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BIOSENSOR GLUCOMETRIC TRANSDUCER

IP.com Disclosure Number: IPCOM000234169D
Publication Date: 2014-Jan-15
Document File: 4 page(s) / 156K

Publishing Venue

The IP.com Prior Art Database

Abstract

Cell Based Biosensors are composed of two major components (1) live cells sensors that detect and respond to stimulant(s) of interest (e.g. toxic molecules, irradiation, motion, force, etc.) and (2) a transducer that relays the cell's response into a measurable/recordable signal. Cell based biosensors are distinct from biosensors in that live cells are used to generate an integrated and biologically relevant response as opposed to simple biochemical quantification.

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BIOSENSOR GLUCOMETRIC TRANSDUCER


Chong Wing Yung

Carsten Carstens

Bernd Buehler

Daniel E Ryan

May Tom Moy

Derek Lee Lindstrom

BACKGROUND

Cell Based Biosensors are composed of two major components (1) live cells sensors that detect and respond to stimulant(s) of interest (e.g. toxic molecules, irradiation, motion, force, etc.) and (2) a transducer that relays the cell’s response into a measurable/recordable signal.  Cell based biosensors are distinct from biosensors in that live cells are used to generate an integrated and biologically relevant response as opposed to simple biochemical quantification.

One of the major disadvantages of cell based biosensors is the speed of detection.  For example, biosensors based on measuring the cellular expression of fluorescent proteins are slow since proper protein expression and folding requires several hours (~ 4 hours at best).  While other cellular response mechanisms may be faster, such as chemilumnescence, they typically require highly sensitive and more costly transducers.  A biosensor that combines fast cellular response with a reliable and low-cost transducer technology may have certain advantages.

DESCRIPTION OF INNOVATION

Proposed herein is a combination of cellular engineering and novel application of existing transducer technology to produce a biosensor system that is both fast and inexpensive.  Starting with the transducer, the biosensor will integrate a blood glucose meter (glucometer), which is the most widely distributed and inexpensive biomedical devices to date.  Utilizing the preexisting biochemistry found in the test strips for glucometers (i.e. glucose dehydrogenase or glucose oxidase and ferric cyanide) one can quantitatively measure glucose concentration (0.1 to 6.0 g/L) in solution.  To interface with the glucometer, cells (e.g. yeast) will be engineered to secrete the α-fragment of the β-galactosidase enzyme, by fusing it to a secretion signal (e.g. pre-pro α factor for S. cerevisiae).  The β-fragment will be provided separately outside the cell, and through α complementation, recombine with the secreted fragment to reconstitute the active holoenzyme.  One advantage of this strategy is that the smaller α-fragment will be easier to secrete.  Additionally, the α-fragment will be inactive until it recombines with the β-fragment outside the cell, to avoid unintended background activity with potential substrates (e.g. lactose or β-galactosidase) that may be inside the cells.   The secretion of the enzymes also obviates the need to lyse the cell for intracellular reactions, which offers a more accelerated and streamlined detection process.  One complication with this strategy, however, is that glucose is a highly preferred carbon source by cells and may be inadvertently consumed before detection by the glucometer.  To circumvent this problem, cells can be conditioned to feed on other carbon sources, such as sucrose or glycerol, which are not reactive with glucos...