Development of a novel immunoassay method determination of hemoglobin A1c

In this thesis, a new assay method based on Time Resolved Fluoroimmunoassay (TR-FIA) for hemoglobin A1c (HbA1c), known as a gold parameter in the diagnosis and treatment of diabetes mellitus (DM) was developed. For this purpose, firstly, the Eu(III) complex of 4,4'-Bis(4,4,5,5,6,6,6-heptafluoro-1,3-dioxyhexyl)-o-terphenyl-4'-sulfonyl chloride (BHHCT) ligand was formed and the complex was tested for use as a label in studies. In the second part of the study, working conditions such as antibody amounts, antibody-antigen interaction times, and dilution ratio of Eu (III) label were optimized and appropriate immunoassay procedure was established. The lineer range was from 0.05 to 9200 ng/mL and 0 to 25% for the HbA1c via the immunoassay procedure prepared in the optimized conditions. In addition, inter assay and intra assay reproducibility were studied and the method was highly reproducible. At the end of the study, the analytical application of the prepared immunoassay procedure was investigated by performing % HbA1c assay on blood samples. The obtained results were compared with those obtained by ELISA, a reference method, and the % RSD and % recovery values were calculated.