OWLS APPLICATION NOTES NO-005

DETERMINATION OF TNFα α CONCENTRATION IN HUMAN SERUM
using OPTICAL WAVEGUIDE LIGHTMODE SPECTROSCOPY (OWLS) detection

Abstract
Immunosensors were constructed for rapid and label-free determination of TNFα concentrations in human serum samples.

Application of OWLS sensors
as selective immun detectors of TNFα
Sample

TNF α Anti-TNFα
Waveguide

1

2

3

4

Laser light

Substrate

X

TNFa
450

Time

anti-TNFa

OWLS records (changes in the effective refractive index) on the TNFa content in various human serum samples. 1: TNF-free control samples; 2, 3 and 4 : sera of different individuals

70
biotinylated anti-mouse IgG avidin bioti n optical waveguide

Deposited material (arb. units)

60 50 40 30 20 10
0 0 0.1

y = 89.328x - 0.5209

Stable, reusable sensors were produced by using biotin-avidin binding for immobilization of anti TNF antibodies
building of a TNFα immunosensor

400
Deposited material l (arb. units)

0.2

0.3

0.4

0.5

0.6

0.7

300

TNFα (ng/ml) α
TNFα concentrations in the calibrating solution ( various samples of human sera ( ) ) and in

200 100
avidin

Conclusion
a

biot-IgG

a-TNF

TNF 2ng/ml

0
0 100 200 300 time (min) 400 500 600

Using appropriately sensitized immunsensors, TNFa can be safely detected in partially purified serum samples in a magnitude of 100 pg/ml

References
1. Vörös J. et al., Optical grating coupler biosensors. Biomaterials 23 (2002) 3699-3710. 2. Barath et al., 2002.International Conf. of Cytokines in Medicine Oslo. 3. www.owls-sensors.com

Sample preparation
Proteins of Mr of 10 to 40 KDa of normal and TNF-free sera were isolated by molecular filtering (Amicon). 100 ml of samples were loaded onto the surface of sensitized sensors. Reproducible assays were carried out on partially purified serum samples

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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