Advanced

Signal amplification using "spot on-a-chip" technology for the identification of proteins via MALDI-TOF MS

Ekstrom, S.; Ericsson, D.; Onnerfjord, P.; Bengtsson, M.; Nilsson, J.; Marko-Varga, György LU and Laurell, Thomas LU (2001) In Analytical Chemistry1949-01-01+01:00 73(2). p.214-219
Abstract
The presented "spot-on-a-chip" technology enables easy enrichment of samples in the low nanomolar (1-5 nM) range and provides a fast and reliable automated sample preparation method for performing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis with high sensitivity and throughput. Through microdispensing, which allows accurate deposition of 60-pL droplets, dilute samples were enriched by making multiple droplet depositions in nanovials. The sample was confined to a defined spot area (300 x 300 mum), and multiple depositions increase the surface density of analyte in the nanovial, thereby providing detection of low attomole levels. The impact of the nanovial geometry with respect to the... (More)
The presented "spot-on-a-chip" technology enables easy enrichment of samples in the low nanomolar (1-5 nM) range and provides a fast and reliable automated sample preparation method for performing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis with high sensitivity and throughput. Through microdispensing, which allows accurate deposition of 60-pL droplets, dilute samples were enriched by making multiple droplet depositions in nanovials. The sample was confined to a defined spot area (300 x 300 mum), and multiple depositions increase the surface density of analyte in the nanovial, thereby providing detection of low attomole levels. The impact of the nanovial geometry with respect to the MALDI-TOF MS resolution for peptides deposited in the microfabricated silicon vials was investigated and the optimal geometry and size were determined. The spot-on-a-chip technology, that is, the combination of microdispensing, micromachined silicon nanovials and on-spot enrichment provides a signal amplification of at least 10-50 times as compared to an ordinary sample preparation. The linearity of the enrichment effect is shown by the analysis of a peptide mixture at the 5 nM level. The signal amplification provided by the spot-on-a-chip enrichment is demonstrated by the analysis of relevant biological samples, interleukin-8 from a spiked cell supernatant, and by successful protein identification of an excised spot from a high-sensitivity silver-stained two-dimensional electrophoresis gel separation. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
IONIZATION MASS-SPECTROMETRY, SAMPLE PREPARATION, HIGH-SENSITIVITY, PEPTIDE, RESOLUTION, PICOLITER, MIXTURES
in
Analytical Chemistry1949-01-01+01:00
volume
73
issue
2
pages
214 - 219
publisher
The American Chemical Society
external identifiers
  • wos:000166366000011
  • scopus:0035863619
ISSN
1520-6882
DOI
10.1021/ac000734u
language
English
LU publication?
yes
id
7f3d915a-b4e9-48a6-95d3-23ae295707c2 (old id 2376304)
date added to LUP
2012-03-23 09:47:09
date last changed
2018-10-03 10:21:49
@article{7f3d915a-b4e9-48a6-95d3-23ae295707c2,
  abstract     = {The presented "spot-on-a-chip" technology enables easy enrichment of samples in the low nanomolar (1-5 nM) range and provides a fast and reliable automated sample preparation method for performing matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis with high sensitivity and throughput. Through microdispensing, which allows accurate deposition of 60-pL droplets, dilute samples were enriched by making multiple droplet depositions in nanovials. The sample was confined to a defined spot area (300 x 300 mum), and multiple depositions increase the surface density of analyte in the nanovial, thereby providing detection of low attomole levels. The impact of the nanovial geometry with respect to the MALDI-TOF MS resolution for peptides deposited in the microfabricated silicon vials was investigated and the optimal geometry and size were determined. The spot-on-a-chip technology, that is, the combination of microdispensing, micromachined silicon nanovials and on-spot enrichment provides a signal amplification of at least 10-50 times as compared to an ordinary sample preparation. The linearity of the enrichment effect is shown by the analysis of a peptide mixture at the 5 nM level. The signal amplification provided by the spot-on-a-chip enrichment is demonstrated by the analysis of relevant biological samples, interleukin-8 from a spiked cell supernatant, and by successful protein identification of an excised spot from a high-sensitivity silver-stained two-dimensional electrophoresis gel separation.},
  author       = {Ekstrom, S. and Ericsson, D. and Onnerfjord, P. and Bengtsson, M. and Nilsson, J. and Marko-Varga, György and Laurell, Thomas},
  issn         = {1520-6882},
  keyword      = {IONIZATION MASS-SPECTROMETRY,SAMPLE PREPARATION,HIGH-SENSITIVITY,PEPTIDE,RESOLUTION,PICOLITER,MIXTURES},
  language     = {eng},
  number       = {2},
  pages        = {214--219},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry1949-01-01+01:00},
  title        = {Signal amplification using "spot on-a-chip" technology for the identification of proteins via MALDI-TOF MS},
  url          = {http://dx.doi.org/10.1021/ac000734u},
  volume       = {73},
  year         = {2001},
}