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Integrated system for high‐throughput protein identification using a microfabricated device coupled to capillary electrophoresis / nanoelectrospray mass spectrometry
Author(s) -
Li Jianjun,
Tremblay TammyLynn,
Wang Can,
Attiya Said,
Harrison D. Jed,
Thibault Pierre
Publication year - 2001
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/1615-9861(200108)1:8<975::aid-prot975>3.0.co;2-h
Subject(s) - capillary electrophoresis , mass spectrometry , chromatography , chemistry , proteomics , capillary electrophoresis–mass spectrometry , throughput , identification (biology) , top down proteomics , tandem mass spectrometry , protein mass spectrometry , computer science , biology , electrospray ionization , biochemistry , gene , telecommunications , botany , wireless
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 facilitating the 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 submicromolar protein digests (< 7 femtomoles/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.