Phage as a molecular recognition element in biosensors immobilized by physical adsorption

Biosens Bioelectron. 2007 Jan 15;22(6):986-92. doi: 10.1016/j.bios.2006.03.025. Epub 2006 May 30.

Abstract

Biosensors based on landscape phages immobilized by physical adsorption on the surface of a quartz crystal microbalance was used for detection of beta-galactosidase from Escherichia coli. The sensor had a detection limit of a few nanomoles and a response time of a approximately 100 s over the range of 0.003-210 nM. The binding dose-response curve had a typical sigmoid shape and the signal was saturated at the beta-galactosidase concentration of about 200 nM. A marked selectivity for beta-galactosidase over BSA was observed in mixed solutions even when the concentration of BSA exceeded the concentration of beta-galactosidase by a factor of approximately 2000. The apparent value of the dissociation constant (K(d)) of the interaction of free phage with beta-galactosidase (9.1+/-0.9 pM) was smaller compared with the one calculated for the bound phage (1.7+/-0.5 nM). The binding was specific with three binding sites needed to bind a single molecule of beta-galactosidase. The K(d) obtained from the enzyme-linked immunosorbent assay (ELISA) for the phage and the monoclonal anti-beta-galactosidase antibodies were 21+/-2 and 26+/-2 nM, respectively. Although the method of physical adsorption is simpler and more economical in comparison with Langmuir-Blodgett and molecular assembling methods the performances of the sensors made by these technologies compare well. This work provides evidence that phage can be used as a recognition element in biosensors using physical adsorption method for immobilization of phage on the sensor surface.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Bacteriophages / drug effects*
  • Bacteriophages / physiology*
  • Biological Assay / instrumentation
  • Biological Assay / methods
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Dose-Response Relationship, Drug
  • Electrochemistry / instrumentation
  • Electrochemistry / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • beta-Galactosidase / analysis*
  • beta-Galactosidase / pharmacology*

Substances

  • beta-Galactosidase