Passive elution

The passive elution was performed at 4°C in different times of incubation, i.e., 24, 48, and 72 h, to optimize the protein release into the elution buffer solution. For passive elution, the excised gels (6 bands for each condition of time incubation) were crushed into small pieces using a sterile razor blade and incubated in 100 μL sterile PBS. The gel debris was centrifugated (3.000 g, 5 min) to isolate the supernatant.