Cloning, expression and characterisation of a single‐chain antibody fragment to the herbicide paraquat
- 1 July 1995
- journal article
- research article
- Published by Wiley in Journal of Chemical Technology & Biotechnology
- Vol. 63 (3) , 279-289
- https://doi.org/10.1002/jctb.280630312
Abstract
New cost effective methods for the detection and removal of pesticides from water samples are required to meet modern safety standards. The recent development of techniques to produce antibody fragments in bacteria has provided the opportunity to exploit antibodies as specialised chemicals for affinity detection/removal technologies. The variable heavy and light polypeptide chains of the anti-paraquat monoclonal antibody PQXB1/2 have been cloned into the single-chain antibody (ScAb) expression vector pBG1. The construct was expressed in Escherichia coli and 0·4 mg functional antibody produced from 1 dm3 of induced culture. Characterisation of ScAb by antigen binding profile and competition ELISA showed it to have a sensitivity one order of magnitude below that of the parent monoclonal. ScAb was purified as a monomer or dimer and analysed by HPLC size exclusion chromatography. When immobilised on polystyrene beads the ScAb could remove 85% of a paraquat–bovine serum albumin conjugate from solution in a single step.Keywords
This publication has 21 references indexed in Scilit:
- Separation and concentration of bacteria with immobilized antibody fragmentsJournal of Applied Bacteriology, 1995
- Spontaneous assembly of bivalent single chain antibody fragments in Escherichia coliMolecular Immunology, 1994
- Cloning, sequencing and expression of the Fab fragment of a monoclonal antibody to the herbicide atrazineProtein Engineering, Design and Selection, 1993
- Miniantibodies: use of amphipathic helixes to produce functional, flexibly linked dimeric FV fragments with high avidity in Escherichia coliBiochemistry, 1992
- Antibody Engineering: Advances From the Use of Escherichia coli Expression SystemsNature Biotechnology, 1991
- The Functional Expression of Antibody Fv Fragments in Ischhuchia coli: Improved Vectors and a Generally Applicable Purification TechniqueNature Biotechnology, 1991
- Single chain antibody variable regionsTrends in Biotechnology, 1991
- A comparison of strategies to stabilize immunoglobulin Fv-fragmentsBiochemistry, 1990
- Recombinant antibodies possessing novel effector functionsNature, 1984
- Enzyme linked immunosorbent assay (ELISA) for paraquatInternational Journal of Immunopharmacology, 1983