We use pepsin for F(ab')2 fragment production and papain for Fab fragment production. Before digesting large amount of antibodies, we perform an analytical digestion to optimise conditions. Undigested antibodies will be removed by protein A columns.
Customers should provide minimum of 10 mg antibodies. Normally 3 mg of Fab or F(ab')2 fragments can be obtained from 10 mg antibodies after absorption with Protein A. The resulted fragments should remain their binding abilities to antigens and show one band (about 100 kDa for F(ab')2 fragments and 50 kDa for Fab fragments) under non-reducing conditions and two bands (both are around 25 kDa) on SDS-PAGE under reducing conditions.
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