Sequential Isoelectric Point Separation of Proteins Using Non-Gel, Microfluidic System

The physiological health of soldiers is at risk due to potential biological agent exposure. Many of these agents are protein-based and can cause alterations in cellular proteins. The ability to rapidly monitor, detect, and analyze proteins will enable earlier identification of exposure and result in quicker intervention potentially saving many lives. A key step in analysis of protein mixtures is protein separation, typically accomplished using two-dimensional gel electrophoresis (2DGE). Although well-established, the process of performing 2DGE is laborious. Furthermore, equipment used in 2DGE is not readily field-deployable. Therefore, a quick and portable method for protein separation is desirable for far-forward military use. To accomplish protein separation without the need for 2DGE, Lynntech proposes to develop a continuous-flow isoelectric point sorting system. In Phase I, Lynntech developed prototype protein separation chips and successfully demonstrated flow of protein sample mixtures through the chips. In Phase II, Lynntech will integrate separation chips with fluid handling and software control to deliver a field-deployable protein separation system to be used in forward medical hospital settings. The molecular weight and isoelectric point (pI) ranges will also be expanded to achieve rapid separation of biologically- and military-relevant proteins in the 16-200 kilodalton range with pI of 3-12.