Engineering recombinant antibodies for polymer biofunctionalization

Abstract

The attachment of recognition elements such as antibody fragments to polymeric substrates can be used to mediate cell‐ or protein‐specific interactions. In this work, single‐chain Fv (scFv) antibody fragments were isolated against two cell types of interest and expressed in an Escherichia coli expression platform. The scFvs were engineered at their C‐terminus to incorporate a cysteine‐containing linker, for reaction with maleimide‐linked polymers, or a heptasaccharide glycan for complexation with surface amine moieties. Antigen binding of the modified scFvs was unchanged, and expression yields of the glyco‐engineered scFvs were similar to the unmodified molecules, while cys‐tagged scFv yields varied between scFv variants. Targeted immobilization of the scFvs via either modification resulted in three‐ to five‐fold higher binding of ligands over adsorbed molecules. The study demonstrates a simple and efficient antibody engineering and modification approach for effective targeted immobilization on polymeric substrates. Copyright © 2015 John Wiley & Sons, Ltd.