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. 2011 Jul;22(2):50–52.

FIGURE 1.

FIGURE 1

Fab purification after papain digest of antibodies using Protein-A or MabSelect SuRe resins. SDS-PAGE, under reducing conditions, was performed on fractions collected during digestion and Fab purification of (A) IH3, (B,i) Herceptin and (B,ii) MabThera, and (C,i) Avastin and (C,ii) Campath. Each gel photo contains SeeBlue Plus2 protein ladder (Invitrogen), and the lanes contain whole antibody (1), papain-digested antibody (2), Protein-A flowthrough (3), Protein-A wash (4), Protein-A eluate (5), MabSelect SuRe flowthrough (6), MabSelect SuRe wash (7), and MabSelect SuRe eluate (8). In each gel, 12 μL of each fraction was loaded, except in gels B and C, where only 2 μL of the whole antibody and digested antibody was loaded. Undigested antibody under reducing conditions produces bands of ∼50 kDa (heavy chain) and 24 kDa (light chain). Papain digest generates bands at ∼30 kDa (Fc fragment) and 24 kDa (reduced Fab fragment and light chain originating from undigested antibody origin). MWs on the heavy chain are estimates only as a result of glycosylation on the Fc region.