Ex12 helper phage improves the quality of a phage-displayed antibody library by ameliorating the adverse effect of clonal variations
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The quality of a phage-displayed antibody library deteriorates with clonal variations, which are caused by differentially expressed Escherichia coli antibody genes. Using the human Fab SP114 against the pyruvate dehydrogenase complex-E2 (PDCE2),we created four E. coli TOP10F' clones with a pCMTG phagemid encoding Fab-pIII (pCMTG-Fab), Fd (VH+CH1)-pIII (pCMTG-Fd), or light chain (L) (pCMTG-L), or the vector only (pCMTG-ΔFab) to investigate the effect of clonal variations in a defined manner. Compared to the others, the E. coli clone with pCMTG-Fab was growth retarded in liquid culture, but efficiently produced phage progenies by Ex12 helper phage superinfection. Our results suggest that an antibody library must be cultured for a short duration before helper phage superinfection,and that the Ex12 helper phage helped to alleviate the detrimental effect of clonal variation, at least in part,by preferentially increasing functional phage antibodies during phage amplification. [BMB reports 2011; 44(4): 244-249]
Abstract
The quality of a phage-displayed antibody library deteriorates with clonal variations, which are caused by differentially expressed Escherichia coli antibody genes. Using the human Fab SP114 against the pyruvate dehydrogenase complex-E2 (PDCE2),we created four E. coli TOP10F' clones with a pCMTG phagemid encoding Fab-pIII (pCMTG-Fab), Fd (VH+CH1)-pIII (pCMTG-Fd), or light chain (L) (pCMTG-L), or the vector only (pCMTG-ΔFab) to investigate the effect of clonal variations in a defined manner. Compared to the others, the E. coli clone with pCMTG-Fab was growth retarded in liquid culture, but efficiently produced phage progenies by Ex12 helper phage superinfection. Our results suggest that an antibody library must be cultured for a short duration before helper phage superinfection,and that the Ex12 helper phage helped to alleviate the detrimental effect of clonal variation, at least in part,by preferentially increasing functional phage antibodies during phage amplification. [BMB reports 2011; 44(4): 244-249]
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