Fig. 5

The effects of signal peptide hydrophobicity on full-length (a) mAb1 and (b) mAb2 production. For all the constructs, light chain was fused to ssSTII0.3 and heavy chain was fused to either ssSTII1, ssSTII1 S-13L, ssDsbA1, or ssDsbA1 L-9S. 64B4 harboring the empty vector (ev) pBR322 was loaded as the negative control. Top panel the non-reduced soluble protein samples from the cell lysates were separated by electrophoresis followed by Western blot probed with HRP-conjugated anti-Fc antibody. Heavy chain-containing species are marked based on the apparent molecular weight. Middle panel Western blot of soluble heavy chain in the periplasm. The periplasmic protein samples were reduced by 0.2 M DTT. Bottom panel Western blot of reduced total heavy chain in the whole cell lysate