The Human Proteome Project (HPP) MS-Pillar Phosphopeptide challenge resource continues to collect new data and invites new participants to also contribute to this knowledgebase. Participants are invited to contribute to both phases of phosphopeptide identification methods development by analyzing a set of phosphopeptides by their favorite method and follow up with a method of affinity purification step using the popular ReSyn HUPO affinity kit to see improvements in their methods. Together with our partners, SynPeptide Co. Ltd in Shanghai (www.synpeptide.com) and Resyn Biosciences Pty Ltd in South Africa (www.resynbio.com), the HPP MS-Pillar will provide the SynPeptide-HUPO phosphopeptide mixtures as well as a comprehensive ReSyn phosphopeptide purification kit to use as the affinity method for the HUPO phosphopeptide mixtures. The SynPeptide phospho peptides and the Resyn MagReSyn® kit including the magnet separator are valued at over US$1000 each are provided free to all interested 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 HPP MS-pillar has combined the peptide sets and affinity purification kit 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, combined with the HUPO 2018 data, and reported at the HUPO 2019 meeting.
The HUPO phosphopeptide mixtures Peptides and iMac kits can be picked up from SynPeptide and Resyn at their booth at HUPO 2018 Orlando.