OWLS APPLICATION NOTES NO-003

LABEL-FREE IMMUNOSENSOR FOR ESCHERICHIA COLI DETECTION
using OPTICAL WAVEGUIDE LIGHTMODE SPECTROSCOPY (OWLS) detection

Abstract
Escherichia Coli is a Gram-negative non-spore forming rod and a representative microorganism in the enteric bacteria. It is also considered as index microorganism in the food industry. OWLS immunosensor offers a real time label-free detection of E.Coli bacteria For biosensing monoclonal antibodies raised against E.Coli bacteria. To form regenerable sensor surface, the waveguide sensor surface was modified with amino group and sensitized by immobilizing antibody/antigen molecules covalently to the surface.

Application of OWLS sensors
as immunosensor for the detection of E.Coli
E. coli monoclonal antibody 100 - 150 nm Laser light EM field distribution waveguide glass support

Calibration curve for E.Coli sensor
Concentration dependent response of E.Coli by OW LS

sample

Surface Chemistry
• • Amino fictionalization of waveguide surface by (γ-aminopropyl)triethoxysilane Immobilization of biomolecules on the amino surface of the OW LS sensors by glutaraldehyde.

Sensitization of the sensor surface
with monochlonal anti-E.Coli
650
S e nsor re sponse (a. u.) anti-E.Coli. 100x HCl, 10 mM GA TRIS

Conclusion
The OW LS Sensor is a rapi d and cost-effective tool for Escherichia Coli detection. W ith the pres ented immunosensor the lowest detectable amount of E. Coli is 3x103 CFU/ml.

550 450 350 250 150 0
(d.w.

References
1.
10 20 30 Time (min) 40 50

Vörös, J. J. Ramsden, G. Csucs, I. Szendrõ, S . M . D e P a u l , M . T e xt o r , N . D . S p e n c e r “Optical Grating Coupler Biosensors” Biomaterials 23 (2002) 3699-3710 Adányi N. Váradi M., Kim N., Szendrı I. “Development of new immunosensors for determination of contaminants in food” Current Applied Physics ( in press) w w w . o wl s - s e n s o r s . c o m

2.

Regeneration cycles of the measurement
Antibodies are covalently attached to the surface a n d l i f e t i m e o f s e n s o r s we r e e x a m i n e d a n d optimized.
560 540

3.

CFU/ml
3E+6 3E+5 3E+3

3E+7

Mass response (a. u.) Sensor (arbitrary unit)

520 500 480 460 440 420 400 380 60 80 100 120

Time (min)

MicroVacuum Ltd. H-1147, Kerékgyártó u. 10., Budapest, Hungary
Web: http://www.owls-sensors.com/; E-mail: info@owls-sensors.com

Phone: + 36 1 252 1991; +36 1 467 0108

Fax: +36 1 221 7996

Copyright @ Microvacuum, 2006

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