Peptide Synthesis

The Proteomics Centre provides custom peptide synthesis services for mass spectrometry standards and antibody production. We use Prelude’s Peptide Synthesizer with parallel reaction vessels, which allows for high through-put synthesis, up to six peptides simultaneously. Current methodologies include Fmoc based chemistry with HBTU activation on a 5 - 20 nM variable synthesis scale. The added availability of a single shot amino acid delivery allows for easy incorporation of amino acids in to custom peptides (e.g. phosphorylated or nitrosylated peptides). The instrument also has an automated on-board cleavage chemistry feature allowing flexible cleavage options for all amino-acid requirements. Isotopically coded peptides are also synthesized at the Centre, which are useful for absolute quantitation of peptides and proteins using mass spectrometric approaches like MRM and or iMALDI (1-3).

Stable isotopically-labeled peptides are synthesized and purified at the Centre, creating exceptionally high quality standards with rapid turn around time. Standard peptides for over 600 proteins (more than 1035 peptides) are currently available. Please contact us for more information.

Refer to Biomarker Validation for further information on absolution quantification.

References
  1. Domanski, D., Murphy, L., Borchers, C. Assay Development for the Determination of Phosphorylation Stoichiometry using MRM methods with and without Phosphatase Treatment: Application to Breast Cancer Signaling Pathways. Anal Chem, 82(13): 5610-20. (2010)
  2. Reid, J.D., Holmes, D.T., Mason, D.R., Shah, B., Borchers, C.H. Towards the Development of an Immuno MALDI (iMALDI) Mass Spectrometry Assay for the Diagnosis of Hypertension JASMS, (2010)
  3. Bordeerat, N.K., Georgieva, N.I., Klapper, D.G., Collins, L.B., Cross, T.J., Borchers, C.H., Swenberg, J.A. and Boysen, G. Accurate quantitation of standard peptides used for quantitative proteomics. Proteomics, 15: 3939-44. (2009)
  4. Kuzyk, M.A., Smith, D., Yang, J., Cross, T.J., Jackson, A.M., Hardie, D.B., Anderson, N.L. and Borchers, C.H. Multiple Reaction Monitoring-based, Multiplexed, Absolute Quantitation of 45 Proteins in Human Plasma. Mol Cell Proteomics, 8(8): 1860-77. (2009)
  5. Warren, E.N., Elms, P.J., Parker, C.E, Borchers, C. H., Development of a Protein Chip: A MS-based Method for Quantitation of Protein Expression and Modification Levels using an Immunoaffinity Approach. Analytical Chemistry, 76: 4082-4092 (2004)