FIGURE 3.

Release of PDBA-modified protein immobilized on SHA-affinity membranes. Versalinx Protein Releasing Reagent (100 μL) was added to spin columns containing immobilized PDBA-AP, and AP and the columns were incubated at 37°C for 30 min. The releasing reagent was passed through the membranes by slow centrifugation. AP activity in the flow-through was analyzed using pNPP and monitoring absorbance at l = 405 nm. Protein concentrations were determined by preparing standard curves of AP at 0 μg/mL, 10 μg/mL, 20 μg/mL, 30 μg/mL and 40 μg/mL.