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Selective extraction of small proteins from biological samples using a novel restricted access column with cation exchange properties

Grimm, C. H.; Boos, K. S.; Apel, Ch; Unger, K. K.; Önnerfjord, P. LU ; Heintz, L.; Edholm, L. E. and Marko-Varga, G. LU (2000) In Chromatographia 52(11-12). p.703-709
Abstract

The determination of proteins utilising a polymer-based restricted access support material with ion exchange properties (IERAM) is outlined. Solid phase extraction coupled on-line with a microbore reversed phase HPLC system for the quantitation of small marker proteins is demonstrated. The cation-exchange restricted access packings were characterised with respect to their adsorption and desorption kinetics. The IERAM material was also investigated by capacity, selectivity, and biocompatibility determinations when applied to the quantification of small molecular weight proteins such as cytochrome C, Lysozyme, Ribonuclease A, Myoglobin, Insulin, human serum albumin, and a Tryptic inhibitor. The integrated system was coupled to mass... (More)

The determination of proteins utilising a polymer-based restricted access support material with ion exchange properties (IERAM) is outlined. Solid phase extraction coupled on-line with a microbore reversed phase HPLC system for the quantitation of small marker proteins is demonstrated. The cation-exchange restricted access packings were characterised with respect to their adsorption and desorption kinetics. The IERAM material was also investigated by capacity, selectivity, and biocompatibility determinations when applied to the quantification of small molecular weight proteins such as cytochrome C, Lysozyme, Ribonuclease A, Myoglobin, Insulin, human serum albumin, and a Tryptic inhibitor. The integrated system was coupled to mass identity of selected proteins by MALDI-TOF mass spectrometry. The chromatographic outlet was interfaced to an "Interplate" fractionation collecter that sampled 1 μL volumes directly onto the MALDI target plate. The fully automated coupled column system was run unattended overnight and applied to protein quantitations in human plasma samples. Recovery data for a selected number of proteins varied between 90-96% (n = 10) with a limit of quantification around 2 μM with an injection volume of 100 μL. The RSD data were typically less than 8% at a 50 μM protein level (n = 7).

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Column liquid chromatography, Extraction of proteins, Restricted access cation exchanger, Sample clean-up
in
Chromatographia
volume
52
issue
11-12
pages
7 pages
publisher
Vieweg Verlag
external identifiers
  • scopus:0034523935
ISSN
0009-5893
language
English
LU publication?
yes
id
7de59417-1e3e-4b64-aadb-6a671aa7408a
date added to LUP
2016-10-14 11:35:04
date last changed
2017-01-01 08:36:49
@article{7de59417-1e3e-4b64-aadb-6a671aa7408a,
  abstract     = {<p>The determination of proteins utilising a polymer-based restricted access support material with ion exchange properties (IERAM) is outlined. Solid phase extraction coupled on-line with a microbore reversed phase HPLC system for the quantitation of small marker proteins is demonstrated. The cation-exchange restricted access packings were characterised with respect to their adsorption and desorption kinetics. The IERAM material was also investigated by capacity, selectivity, and biocompatibility determinations when applied to the quantification of small molecular weight proteins such as cytochrome C, Lysozyme, Ribonuclease A, Myoglobin, Insulin, human serum albumin, and a Tryptic inhibitor. The integrated system was coupled to mass identity of selected proteins by MALDI-TOF mass spectrometry. The chromatographic outlet was interfaced to an "Interplate" fractionation collecter that sampled 1 μL volumes directly onto the MALDI target plate. The fully automated coupled column system was run unattended overnight and applied to protein quantitations in human plasma samples. Recovery data for a selected number of proteins varied between 90-96% (n = 10) with a limit of quantification around 2 μM with an injection volume of 100 μL. The RSD data were typically less than 8% at a 50 μM protein level (n = 7).</p>},
  author       = {Grimm, C. H. and Boos, K. S. and Apel, Ch and Unger, K. K. and Önnerfjord, P. and Heintz, L. and Edholm, L. E. and Marko-Varga, G.},
  issn         = {0009-5893},
  keyword      = {Column liquid chromatography,Extraction of proteins,Restricted access cation exchanger,Sample clean-up},
  language     = {eng},
  number       = {11-12},
  pages        = {703--709},
  publisher    = {Vieweg Verlag},
  series       = {Chromatographia},
  title        = {Selective extraction of small proteins from biological samples using a novel restricted access column with cation exchange properties},
  volume       = {52},
  year         = {2000},
}