Two analysis platforms have been established currently: Capillary Nano-immunoassay (CNIA) system - Simple Western (ProteinSimple, CA) -- Automated capillary immunoassay system, which employ 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. The capillary platform allows sample analysis with nanogram level of proteins, which makes it attractive for extremely small and precious samples, such as stem cells, primary cells, micro-dissected samples, fine needle aspirates, and other patient specimens etc. Luminex multiplexing immunoassays -- Multiplex in-solution ELISA analysis system, based on the Luminex xMAP beads technology, which combines advanced fluidics, optics, and digital signal processing with proprietary microsphere technology to deliver multiplexed assay capabilities. These technologies have been successfully applied in both discovery research and clinical practice for precise and accurate measurement of signaling molecules and their activations, as well as mid-throughput quantitative proteomic analysis. Highly quantitative data, good assay sensitivity and reproducibility have been observed in both analysis platforms. Through collaborations with investigators from different branches/laboratories of CCR/NCI/NIH, we have successfully applied the CNIA assays to profile cell signaling events in a variety of projects including cell signaling profiling, biomarker development, targeted therapy evaluations and investigation on drug resistance mechanism etc. A panel of about two hundred capillary-immunoassays, covering key signaling pathways from receptor activation, down-stream signaling transduction, transcriptional regulation, cell cycle control to apoptosis etc., has been developed / established in our core, providing a novel platform at CCR for comprehensive and quantitative proteomic analysis with minute sample consumption. The IEF analysis platform of the CNIA assays offers the capability to separate, detect and quantify different post-translationally modified states of a protein without using modification-specific antibodies. Assays and protocols have also been developed and validated for patient specimen analysis and applied in several clinical trial projects. The effort greatly facilitated the clinical investigations on monitoring drug response at the protein level, and/or helped the investigators to further modify treatment strategies. Our efforts have demonstrated that the CNIA system provides quantitative and precise measurement of signaling molecules and their activation status, and have led to multiple publications and presented at national and international meetings, as well as established our core as a leader in the field of CNIA technology. Recent implementation of the Luminex multiplexing immunoassay system allows our core to accommodate the demand for affordable, high performance, good-throughput multiplex in-solution ELISA assays. The technology is recently selected to be deployed for biobank profiling research in the Human Protein Atlas (HPA) project, and provides complementary assays to the CNIA analysis (such as cytokine, metabolite, immune response, serum/plasma biomarker analysis etc). In the past months, we have received projects for cytokine analysis, epigenetic drug evaluation etc. Following services have been established and offered at the core: 1) Protein characterization and post-translational modification analysis; 2) Cell signaling profiling and mid-throughput quantitative proteomics; 3) Biomarker development and analysis; 4) Pharmcodynamic and molecular diagnostic analysis, target therapy evaluation etc;

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Scientific Cores Intramural Research (ZIC)
Project #
1ZICBC011434-04
Application #
9154336
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
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