Advertisement

Chromatographia

, Volume 52, Issue 5–6, pp 334–339 | Cite as

Development of a push-pull microdialysis sampling technique for the quantitative determination of proteins

  • S. Kjellström
  • S. Lindberg
  • T. Laurell
  • G. Marko-Varga
Originals Column Liquid Chromatography

Summary

An on-line push-pull sampling technique has been developed for continuous analysis of proteins of molecular weight from 5.7 to 67 kDa. The characteristics of the system include gradient elution with a total cycle time of 21 min, membrane stability, unattended automatic operation, and adjustment of the sampling mode and extraction fraction (the ratio of the concentration of analyte in the dialysate to that in the sample) by varying the effective dialysis length. The push and pull flow rates were adjusted in a manner which enabled three different modes of operation. When push-pull microdialysis was compared with conventional microdialysis sampling, significantly higher extraction fractions were obtained for all five model proteins studied. The technique has been applied to the quantification of proteins in cell samples. On-line fractionation enabled complementary MS identification of the proteins present.

Key Words

Column liquid chromatography Microdialysis sampling Push-pull Proteins 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. [1]
    Anal. Chim. Acta 1999,379.Google Scholar
  2. [2]
    Nakamura, M.; Itano, T.; Yamaguchi, F.; Mizobuchi, M.; Tokuda, M.; Matsui, H.; Hosokawa, S.K.; Ohmoto, T.; Hatase, O.Acta Med. Okayama 1990,44, 1–8.Google Scholar
  3. [3]
    Woodroofe, M.N.; Sarna, G.S.; Wadhwa, M.; Hayes, G.M.; Loughlin, A.J.; Tinker, A.; Cuzner, M.L.J. Neuroimmunology 1991,33, 227–236.CrossRefGoogle Scholar
  4. [4]
    Schneiderheinze, J.M.; Hogan, B.L.Anal. Chem. 1996,68, 3758–3762.CrossRefGoogle Scholar
  5. [5]
    Takahashi, H.; Shintani, Y.; Okauchi, T.; Ishikawa, M.; Bando, H.; Azekawa, T.; Morita, Y.; Saito, S.J. Neuroscience Meth. 1994,52, 33–38.CrossRefGoogle Scholar
  6. [6]
    Asai, S.; Kohno, T.; Ishii, Y.; Ishikawa, K.Anal. Biochem. 1996,237, 182–187.CrossRefGoogle Scholar
  7. [7]
    Kjellström, S.; Emnéus, J.; Laurell, T.; Heintz, L.; Marko-Varga, G.J. Chromatogr. A 1998,823, 489–496.CrossRefGoogle Scholar
  8. [8]
    Kjellström, S.; Appels, N.; Laurell, T.; Marko-Varga, G.Chromatographia 1999,50, 539–546.CrossRefGoogle Scholar
  9. [9]
    Heintz, L.; Österlind, E.; Alkner, U.; Marko-Varga, G.Chromatographia 1997,46, 365–373.CrossRefGoogle Scholar
  10. [10]
    Laurell, T.; Buttler, T.Anal. Methods Instr. 1995,2, 197–201.Google Scholar
  11. [11]
    Önnerfjord, P.; Ekström, S.; Berquist, J.; Nilsson, J.; Laurell, T.; Marko-Varga, G.Rapid Commun. Mass Spectrom. 1999,13, 315–322.CrossRefGoogle Scholar
  12. [12]
    Khramov, A.N.; Stenken, J.S.Anal. Chem. 1999,71, 1257–1264.CrossRefGoogle Scholar
  13. [13]
    Bungay, P.M.; Morrison P.F.; Dedrick R.L.Life Sci. 1990,46, 105–119.CrossRefGoogle Scholar
  14. [14]
    Wages, S.; Church, W.; Justice, J.Anal. Chem. 1986,58, 1649–1656.CrossRefGoogle Scholar
  15. [15]
    Miliotis, T.; Kjellström, S.; Nilsson, J.; Laurell, T.; Edholm, L.-E.; Marko-Varga, G.J. Mass Spec. 2000,35, 369–377.CrossRefGoogle Scholar
  16. [16]
    Miliotis, T.; Kjellström, S.; Önnefjord, P.; Nilsson, J.; Laurell, T.; Edholm, L.-E.; Marko-Varga, G.J. Chromatogr. A. 2000,886, 99–100.CrossRefGoogle Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 2000

Authors and Affiliations

  • S. Kjellström
    • 1
  • S. Lindberg
    • 2
  • T. Laurell
    • 4
  • G. Marko-Varga
    • 3
  1. 1.Department of analytical ChemistryUniversity of LundLundSweden
  2. 2.AstraZeneca R&D Lund, HTS LabCell & Molecular BiologyLundSweden
  3. 3.AstraZeneca R&D LundCell & Molecular BiologyLundSweden
  4. 4.Department of Electrical Measurements, Lund Institute of TechnologyUniversity of LundLundSweden

Personalised recommendations