An experimental study was carried out on the immobilization of antibody via two methods, the covalent binding and the avidin-biotin binding, and on the effect of two blocking agents, bovine serum albumin and ethanolamine. In the covalent binding method, a slide glass was treated with 3-mercaptopropyl-trimethoxysilane (MTS) and γ-maleimidobutyric acid N-hydroxysuccinimide ester (GMBS), then the antibody, Anti-Mouse IgG was covalently attached to GMBS. In the avidin biotin binding method, avidin was attached to GMBS then biotinylated Anti-Mouse IgG was attached to avidin. In both methods, the amount of immobilized antibody increased with increasing antibody concentration in the solution added. However, it was not exactly proportional to the antibody concentration showing a saturated form. When the avidin-biotin binding method was used, about 3 times larger amount of the antibody was immobilized than the covalent binding method. Bovine serum albumin(BSA) was more effective as blocking agent than ethanolamine(EA) in the covalent binding method, while EA was more effective than BSA in the avidin-biotin method.
