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

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Eur. J. Mass Spectrom. 7, 143 - 155 (2001)

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

Jianjun Li, Tammy-Lynn Tremblay and Pierre Thibault^*
Institute for Biological Sciences, 100 Sussex Dr., Ottawa, Ontario, Canada K1A 0R6 E-mail:
Can Wang, Said Attiya and D. Jed Harrison
Department of Chemistry, University of Alberta, Edmonton, AB, Canada T6G 2G2

ABSTRACT:

An integrated microsystem providing rapid analyses of trace-level tryptic digests for proteomics application is presented. This modular microsystem includes an autosampler and a microfabricated device comprising a sample introduction 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 digests was achieved with a throughput of up to 30 samples per hour with less than 3% sample carryover. Replicate injections of peptide mixtures indicated that reproducibility of migration time was typically better than 2.3% relative standard deviation (RSD), whereas RSD values of 3.711.8% were observed on peak height. Mass spectral detection of sub-micromolar protein digests (< 7 fmol per injection) was achieved using a quadrupole/time-of-flight instrument in less than 2 min per sample with peak widths of 1.87.0 s. The analytical potential of this integrated device for the identification of gel-isolated proteins from Neisseria meningitidis immunotype L3 has been demonstrated using both peptide mass-fingerprint database searching and on-line tandem mass spectrometry.

Keywords: microfabricated device, capillary electrophoresis, nanoelectrospray, tandem mass spectrometry, protein digests, Neisseria meningitidis, proteomics

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