PEGylation of Concanavalin A to Decrease Non-Specific Interactions in a Fluorescent Glucose Sensor
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The ability of people with diabetes to both monitor and regulate blood sugar levels is limited by the conventional "finger-pricktest" that provides intermittent, single point measurements. Toward the development of a continuous glucose monitoring (CGM) system, the lectin, Concanavalin A (ConA), has been utilized as a component in a Frster resonance energy transfer (FRET), competitive glucose binding assay. Recently, to avoid reversibility problems associated with ConA aggregation, a suitable competing ligand labeled with 8-aminopyrene-1,3,6-trisulfonic acid trisodium salt (APTS) has been engineered. However, its ability to function as part of a glucose sensing assay is compromised due to the negative charge (at physiological pH) of native ConA that gives rise to non-specific binding with other ConA groups as well as with electrostatically charged assay-delivery carriers. To minimize these undesirable interactions, we have conjugated ConA with monomethoxy-poly(ethylene glycol) (mPEG) (i.e. "PEGylation"). In this preliminary research, fluorescently-labeled ConA was successfully PEGylated with mPEG- Nhydroxylsuccinimide(succinimidyl carbonate) (mPEG-NHS(SC)). The FRET response of APTS-labeled competing ligand (donor) conveyed an increase in the fluorescence intensity with increasing glucose concentrations. 2014 SPIE.
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Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics