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Updated: May 19, 2026
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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
Published on: April 17, 2017
Probing cocaine-antibody interactions in buffer and human serum.
Muthu Ramakrishnan1, Fernando Alves De Melo, Berma M Kinsey
1Veterans Affairs Medical Center, Houston, Texas, United States of America.
Plos One
|August 4, 2012
Summary
Antibody binding to cocaine is significantly affected by human serum, impacting immunotherapy effectiveness. Understanding these interactions is crucial for developing better cocaine vaccines and immunization strategies.
Area of Science:
- Immunology
- Pharmacology
- Biochemistry
Background:
- Cocaine immunotherapy progress is hindered by poorly understood antibody-cocaine interactions.
- The influence of biological matrices like human serum on these interactions remains unclear.
Purpose of the Study:
- To characterize the kinetic and thermodynamic properties of antibodies binding to cocaine and its metabolites.
- To evaluate how these interactions differ in human serum compared to buffer.
- To assess the affinity of monoclonal and polyclonal antibodies in complex biological fluids.
Main Methods:
- Microscale thermophoresis (MST) to measure antibody-serum interactions.
- Isothermal titration calorimetry (ITC) to determine binding affinity.
- Surface plasmon resonance (SPR) to assess binding kinetics and competition.
Main Results:
- Antibody affinity for cocaine decreased significantly in human serum.
- Polyclonal antibodies showed preferential binding to cocaine over its metabolite.
- A binding model was developed to simulate immunotherapy challenges.
Conclusions:
- Calorimetric and MST methods provide valuable insights into antibody-cocaine interactions and thermodynamics.
- Affinity measurements in biological fluids are essential for designing effective cocaine vaccines.
- Wider adoption of these characterization methods is recommended.
