Anthracene boronic acid methacrylate-based fluorescence activation assay for determination of glycated proteins in biological samples

In our manuscript the main highlights are as follows: • Firstly, we present a refined synthesis of the compound anthracene boronic acid methacrylate (ABAM) which is then evaluated as a fluorescence detector of diabetes-specific biomarker proteins in biological samples of human origin • After verifying optimal fluorescence assay conditions, ABAM is shown to be able to detect not only basal concentrations of the diabetes-specific biomarker glycated human serum albumin (gHSA) in human derived plasma samples but is then able to detect reliably the full range of gHSA concentration rises seen in prediabetic and then in diabetic patients. • ABAM is also shown able to detect basal levels of the well known the diabetes-specific biomarker glycated hemoglobin (HbA1c) but is unable to reliably detect the full range of HbA1c concentration rises seen in prediabetic and then in diabetic patients. • Using CE-LIF we then confirm using human derived blood and plasma samples that ABAM can detect at least gHSA, HbA1c and likely other glycated proteins in blood samples but appears selective for gHSA detection only in plasma samples, indicating that ABAM is capable of specific and selective detection of gHSA in human derived plasma samples at concentration levels in line with and appropriate for the monitoring of individuals who might be normal, prediabetic or diabetic. • Finally, we demonstrate that our ABAM-based assay is analytically equivalent to the market leader Fructosamine assay (ab228558, Abcam, Cambridge, UK) but is much easier to use, more specific, more selective and more cost effective. Moreover, our ABAM-based assay is not subject to interference in plasma samples, unlike the Fructosamine assay. . There is a current unmet medical need for the creation and development of point-of-care diagnostic tests/devices that can determine if a given individual is diabetic, pre-diabetic or normal. In pursuit of this objective, we report here on the successful evaluation of a bespoke anthracene boronic acid methacrylate (ABAM) in a rapid and simple fluorescence assay for the detection of glycated proteins known as disease biomarkers in human plasma samples. In so doing, we determine that increases in ABAM fluorescence intensity, as a function of glycated human serum albumin (gHSA) levels, in 50-fold diluted plasma samples, are sufficiently well resolved to suggest that ABAM could form the basis of a potential point-of-care diagnostic device to discriminate between blood plasma samples taken from individuals who are diabetic, pre-diabetic or normal. Indeed, our ABAM-based assay is shown to discriminate clearly in 50-fold diluted plasma samples between increases in gHSA plasma concentrations (from 5 mg L −1 to 65 mg L −1 ) above the normal base line, reaching final concentrations indicative of diabetes. In comparison to the nearest competitor, the fructosamine detection assay, our ABAM-based assay is twelve times faster, more than six times less expensive, requires ten times lower sample volumes, is both gHSA selective and specific, and is much less prone to interference from molecular interferents. Therefore, we would suggest that our ABAM-based assay has real future potential utility in diabetes disease management. .