Recent advances in open tubular capillary liquid chromatography

Lam, Shing Chung; Rodríguez, Estrella Sanz; Haddad, Paul R.; Paull, Brett

doi:10.1039/c9an00329k

Cited by 55 publications

(18 citation statements)

References 122 publications

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“…Therefore, high-efficiency separation, e.g., capillary electrophoresis ,− or LC, is needed to increase the detection sensitivity by ESI-MS. In the past decades, significant efforts were devoted to improving the detection sensitivity, separation performance, and throughput of LC–MS analysis by decreasing the flow rate, downscaling the column diameter, and increasing the column length. − With an increased need for the improved separation efficiency and, therefore, the depth of profiling sensitivity in LC–MS-based proteomics, , the development of nanoLC columns has been rapid, and sub-2-μm bead-packed, , monolithic ,,− and open-tubular ,− capillary columns, as well as pillar array columns, have been developed, as overviewed in several recent reviews. − …”

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confidence: 99%

Improved Sensitivity of Ultralow Flow LC–MS-Based Proteomic Profiling of Limited Samples Using Monolithic Capillary Columns and FAIMS Technology

et al. 2020

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In this work, we pioneered a combination of ultralow flow (ULF) high-efficiency ultranarrow bore monolithic LC columns coupled to MS via a high-field asymmetric waveform ion mobility spectrometry (FAIMS) interface to evaluate the potential applicability for high sensitivity, robust, and reproducible proteomic profiling of low nanogram-level complex biological samples. As a result, ULF LC-FAIMS-MS brought unprecedented sensitivity levels and high reproducibility in bottom-up proteomic profiling. In addition, FAIMS improved the dynamic range, signal-to-noise ratios, and detection limits in ULF LC–MS-based measurements by significantly reducing chemical noise in comparison to the conventional nanoESI interface used with the same ULF LC–MS setup. Two, three, or four compensation voltages separated by at least 15 V were tested within a single LC–MS run using the FAIMS interface. The optimized ULF LC-ESI-FAIMS-MS/MS conditions resulted in identification of 2,348 ± 42 protein groups, 10,062 ± 285 peptide groups, and 15,734 ± 350 peptide-spectrum matches for 1 ng of a HeLa digest, using a 1 h gradient at the flow rate of 12 nL/min, which represents an increase by 38%, 91%, and 131% in respective identifications, as compared to the control experiment (without FAIMS). To evaluate the practical utility of the ULF LC-ESI-FAIMS-MS platform in proteomic profiling of limited samples, approximately 100, 1,000, and 10,000 U937 myeloid leukemia cells were processed, and a one-tenth of each sample was analyzed. Using the optimized conditions, we were able to reliably identify 251 ± 54, 1,135 ± 80, and 2,234 ± 25 protein groups from injected aliquots corresponding to ∼10, 100, and 1,000 processed cells.

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confidence: 99%

Improved Sensitivity of Ultralow Flow LC–MS-Based Proteomic Profiling of Limited Samples Using Monolithic Capillary Columns and FAIMS Technology

et al. 2020

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“…OT columns are another type of nanocolumn. In recent years, OT columns have gained an increasing amount of attention and have been widely used in diverse areas of separation science. − Even though this technique was first reported a number of years ago by Tsuda et al. using an octadecyl modified 30 μm i.d.…”

Section: Miniaturized Lc Technologymentioning

confidence: 99%

Miniaturized LC in Molecular Omics

et al. 2020

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Miniaturized LC has evolved at an exponential rate over the last 50 years. In the past decade, it has received considerable attention in the field of bioanalytical separation science and technology due to the need to measure different classes of biomolecules present in a variety of matrixes on a global scale to gain a deeper understanding of complex biological processes. This field has become a dominant area underpinning the molecular omics research (e.g., proteomics, metabolomics, lipidomics, and foodomics), allowing key insights into the function and mechanism of small to very large biomolecules on a molecular level. This Feature highlights the recent advances in molecular omics focusing on miniaturized LC technology combined with mass spectrometry-based platforms, with a particular emphasis on the strategies adopted and applications using new and sensitive nanoscale analytical methodologies.

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“…Similar columns were applied in studies of stem cell-like cancer cells by the Lundanes group 72,73 . However, despite the advances in open tubular column LC, as reviewed recently by Lam et al 74 , to the authors' knowledge, such columns are not commercially available, yet.…”

Section: Open Tubular Columnsmentioning

confidence: 99%