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

Y. Olguín, P. Villalobos, L. G. Carrascosa, M. Young, E. Valdez, L. Lechuga, R. Galindo

Resultado de la investigación: Contribución a una revistaArtículo

16 Citas (Scopus)

Resumen

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.

Idioma originalInglés
Páginas (desde-hasta)1267-1281
Número de páginas15
PublicaciónAnalytical and Bioanalytical Chemistry
Volumen405
N.º4
DOI
EstadoPublicada - 1 ene 2013

Áreas temáticas de ASJC Scopus

  • Química analítica
  • Bioquímica

Huella Profundice en los temas de investigación de 'Detection of flagellin by interaction with human recombinant TLR5 immobilized in liposomes'. En conjunto forman una huella única.

  • Citar esto

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