EAGER: Collaborative Proposal: Use of Elastin-Like Polymer as an Electrochemical Biosensor

合作项目PI Halpern，Jeffrey M./Balog，Eva Rose M.该项目的目的是开发一种快速测量复杂流体中类胡萝卜素的新方法。这个项目中提出的方法以前没有被研究过，如果成功，随着越来越多的产品进入市场，表明食品和食品补充剂中的类胡萝卜素含量较高，将有可能进行简单的测量，从而造福于整个社会。拟议工作的目标是利用弹性蛋白类聚合物(ELP)和电化学检测来开发一种选择性和灵敏的生物传感器。我们专注于虾青素的检测，虾青素是一种重要的抗氧化剂，在血清中很难选择性地检测出来。这项急切的提议的目标是为开发能够在复杂流体中快速测量的类胡萝卜素传感器获得初步数据。重点是提高对(1)电场下的ELP响应；(2)表面固定化ELP的电化学活性；以及(3)通过模块化融合蛋白成分引入分析物特异性的理解。预期的结果将导致使用工程的电化学标记的类胡萝卜素结合弹性蛋白类聚合物融合蛋白来监测复杂基质中的虾青素。虽然蛋白质-蛋白质相互作用和合成刺激反应聚合物在传感技术方面的利基越来越大，但在拟议的研究中，两者的优点被独特地结合在一起。通过将电化学标签连接到类弹性蛋白聚合物上，它们的刺激响应行为应该可以使用伏安传感器来检测。通过比较传感器表面上电化学标记的类弹性蛋白聚合物的收缩状态和拉伸状态，可以监测刺激响应行为。使用电化学标记的壳聚糖蓝蛋白-类弹性蛋白聚合物融合蛋白，将监测虾青素的结合。最后，将通过测量含有多种类胡萝卜素(即叶黄素和β-胡萝卜素)的溶液中的虾青素来测试特异性。定量了解类胡萝卜素在人类健康中的作用是有益于公众的。需要更好的传感技术来创建准确的基础生化研究，以调查类胡萝卜素的摄取和促进健康的类胡萝卜素水平。此外，在该项目中开发的类胡萝卜素技术可以扩展到其他蛋白质分析物。该研究项目将整合新英格兰大学和新汉普郡大学的研究人员进行跨学科研究，为所有学生提供机会。本科生将在新英格兰大学为普通社区举办公开研讨会，并发表演讲。

Collaborative Project PI Halpern, Jeffrey M./Balog, Eva Rose M. The purpose of this project is to develop a new method of measuring carotenoid rapidly in complex fluids. The approach proposed in this project has not been investigated previously and if successful, easy measurement will be possible benefiting society at large as increasing number of products enter the market place indicating high carotenoid levels in food and food supplements.The objective of the proposed work is to develop a selective and sensitive biosensor using elastin-like polymers (ELP) and electrochemical detection. We focus on the detection of astaxanthin, an important antioxidants that has been difficult to detect selectively in serum. The goal of this EAGER proposal is to obtain preliminary data towards the development of a carotenoid sensor that is capable of rapidly measuring in complex fluids. The focus will be towards improving the understanding of (1) ELP response under electric field; (2) the electrochemical activity of surface-immobilized ELPs, and (3) introduction of analyte specificity via a modular fusion protein component. The expected results will lead to using engineered electrochemically-tagged carotenoid-binding elastin-like polymer fusion proteins to monitor astaxanthin in complex matrix. While protein-protein interactions and synthetic stimuli-responsive polymers have growing niches in sensing technology, the merits of both are uniquely combined in the proposed research. By conjugating an electrochemical tag to elastin-like polymers, their stimuli-responsive behavior should be detectable using voltammetric sensors. Stimuli-responsive behavior will be monitored by comparing the collapsed against the extended state of electrochemically tagged elastin-like polymers located on the sensor surface. Using electrochemically-tagged crustacyanin-elastin-like polymer fusion proteins, binding of astaxanthin will be monitored. Finally, specificity will be tested by measuring astaxanthin in a solution with multiple carotenoids (i.e. lutein and beta-carotene). There is public benefit in quantitative understanding of the role of carotenoids in human health. Better sensing technology is needed to create accurate basic biochemical research to investigate carotenoid uptake and promote healthy carotenoid levels. In addition, techniques developed in this project for carotenoids can be extended to other protein-analytes. The research project will be integrating University of New England and University of New Hampshire researchers in cross-disciplinary research with opportunities for all students. Open seminar at University of New England for the general community will be offered with presentations by the undergraduate students.