DIFFERENTIAL GEL ELECTROPHORESIS (DIGE) SYSTEM: INFECTIOUS DISEASE

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This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): Funding is requested for a 2-dimensional differential (fluorescence) gel electrophoresis (DIGE) system for the Keck Lab. This leased system is being used very successfully to identify differentially expressed proteins/biomarkers in conjunction with an existing nano-LC-ESI-QTOF for protein identification. DIGE is the most cost effective, powerful, and productive protein profiling technology in the Keck Lab and it is being heavily used to analyze a wide range of experimental versus control samples from whole cells, partially purified protein extracts, and biological fluids like sera. Our DIGE system has been used to carry out >350 quantitative analyses for 48 Yale and 32 non-Yale investigators at 19 institutions. Since entering into a lease in December, 2003, demand for DIGE has continued to increase. Improvements made with the requested, newer version image analysis software and our newly installed, high throughput and sensitive MALDI TOF/TOF mass spectrometer will make this technology state-of-the-art, and these improvements will further increase demand by investigators studying fundamental mechanisms of diseases including autism, substance abuse, cerebral palsy, mental retardation, premature birth, asthma, type I diabetes, renal diseases, tumor cell resistance, genesis of myeloid leukemia, molecular bone development, and Lupus. If this grant is not awarded, the 4-fold increase in monthly lease payments (as the lease matures from interest only to interest + capital equipment cost repayments starting in May, 2006) within the next year will result in a very substantial increase in DIGE service charges. Such an increase would very negatively impact on use of this advanced technology by both the Yale and non-Yale scientific community. Strengths of this application include the experience of the co-Pi overseeing large, multi-investigator NIH contracts and Centers; the complementary biotechnological, statistical, and database expertise of the principal investigator and other key personnel; the extensive protein profiling technologies already in use in the Keck Lab, and the strong infrastructure and technological research programs supported by several proteomics and high performance computing centers that are closely associated with the Keck Lab and are funded by NHLBI, NIDA, NIAID, and NCRR. If this grant is funded, the DIGE system would be unique to the Yale Community, and it surely would make a unique contribution to biomedical research that would extend far beyond Yale.