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

36 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

Fingerprint

Arginine
Planar waveguides
Optical waveguides
Antibodies
Assays
Leucine
Immunoglobulin Fragments
tryptophan-leucine
Immunoassay
Surface plasmon resonance
Directed Molecular Evolution
Microcystins
Fluorescence
Single-Chain Antibodies
Surface Plasmon Resonance
microcystin
cyanoginosin LR
cyanobacterial toxin
toxin-LR
Fresh Water

Keywords

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

Cite this

Murphy, Caroline ; Stack, Edwina ; Krivelo, Svetlana ; McPartlin, Daniel A. ; Byrne, Barry ; Greef, Charles ; Lochhead, Michael J. ; Husar, Greg ; Devlin, Shauna ; Elliott, Christopher T. ; O'Kennedy, Richard J. / Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform. In: Biosensors and Bioelectronics. 2015 ; Vol. 67. pp. 708-714.
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title = "Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform",
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.",
keywords = "Cyanobacteria, Fluorescent planar wave-guide, Immunoassay, Microcystin, Recombinant Antibody",
author = "Caroline Murphy and Edwina Stack and Svetlana Krivelo and McPartlin, {Daniel A.} and Barry Byrne and Charles Greef and Lochhead, {Michael J.} and Greg Husar and Shauna Devlin and Elliott, {Christopher T.} and O'Kennedy, {Richard J.}",
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Murphy, C, Stack, E, Krivelo, S, McPartlin, DA, Byrne, B, Greef, C, Lochhead, MJ, Husar, G, Devlin, S, Elliott, CT & O'Kennedy, RJ 2015, 'Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform', Biosensors and Bioelectronics, vol. 67, pp. 708-714. https://doi.org/10.1016/j.bios.2014.10.039

Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform. / Murphy, Caroline; Stack, Edwina; Krivelo, Svetlana; McPartlin, Daniel A.; Byrne, Barry; Greef, Charles; Lochhead, Michael J.; Husar, Greg; Devlin, Shauna; Elliott, Christopher T.; O'Kennedy, Richard J.

In: Biosensors and Bioelectronics, Vol. 67, 15.05.2015, p. 708-714.

Research output: Contribution to journalArticle

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T1 - Detection of the cyanobacterial toxin, microcystin-LR, using a novel recombinant antibody-based optical-planar waveguide platform

AU - Murphy, Caroline

AU - Stack, Edwina

AU - Krivelo, Svetlana

AU - McPartlin, Daniel A.

AU - Byrne, Barry

AU - Greef, Charles

AU - Lochhead, Michael J.

AU - Husar, Greg

AU - Devlin, Shauna

AU - Elliott, Christopher T.

AU - O'Kennedy, Richard J.

PY - 2015/5/15

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KW - Fluorescent planar wave-guide

KW - Immunoassay

KW - Microcystin

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