Papain酶切

Papain digestion of different mouse IgG subclasses as studied by electrospray mass spectrometry

Papain (10 ml, 10 mg / ml suspension) was mixed with 90 ml of freshly prepared activation buffer (1mM EDTA, 10 mM cysteine, 50 mM sodium phosphate buffer, pH 7.0) and was activated by incubation for 10 min at 37 ℃。The digestion was initiated by the addition of papain to the mAb solutions (1–2 mg / ml) to give papain / antibody ratios of 1%。The resulting solutions were incubated at 37 ℃ for 2 h.

The presence of cysteine (thiol) in digestion buffer significantly promoted further digestion of F(ab) to form Fab fragment.