A streptavidin surface on planar glass substrates for the detection of biomolecular interaction.
Based on the requirements of biomolecular interaction analysis on direct optical transducers, a streptavidin surface is examined. A general protocol was developed allowing the immobilization of biotinylated compounds using the rife biotin-streptavidin system. This type of surface modification can be applied to all biosensors using glass surfaces as sensor devices. Reflectometric interference spectroscopy (RIfS), a label-free, direct optical method was used to demonstrate the quality of the transducer surfaces. The surface modification is based on an aminofunctionalized polyethylene glycol layer covalently bound to the silica surface of the transducer and shows very little nonspecific binding. Biotin molecules can be easily coupled on such layers. Streptavidin followed by a biotinylated estrone derivative was immobilized by incubation of the biotinylated transducer surface. For the streptavidin layer we obtained interference signals corresponding to a protein monolayer. Finally, using a surface prepared as described above, biomolecular interaction experiments with an antibody against estrone were carried out to show the quality of the transducer surface. With RIfS all of the affinity-based surface modifications can be detected online and time resolved.
Birkert
O
O
Institute of Physical Chemistry, Institute of Organic Chemistry, University of Tübingen, Auf der Morgenstelle 8, Tübingen, D-72076, Germany.
Haake
H M
HM
Schütz
A
A
Mack
J
J
Brecht
A
A
Jung
G
G
Gauglitz
G
G
D000906:Antibodies; D015374:Biosensing Techniques / Q000379:methods*; D019921:Biotinylation; D003911:Dextrans / Q000737:chemistry; D004970:Estrone / Q000031:analogs & derivatives / Q000737:chemistry* / Q000276:immunology; D005898:Glass; D011092:Polyethylene Glycols / Q000737:chemistry; D012997:Solvents; D013057:Spectrum Analysis / Q000379:methods; D019809:Streptavidin / Q000737:chemistry
D000906:Antibodies; D003911:Dextrans; D012997:Solvents; D004970:Estrone; D011092:Polyethylene Glycols; D019809:Streptavidin