In the nano-immunoassay division of CPTR, we have developed and implemented automated capillary immunoassays (The Simple Western assays) for better characterization of signaling proteins and their post-translational modification (PTM) isoforms. The system performs automated western analysis in capillaries using nanogram level of proteins, and provides digital data quantitation with good assay sensitivity and reproducibility. The technology offers precise and accurate proteomic assessment in limited sample sizes, such as stem cells and fine needle aspirates etc. The Simple Western system can also perform iso-electric focusing (IEF) immunoassays to separate, detect and quantify different post-translationally modified states of a protein without using modification-specific antibodies. In addition, the IEF analysis provides additional level of information on the activation status of signaling molecules that are not accessible by conventional Western. Through collaborations with investigators from different branches/laboratories of CCR/NCI/NIH, we have successfully applied the technology to profile cell signaling events in a variety of projects from basic research to clinical studies. A panel of about two hundred Simple Western assays, covering major key signal 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 noval platform for comprehensive quantitative proteomic analysis. The potential of using Simple Western system for clinically relevant biomarker development and therapeutic target identification is particularly exciting. We have established clinically feasible assays for pharmacodynamic assessment and targeted therapy evaluation. We also developed / validated assays and protocols for patient specimen analysis in several clinical trial projects, including lung cancer, breast cancer, ovarian cancer, melanoma, multiple myeloma etc... Our efforts have led to multiple manuscript preparation and publications, as well as presentations in multiple national and international meetings. We have also established a close working relationship with the technology developer, ProteinSimple, for continuing improvement of the assay systems to be more robust and reliable. The effort has resulted in enhanced assay performance and readiness for clinical applications, and as well as making our core a leader in these novel technologies as we support CCR science. We also recently acquired the Bio-Plex suspenstion array system, which performs in-solution multiplex sandwich ELISA assays based on Luminex xMAP technology. The new system provides complementary assays to the Simple Western system (such as cytokine, metabolite, immune response, serum/plasma biomarker analysis etc.). This further broadens the scope and capability of our core to provide comprehensive quantitative proteomic analysis for cell signaling molecule profiling, biomarker assessment and therapeutic treatment evaluation.
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