Immobilization of thiabendazole-specific monoclonal antibodies to silicon substrates via aqueous silanization

• Published in: Applied biochemistry and biotechnology Vol. 50; no. 3; pp. 265 - 284
• Main Authors: Flounders, A. W., Brandon, D. L., Bates, A. H.
• Format: Journal Article
• Language: English
• Published: 01-03-1995
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/BF02788097

Monoclonal antibodies specific for thiabendazole were immobilized to silicon, silicon dioxide, stoichiometric silicon nitride, and silicon-rich silicon nitride surfaces. This work provides the foundation for the development of a homogeneous sensor system for rapid detection and quantification of thiabendazole residues in produce and animal tissue. Immobilization was performed via aqueous silanization of the substrate followed sequentially by treatment with glutaraldehyde and contact with antibody solution in the presence of detergent. Surfaces were challenged with thiabendazole-horseradish peroxidase conjugate in an ELISA format to estimate immobilized antibody load. A stable and reproducible surface loading of 2 x 10 super(11) antibodies/cm super(2) was obtained only after surfaces received postimmobilization treatments to remove nonspecifically adsorbed antibody. No difference in surface loading was noted when using 30% hydrogen peroxide rather than nitric acid for silanol activation. Little difference was noted among the antibody loadings achieved on the various silicon substrates. Bound antigen-enzyme conjugate was eluted with 0.1N acetic acid and reproducible surface activity was measured for up to four consecutive antigen challenges. Immobilized antibody surfaces were stabilized with 2% sucrose, dehydrated at 37 degree C and stored in vacuum or stored at 4 degree C in phosphate buffered saline containing 0.01% sodium azide without significant loss of activity.