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Downsizing proteolytic digestion and analysis using dispenser-aided sample handling and nanovial matrix-assisted laser/desorption ionization-target arrays.

Ericsson, David LU ; Ekström, Simon LU ; Nilsson, Johan LU ; Bergquist, Jonas; Marko-Varga, György LU and Laurell, Thomas LU (2001) In Proteomics 1(9). p.1072-1081
Abstract
An efficient technique for enzymatic digestion of proteins in nanovial arrays and identification by peptide mass fingerprinting using matrix-assisted laser desorption/ionization (MALDI-MS) is presented in this work. Through dispensing of a protein solution with simultaneous evaporation the protein (substrate) is concentrated up to 300 times in-vial. At higher substrate concentrations the catalytic turnover numbers increase according to the Michaelis-Menten kinetics. Therefore, the dispenser-aided nanodigestion is valuable for identification of low-level proteins (10 nM-500 nM) as well as for automatic high efficiency digestions performed in 0.2-10 min. As an example of low-level protein identification, a 10 nM solution of lysozyme C was... (More)
An efficient technique for enzymatic digestion of proteins in nanovial arrays and identification by peptide mass fingerprinting using matrix-assisted laser desorption/ionization (MALDI-MS) is presented in this work. Through dispensing of a protein solution with simultaneous evaporation the protein (substrate) is concentrated up to 300 times in-vial. At higher substrate concentrations the catalytic turnover numbers increase according to the Michaelis-Menten kinetics. Therefore, the dispenser-aided nanodigestion is valuable for identification of low-level proteins (10 nM-500 nM) as well as for automatic high efficiency digestions performed in 0.2-10 min. As an example of low-level protein identification, a 10 nM solution of lysozyme C was unambiguously identified after 5 min of nanodigestion. Moreover, only 30 s nanodigestion was sufficient to identify hemoglobin (10 microM), exemplifying the fast catalysis of the nanodigestion technique. The developed silicon flow-through piezoelectric dispenser is adapted for low-volume and preconcentrated samples in the nL-microL range and provides fast, accurate and contact-free sample positioning into the nanovials. In this work, the properties of the nanodigestion concept regarding proteins of different characteristics are explored. Furthermore, the potential of automated protein identification using precoated proteolytic nanovial-arrays is demonstrated. (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
Silicon flow-through microdispenser, Nanovial array, On-target proteolytic digestion, Protein identification, Matrix-assisted laser desorption ionization-mass spectrometry
in
Proteomics
volume
1
issue
9
pages
1072 - 1081
publisher
John Wiley & Sons
external identifiers
  • wos:000171480300003
  • scopus:0035468131
ISSN
1615-9861
DOI
10.1002/1615-9861(200109)1:9<1072::AID-PROT1072>3.0.CO;2-W
language
English
LU publication?
yes
id
73f00884-b928-4601-9676-014fa45e6131 (old id 107991)
date added to LUP
2007-06-26 15:02:21
date last changed
2018-01-07 06:27:45
@article{73f00884-b928-4601-9676-014fa45e6131,
  abstract     = {An efficient technique for enzymatic digestion of proteins in nanovial arrays and identification by peptide mass fingerprinting using matrix-assisted laser desorption/ionization (MALDI-MS) is presented in this work. Through dispensing of a protein solution with simultaneous evaporation the protein (substrate) is concentrated up to 300 times in-vial. At higher substrate concentrations the catalytic turnover numbers increase according to the Michaelis-Menten kinetics. Therefore, the dispenser-aided nanodigestion is valuable for identification of low-level proteins (10 nM-500 nM) as well as for automatic high efficiency digestions performed in 0.2-10 min. As an example of low-level protein identification, a 10 nM solution of lysozyme C was unambiguously identified after 5 min of nanodigestion. Moreover, only 30 s nanodigestion was sufficient to identify hemoglobin (10 microM), exemplifying the fast catalysis of the nanodigestion technique. The developed silicon flow-through piezoelectric dispenser is adapted for low-volume and preconcentrated samples in the nL-microL range and provides fast, accurate and contact-free sample positioning into the nanovials. In this work, the properties of the nanodigestion concept regarding proteins of different characteristics are explored. Furthermore, the potential of automated protein identification using precoated proteolytic nanovial-arrays is demonstrated.},
  author       = {Ericsson, David and Ekström, Simon and Nilsson, Johan and Bergquist, Jonas and Marko-Varga, György and Laurell, Thomas},
  issn         = {1615-9861},
  keyword      = {Silicon flow-through microdispenser,Nanovial array,On-target proteolytic digestion,Protein identification,Matrix-assisted laser desorption ionization-mass spectrometry},
  language     = {eng},
  number       = {9},
  pages        = {1072--1081},
  publisher    = {John Wiley & Sons},
  series       = {Proteomics},
  title        = {Downsizing proteolytic digestion and analysis using dispenser-aided sample handling and nanovial matrix-assisted laser/desorption ionization-target arrays.},
  url          = {http://dx.doi.org/10.1002/1615-9861(200109)1:9<1072::AID-PROT1072>3.0.CO;2-W},
  volume       = {1},
  year         = {2001},
}