Electrochemical Immunoassay for Determination of Glycated Albumin using Nanozymes

We developed a new nanozyme-based electrochemical immunoassay method for the monitoring of glycated albumin (GA) known to reflect short-term glycaemic levels. For this study, we synthesized urchin-like Pt nanozymes (uPtNZs) and applied them to colorimetric and electrochemical assays for sensitive determination of GA in total human serum albumin (tHSA) using 3,3 ',5,5 '-tetramethylbenzidine (TMB) and thionine as substrates, respectively. The uPtNZs showed peroxidase-mimic activity in the presence of hydrogen peroxide. Boronic acid (BA)-agarose bead was used to capture GA through specific cis-diol interactions. uPtNZs were modified with GA antibody (GA-Ab) to form sandwich complexes with GA/BA-agarose bead. The amount of Ab-uPtNZ/GA/BA-agarose bead complex increased with increasing percentage of GA in 50mg/mL tHSA. The colorimetric assay exhibited linearity from 0.02 to 10% (10 mu g/mL - 5mg/mL) GA with an LOD of 0.02% (9.2 mu g/mL). For electrochemical assay, GA was detected from 0.01 to 20% (5 mu g/mL - 10mg/mL) with an LOD of 0.008% (3.8 mu g/mL). The recovery values of measured GA in human plasma samples were from 106 to 107%. These results indicate that electrochemical assay using uPtNZs is a promising method for determining GA.