OWLS APPLICATION NOTES NO-008
TEST MEASUREMENT OF LABEL-FREE IMMUNOSENSOR
USING BSA - ANTI BSA MODEL MOLECULE PAIR
using OPTICAL WAVEGUIDE LIGHTMODE SPECTROSCOPY (OWLS) detection
I n t h e c a s e o f g l a s s t y p e m e t a l o xi d e s u r f a c e s s u c h
as SiO2-TiO2, mainly hydroxyl groups are present,
the surface of the waveguide has to be modified by
silanisation using reactive silane reagents for
introducing functional groups of all sorts (e.g.
aliphatic amine, sulfhydryl, aromatic amine and
epoxy) onto the surface of inorganic materials.
In the case of APTS modification:
succinic anhydride is used to change the amino
functionality to carboxyl groups. Carboxyl groups
are usually activated wi t h the EDC/NHS
(1-etil-3-(3-dimetylaminopropyl) carbodiimide \ N-
hydroxysuccinimid) method first forming an easily
attackable reactive succinimid ester that can couple
the proteins to the surface by their amino groups.
MicroVacuum Ltd. H-1147, Kerékgyártó u. 10., Budapest, Hungary
Web: http://www.owls-sensors.com/;
To test the layers formed, immobilisation
e xp e r i m e n t s w e r e u n d e r t a k e n o n t h e m o d i f i e d
surfaces using the bovine serum albumin (BSA) -
anti-BSA antibody model molecule pair.
BSA
Evaluation of layers formed
T o s t u d y t h e a p p l i c a b i l i t y o f t h e f o r m e d l a y e r s we
attached covalently BSA molecules on the surfaces
and measured the specific response of the
sensitized surface to anti-BSA antibody standard
solutions of different concentration.
Immobilisation and measurement on carboxy
functionalised sensor surfaces
References
1. Vörös, J. J. Ramsden, G. Csucs, I. Szendrõ, S.M. De
Paul, M. Textor, N. D. Spencer (2002): Optical Grating
Coupler Biosensors. Biomaterials 23 3699-3710.
2. www.owls-sensors.com
Phone: + 36 1 252 1991; +36 1 467 0108 Fax: +36 1 221 7996
E-mail: info@owls-sensors.com Copyright @ Microvacuum, 2006