Kinetic characterisation of a single chain antibody against the hormone abscisic acid: comparison with its parental monoclonal

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dc.contributor.authorBadescu, George
dc.contributor.authorMarsh, Andrew
dc.contributor.authorSmith, Timothy R.
dc.contributor.authorThompson, Andrew J.
dc.contributor.authorNapier, Richard M.
dc.date.accessioned2016-06-21T12:54:57Z
dc.date.available2016-06-21T12:54:57Z
dc.date.issued2016-03-09
dc.description.abstractA single-chain Fv fragment antibody (scFv) specific for the plant hormone abscisic acid (ABA) has been expressed in the bacterium Escherichia coli as a fusion protein. The kinetics of ABA binding have been measured using surface plasmon resonance spectrometry (BIAcore 2000) using surface and solution assays. Care was taken to calculate the concentration of active protein in each sample using initial rate measurements under conditions of partial mass transport limitation. The fusion product, parental monoclonal antibody and the free scFv all have low nanomolar affinity constants, but there is a lower dissociation rate constant for the parental monoclonal resulting in a three-fold greater affinity. Analogue specificity was tested and structure-activity binding preferences measured. The biologically-active (+)-ABA enantiomer is recognised with an affinity three orders of magnitude higher than the inactive (-)-ABA. Metabolites of ABA including phaseic acid, dihydrophaseic acid and deoxy-ABA have affinities over 100-fold lower than that for (+)-ABA. These properties of the scFv make it suitable as a sensor domain in bioreporters specific for the naturally occurring form of ABA.en_UK
dc.identifier.citationBadescu GO, Marsh A, Smith TR, et al., (2016) Kinetic characterisation of a single chain antibody against the hormone abscisic acid: comparison with its parental monoclonal. PLoS ONE, Volume 11, Issue 3, March 2016, Article number e0152148en_UK
dc.identifier.issn1932-6203
dc.identifier.urihttps://doi.org/10.1371/journal.pone.0152148
dc.identifier.urihttp://dspace.lib.cranfield.ac.uk/handle/1826/9986
dc.language.isoenen_UK
dc.publisherPLOS (Public Library of Science)en_UK
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.titleKinetic characterisation of a single chain antibody against the hormone abscisic acid: comparison with its parental monoclonalen_UK
dc.typeArticleen_UK

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