Cloning, expression and characterization of a single-chain antibody specific for the herbicide atrazine

Fergus R. Byrne, Steven D. Grant, Andrew J. Porter, William J. Harris*

*Corresponding author for this work

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

The antigen-binding domain of an antibody specific for the herbicide atrazine was cloned from hybridoma 4063-21-1 and expressed as a single-chain antibody (scAb) in the vector pPM1-His. The observed heavy and light chain gene sequences were consistent with those in subgroup VH I (B) and VK IV, as defined by the Kabat database of sequences. Subsequent expression in Escherichia coli strain XL-1 Blue produced up to 0.3 μg of functional single-chain antibody, from periplasmic protein preparations, per millilitre of culture. The scAb was purified as a monomer by immobilized metal chelate affinity chromatography via a hexa-histidine tail or as a dimer by antibody affinity chromatography. The functionality and specificity of the recombinant antibody was confirmed by binding to an atrazine-bovine serum albumin conjugate and free atrazine and simazine in a competition ELISA. This is the first example of the production and purification of a recombinant antibody that retains antigen binding for a triazine herbicide.

Original languageEnglish
Pages (from-to)19-29
Number of pages11
JournalFood and Agricultural Immunology
Volume8
Issue number1
DOIs
Publication statusPublished - 1 Jan 1996

Keywords

  • Atrazine
  • Dimerization
  • Pesticide detection
  • scAb
  • Single-chain antibody

ASJC Scopus subject areas

  • Food Science
  • Immunology
  • Agronomy and Crop Science

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