Single-Chain Fv of Anti-idiotype 11-1G10 Antibody Interacts with Antibody NC41 Single-Chain Fv with a Higher Affinity than the Affinity for ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1998-04

AUTHORS

Peter Iliades, David A. Dougan, Geoffrey W. Oddie, Dennis W. Metzger, Peter J. Hudson, Alexander A. Kortt

ABSTRACT

A single-chain Fv (scFv) fragment of anti-idiotype antibody 11-1G10, which recognizes an idiotope of anti-neuraminidase antibody NC41, was constructed by joining VH and VL domains with a (Gly4Ser)3 linker, with a pelB leader sequence, and two C-terminal FLAG tag sequences, and expressed in E. coli (10 mg/L). The 11-1G10 scFv was isolated by affinity chromatography on an anti-FLAG M2 antibody column as a 2:1 mixture of monomer and dimer forms which were separated by Superdex 75 chromatography; monomer (at 100 microg/ml) was stable for 7 days at 21 degrees C and 30 days at 4 degrees C, whereas the dimer slowly dissociated to monomer to yield a 2:1 monomerdimer equilibrium mixture after 30 days at 4 degrees C. The dimer was bivalent, with each combining site binding an NC41 Fab to yield a stable complex of Mr approximately 156,000. Binding affinities, determined in solution using a BIAcore biosensor, showed that the affinity for the interaction of 11-IG10 scFv monomer with NC41 scFv monomer was five- to six-fold higher than the interaction of the parent Fab pair. This is the first example of an scFv derived from a monoclonal antibody with a higher affinity than its parent Fab. More... »

PAGES

245-254

Identifiers

URI

http://scigraph.springernature.com/pub.10.1023/a:1022584702108

DOI

http://dx.doi.org/10.1023/a:1022584702108

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1002699130

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/9588948


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