Noncovalent scFv multimers of tumor-targeting anti-Lewisy hu3S193 humanized antibody

Abstract

Single-chain variable fragments (scFvs) of anti-Lewisy hu3S193 humanized antibody were constructed by joining the VH and VL domains with either +2 residues, +1 residue, or by directly linking the domains. In addition two constructs were synthesized in which one or two C-terminal residues of the VH domain were removed (-1 residue, -2 residue) and then joined directly to the VL domain. An scFv construct in the reverse orientation with the VL joined directly to the VH domain was also synthesized. Upon transformation into Escherichia coli all scFv constructs expressed active protein. Binding activity, multimeric status, and multivalent properties were assessed by flow cytometry, size exclusion c..