Integrated System for High-Throughput Protein Identification Using a Microfabricated Device Coupled to Capillary Electrophoresis/Nanoelectrospray Mass Spectrometry

• Published in: European journal of mass spectrometry (Chichester, England) England), 2001-04, Vol.7 (2), p.143-155
• Main Authors: Li, Jianjun, Tremblay, Tammy-Lynn, Thibault, Pierre, Wang, Can, Attiya, Said, Harrison, D. Jed
• Format: Article
• Language: English
• ISSN: 1469-0667; 1751-6838
• DOI: 10.1255/ejms.399

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 3.7–11.8% 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 1.8–7.0 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.