SPR biosensor for the detection of L. monocytogenes using phage-displayed antibody

Biosens Bioelectron. 2007 Sep 30;23(2):248-52. doi: 10.1016/j.bios.2007.04.007. Epub 2007 Apr 19.

Abstract

Whole cells of Listeria monocytogenes were detected with a compact, surface plasmon resonance (SPR) sensor using a phage-displayed scFv antibody to the virulence factor actin polymerization protein (ActA) for biorecognition. Phage Lm P4:A8, expressing the scFv antibody fused to the pIII surface protein was immobilized to the sensor surface through physical adsorption. A locally constructed fluidics system was used to deliver solutions to the compact, two-channel SPREETA sensor. Specificity of the sensor was tested using common food-borne bacteria and a control phage, M13K07 lacking the scFv fusion on its coat protein. The detection limit for L. monocytogenes whole cells was estimated to be 2 x 10(6)cfu/ml. The sensor was also used to determine the dissociation constant (Kd) for the interaction of phage-displayed scFv and soluble ActA in solution as 4.5 nM.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies, Monoclonal / immunology*
  • Biosensing Techniques / instrumentation*
  • Biosensing Techniques / methods
  • Colony Count, Microbial / instrumentation
  • Colony Count, Microbial / methods
  • Equipment Design
  • Equipment Failure Analysis
  • Immunoassay / instrumentation*
  • Immunoassay / methods
  • Listeria monocytogenes / immunology*
  • Listeria monocytogenes / isolation & purification*
  • Peptide Library*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Surface Plasmon Resonance / methods*

Substances

  • Antibodies, Monoclonal
  • Peptide Library