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Covalent dimerization of camelidae anti‐human TNF‐alpha single domain antibodies by the constant kappa light chain domain improves neutralizing activity
Author(s) -
Giersberg Martin,
Floss Doreen M.,
Kipriyanov Sergey,
Conrad Udo,
Scheller Jürgen
Publication year - 2010
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22653
Subject(s) - immunoglobulin light chain , tumor necrosis factor alpha , avidity , fusion protein , monoclonal antibody , antibody , recombinant dna , microbiology and biotechnology , chemistry , stereochemistry , nicotiana benthamiana , monomer , biology , biochemistry , gene , immunology , organic chemistry , polymer
The tumor necrosis factor‐alpha (TNFα) plays an important role in a number of chronic inflammatory disorders. Monoclonal camelidae variable heavy chain domain‐only antibodies (V H H) have been developed to antagonize the action of human TNFα (anti‐TNF‐V H H). Here we describe a strategy to obtain functional dimeric anti‐TNF‐V H H molecules, based on the C‐terminal fusion of a κ light chain domain to the anti‐TNF‐V H H. The resulting fusion protein was transiently expressed by use of viral vectors in Nicotiana benthamiana (Nb) leaves and purified. Competitive ELISA and cell cytotoxicity assays revealed that the dimerized anti‐ Nb TNF‐V H H Cκ proteins blocked TNFα‐activity more effectively than either the monomeric Escherichia coli (Ec) produced anti‐ Ec TNF‐V H H or the monomeric anti‐ Nb TNF‐V H H Cκ . We suggest that enhanced inhibition shown by dimeric anti‐ Nb TNF‐V H H Cκ proteins is achieved by an increase in avidity. Biotechnol. Bioeng. 2010; 106: 161–166. © 2009 Wiley Periodicals, Inc.