Peptide Synthesis
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Stable Isotope-Labeled Peptide and Protein Reagents/Kits
- Stable Isotope-Labeled Peptide and Protein Reagents/Kits
- Preloaded Resins for Solid-phase Synthesis of Stable Isotope Labeled Tryptic Peptides
- Synthetic Peptides As Internal Standards
Targeted mass spectrometry isotope analysis (i.e. selected reaction monitoring or SRM) is an alternative to antibody-based assays for the validation of clinically relevant biomarkers, but also has been employed for discovery-based quantitative proteomics (see example references below). One obstacle of this strategy is that every peptide possesses unique biochemical characteristics. Its amino acid composition and possible post-translational modifications defines its elution profile from the liquid chromatography column, ionization and fragmentation. For developing a diagnostic clinical MS assay, these peptide properties must be characterized with a synthesized peptide before analyzing the peptide of interest in vivo. Peptides also can be synthesized with heavy stable isotopes for absolute quantitation by spiking in a known amount of the heavy peptide into the biological sample. These strategies are also commonly employed to validate results from large scale quantitative proteomic analyses.
Resources
Stable Isotope-Labeled Peptide and Protein Reagents/Kits
Preloaded Resins for Solid-Phase Tryptic Peptide Synthesis
Frequently Asked Questions
Does CIL provide catalog and custom-labeled peptides?
Stable-isotope labeled peptides are used in biomarker discovery and validation as well as in drug and metabolite monitoring, peptide signaling experiments, metabolomics, and pharmacokinetics. CIL is pleased to supply highly enriched and pure amino acids (both free and protected) to the world’s leading peptide manufacturers for synthesis activities.
Would 0.8 mM of lysine or arginine preloaded resins provide approximately 8 synthesis at 0.1 mM scale?
Yes, theoretically 0.8 mM of resin would provide 8 synthesis at 0.1 mM scale. Final yields are always dependent on actual sequence and the efficiency of the couplings.
Does CIL provide catalog and custom preloaded resins?
Yes. Through collaboration with NE Peptide, Inc. (NEP), CIL is pleased to offer synthesis-ready preloaded resins. For a list of our current offerings, please refer to our preloaded resins webpage, and see our preloaded resins flyer for more information.
References
Lu, B.L.; Loomes, K.M.; Hay, D.L.; et al. 2020. Synthesis of isotopically labelled αCGRP8-37 and its lipidated analogue. J Labelled Comp Radiopharm, in press.
Lau, J.K.-C.; Lam, K.H.B.; Lai, C.-K.; et al. 2019. Imidazolone formation from pronated tetrapeptides: effects of replacing a glycine by an alanine or proline residue. Int J Mass Spec, 435, 69-77. doi.org/10.1016/j.ijms.2018.10.002
Li, F.; Cui, L.; Yu, D.; et al. 2019. Exogenous glutathione improves intracellular glutathione synthesis via the γ-glutamyl cycle in bovine zygotes and cleavage embryos. J Cell Physiol, 234(5), 7384-7394. PMID: 30362550
Orti, V.; Mertens, B.; Vialaret, J.; et al. 2018. Data from a targeted proteomics approach to discover biomarkers in saliva for the clinical diagnosis of perioddontitis. Data Brief, 18, 294-299. PMID: 29900194
Peng, X.; Xu, X.; Wang, Y.; et al. 2018. A-to-I RNA editing contributes to proteomic diversity in cancer. Cancer Cell, 33(5), 817-828. PMID: 29706454
Mertens, B.; Orti, V.; Vialaret, J.; et al. 2018. Assessing a multiplex-targeted proteomics approach for the clinical diagnosis of periodontitis using saliva samples. Bioanalysis, 10(1), 35-45. PMID: 29243487
LeBlanc, A.; Michaud, S.A.; Percy, A.J.; et al. 2017. Multiplexed MRM-based protein quantitation using two different stable isotope-labeled peptide isotopologues for calibration. J Proteome Res, 16(7), 2527-2536. PMID: 28516774
Percy, A.J.; Hardie, D.B.; Jardim, A.; Elliott, M.H.; Zhang, S.; Mohammed, Y.; Borchers, C.H. 2016. Multiplexed panel of precisely quantified salivary proteins for biomarker assessment. Proteomics, 17(6). PMID: 27538354
Delgado, D.A., Doherty, K.; Cheng, Q.; Kim, H.; Xu, D.; Dong, H.; Grewer, C.; Qiang, W. 2016. Distinct membrane disruption pathways are induced by 40-residue β-amyloid peptides. J Biol Chem, 29(23), 12233-12244. PMID: 27056326
Wang, D.; Krilich, J.; Baudys, J.; Barr, J.R.; Kalb, S.R. 2015. Optimization of peptide substrates for botulinum neurotoxin E improves detection sensitivity in the Endopep-MS assay. Anal Biochem, 468, 15-22. PMID: 25232998
Hamid, M.; Theo, K.; Bong, K. 2013. Systematic measurement of transcription factor-DNA interactions by targeted mass spectrometry identifies candidate gene regulatory proteins. Proc Natl Acad Sci U S A, 110(9), 3645-3650. PMID: 23388641
Karlsson, C.; Malmstrom, L.; Aebersold, R. 2012. Proteome-wide selected reaction monitoring assays for the human pathogen Streptococcus pyogenes. Nat Commun, 3, 1301. PMID: 23250431
Chambers, A.G.; Percy, A.J.; Yang, J. 2013. Multiplexed quantitation of endogenous proteins in dried blood spots by multiple reaction monitoring-mass spectrometry. Mol Cell Proteomics, 12(3), 781-91. PMID: 23221968
Altvater, M.; Yiming, C.; Melnik, A. 2012. Targeted proteomics reveals compositional dynamics of 60S pre-ribosomes after nuclear export. Mol Syst Biol, 8, 628. PMID: 23212245
McClatchy, D.B.; Liao, L.; Park, S.K.; Venable, J.D.; Yates, J.R. 2007. Quantification of the synaptosomal proteome of the rat cerebellum during post-natal development. Genome Res, 17(9), 1378-1388. PMID: 17675365
Venable, J.D.; Wohlschlegel, J.; McClatchy, D.B.; Park, S.K.; Yates, J.R. III. 2007. Relative quantification of stable isotope labeled peptides using a linear ion trap-Orbitrap hybrid mass spectrometer. Anal Chem, 79(8), 3056-3064. PMID: 17367114
McClatchy, D.B.; Dong, M.Q.; Wu, C.C.; Venable, J.D.; Yates, J.R. III. 2007. 15N metabolic labeling of mammalian tissue with slow protein turnover. J Proteome Res, 6(5), 2005-2010. PMID: 17375949
Andrew Percy, PhD
Senior Applications Chemist – Mass Spectrometry
Dr. Andrew Percy is the Senior Applications Chemist for Mass Spectrometry and the MS ‘Omics Product Manager at CIL. His responsibilities minimally involve providing technical support, overseeing product development, identifying new product market opportunities, assisting in the analysis of product-related applications, and writing/reviewing marketing literature.
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Kevin Millis, PhD
Senior Scientist, Application Development Manager
Kevin Millis, PhD, is the Senior Scientist and Market Development Manager for all NMR and mass spectrometry product lines. Kevin is responsible for Technical Services both internally and externally for all CIL customers as well as being responsible for the application and market development for the CIL products.
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