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Integrated microanalytical technology enabling rapid and automated protein identification

Ekström, Simon LU ; Önnerfjord, Patrik LU ; Nilsson, Johan LU ; Bengtsson, Martin LU ; Laurell, Thomas LU and Marko-Varga, György LU (2000) In Analytical Chemistry 72(2). p.286-293
Abstract

Protein identification through peptide mass mapping by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become a standard technique, used in many laboratories around the world. The traditional methodology often includes long incubations (6-24 h) and extensive manual steps. In an effort to address this, an integrated microanalytical platform has been developed for automated identification of proteins. The silicon micromachined analytical tools, i.e., the microchip immobilized enzyme reactor (μ-chip IMER), the piezoelectric microdispenser, and the high-density nanovial target plates, are the cornerstones in the system. The μ-chip IMER provides on-line enzymatic digestion of protein samples (1... (More)

Protein identification through peptide mass mapping by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become a standard technique, used in many laboratories around the world. The traditional methodology often includes long incubations (6-24 h) and extensive manual steps. In an effort to address this, an integrated microanalytical platform has been developed for automated identification of proteins. The silicon micromachined analytical tools, i.e., the microchip immobilized enzyme reactor (μ-chip IMER), the piezoelectric microdispenser, and the high-density nanovial target plates, are the cornerstones in the system. The μ-chip IMER provides on-line enzymatic digestion of protein samples (1 μL) within 1-3 min, and the microdispenser enables subsequent on- line picoliter sample preparation in a high-density format. Interfaced to automated MALDI-TOF MS, these tools compose a highly efficient platform that can analyze 100 protein samples in 3.5 h. Kinetic studies on the microreactors are reported as well as the operation of this microanalytical platform for protein identification, wherein lysozyme, myoglobin, ribonuclease A, and cytochrome c have been identified with a high sequence coverage (50-100%).

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author
organization
publishing date
type
Contribution to journal
publication status
published
in
Analytical Chemistry
volume
72
issue
2
pages
286 - 293
publisher
The American Chemical Society
external identifiers
  • scopus:0034650858
ISSN
0003-2700
DOI
10.1021/ac990731l
language
English
LU publication?
yes
id
24d3ddf8-55b4-4843-9578-094a0044b43d
date added to LUP
2016-10-14 11:35:48
date last changed
2017-08-20 05:03:04
@article{24d3ddf8-55b4-4843-9578-094a0044b43d,
  abstract     = {<p>Protein identification through peptide mass mapping by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become a standard technique, used in many laboratories around the world. The traditional methodology often includes long incubations (6-24 h) and extensive manual steps. In an effort to address this, an integrated microanalytical platform has been developed for automated identification of proteins. The silicon micromachined analytical tools, i.e., the microchip immobilized enzyme reactor (μ-chip IMER), the piezoelectric microdispenser, and the high-density nanovial target plates, are the cornerstones in the system. The μ-chip IMER provides on-line enzymatic digestion of protein samples (1 μL) within 1-3 min, and the microdispenser enables subsequent on- line picoliter sample preparation in a high-density format. Interfaced to automated MALDI-TOF MS, these tools compose a highly efficient platform that can analyze 100 protein samples in 3.5 h. Kinetic studies on the microreactors are reported as well as the operation of this microanalytical platform for protein identification, wherein lysozyme, myoglobin, ribonuclease A, and cytochrome c have been identified with a high sequence coverage (50-100%).</p>},
  author       = {Ekström, Simon and Önnerfjord, Patrik and Nilsson, Johan and Bengtsson, Martin and Laurell, Thomas and Marko-Varga, György},
  issn         = {0003-2700},
  language     = {eng},
  month        = {01},
  number       = {2},
  pages        = {286--293},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Integrated microanalytical technology enabling rapid and automated protein identification},
  url          = {http://dx.doi.org/10.1021/ac990731l},
  volume       = {72},
  year         = {2000},
}