Immobilization of antibodies on the self-assembled monolayer by antigen-binding site protection and immobilization kinetic control

Abstract

The orientation of the biological molecule immobi-lized on a solid surface has been critical in devel-opment of various applications. In this study, ori-entation of antibody was retained by protecting the antigen-binding site of the antibody prior to immo-bilization to -functionalized mixed self-assembled monolayer (SAM) of 12-mercaptododecanoic acid and 1-heptanethiol. More importantly, the number of immobilization bonds formed between each an-tigen-binding site protected antibody molecule and the solid surface was controlled by optimizing the mole fraction of the activated carboxyl group of the linker molecules in the mixed SAM. The amount of antibody used in this study was approximately equivalent to the amount for one monolayer surface coverage. The resulting activity of protected immo-bilized antibody was about 10 fold higher than that of random immobilized antibody.