Figure 3: Redirecting cyclonals to new antigens with swapped variable regions.
From: Efficient expression of full-length antibodies in the cytoplasm of engineered bacteria
(a) Antigen-binding activity in SHuffle lysates for anti-MBP cyclonals with swapped VH and VL domains with specificity for antigens as indicated. ELISA signals (Abs492) for mFab/hFc cyclonals (red), parental anti-MBP cyclonal (orange) and empty plasmid control (green) in cell lysates were obtained with anti-human Fc antibodies. Data are expressed as the mean±s.e.m. of biological triplicates. (b) Representative non-reducing western blot of different cyclonals in the mouse Fab-human Fc format following protein-A affinity purification from cell lysates prepared from SHuffle cells. Arrows indicate fully assembled cyclonal and other HC intermediate species. The percentage of fully assembled product (% heterotetrameric) among all products was determined for each cyclonal using densitometry analysis. Percentages are expressed as the mean±s.e.m. of biological triplicates.
