Skip to main content
SpringerLink
Log in
Book cover

BIOACTIVE ANALYTES, Including CNS Drugs, Peptides, and Enantiomers pp 17–36Cite as

Analysis of Protein Growth Factors and Optimization of Their Separation by HPLC

  • Chapter

  • 101 Accesses

Part of the book series: Methodological Surveys in Biochemistry and Analysis ((MSBA,volume 16 A))

Abstract

For polypeptide and protein GF’s*, which may occur in very small amounts and whose bioassay may lack specificity, chromatographic systems are needed which give high resolution and recovery and preserve biological activity. Resolving power in RP-HPLC depends on the nature of the stationary phase, on the pH and ionic strength of the primary solvent, and on the organic modifier used. Hydrophobic ion-pairing agents, anionic or cationic, can enhance selectivity. Protein recovery depends on solubility in the eluent, which must not irreversibly disrupt secondary structure and so cause inactivation.

Milder conditions are possible using ‘hydrophobic interaction’ HPLC — with packings still to be fully tested — or ion-exchange HPLC as in conventional operation but with enhanced speed and resolution, extendable by use of hydrophobic ion-pairing agents. Size-exclusion HPLC resolves no better than conventional operation.

This is a preview of subscription content, 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   39.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   54.99
Price excludes VAT (USA)
  • Compact, lightweight 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

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Smith, J.A., Ham, J., Winslow, D.P., O’Hare, M.J. & Rudland, P.S. (1984) J. Chromatog.305, 295–308.

    CAS  Google Scholar 

  2. Simpson, A.J., Smith, J.A., Moritz, R.L., O’Hare, M.J., Rudland, P.S., Morrison, J.R., Lloyd, C.J., Grego, B., Burgess, A.W. & Nice, E.C. (1985) Eur. J. Biochem.153, 629–637.

    Article  CAS  Google Scholar 

  3. Unger, K.K. (1979) Porous Silica: its Properties and Use as a Support in Column Liquid Chromatography, Elsevier, Amsterdam.

    Google Scholar 

  4. O’Hare, M.J. & Nice, E.C. (1979) J. Chromatog.171, 209–226.

    Article  Google Scholar 

  5. Nice, E.G., Capp, M., Cooke, N. & O’Hare, M.J. (1981) J. Chromatog.218, 569–580.

    Article  CAS  Google Scholar 

  6. Meek, J.L. (1980) Proc. Nat. Acad. Sci.77, 1632–1636.

    Article  CAS  Google Scholar 

  7. Browne, C.A., Bennett, H.P.J. & Solomon, S. (1982) Anal. Biochem.124, 201–208.

    Article  CAS  Google Scholar 

  8. Nice, E.C., Capp, M. & O’Hare, M.J. (1979) J. Chromatog.185, 413–427.

    Article  CAS  Google Scholar 

  9. Cooke, N., Archer, B.G., O’Hare, M.J., Nice, E.C. & Capp M. (1983) c7. Chromatog. 225, 115–123.

    Google Scholar 

  10. O’Hare, M.J., Capp, M., Nice, E.C., Cooke, N. & Archer, B.G. (1982) Anal.. Biochem.126, 17–28.

    Article  Google Scholar 

  11. O’Hare, M.J., Smith, J.A., Archer, B.G., Cooke, N. & Nice, E.C. (1983) in Protides of Biological Fluids Colloquium30 (Peeters, H., ed.), Pergamon, Oxford, pp.723–726.

    Google Scholar 

  12. Smith, J.A. & O’Hare, M.J. (1984) J. Chromatog.299, 13–28.

    Article  CAS  Google Scholar 

  13. D’Agostino, G., Mitchell, F., Castagnetta, L. & O’Hare, M.J. (1984) J. Chromatog.305, 13–26.

    Google Scholar 

  14. D’Agostino, G., Castagnetta, L., Mitchell, F. & O’Hare, M.J. (1985) J. Chromatog.338, 1–23.

    Google Scholar 

  15. Hearn, M.T.W. (1983) in High Performance Liquid Chromatography-Advances and Perspectives, Vol. 3 (Horvath, C.S., ed.), Academic Press, New York, pp. 87–155.

    Google Scholar 

  16. Smith, J.A., Winslow, D.P., O’Hare, M.J. & Rudland, P.S.(1984) Biochem. Biophys. Res. Cornm.119, 311–318.

    Article  CAS  Google Scholar 

  17. Smith, J.A. & O’Hare, M.J. (1985) J. Chromatog.345, 168–172.

    CAS  Google Scholar 

  18. Nice, E.C., Lloyd, C. & Burgess, A.W. (1984) J. Chromatog.296, 153–170.

    Article  CAS  Google Scholar 

  19. Cohen, S.A., Benedek, K.P., Dong, S., Tapuhi, Y. & Karger, B.L. (1984) Anal. Chem.56, 217–221.

    Article  CAS  Google Scholar 

  20. Cowley, G., Smith, J.A., Ellison, M. & Gusterson, B. (1985) Internat. J. Cancer.35, 575–579.

    Article  CAS  Google Scholar 

  21. Grego, B. & Hearn, M.T.W. (1984) J. Chromatog.336, 25–40.

    CAS  Google Scholar 

  22. Fausnaugh, J.L., Pfannkoch, E., Gupta, S. & Regnier, F.G. (1984) Anal. Biochem.137, 464–472.

    Article  CAS  Google Scholar 

  23. Kato, Y., Kitamura, T. & Hashimoto, T. (1984) J. Chromatog.292, 418–426.

    Article  CAS  Google Scholar 

  24. Regnier, F.G. (1984) Meths. Enzymol.104, 170–189.

    Article  CAS  Google Scholar 

  25. Hofstee, B.H.J. (1973) Biochem. Biophys. Res. Comm.50, 751–757.

    Article  CAS  Google Scholar 

  26. Kopaciewicz, W., Rounds, M.A. & Regnier, F.G. (1985) J. Chromatog.318, 157–172.

    Article  CAS  Google Scholar 

  27. Schmidt, D.E., Giese, R.W., Conron, D. & Karger, B.L. (1980) Anal. Chem.52, 177–182.

    Article  CAS  Google Scholar 

  28. Unger, K. (1984) Meths. Enzymol.104, 154–169.

    Article  CAS  Google Scholar 

  29. Bohlen, P., Baird, A., Esch, F., Ling, N. & Gospodarowicz, D. (1984) Proc. Nat. Acad. Sci. 01, 5364–5368.

    Article  Google Scholar 

  30. Stroobant, P. & Waterfield, M.D. (1984) EMBO J. 3, 2963–2967.

    CAS  Google Scholar 

  31. Nicola, N.A., Metcalf, D., Matsumoto, M. & Johnson, G.R. (1983) J. Biol. Chem.258, 9017–9023.

    CAS  Google Scholar 

  32. Kimball, E.S., Bohn, W.H., Cockley, K.D., Warren, T.C & Sherwin, S.A. (1984) Cancer Res. 44, 3613–3619.

    CAS  Google Scholar 

  33. Twardzik, D.R., Kimball, E.S., Sherwin, S.A., Ranchalis, J.E. & Todaro, G. (1985) Cancer Res. 45, 1934–1939.

    CAS  Google Scholar 

  34. Twardzik, D.R., Brown, J.P., Ranchalis, J.E., Todaro, G.J. & Mors, B, (1985) Proc. Nat. Acad. Sci.82, 5300–5304.

    Article  CAS  Google Scholar 

  35. Libby, P., Raines, E.W., Cullinane, P.M. & Ross, R. (1985) J. Cell Physiol.125, 357–366.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Ludwig Institute for Cancer Research, London Branch, Royal Marsden Hospital, Sutton, Surrey, SM2 5PX, UK

    John A. Smith & Michael J. O’Hare

Authors
  1. John A. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael J. O’Hare
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Guildford Academic Associates, Guildford, UK

    Eric Reid

  2. ICI Pharmaceuticals Division, Alderley Park, UK

    Bryan Scales  & Ian D. Wilson  & 

Rights and permissions

Reprints and permissions

Copyright information

© 1986 Springer Science+Business Media New York

About this chapter

Cite this chapter

Smith, J.A., O’Hare, M.J. (1986). Analysis of Protein Growth Factors and Optimization of Their Separation by HPLC. In: Reid, E., Scales, B., Wilson, I.D. (eds) BIOACTIVE ANALYTES, Including CNS Drugs, Peptides, and Enantiomers. Methodological Surveys in Biochemistry and Analysis, vol 16 A. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-1892-8_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4757-1892-8_3

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4757-1894-2

  • Online ISBN: 978-1-4757-1892-8

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation