Next Generation Western for high-throughput and high-content protein analysis

Abstract The identification and quantification of specific proteins in complex samples are critical processes in discoveringunderstandingand ultimately addressingthe underlying nature of human health and diseaseThe western blot is a fundamental method used for these processes due to the sensitivity and specificity enabled by combining protein sizing with immuno detectionDespite being widelyusedthe western blot has limits related to required sample sizethroughputinformation contentquantificationand repeatabilitywhich hinder many research effortsNext Generation WesternNGWis an innovative approach to alleviate limitations of traditional western blotting using a novel method that automates and miniaturizes the process of electrophoresis followed by ink jet blotting onto a moving capture medium for subsequent immuno probing and detectionTo develop the NGW prototype systemthe specific aims are todevelop microchips and reagents that enableminute separation blotting with performance superior to traditional methodsdevelop instrument to control automate microchip functions and interface with blotting membranesdevelop automated data analysis and visualization softwareandconduct intraand inter laboratory reproducibility studies and deploy the prototype system for performance verification and market validationLI COR Biosciences is well established in the western blot market with worldwide sales and support of instrumentation and reagents Narrative The western blot is a fundamental technique used to identify and quantify specific proteins in complex sampleshoweverthe technique has significant limitations that hinder progress in research efforts aimed at understanding and diagnosing various human health issues and diseasesThis proposal focuses on development of a novelmicrofluidicink jet blotting technique to automate and miniaturize western blottingThis technology will enable throughput and content beyond the capabilities of traditional western blottingincluding quantification of multiple proteins in small tissue samples used in research related to human health and disease