Immobilization of immunoglobulins on silica surfaces. Stability.
The development of new immunosensors based on surface-concentration-measuring devices requires a stable and reproducible immobilization of antibodies on well-characterized solid surfaces. We here report on the immobilization of immunoglobulin G (IgG) on chemically modified silica surfaces. Such surfaces may be used in various surface-oriented analytical methods. Reactive groups were introduced to the silica surfaces by chemical-vapour deposition of silane. The surfaces were characterized by ellipsometry, contact-angle measurements and scanning electron microscopy. IgG covalently bound by the use of thiol-disulphide exchange reactions, thereby controlling the maximum number of covalent bonds to the surface, was compared with IgG adsorbed on various silica surfaces. This comparison showed that the covalently bound IgG has a superior stability when the pH was lowered or incubation with detergents, urea or ethylene glycol was carried out. The result was evaluated by ellipsometry, an optical technique that renders possible the quantification of amounts of immobilized IgG. The results outline the possibilities of obtaining a controlled covalent binding of biomolecules to solid surfaces with an optimal stability and biological activity of the immobilized molecules.
Jönsson
U
U
Malmqvist
M
M
Rönnberg
I
I
D003432:Cross-Linking Reagents; D004355:Drug Stability; D007074:Immunoglobulin G; D007525:Isoelectric Focusing; D008722:Methods; D008956:Models, Chemical; D011485:Protein Binding; D012822:Silicon Dioxide; D013388:Succinimides; D013499:Surface Properties
D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D013388:Succinimides; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D012822:Silicon Dioxide