Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces. Academic Article uri icon

abstract

  • A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional cross-linkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG antibody immobilized was determined to be in the range of 0.66 to 0.96 ng/mm2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2-6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immobilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components.

published proceedings

  • Anal Biochem

altmetric score

  • 3.75

author list (cited authors)

  • Bhatia, S. K., Shriver-Lake, L. C., Prior, K. J., Georger, J. H., Calvert, J. M., Bredehorst, R., & Ligler, F. S.

citation count

  • 331

complete list of authors

  • Bhatia, SK||Shriver-Lake, LC||Prior, KJ||Georger, JH||Calvert, JM||Bredehorst, R||Ligler, FS

publication date

  • May 1989