Fig. 2
From: Systematic screening of soluble expression of antibody fragments in the cytoplasm of E. coli
Purification of antibodies and antibody fragments expressed in DWP in the cytoplasm of E. coli. Representative Coomasie stained non-reducing SDS-PAGE analysis of NEM treated IMAC purified antibody fragments in the cytoplasm of a K12 E. coli strain with (+) and without (−) expression of CyDisCo components Erv1p and PDI. Expression in 24 deep well plates, EnPressoB media at 30 °C. a scFv. The position of a Herceptin scFv disulfide linked dimer is marked with an arrow; b Fab. The position of the Fab dimer and light chain and heavy chain monomers are marked. In both panels the order is molecular weight markers (1) Humira (IgG1), (2) Maa48 (IgG1), (3) K2 (IgG1), (4) Avastin (IgG1 humanized), (5) Herceptin (IgG1 humanized), (6) 3211 (IgG1 mouse), (7) 1IGT (IgG2 mouse), (8) Tysabri (IgG4 humanized), (9) 3M8O (IgA1), (10) 2R56 (IgE) and (11) 1QLR (IgM). All antibodies are human unless otherwise indicated. In both panels an E. coli protein which is occasionally seen co-eluting is marked with *. Treatment with NEM results in a slight smearing and laddering of the protein band. This is not seen in the absence of NEM (see Fig. 3a as an example)