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

In English

Författare

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.

Publiceringsår

2004

Språk

Engelska

Sidor

124-128

Publikation/Tidskrift/Serie

Journal of Separation Science

Volym

27

Issue

1-2

Dokumenttyp

Artikel i tidskrift

Förlag

John Wiley & Sons Inc.

Ämne

  • Analytical Chemistry

Nyckelord

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

Status

Published

ISBN/ISSN/Övrigt

  • ISSN: 1615-9314