemistry) (Professor of Analytical Chemistry)

Rights and permissions

Reprints and permissions

Copyright information

© 1997 Chapman & Hall

About this chapter

Cite this chapter

Chen, FT.A. (1997). Analysis of erythropoietin by capillary electrophoresis. In: Shintani, H., Polonský, J. (eds) Handbook of Capillary Electrophoresis Applications. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-1561-9_16

Download citation

  • DOI: https://doi.org/10.1007/978-94-009-1561-9_16

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-7197-0

  • Online ISBN: 978-94-009-1561-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation