Surface Plasmon Resonance (SPR) as a Tool for Antibody Conjugate Analysis

Abstract

Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin (4) and anti-fluorescein (5) monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1. Only minor changes in the apparent equilibrium dissociation constants of the antibody conjugates for their ligands resulted from the conjugation process. However, comparison of the initial binding rate of the conjugates with their ligands with those of the unmodified antibodies over a range of concentrations showed that the antibody conjugates were partially inactivated. The anti-fluorescein conjugates retained at least 90% of their immunoreactivity over the range of modification tested, while anti-biotin conjugates showed a progressive loss of reactivity with increased substitution by the label.

Molecular Recognition of Avidin on Biotin-Functionalized Gold Surfaces Detected by FT-IRRAS and Use of Metal Carbonyl Probes

“…Previously described methods of formation of SAMs of biotin molecules on gold or silicon substrates consisted either in the covalent coupling to previously chemisorbed ω-functionalized thiolates (19)(20)(21)(22) or in the direct chemisorption of biotinylated thiols or disulfides (23,24). Mixed biotinylated SAMs, consisting in a biotinylated alkylthiol coadsorbed with an hydroxy-terminated second thiol, have also been successfully utilized to monitor the structure and order of the reactive adsorbed layer (25)(26)(27).…”

Polyelectrolyte nanogels decorated with monoclonal antibody for targeted drug delivery

“…The SPR technique has been used extensively to analyze antibody-antigen binding [51]. To determine the specificity of CC49-nanogel interaction, BSA coated sensor chip and IgG-nanogels were used as controls [31].…”

Quantitative study of stereospecific binding of monoclonal antibody to anti-benzo(a)pyrene diol epoxide-N2-dG adducts by capillary electrophoresis immunoassay

“…As B binds independently to P 1 and P 2 , the binding constants (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14) and (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15) can be obtained: …”

“…(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) shows that Eq. (3-17) is consistent with the definition of binding constant of B and P 1 , as shown in the Eq.…”

“…Qualitative and quantitative study of such binding is essential not only to the understanding of the molecular basis for immune functions, and also important to the method development of immunoassays. Various techniques have been developed for binding study, such as enzyme-linked immunosorbent assay (ELISA) [2][3][4], surface plasmon resonance (SPR) [5][6][7], gel electrophoresis mobility shift assay (EMSA) [8,9], and affinity chromatography [10,11]. Most immunoassays heavily rely on the adsorption of antigen or antibody on solid/liquid surface, e.g.…”