Accurate peptide retention data collection for retention modeling studies using the Q Exactive HF-X Orbitrap mass spectrometer
This presentation provides step-by-step guide for construction of peptide retention library using 2D LC-MS/MS analysis of whole cell tryptic digests.
Introduction of proteomics provided separation scientists with virtually unlimited source of retention data: identification and assignment of retention properties of tens of thousands peptides in a few hours of instrument time has become a routine task. Nevertheless, accurate retention data collection for retention modeling or construction of comprehensive retention libraries for data independent acquisition protocols still represents a formidable task. This presentation provides step-by-step guide for construction of peptide retention library using 2D LC-MS/MS (high pH RP – low pH RP separation scheme, Q Exactive HF-X Orbitrap mass spectrometer) analysis of whole cell tryptic digests.
Learning Objectives of Webinar
- To introduce the concept of “chromatographic quality control” for accurate retention data collection via application of peptide retention standards;
- To learn about major parameters of separation system affecting separation selectivity, i.e. accuracy of retention data collection;
- To estimate expected output from a typical 2D LC-MS/MS acquisition using Q Exactive HF-X platform.
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