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Orthogonality of Separation in Two-Dimensional Liquid Chromatography
Abstract
Two-dimensional liquid chromatography is often used to reduce the proteomic sample complexity prior to tandem mass spectrometry analysis. The 2D-LC performance depends on the peak capacity in both chromatographic dimensions, and separation orthogonality. The peak capacity and selectivity of many LC modes for peptides is not well known, and mathematical characterization for orthogonality is underdeveloped. Consequently, it is difficult to estimate the performance of 2D-LC for peptide separation. The goal of this paper was to investigate a selectivity of common LC modes and to identify the 2D-LC systems with a useful orthogonality. A geometric approach for orthogonality description was developed and applied for estimation of a practical peak 2D-LC capacity. Selected LC modes including various RP, SCX, SEC, and HILIC were combined in 2D-LC setups. SCX-RP, HILIC-RP, and RP-RP 2D systems were found to provide suitable orthogonality. The RP-RP system (employing significantly different pH in both RP separation dimensions) had the highest practical peak capacity of 2D-LC systems investigated.
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Cited by
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Gadd, M. S., Testa, A., Lucas, X., Chan, K.-H., Chen, W., Lamont, D. J., Zengerle, M., & Ciulli, A. (2017) · Nature Chemical Biology · doi:10.1038/nchembio.2329 · Cited by 1,242
A Multidimensional Chromatography Technology for In-depth Phosphoproteome Analysis
Albuquerque, C. P., Smolka, M. B., Payne, S. H., Bafna, V., Eng, J., & Zhou, H. (2008) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m700468-mcp200 · Cited by 504
Discovery and mass spectrometric analysis of novel splice-junction peptides using RNA-Seq
Sheynkman, G. M., Shortreed, M. R., Frey, B. L., & Smith, L. M. (2013) · Molecular & Cellular Proteomics · doi:10.1074/mcp.o113.028142 · Cited by 128
Wu, Q., Yuan, H., Zhang, L., & Zhang, Y. (2012) · Analytica Chimica Acta · doi:10.1016/j.aca.2012.04.010 · Cited by 94
Looking for Missing Proteins in the Proteome of Human Spermatozoa: An Update
Vandenbrouck, Y., Lane, L., Carapito, C., Duek, P., Rondel, K., Bruley, C., Macron, C., Gonzalez de Peredo, A., Couté, Y., Chaoui, K., et al. (2016) · Journal of Proteome Research · doi:10.1021/acs.jproteome.6b00400 · Cited by 77
Capillary HILIC-MS: A New Tool for Sensitive Top-Down Proteomics
Gargano, A. F. G., Roca, L. S., Fellers, R. T., Bocxe, M., Domínguez-Vega, E., & Somsen, G. W. (2018) · Analytical Chemistry · doi:10.1021/acs.analchem.8b00382 · Cited by 52
Yu, D., Wang, Z., Cupp-Sutton, K. A., Liu, X., & Wu, S. (2019) · Journal of the American Society for Mass Spectrometry · doi:10.1007/s13361-019-02315-2 · Cited by 39
Baghdady, Y. Z., & Schug, K. A. (2019) · Analytical Chemistry · doi:10.1021/acs.analchem.9b01665 · Cited by 36
(2022) · doi:10.3390/proteomes10030026 · Cited by 20
Metadata
- Authors
- Gilar, M., Olivova, P., Daly, A. E., & Gebler, J. C.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2005-08-26
- Volume
- 77
- Issue
- 19
- Pages
- 6426-6434
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2034356373
- Cited by
- 816
- References
- 0
- FWCI
- 10.41
- Type
- journal article
- Language
- en
- Open access
- closed
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