ELISA Sisteminde Protein-İnorganik Hibrit Konjugatın Kullanılabilirliği

Abstract

In the Enzyme-linked immunosorbent assay method (ELISA), it is important to increase the immobilization efficiency of the functional molecules on the microplate and increase the signal-to-noise ratio in order to detect the target molecules with high sensitivity and selectivity. For this purpose, antibody functionalized materials can generate remarkable signal amplification with high enzyme capacity by using conventional immobilization methods. Recently, hybrid structures containing protein/enzyme and Cu3(PO4)2, which is a different enzyme immobilization method, have higher catalytic activity compared to the free form of protein-containing molecules as they have hierarchical microstructures and form large active surface areas. In this study; using protein-inorganic hybrid structure synthesis method, hybrid functionalized conjugate systems with enzyme, antibody and Cu3(PO4)2 were synthesized all together and characterization of the resulting structures was performed by SEM, EDX, XRD and FTIR analysis. When the findings obtained from the researches were evaluated, the hybrid nano structure showed 183.5 EU/mg peroxidase activity and free HRP enzyme showed 59.01 EU/mg activity. The performance of the ELISA system was measured using hybrid conjugate constructs prepared using various TNF-alpha specific antibodies at a -1 concentration of 5-1000 g mL . The performance of hybrid conjugate structure containing multiple organic molecules in ELISA system was higher than other structures. This method is a highly practical method that can replace enzyme-labeled antibody method in ELISA.