Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform

Caroline Murphy*, Edwina Stack, Svetlana Krivelo, Daniel A. McPartlin, Barry Byrne, Charles Greef, Michael J. Lochhead, Greg Husar, Shauna Devlin, Christopher T. Elliott, Richard J. O'Kennedy

*Corresponding author for this work

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Microcystins are a major group of cyanobacterial heptapeptide toxins found in freshwater and brackish environments. There is currently an urgent requirement for highly-sensitive, rapid and in-expensive detection methodologies for these toxins. A novel single chain fragment variable (scFv) fragment was generated and is the first known report of a recombinant anti-microcystin avian antibody. In a surface plasmon resonance-based immunoassay, the antibody fragment displayed cross-reactivity with seven microcystin congeners (microcystin-leucine-arginine (MC-LR) 100%, microcystin-tyrosine-arginine (MC-YR) 79.7%, microcystin-leucine-alanine (MC-LA) 74.8%, microcystin-leucine-phenylalanine (MC-LF) 67.5%, microcystin-leucine-tryptophan (MC-LW) 63.7%, microcystin-arginine-arginine (MC-RR) 60.1% and nodularin (Nod) 69.3%, % cross reactivity). Following directed molecular evolution of the parental clone the resultant affinity-enhanced antibody fragment was applied in an optimized fluorescence immunoassay on a planar waveguide detection system. This novel immuno-sensing format can detect free microcystin-LR with a functional limit of detection of 0.19ngmL-1and a detection range of 0.21-5.9ngmL-1. The assay is highly reproducible (displaying percentage coefficients of variance below 8% for intra-day assays and below 11% for inter-day assays), utilizes an inexpensive cartridge system with low reagent volumes and can be completed in less than twenty minutes.

Original languageEnglish
Pages (from-to)708-714
JournalBiosensors and Bioelectronics
Volume67
Early online date23 Oct 2014
DOIs
Publication statusPublished - 15 May 2015

Keywords

  • Cyanobacteria
  • Fluorescent planar wave-guide
  • Immunoassay
  • Microcystin
  • Recombinant Antibody

ASJC Scopus subject areas

  • Biophysics
  • Biomedical Engineering
  • Biotechnology
  • Electrochemistry
  • Medicine(all)

Fingerprint Dive into the research topics of 'Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform'. Together they form a unique fingerprint.

Cite this