scFv by Two-Step Cloning

The scFv-chain Fv (scFv) corresponds to the smallest functional binding unit of a classical antibody and constitutes a favourite format for the comparative first evaluation of the recombinant potential of an antibody. In this protocol, a two-step cloning strategy is described following the modular exchange principle as a general guideline for converting antibodies into an scFv format suitable for periplasmatic expression in E. coli .