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

亚利桑那州立大学和NASA格伦研究中心建议结合两种相对较新的技术，以改变现有的如何从复杂的生物流体中分离蛋白质的范例。为了探索通过在单个液滴中进行光散射检测来控制液滴分裂的早期想法，来自早期概念拨款探索性研究计划的资金非常合适，因为它是一个高风险和高回报的项目。作为在单个液滴中控制分离的手段的光散射可视化范例将在许多方面代表与当前生物分离技术状态的根本变化：(1)液体样品不受毛细管、凝胶板或管内的限制；(2)可以使用复杂的生物样品；(3)与分离过程一致的蛋白质的同时分子量和等电点检测；(4)利用液体处理的数字(例如，滴状)性质，可以在几分钟内编程并完成分级序列；以及(5)改变聚焦条件的能力，以更好地识别在分离过程中检测到的蛋白质。研究的预期结果是生成数据和增强建模能力，以便能够分离和分析生物液体中的多种蛋白质，如尿液、唾液或血液。我们预计，等电聚焦和光散射扫描的独特组合将产生以前在快速、便携系统中无法获得的丰富信息。为了了解灵敏度、分辨率和动态电场如何控制一滴蛋白质的分离，需要进行探索性工作。所开展的工作将极大地有助于制定利用光散射控制和检测的数字等电聚焦对体液进行大规模蛋白质组分析的研究计划。拟议的EIGER将扩大代表不足的群体的参与，并加强研究和教育的基础设施。亚利桑那州立大学的两名西班牙裔女学生(一名毕业生安娜·戈麦斯和一名本科生丽莎·加西亚)将直接参与等电聚焦和水滴分裂的探索性工作。我们还希望招募另一名代表性不足的学生来进行光散射研究，这将指导蛋白质的生物分离。这笔急切的资金将促进亚利桑那州立大学和NASA约翰·格伦研究中心之间的多学科合作，为亚利桑那州立大学哈林顿生物工程系和NASA生命系统小组未来的其他联合努力打开大门。

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.