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

EAGER：合作提案：使用类弹性蛋白聚合物作为电化学生物传感器

• 批准号: 1638896
• 负责人: Jeffrey Halpern
• 金额: $ 11.17万
• 依托单位: University of New Hampshire
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1638896&HistoricalAwards=false
• 关键词: EAGER; Collaborative; Proposal; Use; Elastin

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.

合作项目PI Halpern，Jeffrey M.伊娃·罗斯·贝奇本课题旨在建立一种快速测定复杂流体中类胡萝卜素的新方法。在这个项目中提出的方法还没有被调查以前，如果成功的话，容易测量将是可能的造福社会，越来越多的产品进入市场，表明高类胡萝卜素水平的食品和食品supplements. Objective的建议的工作是开发一个选择性和敏感的生物传感器使用弹性蛋白样聚合物（ELP）和电化学检测。我们专注于检测虾青素，一种重要的抗氧化剂，一直难以在血清中选择性检测。这个EAGER建议的目标是获得初步的数据，发展的类胡萝卜素传感器，能够在复杂的流体中快速测量。重点将是提高对以下方面的理解：（1）电场下ELP响应;（2）表面固定ELP的电化学活性;（3）通过模块化融合蛋白组分引入分析物特异性。预期的结果将导致使用工程电化学标记的类胡萝卜素结合弹性蛋白样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的刺激响应聚合物在传感技术中具有越来越大的优势，但两者的优点在拟议的研究中独特地结合在一起。通过将电化学标记物与弹性蛋白样聚合物缀合，它们的刺激响应行为应该可以使用伏安传感器检测到。通过比较位于传感器表面上的电化学标记的弹性蛋白样聚合物的塌陷与伸展状态，监测刺激响应行为。使用电化学标记的甲壳青素-弹性蛋白样聚合物融合蛋白，将监测虾青素的结合。最后，将通过测量具有多种类胡萝卜素（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，本项目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罕布什尔州大学的研究人员在跨学科的研究与机会，为所有学生。在新英格兰大学为一般社区举办的公开研讨会将由本科生进行演讲。

项目成果

Electrochemical characterization of the stimuli-response of surface-immobilized elastin-like polymers

表面固定弹性蛋白类聚合物刺激响应的电化​​学表征

• DOI: 10.1039/c9sm01681c
• 发表时间: 2019
• 期刊: Soft Matter
• 影响因子: 3.4
• 作者: Morales, Marissa A.;Paiva, Wynter A.;Marvin, Laura;Balog, Eva Rose;Halpern, Jeffrey Mark
• 通讯作者: Halpern, Jeffrey Mark

Flow imaging microscopy as a novel tool for high-throughput evaluation of elastin-like polymer coacervates

流式成像显微镜作为类弹性蛋白聚合物凝聚层高通量评估的新工具

• DOI: 10.1371/journal.pone.0216406
• 发表时间: 2019
• 期刊: PLOS ONE
• 影响因子: 3.7
• 作者: Marvin, Laura;Paiva, Wynter;Gill, Nicole;Morales, Marissa A.;Halpern, Jeffrey Mark;Vesenka, James;Balog, Eva Rose;Zelikin, Alexander N.
• 通讯作者: Zelikin, Alexander N.