The identification and quantification of specific proteins in complex samples are critical processes in discovering, understanding, and ultimately addressing, the underlying nature of human health and disease. The western blot is a fundamental method used for these processes due to the sensitivity and specificity enabled by combining protein sizing with immuno-detection. Despite being widely- used, the western blot has limits related to required sample size, throughput, information content, quantification, and repeatability, which hinder many research efforts. Next Generation Western (NGW) is 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 detection. To develop the NGW prototype system, the specific aims are to (1) develop microchips and reagents that enable 10 minute separation/blotting with performance superior to traditional methods, (2) develop instrument to control/automate microchip functions and interface with blotting membranes, (3) develop automated data analysis and visualization software, and (4) conduct intra- and inter-laboratory reproducibility studies and deploy the prototype system for performance verification and market validation. LI-COR Biosciences is well established in the western blot market with worldwide sales and support of instrumentation and reagents.
The western blot is a fundamental technique used to identify and quantify specific proteins in complex samples; however, the technique has significant limitations that hinder progress in research efforts aimed at understanding and diagnosing various human health issues and diseases. This proposal focuses on development of a novel, microfluidic, ink-jet blotting technique to automate and miniaturize western blotting. This technology will enable throughput and content beyond the capabilities of traditional western blotting, including quantification of multiple proteins in small tissue samples used in research related to human health and disease.