A functional antibody mutant with an insertion in the framework region 3 loop of the VH domain: implications for antibody engineering

We have studied the effects of a four residue insertion intothe FR3 loop of the heavy chain variable region from the anti-NPantibody Bl-8. The insertion mutant is obtained as secretedantibody without major defects in biosynthesis, indicating thatantibody variable domains can accommodate length variation notonly in complementarity determining regions (CDRs), but alsoin framework region (FR) loops. The Bl-8 antigen binding siteis not affected by the change in a neighbouring loop. FR3 insertionsrepresent a new method of antibody engineering with a potentialto obtain strong antigen binding by designing additional antigencontacting residues.