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

Palm, A; Zaragoza-Sundqvist, M and Marko-Varga, György LU (2004) In Journal of Separation Science 27(1-2). p.124-128
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
capillary isoelectric focusing (CIEF), proteins, capillary coatings, reproducibility
in
Journal of Separation Science
volume
27
issue
1-2
pages
124 - 128
publisher
John Wiley & Sons
external identifiers
  • wos:000188551400018
  • pmid:15335068
  • scopus:0442294234
ISSN
1615-9314
DOI
10.1002/jssc.200301596
language
English
LU publication?
yes
id
476fc17e-ca28-41d0-97e1-5b882176b7c1 (old id 288640)
date added to LUP
2007-10-29 13:50:42
date last changed
2017-01-01 04:27:22
@article{476fc17e-ca28-41d0-97e1-5b882176b7c1,
  abstract     = {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.},
  author       = {Palm, A and Zaragoza-Sundqvist, M and Marko-Varga, György},
  issn         = {1615-9314},
  keyword      = {capillary isoelectric focusing (CIEF),proteins,capillary coatings,reproducibility},
  language     = {eng},
  number       = {1-2},
  pages        = {124--128},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Separation Science},
  title        = {Capillary isoelectric focusing of proteins utilizing poly(vinylpyrrolodone)- and plexigla-coated columns},
  url          = {http://dx.doi.org/10.1002/jssc.200301596},
  volume       = {27},
  year         = {2004},
}