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Engineering recombinant antibodies for polymer biofunctionalization
Author(s) -
Hortigüela María J.,
Aumailley Lucie,
Srivastava Akshay,
Cunningham Claire,
Anandakumar Soshee,
Robin Sylvain,
Pandit Abhay,
Hu Xuejun,
Wall J. Gerard
Publication year - 2015
Publication title -
polymers for advanced technologies
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.61
H-Index - 90
eISSN - 1099-1581
pISSN - 1042-7147
DOI - 10.1002/pat.3619
Subject(s) - glycan , linker , combinatorial chemistry , protein engineering , surface modification , recombinant dna , escherichia coli , chemistry , antibody , cysteine , bioconjugation , amine gas treating , covalent bond , polymer , maleimide , biochemistry , biophysics , polymer chemistry , biology , enzyme , glycoprotein , organic chemistry , gene , computer science , immunology , operating system
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.