Accurate peptide retention data collection for retention modeling studies using the Q Exactive HF-X Orbitrap mass spectrometer

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

1. To introduce the concept of “chromatographic quality control” for accurate retention data collection via application of peptide retention standards;
2. To learn about major parameters of separation system affecting separation selectivity, i.e. accuracy of retention data collection;
3. To estimate expected output from a typical 2D LC-MS/MS acquisition using Q Exactive HF-X platform.

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