To address the U.S. Army CBD need for a low-cost, universal, robust, and fieldable device for rapid, reproducible identification of protein variants from biological samples with high throughput, Physical Optics Corporation (POC) proposes to develop a new Dielectrophoretic Microfluidic Protein Analysis (DiMiPA) system based on an electrodeless dielectrophoresis-based, continuous-flow protein separation technique followed by protein variants quantification and identification via detection of the separated protein bands by UV absorption spectrometry and enhanced Raman spectrometry with POC-designed SERS substrates. The DiMiPA system will be composed of a microfluidic chip, an AC power supply, a syringe pump, and a readout unit. The DiMiPA tool will rapidly (10 min) and with high resolution and reproducibility separate protein variants, preparing them for identification by the readout unit. The DiMiPA system will be an inexpensive, portable, and easy-to-use automated instrument. In Phase I, POC demonstrated the feasibility of the DiMiPA device by fabricating a prototype and demonstrating its capability to separate allozymes of sulfotransferase (SULT1A1), SULT1A1*1, and SULT1A1*3, which differ from each other by a single amino acid. In Phase II, DiMiPA will be optimized to enhance system throughput and identification resolution to identify allozymes of SULT1A1 and heat shock protein 70, HSP70-1A, and HSP70-1B.
Keywords: Polymorphism, Protein Variants Separation, Microfluidics, Dielectrophoresis, Raman Spectroscopy, Sers Substrate
