| Abstract [eng] |
This doctoral dissertation investigates the application of surface plasmon resonance (SPR) spectroscopy for bioconjugate analysis and direct antibody detection. The aim was to apply SPR to evaluate the antibody functionalization efficiency of quantum dots and to detect antibodies against the SARS-CoV-2 nucleocapsid protein. The study comprises two parts. In the first, amino-functionalized quantum dots (QD-NH2) were conjugated with anti-CD44 using BS3 crosslinker. SPR was employed to evaluate the interaction of bioconjugates with immobilized CD44, and the optimal QD-NH2 to anti-CD44 molar ratio (1:10) was established. The conjugates demonstrated specific binding to CD44. In the second part, a direct-format SPR immunosensor for anti-SCoV2-rN detection was developed. The SCoV2-rN protein was immobilized on a gold surface modified with 11-mercaptoundecanoic acid via carbodiimide coupling. Optimal immobilization conditions were 500 nmol L-1 in acetate buffer (pH 5.3), and regeneration was achieved using 10 mmol L-1 NaOH with 0.5% SDS. The sensor enables detection within 0.5-50 nmol L-1, shows good repeatability (RSD 4.5%), high selectivity, and low detection (0.057 nmol L-1) and quantification (0.19 nmol L-1) limits. The results confirm the suitability of SPR for bioanalytical and diagnostic applications. |
