VOLUME 64 : 2016

VOLUME 64 : 2016

ACTA MANILANA publishes research and innovation in the different branches of the natural and applied sciences. It reports significant development in the discipline, and novel applications, unconfined by the traditional coverage of the disciplines.

Fluorophore-labeled bioengineered glucose binding protein for measurement of transdermal glucose

Page 25-32

Cristina Tiangco, Sheniqua Brown, Fortunato Sevilla III, Govind Rao, & Leah Tolosa

Graphical Abstract

Abstract

Measurement of glucose is an important component of care in the intensive care unit. Current methodologies for glucose monitoring include enzymatic-based laboratory analyzers, point of care testing and continuous glucose monitoring systems that all require breaking the skin. This study focuses on the development of a glucose biosensor to measure transdermal glucose (TG) by noninvasive passive diffusion. The H152C glucose binding protein (GBP) labeled with the polarity-sensitive dye BADAN (6-bromoacetyl-2-dimethylaminonaphthalene) was prepared and characterized with the aim of producing a biosensor with an operating range at micromolar levels compatible with TG concentrations. The BADAN-labeled GBP exhibited a large fluorescence intensity increase (228%) upon addition of a saturating concentration of glucose. The glucose binding constant was calculated to be (Kd) 1.124 ± 0.2361 mM. The biosensor has a linear operating range of 0.030–0.460 mM, making it suitable for monitoring transdermal glucose. The use of the biosensor was demonstrated by measuring TG collected by passive diffusion of glucose through the skin of healthy adult subjects. Results showed that the H152C GBP labeled with BADAN is capable of measuring TG and can be used for noninvasive glucose sensing.

Keywords: glucose binding protein, biosensor, BADAN, transdermal glucose

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