A real-time biosensor with 1,1’-oxalyldiimidazole chemiluminescence (ODI-CL) detection was developed to consecutively or simultaneously quantify triple biomarkers (e.g., glucose, hemoglobin A1c (HbA1c), glycated blood proteins) for the diagnosis and management of diabetes. Using the conventional enzyme assay for the analysis of glucose, we found for the first time that the glucose of HbA1c and the glucose of glycated blood proteins have the same chemical properties as pure glucose. This is because glucose bound at the N-terminal of hemoglobin (or blood proteins), formed by the non-enzymatic interaction, reacts with glucose oxidase (GOx) to produce H2O2. The reaction between HbA1c (or glycated blood protein) and GOx is as rapid as that between our glucose and GOx. Based on the preliminary research result, first, H2O2 was formed from the enzyme reaction occurring in each well containing GOx and one of the triple biomarkers. H2O2 formed from the enzyme reaction also consecutively reacts with Amplex Red, non-fluorescent dye, to produce resorufin, fluorescent dye, in the well during the 3-min incubation at room temperature. After the reaction, resorufin formed in the each well emits light when H2O2 and ODI were added in the well. The relative CL intensity of light emitted in the well was proportionally enhanced with the increase of biomarker concentration. Thus, the real-time biosensor can accurately diagnose and precisely monitor diabetes at any time of day, regardless of before or after food intake. In conclusion, we expect that the real-time biosensor can be used as a new diagnostic device for diabetes instead of the conventional tools such as glucose meters and HPLC for determining glycated blood proteins and HbA1c. Also, the real-time biosensor will be applied to quantify trace levels of various biomarkers existing in whole blood for diagnosing human diseases.

Real-time biosensor capable of quantifying triple biomarkers for diagnosing diabetes

256th National Meeting (2018)

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