AVS 50th International Symposium
    Biomaterial Interfaces Monday Sessions
       Session BI-MoM

Paper BI-MoM5
Limitations of Molecular Streptavidin/Anti-biotin Antibody Architectures using Micro-contact Printed Biotinylated Thiols

Monday, November 3, 2003, 9:40 am, Room 307

Session: Protein-Surface Interactions
Presenter: Ch. Grunwald, Ruhr-University Bochum, Germany
Authors: Ch. Grunwald, Ruhr-University Bochum, Germany
N. Opitz, Max-Planck Institute for Molecular Physiology, Germany
S. Herrwerth, University of Heidelberg, Germany
W. Eck, University of Heidelberg, Germany
J. Kuhlmann, Max-Planck Institute for Molecular Physiology, Germany
Ch. Woell, Ruhr-University of Bochum, Germany
Correspondent: Click to Email
Atomic force microscopy (AFM) and confocal fluorescence microscopy have been used to study the interaction of streptavidin and anti-biotin antibodies with a patterned, biotinylated organic surface. This system presently attracts considerable interest because of its potential for molecular architectures employing protein-protein interactions. The substrates were prepared by first using the µCP technique to print a periodic pattern of an oligoethylenglycol (OEG) self-assembled monolayer (SAM) on clean gold surfaces. The pattern consists of squares (40 µm x 40 µm) which are separated from each other in each direction by 10 µm. By immersing the stamped substrates into a mixture of OH-terminated and biotinylated organothiols a patterned SAM is obtained. These 2D-SAM patterns have been imaged via contact atomic force and lateral force microscopy as well as with tapping-mode AFM. The patterned SAMs were then incubated with two fluorescence-labelled proteins exhibiting a strong affinity towards biotin, streptavidin and anti-biotin antibody. Incubation time, temperature and concentration of the protein solution as well as the biotin surface concentration were varied systematically. The comparison of the AFM-data with the results of the fluorescence microscopy allows for important conclusion on the protein-protein binding, in particular concerning reproducibility, unspecific binding and protein resistance.