A master thesis from Aalborg University
Characterization of Antibody Interaction Using Surface Plasmon Resonance for the Application in Nanoparticle-Based Immunoassays
Author(s)
Term
4. term
Education
Publication year
2017
Submitted on
2017-06-12
Pages
102 pages
Abstract
During this study recombinant dengue type 2 NS1 interactions with a pair of free and nanoparticle conjugated monoclonal antibodies were investigated through kinetic and sandwich analysis using surface plasmon resonance. The antibodies were covalently immobilized using amine coupling method to measure binding events. NS1 association, dissociation kinetics and affinities were evaluated with both antibodies. The inconsistent results indicated the non optimal conditions for the analysis. Kinetics of one antibody, Mer 39, were estimated in a sandwich type analysis, resulting in association constant of 1.13e5 1/M s, dissociation constant of 1.23e-4 1/s and affinity constant of 1.09e-9 M. This type of analysis was used to test NS1 epitope overlap to the said couple of antibodies as well. The investigated antigen did not have the overlapping epitopes to the particular pair. However, the interaction was possible only under specific complex formation sequence. Further research needs to be done to explore the suitability of surface plasmon resonance technique for a comprehensive characterization of nanoparticle conjugated antibodies interaction with the antigen.
Documents
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