Automated Sample Preparation for a More Efficient and Reproducible Phosphopeptide Enrichment Workflow

Jun 25, 2018

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Protein phosphorylation is a central regulatory mechanism of cell signaling pathways. Proper function of protein kinases and phosphatases is important in biological processes, which means it is important to understand phosphorylation-mediated signaling networks. Analytical strategies combined with mass spectrometry have been developed to identify and quantify the dynamics of phosphorylation under different biological conditions. Global phosphopeptide enrichment prior to LC–MS is one of the most challenging sample preparation workflows to carry out in a reproducible manner.

In this webcast we present results of a study in which we compared automated phosphopeptide enrichment workflow using Fe(III)-IMAC cartridges on an automated protein sample prep platform with our manual phosphopeptide enrichment workflow and applied them to standard Hela cell digests and biological sample digests. Compared to our traditional manual workflow, the automated workflow resulted in an efficient, sensitive, and reproducible enrichment of thousands of phosphopeptides. The automated phosphoproteomics workflow applied to WT and SHANK3-deficient rat primary neurons or mouse brain allowed the identification of many phosphosites downregulated and involved in Akt-mTORC1 signaling pathways.


Key Learning Objectives:

  • Understand the capabilities of automated protein sample preparation to enable turnkey workflows
  • See how automated sample preparation compares to manual sample preparation for reproducibility
  • Find out the benefits of automated workflow for the enrichment of thousands of phosphopeptides to improve selectivity and efficiency



Dr. Debora Bonenfant, Senior Investigator, Novartis Institutes for BioMedical Research


Date and Time:

Live: Europe: Monday, 25 June, 2018 at 1400 BST | 1500 CEST

North America: Monday, June 25, 2018 at 11 am EDT | 10 am CDT | 8 am PDT

Asia Pacific: Tuesday, 26 June, 2018 at 1100 CST | 1200 JST | 1300 AEST

After the final airing of the webcast on 26 June, 2018 it will be available on demand until 26 June, 2019. 

Sponsor: Agilent Technologies, Inc.  

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