Figure 3.

Catalytic activities of enzymes coupled to different immobilized adapters. (A) Different carrier scaffolds (adapters) immobilized on a solid support (in a high binding microtiter plate) were analyzed for the ABTS turnover rates achieved with equal input amounts of enzymes. To investigate their binding capacity, wells were functionalized with different quantities (between 0 and 9500 ng) TMV_Cys/Bio sticks, CP_Cys/Bio aggregates or the equivalent amounts of biotin linkers. 0.9 μg [SA]-enzymes per well were added and coupling to the biotin, or the non-treated plate support, allowed. Catalytic activities of immobilized [SA]-enzymes were determined spectroscopically using ABTS as substrate, and depicted in the histogram for the different assay layouts, as specified in the legend. The experiment indicated a maximum ABTS turnover rate upon application of 3.5 μg TMV_Cys/Bio adapter sticks per well. (B) Percental turnover rates of ABTS reached with equal amounts of [SA]-enzyme conjugates and different adapters, normalized to the turnover achieved on bare plate support. Left: application of 1500 ng adapters (or molar equivalent of linker) per well (non-saturated conditions), right: 3500 ng adapters (or molar linker equivalent) per well, respectively, as indicated. For details, refer to text.