EAGER: Separating Proteins Via Dynamic Light Scattering Guidance of Drop Splitting

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ASU and the NASA Glenn Research Center propose to combine two relatively new techniques in order to change the existing paradigm on how proteins can be separated from a complex biological fluid. In order to explore the early-stage idea of controlling drop splitting via light scattering detection in a single drop, funding from the Early- Concept Grants for Exploratory Research program is well suited because it is a high risk and high payoff project. The paradigm of light scattering visualization as a means of controlling separation in a single drop would represent a fundamental change from current state of the art bioseparations in many ways: (1) there is no confinement of the liquid samples within a capillary, gel plate, or tube; (2) complex biological samples can be used; (3) simultaneous molecular weight and isoelectric point detection of proteins coincident with the separation process; (4) sequences of fractionations can be programmed and accomplished within minutes using the digital (e.g., drop-wise) nature of the liquid handling; and (5) the ability to modify focusing conditions on-thefly to better identify the proteins being detected during separation. The desired outcome of the research is the generation of data and the enhancement of modeling capabilities so that multiple proteins in biological fluids such as urine, saliva, or blood can be separated and analyzed. We expect that the unique combination of isoelectric focusing and scanning by lght scattering will generate a wealth of information not previously available in a rapid, portable system. The exploratory work is needed in order to understand how sensitivity, resolution, and the role of dynamic electric fields control protein separation in a drop. The work conducted will greatly help to formulate a research plan on wide-scale proteomics analysis of bodily fluids using digital isoelectric focusing with light scattering control and detection. The proposed EAGER will broaden participation by underrepresented groups and enhance infrastructure for research and education. Two Hispanic female students at Arizona State University (one graduate, Ana Gomez, and one undergraduate, Risa Garcia) will be directly involved with the exploratory work on isolectric focusing and splitting of a drop. We also hope to recruit another underrepresented student to conduct the light scattering research that will guide the protein bioseparation. The EAGER funding will catalyze a multidisciplinary collaboration between Arizona State University and NASA John Glenn Research Center opening the door to other future joint efforts between the Harrington Department of Bioengineering at ASU and the life systems group at NASA.