IUVSTA 15th International Vacuum Congress (IVC-15), AVS 48th International Symposium (AVS-48), 11th International Conference on Solid Surfaces (ICSS-11)
    Biomaterials Wednesday Sessions
       Session BI+SS-WeM

Invited Paper BI+SS-WeM1
Hybridization Reactions between Surface Attached Oligonucleotides and Complements from Solution

Wednesday, October 31, 2001, 8:20 am, Room 102

Session: Biological Interface & Surface Science
Presenter: W. Knoll, Max-Planck-Institut für Polymerforschung, Germany
Authors: W. Knoll, Max-Planck-Institut für Polymerforschung, Germany
D. Kambhampati, Max-Planck-Institut für Polymerforschung, Germany
T. Neumann, Max-Planck-Institut für Polymerforschung, Germany
M. Chen, Max-Planck-Institut für Polymerforschung, Germany
Correspondent: Click to Email
The quantitative evaluation of hybridization reactions between surface-attached 15mer oligonucleotides and their complements from solution will be described. Reaction kinetics, as well as equilibrium binding studies are conducted in order to reveal the association/dissociation mechanism. Different strategies to prepare the interfacial probe layers are tested and compared to each other: These are 1) direct coupling of the catcher oligonucleotides to a gold substrate by thiol groups, 2) a streptavidin monolayer-based coupling scheme via biotinylated probe oligos, 3) a similar approach but based on a commercial dextran-streptavidin structure, 4) 15mers attached to polymer brushes grown by a "grafting-from" approach, and finally 5) a layer prepared by electropolymerization of hydroxyphenol-derivatized oligonucleotides. In cases where surface plasmon spectroscopy was not sensitive enough for label-free detection of the hybridization we employed our recently developed surface-plasmon field-enhanced fluorescence spectroscopy. Different versions based on having either the catcher strand labeled, or the target, or both (e.g., for energy transfer studies) will be discussed. Parameters that are studied include the effect of temperature, ionic strength, mismatch (number of bases, position) length of complement, charge density (DNA versus PNA) etc.