Immobilization of NTPDase-1 from Trypanosoma cruzi and Development of an Online Label-Free Assay

Figure 2

Schematic diagram of the two-dimensional system and the switch valve positions. Position B: column conditioning; Position A: coupled columns. In the chromatographic conditions optimized for the two-dimensional system (Figure 1), Phenomenex Luna C18 (100 Å 250 × 4.6 mm, 5 μm) analytical column conditioned with Buffer C : MeOH (84 : 16 v/v) at a flow rate of 1.0 mL⋅min⁻¹ and UV detection at  nm led to the best analytical separation of ATP, ADP, and AMP (Figure 2). Table 1 summarizes all the optimized conditions.