3/21/2023 0 Comments Igg starmancerThus, there is no need to develop an ion exchange method for separating the fragments from the enzyme. Immobilized papain (i.e., Papain Agarose Resin) is the preferred reagent because it allows for easy control of the digestion reaction and quick removal of enzyme from the digestion products following incubation. Mercuripapain, which is less prone to autodigestion than crystalline papain, can be used however, both of these non-immobilized enzymes require an oxidant to terminate digestion. If the cysteine is not completely removed, over digestion can be a problem (2).Ĭrystalline papain is often used for the digestion of IgG however, it is prone to autodigestion. These fragments are often inconsistent, and reproducibility can be a problem. If no cysteine is present during papain digestion, F(ab')2 fragments can be generated. The excess cysteine is then removed by gel filtration. To prepare F(ab')2 fragments, the papain is first activated with 10mM cysteine. Papain is primarily used to generate Fab fragments, but it also can be used to generate F(ab')2 fragments (2). Learn more: Tech Tip #59: Secondary antibodies: A guide to fragment specificity
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