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Guide to Peptide Quantitation


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Guide to Peptide Quantitation - 1

Agilent ' Trusted Answers Guide to Peptide Quantitation Agilent clinical research

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Peptide Quantitation for the Clinical Research Laboratory Peptide quantitation is rapidly growing in clinical research as scientists are translating peptide biomarkers identified in basic research to clinical research and utility. This quantitation approach measures unique peptides from the proteins that are expected to be present in clinical research samples based on prior knowledge. That is, from literature, genomic and proteomic studies, or combined proteogenomic analysis. As you develop peptide quantitation methods for your clinical research, we can offer expert advice at every step:...

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Small Versus Large Molecule Analysis Analyzing small molecules, and analyzing larger molecules such as peptides, are fundamentally different undertakings. Sample preparation for traditional small molecules involves a variety of techniques such as extraction and protein precipitation, depending on the analyte of interest. In contrast, peptide analysis follows a relatively standard protocol. Proteins in solution are first denatured, then reduced and alkylated before enzymatic digestion. While the same type of liquid chromatography and similar mobile phases are used to analyze peptides, the...

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Automated Peptide Sample Preparation for Mass Spectrometric Analysis Reproducibility in sample preparation has always been critical in clinical research. Preparing peptide samples involves many timed steps and reagent additions that can impact analytical reproducibility. Automating this process reduces variability and provides error-free operation with sample tracking. The Agilent AssayMAP Bravo platform efficiently prepares protein and peptide samples for LC/MS analysis. This platform offers a simplified user interface with laboratory-tested peptide and protein sample preparation protocols...

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Chromatographic Separation After the samples have been prepared and are ready for analysis, the next step is developing an efficient chromatographic method for the target peptides. The Agilent 1290 Infinity II liquid chromatography system gives you the ability to perform demanding separations with exceptional reliability and robustness. The UHPLC capability of the Agilent 1290 Infinity II LC system offers a variety of flow rates and column choices so that you can optimize your chromatographic methods for target peptides in complex matrices such as biofluids. “The flexibility of being able...

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The chromatographic columns used for peptide separations typically have larger pores to accommodate the larger molecules and include chromatographic media that minimizes non-specific binding of analytes for improved recovery. Agilent AdvanceBio Peptide Mapping columns deliver consistent, exceptional performance for separating and characterizing peptides. This column is superficially porous and has a 120 A pore size, which is optimal for analyzing the peptides produced by enzymatic digestion of proteins. The 2.7 pm particle size enables higher flow rates and better resolution of the target...

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Mass Spectrometric Detection For peptide quantitation, sensitivity and a wide dynamic range can be critical because the protein concentrations in biofluids may span many orders of magnitude. The Agilent 6495 Triple Quadrupole LC/MS system incorporates iFunnel technology. When combined with the Agilent Jet Stream ion source, iFunnel technology dramatically increases ion sampling, which results in lower detection limits. The 6495 is the proven choice for quantitative applications requiring superior analytical sensitivity, reliability, and overall system robustness. When maintenance is needed,...

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Agilent’s unique Jet Stream ion source provides an interface that is three to five times more sensitive than standard electrospray ionization, and is not concentration dependent. This enables your lab to use standard 2.1 mm id columns and higher LC flow rates without the loss of sensitivity typically associated with such an approach. For peptide quantitation, clinical researchers that are not sample-limited can achieve near-nanoflow HPLC analytical sensitivity using conventional-flow HPLC with the Jet Stream source. As a result, your LC/MS analysis of complex proteomics samples will be...

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MassHunter Software Agilent MassHunter Workstation software includes an acquisition module for instrument control as well as both quantitative and qualitative data analysis. Data processing can be automated with MassHunter Quantitative Analysis tools to rapidly provide peptide quantitation results. These tools enable you to customize the results criteria that you use and will highlight the results that warrant further review. MassHunter Workstation software can also integrate directly with your Laboratory Information System (LIS) so you can import sample lists for analysis, and ultimately...

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Skyline Software Agilent MassHunter software provides tight integration with Skyline software. Designed for proteomics researchers, Skyline software (from the MacCoss Group at the University of Washington) supports MRM, dynamic MRM (dMRM), and triggered MRM (tMRM) modes in MassHunter so you can acquire the desired quantifier and qualifier signals in increasingly complex samples. Automated MRM in three easy steps using the Agilent Skyline Automation tool 1 Determine RT 3 Create final method –– Create Skylinedocument –– Import CE optimization results –– Export CE optimize method –– Export...

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Agilent offers solutions for all aspects of the peptide quantitation workflow, including sample preparation, LC/MS analysis, and data processing. Agilent is proud to be your company of choice in peptide quantitation for clinical research. Learn more: Buy online: Find a local Agilent customer center: U.S. and Canada 1-800-227-9770 Europe Asia Pacific For Research Use Only. Not for use in diagnostic procedures. This...

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