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Kinetic characterisation of a single chain antibody against the Hormone Abscisic Acid : comparison with its parental monoclonal
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Badescu, George O., Marsh, Andrew, Smith, Timothy R., Thompson, Andrew J. and Napier, R. (Richard) (2016) Kinetic characterisation of a single chain antibody against the Hormone Abscisic Acid : comparison with its parental monoclonal. PLoS One, 11 (3). pp. 1-14. e0152148. doi:10.1371/journal.pone.0152148 ISSN 1932-6203.
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Official URL: http://dx.doi.org/10.1371/journal.pone.0152148
Abstract
A 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.
Item Type: | Journal Article | ||||||||
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Subjects: | Q Science > QR Microbiology | ||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Library of Congress Subject Headings (LCSH): | Immunoglobulins, Biosensors, Abscisic acid | ||||||||
Journal or Publication Title: | PLoS One | ||||||||
Publisher: | Public Library of Science | ||||||||
ISSN: | 1932-6203 | ||||||||
Official Date: | 29 March 2016 | ||||||||
Dates: |
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Volume: | 11 | ||||||||
Number: | 3 | ||||||||
Number of Pages: | 14 | ||||||||
Page Range: | pp. 1-14 | ||||||||
Article Number: | e0152148 | ||||||||
DOI: | 10.1371/journal.pone.0152148 | ||||||||
Status: | Peer Reviewed | ||||||||
Publication Status: | Published | ||||||||
Access rights to Published version: | Restricted or Subscription Access | ||||||||
Date of first compliant deposit: | 11 April 2016 | ||||||||
Date of first compliant Open Access: | 11 April 2016 | ||||||||
Funder: | Seventh Framework Programme (European Commission) (FP7), Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Kyōto Daigaku‏. Nōgaku Kenkyūka [Kyoto University. Graduate School of Agriculture], University of Saskatchewan. Structural Sciences Centre | ||||||||
Grant number: | HPRN-CT-2002-00334 (FP7), BB/L009366 |
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