Glycosylated protein-functionalized gold nanoparticle-based detection of heat-labile enterotoxin from complex samples

Karuppuchamy Selvaprakash, Yu-Chie Chen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Enterotoxigenic Escherichia coli O78:H11 is a food-borne pathogen that can cause "traveler's diarrhea." The pathogenicity of E. coli O78:H11 is attributed to the expression of enterotoxin, particularly heat-labile enterotoxin (LT). LT is a pentameric protein composed of one A and five B subunits. The B subunit of LT (LT-B) contains the binding moieties toward Gal beta(1 -> 4)Glc. Chicken ovalbumin (COA) is a glycoprotein composed of Gal beta(1 -> 4) Glc termini and abundant (similar to 54%) in chicken egg white (CEW) proteins. COA decorated with similar to 17 additional galactosides, obtained by reacting CEW proteins with lactose (Gal beta[1 -> 4]Glc) through the Maillard reaction, was used as a probe molecule against LT-B. The generated CEW derivatives, mainly dominated by glycated COA, were immobilized on gold nanoparticles (Au@Lac-CEW NPs) by reacting with aqueous tetrachloroaurate through onepot reactions. Au@Lac-CEW NPs had a good binding affinity toward LT-B with a dissociation constant of similar to 1.86 x 10(-7) M. A colorimetric method by using Au@Lac-CEW NPs as sensing probes against LT-B (>= 31 nM) that can be visualized was demonstrated. The limit of detection against LT-B was reduced to similar to 4 nM as revealed by matrix-assisted laser desorption/ionization mass spectrometry as a detection tool.

Original languageEnglish
Article number128640
Number of pages8
JournalSensors and actuators b-Chemical
Volume322
DOIs
StatePublished - 1 Nov 2020

Keywords

  • Enterotoxigenic Escherichia coli
  • Heat-labile enterotoxin
  • Colorimetric
  • Chicken egg whites
  • Glycoproteins
  • ESCHERICHIA-COLI
  • CHOLERA-TOXIN
  • MASS-SPECTROMETRY
  • NANOCLUSTERS
  • BINDING
  • IDENTIFICATION
  • EPIDEMIOLOGY
  • PURIFICATION
  • OVALBUMIN
  • MECHANISM

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