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

Simple item page
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.citationGeorge O. Badescu, Andrew Marsh, Timothy R. Smith, Andrew J. Thompson, and Richard M. Napier. Kinetic characterisation of a single chain antibody against the hormone abscisic acid: comparison with its parental monoclonal, PLoS ONE. Volume 11, Issue 3, e0152148.en_UK
dc.identifier.issn1932-6203
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0152148
dc.identifier.urihttp://dspace.lib.cranfield.ac.uk/handle/1826/9986
dc.language.isoenen_UK
dc.publisherPublic Library of Scienceen_UK
dc.rightsCopyright: © 2016 Badescu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Attribution 4.0 International (CC BY 4.0) You are free to: Share — copy and redistribute the material in any medium or format, Adapt — remix, transform, and build upon the material for any purpose, even commercially. The licensor cannot revoke these freedoms as long as you follow the license terms. Under the following terms: Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. Information: No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.
dc.titleKinetic characterisation of a single chain antibody against the hormone abscisic acid: comparison with its parental monoclonalen_UK
dc.typeArticleen_UK

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Kinetic_Characterisation_of_a_Single_Chain-2016.pdf
Size:
602.39 KB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.79 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

Staff publications (SWEE)