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

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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.