title: Binding characteristics of anti-atrazine monoclonal antibodies and their fragments synthesised in bacteria and plants.
abstract: Single-chain antibody fragments (scAb), specific for the herbicide atrazine, have been expressed in the bacterium Escherichia coli and in transgenic tobacco plants. The scAb could be purified as a monomer (monovalent) via a hexa-histidine tail or as a dimer (divalent) by antibody affinity chromatography. In competition ELISA, the bacterial scAb showed the same specificity for atrazine and related triazine herbicides as the parental mAb cell line, but both plant and bacterial monomeric scAbs showed increased sensitivity to free atrazine. Surface plasmon resonance (BIAcore 2000) analysis confirmed that purified scAb, derived from plant or bacteria, retained similar association rates as the mAb. However, the monomeric plant and bacterial scAbs showed a lower affinity for immobilised antigen, than the equivalent dimeric scAbs or mAb. This decrease in affinity was due to a 10 fold slower dissociation rate and is likely due to loss of the avidity contribution of dimeric molecules.
authors.affiliation: Department of Molecular and Cell Biology, University of Aberdeen, Foresterhill, UK.
authors.forename: G
authors.initials: G
authors.lastname: Strachan
authors.forename: S D
authors.initials: SD
authors.lastname: Grant
authors.forename: D
authors.initials: D
authors.lastname: Learmonth
authors.forename: M
authors.initials: M
authors.lastname: Longstaff
authors.forename: A J
authors.initials: AJ
authors.lastname: Porter
authors.forename: W J
authors.initials: WJ
authors.lastname: Harris
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001280:Atrazine / Q000276:immunology*; D001667:Binding, Competitive; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000276:immunology; D015202:Protein Engineering
chemical_list: D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D001280:Atrazine