The interaction of proteins and cells with affinity ligands covalently coupled to silicon surfaces as monitored by ellipsometry.
Two methods for the chemical binding of biomolecules to silicon surfaces are described. The first method utilizes an alkyl silane and a nucleophilic reagent to join the biomolecule to the silicon surface; the second method involves crosslinking with glutaraldehyde in order to couple the biomolecule and albumin molecules, which have first been physically adsorbed. The course of binding to the silicon surface has been followed with the aid of ellipsometry. This optical measuring technique estimates the thicknesses of, e.g., organic layers, by measuring the polarization properties of a light beam before and after reflection at surfaces. The method by which the binding of a biomolecule to its corresponding affinity ligand on silicon wafers can be followed with this technique is reported. The systems studied are concanavalin A-Saccharomyces cerevisiae cells, immunoglobulin G-Staphylococcus aureus cells, and an NAD-analog-lactate dehydrogenase. With ellipsometry it was possible to assess how the incubation time and the concentration of the cells and the biomolecules added influenced the results. It was found that an increasing time of incubation and higher concentration resulted in a more complete coverage of the silicon wafer surfaces.
D001419:Bacteria / Q000302:isolation & purification*; D001671:Biochemistry / Q000379:methods*; D003208:Concanavalin A; D004798:Enzymes / Q000302:isolation & purification*; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D008855:Microscopy, Electron, Scanning; D009243:NAD; D055095:Optics and Photonics / Q000295:instrumentation; D012821:Silanes; D012825:Silicon; D013450:Sulfones; D013499:Surface Properties
D004798:Enzymes; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D012821:Silanes; D013450:Sulfones; D009243:NAD; D003208:Concanavalin A; C032605:2,2,2-trifluoroethanesulfonyl chloride; D012825:Silicon