Cloning, expression and characterisation of a single‐chain antibody fragment to the herbicide paraquat

Barbara M. Graham, Andy J.R. Porter, William J. Harris*

*Corresponding author for this work

Research output: Contribution to journalArticle

41 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)279-289
Number of pages11
JournalJournal of Chemical Technology & Biotechnology
Volume63
Issue number3
DOIs
Publication statusPublished - 1 Jan 1995

Keywords

  • immunoaffinity
  • paraquat
  • pesticide analysis
  • pesticide removal
  • ScAb
  • single‐chain antibodies

ASJC Scopus subject areas

  • Biotechnology
  • Chemical Engineering(all)
  • Renewable Energy, Sustainability and the Environment
  • Fuel Technology
  • Waste Management and Disposal
  • Pollution
  • Organic Chemistry
  • Inorganic Chemistry

Fingerprint Dive into the research topics of 'Cloning, expression and characterisation of a single‐chain antibody fragment to the herbicide paraquat'. Together they form a unique fingerprint.

  • Cite this