Anti-mesotrione single chain antibody fragments derived using either phage display or via hybridoma technology: Selection, characterization and stabilization

G Strachan, P Molloy, J Lamont, R Claissie, L Todd, J Vincent, D Holt, A Dinsmore, A Porter

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Single-chain antibody fragments (scAb), specific for the triketone herbicide mesotrione, have been expressed and purified from the bacterium Escherichia coli. ScAbs were identified following the cloning of variable heavy and light chains from anti-mesotrione monoclonal antibodies (mAbs) and from panning an immunized phage display library. Both the monoclonal and phage derived scAbs were produced from the same immunized mouse spleen. In competition ELISA, the mAb derived scAbs (35 and 36) showed similar specificity for mesotrione compared with the parental mAbs. MAb 35 was the most sensitive with a limit of detection (IC20 value) within EC legislative limits for drinking water at 0.04 mug l(-1). The best phage scAb (H6) had sensitivities approximately 20 x less than scAb 35. In competition ELISA, scAb 35 showed reduced sensitivity to sulcotrione, a structurally related triketone, with IC50 values in the muM range. The introduction of an interdomain disulphide bond into scAb 35 resulted in greater stability in non-physiological environments. Immunoassay may provide a simple, cheap and rapid test for detection of mesotrione in a range of environments including soil and food.

Original languageEnglish
Pages (from-to)115-125
Number of pages11
JournalFood and Agricultural Immunology
Volume15
DOIs
Publication statusPublished - 2003

Keywords

  • antibody fragments
  • pesticide
  • phage display
  • ELISA
  • ESCHERICHIA-COLI
  • ANTIHAPTEN ANTIBODIES
  • GAS-CHROMATOGRAPHY
  • WATER
  • IMMUNOASSAY
  • HERBICIDES
  • EXPRESSION
  • BINDING
  • SOIL

Cite this

Anti-mesotrione single chain antibody fragments derived using either phage display or via hybridoma technology: Selection, characterization and stabilization. / Strachan, G ; Molloy, P ; Lamont, J ; Claissie, R ; Todd, L ; Vincent, J ; Holt, D ; Dinsmore, A ; Porter, A .

In: Food and Agricultural Immunology, Vol. 15, 2003, p. 115-125.

Research output: Contribution to journalArticle

Strachan, G ; Molloy, P ; Lamont, J ; Claissie, R ; Todd, L ; Vincent, J ; Holt, D ; Dinsmore, A ; Porter, A . / Anti-mesotrione single chain antibody fragments derived using either phage display or via hybridoma technology: Selection, characterization and stabilization. In: Food and Agricultural Immunology. 2003 ; Vol. 15. pp. 115-125.
@article{b2fd67a07f0d4d8fa4dc44d7f8a0b16a,
title = "Anti-mesotrione single chain antibody fragments derived using either phage display or via hybridoma technology: Selection, characterization and stabilization",
abstract = "Single-chain antibody fragments (scAb), specific for the triketone herbicide mesotrione, have been expressed and purified from the bacterium Escherichia coli. ScAbs were identified following the cloning of variable heavy and light chains from anti-mesotrione monoclonal antibodies (mAbs) and from panning an immunized phage display library. Both the monoclonal and phage derived scAbs were produced from the same immunized mouse spleen. In competition ELISA, the mAb derived scAbs (35 and 36) showed similar specificity for mesotrione compared with the parental mAbs. MAb 35 was the most sensitive with a limit of detection (IC20 value) within EC legislative limits for drinking water at 0.04 mug l(-1). The best phage scAb (H6) had sensitivities approximately 20 x less than scAb 35. In competition ELISA, scAb 35 showed reduced sensitivity to sulcotrione, a structurally related triketone, with IC50 values in the muM range. The introduction of an interdomain disulphide bond into scAb 35 resulted in greater stability in non-physiological environments. Immunoassay may provide a simple, cheap and rapid test for detection of mesotrione in a range of environments including soil and food.",
keywords = "antibody fragments, pesticide, phage display, ELISA, ESCHERICHIA-COLI, ANTIHAPTEN ANTIBODIES, GAS-CHROMATOGRAPHY, WATER, IMMUNOASSAY, HERBICIDES, EXPRESSION, BINDING, SOIL",
author = "G Strachan and P Molloy and J Lamont and R Claissie and L Todd and J Vincent and D Holt and A Dinsmore and A Porter",
year = "2003",
doi = "10.1080/09540100400003212",
language = "English",
volume = "15",
pages = "115--125",
journal = "Food and Agricultural Immunology",
issn = "0954-0105",
publisher = "Taylor and Francis Ltd.",

}

TY - JOUR

T1 - Anti-mesotrione single chain antibody fragments derived using either phage display or via hybridoma technology: Selection, characterization and stabilization

AU - Strachan, G

AU - Molloy, P

AU - Lamont, J

AU - Claissie, R

AU - Todd, L

AU - Vincent, J

AU - Holt, D

AU - Dinsmore, A

AU - Porter, A

PY - 2003

Y1 - 2003

N2 - Single-chain antibody fragments (scAb), specific for the triketone herbicide mesotrione, have been expressed and purified from the bacterium Escherichia coli. ScAbs were identified following the cloning of variable heavy and light chains from anti-mesotrione monoclonal antibodies (mAbs) and from panning an immunized phage display library. Both the monoclonal and phage derived scAbs were produced from the same immunized mouse spleen. In competition ELISA, the mAb derived scAbs (35 and 36) showed similar specificity for mesotrione compared with the parental mAbs. MAb 35 was the most sensitive with a limit of detection (IC20 value) within EC legislative limits for drinking water at 0.04 mug l(-1). The best phage scAb (H6) had sensitivities approximately 20 x less than scAb 35. In competition ELISA, scAb 35 showed reduced sensitivity to sulcotrione, a structurally related triketone, with IC50 values in the muM range. The introduction of an interdomain disulphide bond into scAb 35 resulted in greater stability in non-physiological environments. Immunoassay may provide a simple, cheap and rapid test for detection of mesotrione in a range of environments including soil and food.

AB - Single-chain antibody fragments (scAb), specific for the triketone herbicide mesotrione, have been expressed and purified from the bacterium Escherichia coli. ScAbs were identified following the cloning of variable heavy and light chains from anti-mesotrione monoclonal antibodies (mAbs) and from panning an immunized phage display library. Both the monoclonal and phage derived scAbs were produced from the same immunized mouse spleen. In competition ELISA, the mAb derived scAbs (35 and 36) showed similar specificity for mesotrione compared with the parental mAbs. MAb 35 was the most sensitive with a limit of detection (IC20 value) within EC legislative limits for drinking water at 0.04 mug l(-1). The best phage scAb (H6) had sensitivities approximately 20 x less than scAb 35. In competition ELISA, scAb 35 showed reduced sensitivity to sulcotrione, a structurally related triketone, with IC50 values in the muM range. The introduction of an interdomain disulphide bond into scAb 35 resulted in greater stability in non-physiological environments. Immunoassay may provide a simple, cheap and rapid test for detection of mesotrione in a range of environments including soil and food.

KW - antibody fragments

KW - pesticide

KW - phage display

KW - ELISA

KW - ESCHERICHIA-COLI

KW - ANTIHAPTEN ANTIBODIES

KW - GAS-CHROMATOGRAPHY

KW - WATER

KW - IMMUNOASSAY

KW - HERBICIDES

KW - EXPRESSION

KW - BINDING

KW - SOIL

U2 - 10.1080/09540100400003212

DO - 10.1080/09540100400003212

M3 - Article

VL - 15

SP - 115

EP - 125

JO - Food and Agricultural Immunology

JF - Food and Agricultural Immunology

SN - 0954-0105

ER -