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MS-Pillar Phosphopeptide Challenge



HPP Phosphopeptide Challenge

01 Jun 2018 2:25 PM

The Human Proteome Project (HPP) MS-Pillar now extends is Phosphopeptide challenge resource to include affinity capture. Participants are invited to contribute to both phases of phosphopeptide identification methods development by analyzing a set of phosphopeptides by their favorite method and now follow up with an affinity purification step to see improvements in their methods. Together with our launch partner, SynPeptide Co. Ltd in Shanghai (, the MS Resource Pillar has partnered with Resyn Biosciences Pty Ltd in South Africa ( to provide a comprehensive phosphopeptide purification kit to use on the SynPeptide-HUPO phosphopeptide mixtures. The Resyn MagReSyn® kit including the magnet separator valued at over US$1000 each are provided free to HUPO members. 

The MS-Pillar Phosphopeptide challenge samples are a complex set of human phosphopeptides (Ser, Thr or Tyr) singly and multiply phosphorylated) and their unphosphorylated counterparts that can be used for method development and verification for phosphopeptide enrichment, sequence analysis by mass spectrometry and bioinformatic evaluation.

The first stage of this initiative is for interested members of the HUPO community to obtain a set of the peptides and apply their own methods and bioinformatic analysis to fully characterize the peptides as a neat mixture and in a tryptic digest background. Samples are still available by contacting the email below.

The second stage is now to compare your methods used with the Resyn purification kit for phospho-enrichment and peptide clean-up. The kit contains 2mL of each MagReSyn® Ti-IMAC, Zr-IMAC, and HILIC, and 4-place magnetic separator. As a result of this collaborative endeavor, multiple purification schemes, analytical protocols and data processing strategies will be evaluated, making it possible to determine the approach(es) that provide the highest coverage of phosphopeptides in the mixture.

By partnering with Resyn and SynPeptide the peptide sets and purification kit are being provided free-of-charge to all HUPO members, with the understanding that the analytical and data processing methods and results will be returned to the HPP MS Resource Pillar committee so they can be collated and reported at the HUPO 2018 meeting.

Free aliquots of the peptide mixtures and MagReSyn k it can be obtained by sending an email to


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