Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Orban, M; Katerkamp, A; Renneberg, R; Spener, F; Cammann, K.
Kinetic analysis of immunointeractions with covalently immobilized fatty acid-binding protein using a grating coupler sensor.
J Immunol Methods. 1998; 215(1-2):17-26
Doi: 10.1016/S0022-1759(98)00042-8
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- Co-Autor*innen der Med Uni Graz
- Spener Friedrich
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- Abstract:
- Application of a grating coupler sensor (GCS) to the real time investigation of the interaction kinetics of covalently immobilized recombinant bovine heart-type fatty acid-binding protein (H-FABP) and corresponding antibody is described. The immobilization of the antigen is performed by activating the matrix hydroxyl groups with p-toluenesulfonyl chloride (TSC) and afterwards coupling the protein by reaction with its nucleophilic aminogroups. Covalent coupling via TSC permits reproducible measurements of immunointeractions on the same grating coupler sensor chip and complete regeneration after each binding cycle with glycine-hydrochloride. We demonstrate the analysis of binding data obtained on a GCS by linearization as well as direct curve fitting using the integrated rate equation for the determination of apparent rate and affinity constants. With both analysis methods we studied H-FABP/monoclonal anti-H-FABP-antibody interactions and obtained an average apparent association rate constant ka = 4.2 X 10(3) M(-1) s(-1) a dissociation rate constant of kd=1.3 X 10(-4) s(-1) and an equilibrium constant of KD=3 X 10(-8) M.
- Find related publications in this database (using NLM MeSH Indexing)
- Animals -
- Antibodies, Monoclonal - immunology Antibodies, Monoclonal - metabolism
- Antigen-Antibody Reactions -
- Biosensing Techniques -
- Carrier Proteins - immunology Carrier Proteins - metabolism
- Cattle -
- Fatty Acid-Binding Proteins -
- Kinetics -
- Myelin P2 Protein - immunology Myelin P2 Protein - metabolism
- Neoplasm Proteins -
- Find related publications in this database (Keywords)
- grating coupler sensor
- heart fatty acid-binding protein
- antigen-antibody interaction
- biosensor
- kinetic analysis