Publication details

Biointerfaces based on immobilized boronic acid with specifity to glycated proteins

Authors

PŘIBYL Jan SKLÁDAL Petr

Year of publication 2008
Type Article in Proceedings
Conference BIODEVICES 2008: PROCEEDINGS OF THE FIRST INTERNATIONAL CONFERENCE ON BIOMEDICAL ELECTRONICS AND DEVICES, VOL 2
MU Faculty or unit

Faculty of Science

Citation
Keywords glycated hemoglobin; aminophenylboronic acid; biosensors; quartz crystal microbalance; heterogeneous affinity assay; microtitre plate
Description Development of bioanalytical assays for determination of glycated hemoglobin content in blood samples is reported. First, a combined biosensor setup for determination of total and glycated hemoglobin content was successfully developed and tested. The effect of various operating parameters, such as ionic strength, flow rate and instrumental set-up was optimized. The total hemoglobin content was analyzed by measuring of absorbance of the hemoglobin-cyanide derivative at 540 nm. Only one standard (calibrator), diluted in various proportions, was necessary for the method calibration. The full range of HbA(1c) content (4 to 15 %) presented in blood can be analyzed. Only 1 mu l of blood was required for analysis. The developed method was successfully evaluated for analysis of blood samples collected from diabetic patients. Next, the heterogeneous affinity assay performed in a microtitre plate with an immobilized boronic acid is described. This assay is based on ELISA (Enzyme-Linked Immunosorbent Assay) principle; however stable chemiselective ligand is used in this case. The content of glycated hemoglobin is determined according to its peroxidase activity after attachment to immobilized boronic acid derivative; the total hemoglobin concentration is measured as an absorbance at 405 nm.

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