Advanced nanoscale separations and mass spectrometry for sensitive high-throughput proteomics
We review recent development in separations and mass spectrometric instrumentation for sensitive and high-throughput proteomic analyses. These efforts have been primarily focused on the development of high-efficiency (separation peak capacity of ~103) nanoscale liquid chromatography (nanoLC; e.g., flow rates extending down to ~20 nL/min at optimal separation linear velocities through narrow packed capillaries) in combination with advanced mass spectrometry (MS), including high sensitivity and high resolution Fourier transform ion cyclotron resonance (FTICR) MS. This technology enables MS analysis of low nanogram-level proteomic samples (i.e., nanoscale proteomics) with individual protein identification sensitivity at the low zeptomole-level. The resultant protein measurement dynamic range can reach 106 for nanogram-sized proteomic samples, while more abundant proteins can be detected from complex sub-picogram size proteome samples. The average proteome identification throughput using MS/MS is >200 proteins/h for a ~3 h analysis. These qualities provide the foundation for proteomics studies of single or small populations of cells. The instrumental robustness required for automation and providing high quality routine performance nanoscale proteomic analyses is also discussed.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 15020630
- Report Number(s):
- PNNL-SA-44169; 400412000; TRN: US200521%%203
- Journal Information:
- Expert Review of Proteomics, 2(3):431-447, Vol. 2, Issue 3
- Country of Publication:
- United States
- Language:
- English
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