Fig. 3.

Generation of scFv-Fc library by CPEC. Amplified VH and VL fragments are cloned in frame into a vector between secretion leader and Fc region cDNAs using overlapping regions added at the ends of the PCR products. The overlapping sequences added to variable domains formes a flexible linker (GGGGS)₃ between VH and VL. In each of 25 CPEC cycle, the denaturated inserts and vector are hybridized and extended until completing full circles. The plasmid library is propagated and recovered by transformation into competent E. coli cells