Pepsin is a proteolytic enzyme that is routinely used for the generation of F(ab)2 fragments from immunoglobulin G (IgG). The pepsin has the ability to cleave the heavy chains near the hinge region. One or more of the disulfide bonds that join the heavy chains in the hinge region are preserved, so the two Fab regions of the antibody remain joined together, yielding a divalent molecule (containing two antibody binding sites), hence the designation F(ab)2. The light chains remain intact and attached to the heavy chain, whereas the Fc fragment is digested into small peptides.
The Immobilized Pepsin offers the distinct advantage of eliminating enzyme contamination of the F(ab)2 fragments.
The F(ab)2 fragments can be purified from undigested IgG with Immobilized Protein A and can be further purified from the small Fc fragments by dialyze with a 50kDa MWCO membrane (Tube-O-DIALYZER™) or by gel filtration.
Supplied as a 50% slurry in 50% glycerol, 0.1M sodium acetate, pH4.4 with sodium azide as a preservative.
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Features
- Generate F(ab)2 fragments
- Eliminate contaminating pepsin enzyme
- Can be used in virtually all scenarios using free pepsin
