Shu-Bai Liu, Sylvia Sardi, Boldbaatar Sonom, Davide Zocco, Russel McSweeney, Andrew D. Fraser, Allison E. Halleck, Haotian Li, Gary B. Smejkal, Steven Munevar, Jason Gang Jin, Toshi Kawai, Ionita Ghiran, John P. McGrath, Malcolm Whitman, Shu-Wing Ng, and Winston Patrick Kuo
There is increasing evidence that abnormal protein synthesis and modification are associated with a variety of human diseases. In the coming era of personalized/precision medicine, it will be required to utilize a rapid, highly sensitive and quantitative method to analyze the proteins and related post-translational modifications in clinical specimens in order to better define specific therapies for patients. However, the current gold standard in proteomic analysis is still the traditional Western blot, which requires many manual steps with lower sensitivity and provides a semi-quantitative read-out. Here, in this manuscript, we present the first report of a novel fully automated Capillary Electrophoresis (CE)-based immunodetection technology, called the Simple Western size assay, which is run on
the instrument, called the SimonTM. This technology is based on nanovolume size-based protein separation that can be used to quantify proteomic profiles of clinical specimens for both biomarker discovery and diagnostics. Our results demonstrated that the Simple Western has higher sensitivity of target protein detection, a greater linear dynamic range of different molecular weight proteins, high reproducibility and the capacity for the higherthroughput screening of samples using small sample input volumes compared to traditional Western blot analysis. In addition, the quantitativeness and accuracy, the exquisite sensitivity and reduced background noise, has made the Simon Western highly versatile. This technology can quantitative the level of protein and related post-translational modifications in translational medicine research, such as specific biomarkers for diabetes and cancer research. These results based on several broad applications in this study suggest the Simple Western size assay will be a novel potential protein detection accelerator in the personalized and translational medicine era.
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