Phage as a molecular recognition element in biosensors immobilized by physical adsorption
| Autor: | Iryna Sorokulova, Aleksandr Simonian, Vitaly Vodyanoy, Viswaprakash Nanduri, Alexandre M. Samoylov, Valery A. Petrenko |
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| Rok vydání: | 2007 |
| Předmět: |
Detection limit
Chromatography Phage display Dose-Response Relationship Drug biology Chemistry Biomedical Engineering Biophysics Reproducibility of Results Biosensing Techniques General Medicine Quartz crystal microbalance beta-Galactosidase biology.organism_classification Sensitivity and Specificity Dissociation constant Bacteriophage Adsorption Molecular recognition Electrochemistry Bacteriophages Biological Assay Biosensor Biotechnology |
| Zdroj: | Biosensors and Bioelectronics. 22:986-992 |
| ISSN: | 0956-5663 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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