Simultaneous binding of mouse monoclonal antibody and streptavidin to heterobifunctional dendritic l-lysine core bearing T-antigen tumor marker and biotin Original Research Article

Thiolated T-antigen [Galβ-(1-3)-GalNAcα, T-Ag] (), derived in situ from thioacetate was coupled to N-chloroacetylated glycylglycyl l-lysine dendritic cores () using high yielding substitution reactions to afford di- (), tetra- (), and octa-valent (12) glycodendrimers in good yields (76–86%). Heterobifunctional conjugate was prepared as a biosensor from tetravalent conjugate and biotin hydrazide using TBTU strategy. In a solid-phase double sandwich enzyme linked immunosorbent assays (ELISA), biotinylated conjugate was shown to bind to streptavidin used as a coating material. Mouse monoclonal anti T-Ag antibody (IgG3) and horseradish peroxydase-labeled goat anti mouse IgG, used for quantification, were found to bind T-Ag tetramer immobilized on the surface of the streptavin layer. A typical saturation curve was observed for while non-biotinylated tetramer showed no binding in the entire concentration range. These results demonstrate the availability of both haptens toward the T-Ag antibody and streptavidin receptors.