+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Improving Proteome Coverage by Reducing Sample Complexity via Chromatography

Improving Proteome Coverage by Reducing Sample Complexity via Chromatography

Advances in Experimental Medicine and Biology 919: 83-143

High performance liquid chromatography (HPLC) is currently one of the most powerful analytical tools that has revolutionized the field of proteomics. Formerly known as high pressure liquid chromatography, this technique was introduced in the early 1960s to improve the efficiency of liquid chromatography separations using small stationary phase particles packed in columns. Since its introduction, continued advancements in column technology, development of different stationary phase materials and improved instrumentation has allowed the full potential of this technique to be realized. The various modes of HPLC in combination with mass spectrometry has evolved into the principal analytical technique in proteomics. It is now common practice to combine different types of HPLC in a multidimensional workflow to identify and quantify peptides and proteins with high sensitivity and resolution from limited amounts of samples. More recently, the introduction of Ultra High Performance Liquid Chromatography (UHPLC) has further raised the level of performance of this technique with significant increases in resolution, speed and sensitivity. The number of applications of HPLC and UHPLC in proteomics has been rapidly expanding and will continue to be a pivotal analytical technique. The aim of the following sections is to familiarize the beginner with the various HPLC methods routinely used in proteomics and provide sufficient practical knowledge regarding each of them to develop a separation and analytical protocol.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 059845188

Download citation: RISBibTeXText

PMID: 27975214

DOI: 10.1007/978-3-319-41448-5_5

Related references

Reducing the complexity of the Escherichia coli proteome by chromatography on reactive dye columns. Methods in Molecular Biology 424: 167-185, 2008

Improving Proteome Coverage and Sample Recovery with Enhanced FASP (eFASP) for Quantitative Proteomic Experiments. Methods in Molecular Biology 1550: 11-18, 2017

In-Depth Proteome Coverage by Improving Efficiency for Membrane Proteome Analysis. Analytical Chemistry 89(10): 5179-5185, 2017

Improving proteome coverage on a LTQ-Orbitrap using design of experiments. Journal of the American Society for Mass Spectrometry 22(4): 773-783, 2011

Cross-sample validation provides enhanced proteome coverage in rat vocal fold mucosa. Plos One 6(3): E17754-E17754, 2011

Improvements in protein identification confidence and proteome coverage for human liver proteome study by coupling a parallel mass spectrometry/mass spectrometry analysis with multi-dimensional chromatography separation. Analytica Chimica Acta 566(2): 147-156, 2006

Improving protein and proteome coverage through data-independent multiplexed peptide fragmentation. Journal of Proteome Research 9(7): 3621-3637, 2010

Enhanced FASP (eFASP) to increase proteome coverage and sample recovery for quantitative proteomic experiments. Journal of Proteome Research 13(4): 1885-1895, 2014

Displacement chromatography as first separating step in online two-dimensional liquid chromatography coupled to mass spectrometry analysis of a complex protein sample--the proteome of neutrophils. Journal of Chromatography. A 1232: 288-294, 2012

Reducing sample complexity by RP-HPLC: beyond the tip of the protein expression iceberg. Methods in Molecular Biology 424: 147-156, 2008

Reducing sample complexity of polyclonal human autoantibodies by chromatofocusing. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 878(24): 2249-2254, 2010

Novel parallelized quadrupole/linear ion trap/Orbitrap tribrid mass spectrometer improving proteome coverage and peptide identification rates. Analytical Chemistry 85(24): 11710-4, 2014

Reducing sample complexity in proteomics by chromatofocusing with simple buffer mixtures. Methods in Molecular Biology 424: 187-203, 2008

Glycosylation as means of reducing sample complexity to enable quantitative proteomics. Proteomics 9(6): 1488-1491, 2009

Dual-purpose sample trap for on-line strong cation-exchange chromatography/reversed-phase liquid chromatography/tandem mass spectrometry for shotgun proteomics. Application to the human Jurkat T-cell proteome. Journal of Chromatography. A 1070(1-2): 193-200, 2005