Nanowell-mediated multidimensional separations combining nanoLC with SLIM IM-MS for rapid, high-peak-capacity proteomic analyses

Dou, Maowei; Chouinard, Christopher D.; Zhu, Ying; Nagy, Gavril; Liyu, Andrey V.; Ibrahim, Yehia M.; Smith, Richard D.; Kelly, Ryan T.

doi:10.1007/s00216-018-1452-5

Title: Nanowell-mediated multidimensional separations combining nanoLC with SLIM IM-MS for rapid, high-peak-capacity proteomic analyses

Journal Article · Thu Aug 01 00:00:00 EDT 2019 · Analytical and Bioanalytical Chemistry

DOI:https://doi.org/10.1007/s00216-018-1452-5· OSTI ID:1572460

Dou, Maowei ^[1];

^[1];

^[1]; Nagy, Gavril ^[1]; Liyu, Andrey V. ^[1];

^[1];

^[2]

BATTELLE (PACIFIC NW LAB)
BRIGHAM YOUNG UNIVERSITY

Mass spectrometry (MS)–based analysis of complex biological samples is essential for biomedical research and clinical diagnostics. The separation prior to MS plays a key role in the overall analysis, with separations having larger peak capacities often leading to more identified species and improved confidence in those identifications. High-resolution ion mobility (IM) separations enabled by Structures for Lossless Ion Manipulation (SLIM) can provide extremely rapid, high-resolution separations and are well suited as a second dimension of separation following nanoscale liquid chromatography (nanoLC). However, existing sample handling approaches for offline coupling of separation modes require microliter-fraction volumes and are thus not well suited for analysis of trace biological samples. We have developed a novel nanowell-mediated fractionation system that enables nanoLC-separated samples to be efficiently preconcentrated and directly infused at nanoelectrospray flow rates for downstream analysis. When coupled with SLIM IM-MS, the platform enables rapid and high-peak-capacity multidimensional separations of small biological samples. In this study, peptides eluting from a 100 nL/min nanoLC separation were fractionated into ~?60 nanowells on a microfluidic glass chip using an in-house–developed robotic system. The dried samples on the chip were individually reconstituted and ionized by nanoelectrospray for SLIM IM-MS analysis. Using model peptides for characterization of the nanowell platform, we found that at least 80% of the peptide components of the fractionated samples were recovered from the nanowells, providing up to ~tenfold preconcentration for SLIM IM-MS analysis. The combined LC-SLIM IM separation peak capacities exceeded 3600 with a measurement throughput that is similar to current one-dimensional (1D) LC-MS proteomic analyses.

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Research Organization:: Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

Sponsoring Organization:: USDOE

DOE Contract Number:: AC05-76RL01830

OSTI ID:: 1572460

Report Number(s):: PNNL-SA-146403

Journal Information:: Analytical and Bioanalytical Chemistry, Vol. 411, Issue 21

Country of Publication:: United States

Language:: English

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