High-speed, high-resolution monolithic capillary LC-MALDI MS using an off-line continuous deposition interface for proteomic analysis

Hsuan Shen Chen, Tomas Rejtar, Viktor Andreev, Eugene Moskovets, Barry L. Karger

Research output: Contribution to journalArticlepeer-review

71 Scopus citations

Abstract

High-speed, high-resolution LC separations, using a poly-(styrene- divinylbenzene) monolithic column, have been coupled to MALDI MS and MS/MS through an off-line continuous deposition interface. The LC eluent was mixed with α-cyano-4-hydroxycinnamic acid matrix solution and deposited on a MALDI plate that had been precoated with nitrocellulose. Deposition at subatmospheric pressure (80 Torr) formed a 250-μm-wide serpentine trace with uniform width and microcrystalline morphology. The deposited trace was then analyzed in the MS mode using a MALDI-TOF/TOF MS instrument. Continuous deposition allowed interrogation of the separation with a high data sampling rate in the chromatographic dimensions, thus preserving the high resolution of narrow peaks (3-5-s peak width at half-height) of the fast monolithic LC. No extracolumn band broadening due to the deposition process was observed. Over 2000 components were resolved in a 10-min linear gradient separation of the model sample, and 386 unique peptides were identified in the subsequent MS/MS analysis. The continuous deposition interface allows the coupling of high-resolution separations to MALDI MS without degradation in separation efficiency, thus enabling high-throughput proteome analysis.

Original languageEnglish (US)
Pages (from-to)2323-2331
Number of pages9
JournalAnalytical Chemistry
Volume77
Issue number8
DOIs
StatePublished - Apr 15 2005

ASJC Scopus subject areas

  • Analytical Chemistry

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