Master of Engineering Science
Chemical and Biochemical Engineering
Dr. Jin Zhang
Bioluminescence Resonance Energy Transfer (BRET), a sensitive, non-destructive and self-illuminated method, has been now commonly used to test protein interactions. Here, we describe a BRET sensor for non-invasively detecting glucose molecules. The sensor is made by the bioconjugation of quantum dots and recombinant protein. The recombinant protein contains the bacterial glucose binding protein (GBP) and a bioluminescent protein, Renilla luciferase (Rluc), used as the donor with the emission peak at 470 nm, which is able to excite the acceptor of BRET sensor made of cadmium tellurium quantum dots ( CdTe QDs) with the emission peak at 570 nm. The distance between the BRET pair depends on the shape of GBP. In the presence of glucose, the conformational change of GBP leads to the enhanced BRET phenomenon due to the short distance between the BRET pair. To optimal the BRET sensor, the ratios of the acceptor (QDs) to the donor (GBP-Rluc), the bioconjugation process, and the performance of recombinant protein have been thoroughly investigated. The results indicate a linear relationship between luminescence intensity ratio and the concentration of glucose in the two ranges, from 0 mM to 0.1 mM and from 0.2mM to 0.8mM, respectively. The limit of detection (LOD) is calculated as15 nM. Meanwhile, tear glucose of animal model were measured by this designed sensor. This study demonstrates a proof-of-concept of a non-invasive measurement system for monitoring glucose.
Zhan, Denghuang, "Nanostructured biosensor for tear glucose detection based on bioluminescence resonance energy transfer (BRET) mechanism." (2017). Electronic Thesis and Dissertation Repository. 4786.