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 1 2 3 4
Waveguide
Laser light

Substrate

X
TNFa
450
Time

OWLS records (changes in the effective refractive index) on the

anti-TNFa TNFa content in various human serum samples. 1: TNF-free
control samples; 2, 3 and 4 : sera of different individuals

70
Deposited material (arb. units)

biotinylated
anti-mouse IgG 60
y = 89.328x - 0.5209
50
avidin
bioti 40
n optical waveguide
30
Stable, reusable sensors were produced by using biotin-avidin
binding for immobilization of anti TNF antibodies 20

building of a TNFα immunosensor 10

400
0
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
Deposited material l (arb. units)

300
TNFα
α (ng/ml)

TNFα concentrations in the calibrating solution ( ) and in

200 various samples of human sera ( )

Conclusion
100 a biot-IgG a-TNF TNF 2ng/ml Using appropriately sensitized immunsensors, TNFa can be safely
avidin detected in partially purified serum samples in a magnitude of 100
0 pg/ml
0 100 200 300 400 500 600

time (min)
References
1. Vörös J. et al., Optical grating coupler biosensors. Biomaterials
23 (2002) 3699-3710.
Sample preparation
2. Barath et al., 2002.International Conf. of Cytokines in Medicine
Proteins of Mr of 10 to 40 KDa of normal and TNF-free sera were Oslo.
isolated by molecular filtering (Amicon). 100 ml of samples were 3. www.owls-sensors.com
loaded onto the surface of sensitized sensors.

Reproducible assays were carried out on partially purified serum

samples

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