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Capillary isoelectric focusing of proteins utilizing poly(vinylpyrrolodone)- and plexigla-coated columns

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Author

Summary, in English

Fused-silica capillaries chemically derivatized with silane/poly(vinylpyrrolidone) (PVP) or dynamically modified with plexiglas [poly(methyl methacrylate)] were prepared and evaluated with regard to column stability and separation performance for capillary isoelectric focusing of standard proteins. The PVP coating showed the better stability and was good for at least 100 runs while the plexiglas coating started to deteriorate after about 30 runs. The time spent for the plexiglas coating is about 40 minutes while the PVP coating requires two days. The migration time reproducibility was better with the PVP capillary (RSD 0.7-1.6%, n = 5) compared to the plexiglas-coated column (RSD 1.2-2.9%, n = 5) while peak area and height varied over a similar interval (RSD 2-28.1% area; 0.9-22.7% height, n = 5). The two most consistent proteins in this evaluation, viz. myoglobin A and carbonic anhydrase II, showed linear dynamic ranges between 5-150 and 5-50 mug/mL, and limits of detections at 2 and 1 mug/mL, respectively, employing UV detection at 280 nm.

Publishing year

2004

Language

English

Pages

124-128

Publication/Series

Journal of Separation Science

Volume

27

Issue

1-2

Document type

Journal article

Publisher

John Wiley & Sons Inc.

Topic

  • Analytical Chemistry

Keywords

  • capillary isoelectric focusing (CIEF)
  • proteins
  • capillary coatings
  • reproducibility

Status

Published

ISBN/ISSN/Other

  • ISSN: 1615-9314