Collaborative protein technology resource nanoscale protein analysis

Overview

The Nanoscale Protein Analysis Section of the CCR Collaborative Protein Technology Resource (CPTR) provides expertise and service in state-of-the-art protein analysis technologies to advance CCR research in basic discovery and translation/clinical studies. Applications include:

Quantitative profiling of cell signaling and post-translational modification
Measurement of cytokine, metabolite and serum/plasma markers
Identification and validation of biomarkers & therapeutic targets
Assessment of on‐ and off‐target drug activity and pharmacodynamic evaluation
Study of tumor microenvironment and single-cell signaling

Established Technologies

Automated capillary immunoassay system – Simple Western technology

High-resolution MW (size-based) or isoelectric-focusing (IEF, charge-based) separation, followed by target-specific immunoprobing to profile proteins and respective post-translational modification isoforms
Nanogram level of protein analysis for extremely small and precious samples, e.g. stem cells, primary cells, micro-dissected samples, fine needle aspirates, non- or minimally- invasive collected patient specimens etc.
Provides precise and accurate measurement of protein expression level and activation status, highly quantitative data, good assay sensitivity, dynamic range and reproducibility
About three hundred assays covering key signaling pathways have been established and successfully applied for functional proteomic research, will validate assays for other targets as needed
Assay readiness for bench to bed transfer

In-solution multiplex ELISA assays – Luminex xMAP Technology

Up to 50-plex simultaneous protein target analysis with small sample consumption
Analysis of cytokine, metabolite, immune response, serum/plasma biomarkers etc.
Clinical applicable assay performance

Immunofluorescence imaging platform – CODEX (CO-Detection by indEXing) technology

Provides full spatial context and is not limited to just regions of interest (ROI)
Provides single-cell resolution down to 600 nm or 250 nm depending on microscope objective used (20X and 40X respectively)
Single-step staining and gentle fluorophore removal preserves the sample for ROI analysis
Simple, benchtop fluidics system that is cost-effective and simple to implement in any research lab

Developing Technologies

Single-cell Western system

Western-based protein analysis on 1000-2000 single-cells in parallel
Measures proteins hard-to-detect by FACS, such as isoforms, phosphorylated proteins, transcription factors, intracellular proteins etc.
Single cell signaling profiling and population heterogeneity study

We actively evaluate, develop, acquire and implement innovative protein analysis technologies to accommodate the demands from the CCR/NCI/NIH community.

Digital Spatial Profiling

NanoString GeoMx Digital Spatial Profiling (GeoMx DSP)
Offered in collaboration with the CCR Genomics Core
FFPE or fresh frozen
Profiling data for 100s of validated proteins
Cancer Trascriptome Atlas (CTA) > 1,800 gene
Whole Transcriptome Atlas (WTA) > 18,000 genes

User Guidelines

Projects are collaborative and subject to the approval of resource advisory committee.
Investigators are expected to share the cost of certain consumable reagents, for which CCR investigators are eligible for a 50% subsidy from the CCR Office of Science and Technology Resource (OSTR).
Staff are available to assist on project design, data interpretation and study milestone evaluation, as well as general consultation on proteomic analysis questions.

Keywords

4E-BP1 ABCG2 AKT ALAS1 AMPK BIM BLM Bmi1 CIAP CODEX CPTR Caspase Chk1 Cofilin Cyclin DDB1 DNA damage DNMT1 DUSP3 E2F-1 EGFR EGR1 ELISA ERB2 ERK FADD FHL1 FHL2 FLIP Flag FosB Fox03A Fra1 GAB1 GAPDH GST Glucose-6-phosphate dehydrogenase (G6PD)HSP70 HSP70s Integrin IκBα Iκκα JAK2 JNK JunB KID LIMK2 Luminex MAGPIX MAP kinase MCM2 MCM4 MCM5 MEK Milo Multiplex Tissue Imaging NFκB NFκB p65 NFκB1 p105/p50 NFκB2 p100/p52 NanoProTM PARP PI3 Kinase p110 α/β PI3 kinase PKC PLCγ1 PLK1 PTEN Peggy Sue Peggy SueTM PeggyTM ProteinSimple Proteins Proteomics RIP ROCK Raf RasGRP Rb RelB S6 Ribosomal protein SHC1 SMAC/Diablo SMAD SMARCB1 SRC STAG2 STAT SallyTM Simple Western Spatial Transcriptomics TACC3 TRAF1 Thioredoxin 1 Vinculin WesTM XIAP analyte apoptosis biomarker c-Jun c-Myc c-Rel cFos capillary immunoassay cell cycle cell signaling chemokine clinical trial core biopsy cultured mammalian cells cytokine drug development epigenetics Proteins and Proteomics Single Cell Analysis fine needle aspirates heterogeneity immunoassay inflammation loading controls mTOR micro-dissection molecular profiling multiplex ELISA multiplex immunoassay nano-fluidic immunoassay nci-core p-glycoprotein p53 p70 S6 peripheral blood mononuclear cells (PMBCs)pharmcodynamic analysis phospho-Histone H2A.X (S139)phosphorylation; post-translational modification plasma cells preclinic and clinic sample analysis proliferation protein characterization protein complex protein quantification single cell subcellular fractionation target therapy tissue samples transcription factor tyrosine kinase western western blot xMAP α-Tubulin β-Actin β-Catenin

Contact Information

Name:

Noemi Kedei, MD

Title:

Facility Head

Address:

Building 41, Room D310
Bethesda, MD 20892

Phone:

(240) 760-6804

E-Mail:

kedein@nih.gov

Visit Website

Additional Core Staff

Name:

Maria O. Hernandez, PhD

Title:

Scientist

Phone:

(240) 760-7774

E-Mail:

maria.hernandez2@nih.gov

Name:

Xiaoling Luo

Title:

Scientist

Phone:

(240) 760-6922

E-Mail:

luox2@mail.nih.gov