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NATIONAL CANCER INSTITUTE - CANCER.GOV

Contact Information

Primary Contact

Thorkell Andresson
Director

Location

8560 Progress Drive
Frederick, MD 21701

Overview

Protein and Metabolite Characterization Core (PMCC), formerly known as the Protein Characterization Lab (PCL),  offers various technologies to CCR investigators to characterize proteins and metabolites. The core develops and applies state-of-the-art analytical technologies, primarily mass spectrometry, and liquid chromatography, to advance the understanding of cellular function at the protein, proteomics, metabolite, and metabolomic levels. It engages in both short and long-term collaborations based on the need of each project. It is operated by Leidos Biomedical Research Inc. on behalf of NCI as part of the Frederick National Laboratory.

Expertise and Technology
  • Protein identification and characterization from different sample types using a variety of proteases, separation, and mass spectrometry methods.
  • Characterization of complex proteomes (e.g., serum, tissue, cell lysates)
  • Global comparative proteomic quantitation using TMT, SILAC, dimethyl labeling and/or LFQ
  • Identification and characterization of protein modifications – PTM site mapping
  • Global comparative Posttranslational Modification (PTM) quantitation using TMT, SILAC, dimethyl labeling, LFQ and affinity enrichment for modifications such as phosphorylation, ubiquitination, and acetylation
  • MHC I and II peptide identification and global immunopeptidome analysis.
  • Protein complex isolation and characterization
  • Validation of protein-protein interaction
  • Identification of protein-nucleic acid (DNA, RNA) interaction
  • Protein contact mapping using mass spectrometry cleavable cross linkers
  • Intact protein analysis, protein-drug conjugate analysis
  • Analytical protein and peptide fractionation using different methods such as size exclusion chromatography (SEC), reverse phase (RP) and ion exchange (SCX).
  • Relative and absolute quantification of metabolites using MRM based targeted assay incorporating isotopic standards for increased identification and quantification accuracy
  • Develops and establishes custom targeted metabolite assays for small molecules and metabolites.
  • Identification and quantification of modified nucleotides (DNA and RNA).
  • Pharmacokinetics (PK) analysis, develop quantitative MRM assay for known and novel drugs and small molecules. Analyzing drug metabolites in relevant biofluids and tissues.
  • Purity determination and structural analysis of small molecules
  • Isotope tracing to monitor metabolite pathway activities

Protein and proteomics analysis:

The core specializes in the identification and characterization of individual proteins, global proteomics analysis from a variety of different starting materials, targeted and global analysis of macromolecular post-translational modifications, characterization of entire protein complexes and other macromolecular interactions, protein cross-linking mapping, and intact protein mass analysis.

Metabolite and metabolomics analysis:

The core specializes in assay development for the identification and quantification of small molecules and metabolites utilizing targeted MRM assays in complex biological samples. For projects requiring 'Routine' metabolomics support, please visit the NIH Metabolomics Consortium (NIH-MC).

Major Instrumentation

  • Orbitrap Eclipse
  • Orbitrap Fusion Tribrid
  • Q Exactive HF
  • Nano flow liquid chromatography HPLC
  • 3x TSQ Quantiva
  • TSQ Altis from Thermo
  • Biacore T200

User Guidelines

Excess capacity can be made available to other NIH Investigators on a case-by-case basis. To request services from this CCR dedicated core facility, you must submit your requisition through NAS.

Keywords

BiaCoreDNA and RNA nucleotide modificationHPLCLTQ Velos ProMRMOrbitrap Fusion hybrid mass spectrometerPK assay developmentPRM parallel reaction monitoringProtein Characterization LabQ ExactiveSECSILAC analysisSILAC labeling of cellsSNP protein interactionSRMSeparation of analytesSize Exclusion ChromatographyTMTabsolute quantitation of proteinsacetylationaffinity purification of protein complexesanalysis of post-translational modificationsanalyte detection by MSantibody affinityantibody pairingassay developmentbinding affinitycell lysateclinical samplesconditioned mediacustomized assay developmentdrug metabolitesMetabolomicsProteins and Proteomicsglobal protein quantitationglobal relative protein quantitationhigh-performance liquid chromatography (HPLC)iTRAQidentification of phosphatase substratesidentification of post-translational modificationsidentification of protein modification siteimmunofluorescenceimmunoprecipitationintact molecular masskinetic constantlabel-free relative protein quantitationlong non coding RNA protein interactionmacromolecular interactionmetabolite analysismetabolomicsmethylationmouse samplesmultiple reaction monitoringmultiplexed quantitationnatural productsnci-corepharmaco kineticpharmocokinetic analysisphosphorylationprotein DNA interactionprotein RNA interactionprotein complex characterizationprotein expression in cell linesprotein identificationprotein interaction validationprotein ligand interactionprotein mass spectrometryprotein separationprotein small molecular interactionprotein-protein interactionpurity assessment of biological moleculesquantitation of analytesquantitative assays developmentrelative target protein quantitationselected reaction monitoringseparationserumsingle nucleotide polymorphism protein interactionsubcellular fractionssumolationtargeted metabolite profilingubiquitylationwestern blottingmass spectrometry