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Improved Protein Identification by NanoLC/MS/MS using Chip-Based Columns with Integrated Post-Column Addition of DMSO for Increased Sensitivity

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Improved Protein Identification by NanoLC/MS/MS using Chip-Based Columns with Integrated Post-Column Addition of DMSO for Increased Sensitivity

28 Sept 2015

NanoLC/MS/MS is the current method of choice for high sensitivity identification of proteins, e.g. for the discovery of biomarkers in plasma or other biological fluids. The solvent system typically used is determined by the requirements for both the chromatography and the electrospray processes. While it has been demonstrated that solvents like DMSO can increase electrospray ionization efficiency and improve sensitivity, the potential adverse effect on chromatographic separation has prevented researchers from adding these to the mobile phases. Postcolumn addition of these solvents is a good alternative, but not easy to achieve in nanoLC because of the unavoidable introduction of dispersion using a post-column Tee and additional connections. This poster introduces an extension to a chip-based nanoLC separation device that includes a Tee that is directly integrated into the chip post-column.

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ProteinPilot software

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ProteinPilot software is a powerful, robust and intuitive software tool for protein identification and quantification for discovery research and protein characterization.  A paradigm shift in protein identification and relative protein expression analysis for protein research, ProteinPilot software streamlines protein identification and quantitation, enabling you to identify hundreds of peptide modifications and non-tryptic cleavages simultaneously. Powered by sophisticated algorithms, you can now easily and confidently distinguish protein isoforms, protein subsets, and suppress false positives, as well as visualize peptide-protein associations and relationships.  

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.NanoLCNano LC is a nano scale liquid chromatography technique typically used for proteomic applications. The nano refers to the very low flow rate of the mobile phase in the system. The advantage of nanoLC is its high sensitivity. Often the system is integrated with a mass spectrometer in nano LC-MS or nano LC-MS/MS. When using nanoLC equipment consider column choice, software and accurate and precise flow rate control.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.BiomarkersBiomarkers are biological markers which can be measured and evaluated to indicate a biological state. The use of biomarkers in research and diagnosis can indicate a normal or disease state or drug response of cells / tissues. Biomarkers include genetic markers, cell surface markers such as antigens, antibodies or receptors and secreted molecules such as cytokines. An assay system is required for identification of biomarkers. :Protein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.ChromatographyChromatography is a powerful technique used to separate and analyze components of mixtures based on their chemical properties. It is widely used in fields like biochemistry, pharmaceuticals, and environmental analysis. By exploiting differences in the interaction of substances with a stationary phase and a mobile phase, chromatography enables precise purification and quantification of compounds. Whether you're working with complex biological samples or industrial chemicals, chromatography is essential for obtaining high-quality results. Browse our peer-reviewed product directory to find the best chromatography systems, compare products, read customer reviews, and get pricing directly from manufacturers.Biomarker DiscoveryClinical biomarkers refer to substances related to known medical conditions that can be accurately measured <i>in vitro</i>. Biomarkers can be used to diagnose presence of a disease and indicate disease severity. The discovery of new biomarkers is incredibly valuable in the field of diagnostics.Human PlasmaHuman plasma is the liquid component of blood, which carries blood components - nutrients, hormones, proteins, and waste - throughout the body. Plasma-based studies provide insights into biomarkers, immune responses, and disease mechanisms. Compare plasma handling tools and systems in our peer-reviewed product directory.DMSOElectrospray Ionization

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Improved Protein Identification by NanoLC/MS/MS using Chip-Based Columns with Integrated Post-Column Addition of DMSO for Increased Sensitivity