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Capillary Isoelectric Focusing

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Electromigration Techniques

Part of the book series: Springer Series in Chemical Physics ((CHEMICAL,volume 105))

Abstract

Capillary isoelectric focusing (CIEF) is a widespread technique for the analysis of peptides and proteins in biological samples. CIEF is used to separate mixtures of compounds on the basis of differences in their isoelectric point. Aspects of sample preparation, capillary selection, zone mobilization procedures as well as various detection modes used have been described and discussed. Moreover CIEF, coupled to various types of detection techniques (MALDI or LIF), has increasingly been applied to the analysis of variety different high-molecular compounds. CIEF is considered as a highly specific analytical method which may be routinely used in the separation of rare hemoglobin variants. In addition, the application of CIEF in proteomic field have been discussed on the examples of analyses of glycoproteins and immunoglobins due to the meaning in clinical diagnostic.

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References

  1. R.A. Mosher, W. Thorman, Experimental and theoretical description of the isotachophoretic behavior of serum albumins. Electrophoresis 11, 292–298 (1990)

    Article  Google Scholar 

  2. J.R. Mazzeo, I.S. Krull, Improvements in the method developed for performing isoelectric focusing in uncoated capillaries. J. Chromatogr. A 606, 291–296 (1992)

    Article  Google Scholar 

  3. S.M. Chen, J.E. Wiktorowicz, Isoelectric focusing by free solution capillary electrophoresis. Anal. Biochem. 206, 84–90 (1992)

    Article  Google Scholar 

  4. W. Thorman, J. Caslavska, S. Molteni, Isoelectric focusing with electroosmotic zone displacement and on-column multichannel detection. J. Chromatogr. A 589, 321–327 (1992)

    Article  Google Scholar 

  5. M. Zhu, R. Rodriquez, D. Hansen, T. Wehr, Capillary electrophoresis of proteins under alkaline conditions. J. Chromatogr. A 516, 123–131 (1990)

    Article  Google Scholar 

  6. S. Hjertén, M. Zhu, Adaptation of the equipment for high-performance electrophoresis to isoelectric focusing. J. Chromatogr. A 346, 265–270 (1985)

    Article  Google Scholar 

  7. R. Rodriquez, C. Siebert (1994) Poster presentation 6th international symposium on capillary electrophoresis (San Diego, CA, 1994)

    Google Scholar 

  8. J.R. Mazzeo, I.S. Krull, Peptide mapping using EOF-driven capillary isoelectric focusing. Anal. Biochem. 208, 323–329 (1993)

    Article  Google Scholar 

  9. S. Hjertén, K. Kubo, A new type of pH-and detergent-stable coating for elimination of electroendosmosis and adsorption in (capillary) electrophoresis. Electrophoresis 14, 390–395 (1993)

    Article  Google Scholar 

  10. J. Wu, J. Pawliszyn, Fluorescence imaging detection for capillary isoelectric focusing. Electrophoresis 16, 1474–1478 (1995)

    Article  Google Scholar 

  11. M.S. Jenkins, Three methods of capillary electrophoresis compared with high-resolution a garowe gel electrophoresis for serum protein electrophoresis. J. Chromatogr. A 720, 49–58 (1999)

    Google Scholar 

  12. X. Liu, Z. Sosic, I.S. Krull, Capillary isoelectric focusing as a toll in the examination of antibodies, peptides and proteins of pharmaceutical interest. J. Chromatogr. A 735, 165–190 (1996)

    Article  Google Scholar 

  13. X.W. Yao, D. Wu, F.E. Regnier, Manipulation of electroosmotic flow in capillary electrophoresis. J. Chromatogr. A 636, 21–29 (1993)

    Article  Google Scholar 

  14. J. Wang, S. Zhou, W. Huang, Y. Liu, C. Cheng, X. Lu, J. Cheng, CE-based analysis of hemoglobin and its applications in clinical analysis. Electrophoresis 27, 3108–3124 (2006)

    Article  Google Scholar 

  15. J.M. Hempe, R.D. Craver, Separation of hemoglobin variants with similar charge by capillary isoelectric focusing: value of isoelectric point for identification of common and uncommon hemoglobin variants. Electrophoresis 21, 743–748 (2000)

    Article  Google Scholar 

  16. T.A. Crowley, M.A. Hayes, Analysis of human blood serum using the off-line coupling of capillary isoelectric focusing to matrix-assisted laser desorption/ionization time of flight mass spectrometry. Proteomics 5, 3798–3804 (2005)

    Article  Google Scholar 

  17. Z. Liu, J. Pawliszyn, Coupling of solid-phase micro extraction and capillary isoelectric focusing with laser-induced fluorescence whole column imaging detection for protein analysis. Anal. Chem. 77, 165–171 (2005)

    Article  Google Scholar 

  18. R. Hackler, T. Arndt, A. Helwig-Rolig, J. Kropf, A. Steinmetz, J.R. Schaefer, Investigation by isoelectric focusing of the initial carbohydrate-deficient transferrin (CDT) and non-CDT transferrin isoform fractionation strep involved in determination of CDT by the CronAlcolD assay. Clin. Chem. 46, 483–492 (2000)

    Google Scholar 

  19. L. Dai, C. Li, K.A. Shedden, D.E. Misek, D.M. Lubman, Comparative proteomic study of two closely related ovarian endometrioid adenocarcinoma cell lines using cIEF fractionation and pathway analysis. Electrophoresis 30, 1119–1131 (2009)

    Article  Google Scholar 

  20. W. Wang, T. Guo, T. Song, C.S. Lee, B.M. Balgley, Comprehensive yeast proteome analysis using a capillary isoelectric focusing-based multidimensional separation platform coupled with ESI-MS/MS. Proteomics 7, 1178–1187 (2007)

    Article  Google Scholar 

  21. S. Ongay, S. Puerta, J.C. Diez-Masa, J. Bergquist, M. Frutos, CIEF and MALDI-TOF MS methods for analyzing forms of the glycoprotein VEGF 165. Electrophoresis 30, 1198–1205 (2009)

    Article  Google Scholar 

  22. P. Dou, Z. Liu, J. He, J.J. Xu, H.Y. Chen, Rapid and high-resolution glycoform profiling of recombinant human erythropoietin by capillary isoelectric focusing with whole column imaging detection. J. Chromatogr. A 1190, 372–376 (2008)

    Article  Google Scholar 

  23. S. Tang, D.P. Nesta, L.R. Maneri, K.R. Anumula, A method for routine analysis of recombinant immunoglobulins (rIgGs) by capillary isoelectric focusing (CIEF). J. Pharm. Biomed. Anal. 19, 569–583 (1999)

    Article  Google Scholar 

  24. G. Janini, N. Saptharishi, M. Waselus, G. Soman, Element of a validation method for MU-BS monoclonal antibody using an imaging capillary isoelectric focusing system. Electrophoresis 23, 1605–1611 (2002)

    Article  Google Scholar 

  25. C.D. Meert, L.J. Brady, A. Guo, A. Balland, Characterization of antibody charge heterogeneity resolved by preparative immobilized pH gradients. Anal. Chem. 82, 3510–3518 (2010)

    Article  Google Scholar 

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Authors and Affiliations

  1. Faculty of Pharmacy, Medical University of Gdańsk, Gdańsk, Poland

    Michał J. Markuszewski

  2. Faculty of Pharmacy, Ludwig Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, Toruń, Poland

    Renata Bujak & Emilia Daghir

Authors
  1. Michał J. Markuszewski
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  2. Renata Bujak
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  3. Emilia Daghir
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Corresponding author

Correspondence to Michał J. Markuszewski .

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Editors and Affiliations

  1. Fac. Chemistry, Dept. Environmental Chemistry &, Nicolaus Copernicus University, Gagarin 7, Torun, 87-100, Poland

    Boguslaw Buszewski

  2. Faculty of Chemistry, Department of Environmental Chemistry, Nicolaus Copernicus University, Gagarin 7, Toruń, 87-100, Poland

    Ewelina Dziubakiewicz

  3. Faculty of Chemistry, Department of Environmental Chemistry, Nicolaus Copernicus University, Gagarin 7, Torun, 87-100, Poland

    Michal Szumski

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Markuszewski, M.J., Bujak, R., Daghir, E. (2013). Capillary Isoelectric Focusing. In: Buszewski, B., Dziubakiewicz, E., Szumski, M. (eds) Electromigration Techniques. Springer Series in Chemical Physics, vol 105. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-35043-6_7

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  • DOI: https://doi.org/10.1007/978-3-642-35043-6_7

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  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-35042-9

  • Online ISBN: 978-3-642-35043-6

  • eBook Packages: Physics and AstronomyPhysics and Astronomy (R0)

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