Detection of flagellin by interaction with human recombinant TLR5 immobilized in liposomes

Olguin, Y., Villalobos, P., Carrascosa, L. G., Young, M., Valdez, E., Lechuga, L. and Galindo, R. (2013) Detection of flagellin by interaction with human recombinant TLR5 immobilized in liposomes. Analytical and Bioanalytical Chemistry, 405 4: 1267-1281. doi:10.1007/s00216-012-6523-4

Author Olguin, Y.
Villalobos, P.
Carrascosa, L. G.
Young, M.
Valdez, E.
Lechuga, L.
Galindo, R.
Title Detection of flagellin by interaction with human recombinant TLR5 immobilized in liposomes
Journal name Analytical and Bioanalytical Chemistry   Check publisher's open access policy
ISSN 1618-2642
Publication date 2013-02
Year available 2012
Sub-type Article (original research)
DOI 10.1007/s00216-012-6523-4
Volume 405
Issue 4
Start page 1267
End page 1281
Total pages 15
Place of publication Heidelberg, Germany
Publisher Springer
Collection year 2014
Language eng
Formatted abstract
Digestive diseases caused by flagellated bacteria are a huge public health problem worldwide and rapid detection methods are needed for contaminated environments. In this study, we propose a method to detect patterns associated with pathogens based on the properties of the innate immune system. Specifically, we use Toll-like receptor 5 (TLR5), a transmembrane protein that specifically recognizes flagellin (the structural protein of bacterial flagella). TLR5, which was obtained by recombinant production in insect cells, was immobilized into liposomes to form TLR5-proteoliposomes. Through surface plasmon resonance (SPR) and competition flow cytometry assays, the sensitivity of proteoliposomes to recognize Escherichia coli and Salmonella typhimurium flagellin was evaluated. In addition, we compared the results obtained by immobilizing anti-flagellin antibodies into liposomes. The results of the flagellin-affinity tests, expressed as an SPR kinetic rate constant ratio in the equilibrium equation K D = k d/k a, showed values of 13.8 × 10−9 and 7.73 × 10−9 M for the TLR5-proteoliposomes and anti-flagellin antibodies, respectively, against S. typhimurium. The anti-flagellin affinity results for E. coli showed K D of 84.1 × 10−8 M for SPR assays and K D of 3.5 × 10−8 M for competitive flow cytometry, which was used as a detection system without the immobilization of proteoliposomes. This research demonstrates the practical possibility of using proteoliposomes as recognition elements in the generation of systems for the rapid detection of flagellated bacteria, which could help avoid consumption of contaminated food by humans and thereby prevent intestinal infections.
Keyword Toll-like receptor 5
Surface plasmon resonance
Competitive flow cytometry
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status Non-UQ
Additional Notes Published online 22 November 2012.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Non HERDC
Australian Institute for Bioengineering and Nanotechnology Publications
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Citation counts: TR Web of Science Citation Count  Cited 6 times in Thomson Reuters Web of Science Article | Citations
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