The sensitive immunoassay methods development for detection of amyloid protein

The aim of this thesis is to develop a new time delay immunoassay (TR-FIA) method for amyloid β (1-42) protein used as biomarker in the determination of Alzheimer's disease which is a neurodegenerative disease. For this, firstly, commercially available 3,4bis [4 (4,4,5,5,6,6,6heptafluoro, 3dioxohexyl) phenyl] benzenesulfonylchloride (BHHCT) and synthesized 10,10 '- (4,4 Fluorescence labels were generated using 5,56,6,7,7-octafluoro-3,8-dioxodecanediol) bis (anthracine-9- sulfonyl chloride) (SD1) ligands. The appropriate immunoassay procedure was established and various experimental parameters were optimized. In the second part of the study, in the light of optimum data, calibration studies of amyloid β (1-42) protein were prepared by using labels prepared TR-FIA technique in Micro Plate Fluorescence Reader. The linear working range for the amyloid β (1-42) biomolecule was 0.5-10000 pg / mL for BHHCT and 0.5-4000 pg / mL for SD1 labels. Finally, when using assay analysis, synthetic serum samples were calculated for examining amyloid β (1-42) protein and % recovery. Keywords: Amyloid β (1-42), Eu (III) Complex, Fluorescence labels, Immunoassay, Time Resolved Fluoro Immunoassay (TR-FIA)