New economically viable methods for the detection and removal of organic pollutants from water samples are needed to meet modern safety requirements. Development of techniques to produce antibody fragments in Escherichia coli (E. coli) have provided the opportunity to exploit antibodies as specialised chemicals for affinity detection and removal technologies. The variable heavy and light antibody domains of the anti-paraquat monoclonal antibody PQXB1/2 have been cloned into the single-chain antibody (scAb) expression vector, pBG1. Anti-paraquat scAb (0.1-0.4 mg/L bacterial culture) was expressed in E. coli. 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 paraquat-bovine serum albumin conjugate from solution in a single step.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:295567 |
| Date | January 1995 |
| Creators | Graham, Barbara Monica |
| Publisher | University of Aberdeen |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://digitool.abdn.ac.uk/R?func=search-advanced-go&find_code1=WSN&request1=AAIU078323 |
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