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High-performance capillary electrophoresis using open tubes and gels

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Summary

This paper overviews several aspects of high performance capillary electrophoresis (HPCE), a promising new method of analytical and micropreparative separation of biochemically important samples. The basic migration equations of electrophoresis are first presented and the benefit of high fields for rapid analysis and high performance emphasized. Since power is generated with high voltages, Joule heating results and this heat must be dissipated. The use of capillary columns is shown to be important in efficient heat removal and in minimizing the temperature differences within the column. The various factors influencing band broadening are next described, and it is shown how plate counts close to 106 can be achieved. Various results from our laboratory on open tube and gel columns are then presented to illustrate the potential of this method. Chiral resolution of dansylated amino acids using a chiral metal chelate micelle in open tube HPCE is shown. With the gel columns, the baseline separation of a 2-chain variant from methionine growth hormone (met-hGH) under non-denaturing conditions at fields close to 1000 V/cm is presented. Finally, the micropreparative purification of a 20-mer oligonucleotide using the gel column is described.

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References

  1. DNA Sequencing: Laboratory Techniques in Biochemistry and Molecular Biology;T. S. Work, R. H. Burdon (Editors); Elsevier Biomedical Press: Amsterdam, New York, Oxford (1986).

    Google Scholar 

  2. Gel Electrophoresis of Proteins: A Practical Approach;B. D. Hames, D. Rirkwood (Editors); IRL Press, Washington, D.C. 1983.

    Google Scholar 

  3. Isoelectric Focusing. Theory, Methodology and Application,P. G. Righetti (ed.), Elsevier Biomedical Press, New York, (1983).

    Google Scholar 

  4. J. C. Giddings, Sep. Sci.4, 181–189 (1969).

    Article  CAS  Google Scholar 

  5. S. Bowman, Anal. Chem.59, 969A (1987).

  6. J. C. Bennett, Methods in Enzymology, Vol. XI, S. P. Colowick and N. O. Kaplan (Editors), Academic Press, New York 330 (1967).

    Google Scholar 

  7. S. Raymond, L. Weintraub, Science,130, 711 (1959).

    Article  CAS  Google Scholar 

  8. V. Neuhoff, Electrophoresis5, 251 (1984).

    Article  CAS  Google Scholar 

  9. F. E. P. Mikkers, F. M. Everaerts, T. P. E. M. Verheggen, J. Chromatogr.169, 11–20 (1979).

    Article  CAS  Google Scholar 

  10. J. W. Jorgenson, K. D., Lukacs, Science222, 266–272 (1983).

    Article  CAS  Google Scholar 

  11. A. S. Cohen, S. Terabe, J. A. Smith, B. L. Karger, Anal. Chem.59, 1021–1028 (1987).

    Article  CAS  Google Scholar 

  12. A. S. Cohen, B. L. Karger, J. Chromatogr.397, 409–417 (1987).

    Article  CAS  Google Scholar 

  13. S. Terabe, K. Otsuka, T. Ando, Anal. Chem.57, 834–841 (1985).

    Article  CAS  Google Scholar 

  14. E. Gassmann, J. E. Kno, R. N. Zare, Science230, 813–814 (1985).

    Article  CAS  Google Scholar 

  15. T. Tsuda, G. Hakagawa, M. Tato, K. Yagi, J. Appl. Biochem.5, 330–336 (1983).

    CAS  Google Scholar 

  16. H. H. Lauer, D. McManigill, Anal. Chem.58, 166–170 (1986).

    Article  CAS  Google Scholar 

  17. R. A. Wallingford, A. G. Ewing, Anal. Chem.59, 1762–1766 (1987).

    Article  CAS  Google Scholar 

  18. F. E. P. Mikkers, F. M. Everaerts, T. P. E. M. Verheggen, J. Chromatogr.169, 11–20 (1979).

    Article  CAS  Google Scholar 

  19. V. Horejsi, J. Tickam, J. Chromatogr.376, 49–67 (1986).

    Article  CAS  Google Scholar 

  20. T. Tsuda, T. Mizuno, J. Akiama, Anal. Chem.59, 799–800 (1987).

    Article  CAS  Google Scholar 

  21. J. W. Jorgenson, K. D. Lukacs, J. Chromatogr.218, 209–216 (1981).

    Article  CAS  Google Scholar 

  22. X. H. Huang, T. K. J. Pang, M. J. Gordon, R. N. Zare, Anal. Chem.,59, 2747–2749 (1987).

    Article  CAS  Google Scholar 

  23. J. A. Olivares, N. T. Nguyen, C. R. Yonker, R. D. Smith, Anal. Chem.59, 1232–1236 (1987).

    Article  Google Scholar 

  24. S. Hjerten, D. M. Zhu, J. Chromatogr.327, 151–164 (1985).

    Article  Google Scholar 

  25. R. J. Wierne, Chromatography: A Laboratory Handbook of Chromatography and Electrophoresis Methods, 3rd, ed., Heftmann, E. (Editor), Van Norstrand: New York (1975).

    Google Scholar 

  26. S. Hjerten, M. D. Zhu, J. Chromatogr.347, 191–198 (1985).

    Article  CAS  Google Scholar 

  27. A. S. Cohen, B. L. Karger, unpublished results.

  28. S. Terabe, K. Otsuka, T. Ando, Anal. Chem., submitted for publication.

  29. S. Hjerten, K. Elbering, F. Kilar, J. L. Liao, A. J. C. Chen, C. J. Siebert, M. D. Zhu, J. Chromatogr.403, 47–61 (1987).

    Article  CAS  Google Scholar 

  30. K. D. Lukacs, J. W. Jorgenson, HRC & CC8, 407, 411 (1985).

    CAS  Google Scholar 

  31. B. L. Karger, R. W. Giese, Anal. Chem.,50, 1048A (1978).

  32. B. L. Karger, P. Vouros, J. Chromatogr.,323, 13–32 (1985).

    Article  CAS  Google Scholar 

  33. P. G. Righetti, E. Gianazza, Electrophoresis 1981,R. C. Allen, P. Arnand (Editor), de Gruyther, Berlin, 655–665.

    Google Scholar 

  34. New Directions in Electrophoresis Methods,J. W. Jorgenson, M. Phillips (eds.), American Chemical Society, Washington, D.C. (1987).

    Google Scholar 

  35. Metal Ion in Biological Systems, H. Sigel (Editor), Marcel Dekker: New York (1979), Vol. 9.

    Google Scholar 

  36. M. V. Oko, R. L. Venable, J. Colloid. Interface. Sci.35, 53–59 (1971).

    Article  CAS  Google Scholar 

  37. B. Feibush, M. J. Cohen, B. L. Karger, J. Chromatogr.282, 3–26 (1983).

    Article  CAS  Google Scholar 

  38. P. Gozel, E. Gassmann, H. Michelsen, R. N. Zare, Anal. Chem.59, 44–49 (1987).

    Article  CAS  Google Scholar 

  39. J. M. Prober, G. L. Trainor, R. J. Dam, F. W. Hobbs, C. W. Robertson, R. J. Zagursky, A. J. Cocuzza, M. A. Jensen, K. Baumeister, Science,238, 336–341 (1987).

    Article  CAS  Google Scholar 

  40. V. J. Lewis, J. T. Dunn, L. F. Benewald, B. K. Seavey, W. P. Vanderlaan, J. Biol. Chem.263, 2679–2687 (1987).

    Google Scholar 

  41. A. E. Wilhelmi, Hormone Drugs,J. L. Guerigriam, E. D. Bransonk, A. S. Outskoern (Editors), 369–381, Convection, Rockville, MD 1982.

    Google Scholar 

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Cohen, A.S., Paulus, A. & Karger, B.L. High-performance capillary electrophoresis using open tubes and gels. Chromatographia 24, 15–24 (1987). https://doi.org/10.1007/BF02688463

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