Integrated system for high-throughput protein identification using a microfabricated device coupled to capillary electrophoresis/nanoelectrospray mass spectrometry

Citation
Jj. Li et al., Integrated system for high-throughput protein identification using a microfabricated device coupled to capillary electrophoresis/nanoelectrospray mass spectrometry, EUR J MASS, 7(2), 2001, pp. 143-155
Citations number
35
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Spectroscopy /Instrumentation/Analytical Sciences
Journal title
EUROPEAN JOURNAL OF MASS SPECTROMETRY
ISSN journal
1469-0667 → ACNP
Volume
7
Issue
2
Year of publication
2001
Pages
143 - 155
Database
ISI
SICI code
1469-0667(2001)7:2<143:ISFHPI>2.0.ZU;2-5
Abstract
An integrated microsystem providing rapid analyses of trace-level tryptic d igests for proteomics application is presented. This modular microsystem in cludes an autosampler and a microfabricated device comprising a sample intr oduction port and an array of separation channels together with a low dead- volume facility enabling an interface to nanoelectrospray mass spectrometry . Sequential injection and separation of peptide standards and tryptic dige sts was achieved with a throughput of up to 30 samples per hour with less t han 3% sample carryover. Replicate injections of peptide mixtures indicated that reproducibility of migration time was typically better than 2.3% rela tive standard deviation (RSD), whereas RSD values of 3.7-11.8% were observe d on peak height. Mass spectral detection of sub-micromolar protein digests (< 7 fmol per injection) was achieved using a quadrupole/time-of-flight in strument in less than 2 min per sample with peak widths of 1.8-7.0 s. The a nalytical potential of this integrated device for the identification of gel -isolated proteins from Neisseria meningitidis immunotype L3 has been demon strated using both peptide mass-fingerprint database searching and on-line tandem mass spectrometry.