Simultaneous binding of mouse monoclonal antibody and streptavidin to heterobifunctional dendritic L-lysine core bearing T-antigen tumor marker and biotin.
Thiolated T-antigen [Galbeta-(1-3)-GalNAcalpha, T-Ag] (6), derived in situ from thioacetate 5 was coupled to N-chloroacetylated glycylglycyl L-lysine dendritic cores (7-9) using high yielding substitution reactions to afford di- (10), tetra- (11), and octa-valent (12) glycodendrimers in good yields (76-86%). Heterobifunctional conjugate 14 was prepared as a biosensor from tetravalent conjugate 11 and biotin hydrazide 13 using TBTU strategy. In a solid-phase double sandwich enzyme linked immunosorbent assays (ELISA), biotinylated conjugate 14 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 14 immobilized on the surface of the streptavin layer. A typical saturation curve was observed for 14 while non-biotinylated tetramer 11 showed no binding in the entire concentration range. These results demonstrate the availability of both haptens toward the T-Ag antibody and streptavidin receptors.
Baek
M G
MG
Department of Chemistry, Centre for Research in Biopharmaceuticals, University of Ottawa, Ottawa, K1N 6N5, Ontario, Canada.
Roy
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000957:Antigens, Viral, Tumor / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D001665:Binding Sites; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006001:Glycoconjugates / Q000737:chemistry* / Q000378:metabolism; D008239:Lysine / Q000737:chemistry; D051379:Mice; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000037:antagonists & inhibitors; D010980:Platelet Membrane Glycoproteins; D019809:Streptavidin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000957:Antigens, Viral, Tumor; D006001:Glycoconjugates; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; C032193:glycoprotein receptor GPIb-IX; D001710:Biotin; D019809:Streptavidin; D008239:Lysine