Explore how immobilization strategies affected immunosensor performance by comparing four methods for antibody immobilization on electrode surfaces

Abstract Among the common methods used for antibody immobilization on electrode surfaces, which is the best available option for immunosensor fabrication? To answer this question, we first used graphene-chitosan-Au/Pt nanoparticle (G-Chi-Au/PtNP) nanocomposites to modify a gold electrode (GE). Second, avian reovirus monoclonal antibody (ARV/MAb) was immobilized on the GE surface by using four common methods, which included glutaraldehyde (Glu), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide (EDC/NHS), direct incubation or cysteamine hydrochloride (CH). Third, the electrodes were incubated with bovine serum albumin, four different avian reovirus (ARV) immunosensors were obtained. Last, the four ARV immunosensors were used to detect ARV. The results showed that the ARV immunosensors immobilized via Glu, EDC/NHS, direct incubation or CH showed detection limits of 100.63 EID50 mL−1, 100.48 EID50 mL−1, 100.37 EID50 mL−1 and 100.46 EID50 mL−1 ARV (S/N = 3) and quantification limits of 101.15 EID50 mL−1, and 101.00 EID50 mL−1, 100.89 EID50 mL−1 and 100.98 EID50 mL−1 ARV (S/N = 10), respectively, while the linear range of the immunosensor immobilized via CH (0–105.82 EID50 mL−1 ARV) was 10 times broader than that of the immunosensor immobilized via direct incubation (0–104.82 EID50 mL−1 ARV) and 100 times broader than those of the immunosensors immobilized via Glu (0–103.82 EID50 mL−1 ARV) or EDC/NHS (0–103.82 EID50 mL−1 ARV). And the four immunosensors showed excellent selectivity, reproducibility and stability.