Proteolytic digestion of mouse IgE.
Conditions are described for the preparation of F(ab')2 and Fab fragments of mouse IgE. Papain, pepsin or trypsin each produced F(ab')2 fragments with Mr approximately equal to 130,000 which yielded Fab fragments on further digestion. The release of Fab fragments from F(ab')2 resulted from further cleavage of the H chain. Pepsin, and especially trypsin appear more suitable for the preparation of F(ab')2 because of the difficulty of separating a 93 kDa by-product from the F(ab')2 produced by papain. The best yields of purified Fab were obtained with papain. Rates of digestion were in the order, pepsin approximately equal to trypsin much greater than papain.
Haba
S
S
Department of Biology, Brandeis University, Waltham, MA 02254.
Nisonoff
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D007073:Immunoglobulin E / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010206:Papain / Q000494:pharmacology; D010434:Pepsin A / Q000494:pharmacology; D010446:Peptide Fragments / Q000032:analysis; D014357:Trypsin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D007073:Immunoglobulin E; D014357:Trypsin; D010206:Papain; D010434:Pepsin A