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Updated: Dec 21, 2025
Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
Published on: April 17, 2017
Kinetic and thermodynamic study of c-Met interaction with single chain fragment variable (scFv) antibodies using
Farzaneh Ghorbani1, Farzaneh Fathi2, Leili Aghebati-Maleki3
1Stem Cell and Regenerative Medicine Institute (SCARM), Tabriz University of Medical Sciences, Tabriz, Iran; Student research committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract:
Mesenchymal epithelial transition factor (c-Met) has been recently regarded as an attractive target for the treatment of cancer. Our previous study showed that c-Met-specific single chain fragment variables (scFvs) can be considered as a promising therapy for cancer, however, their molecular interaction with c-Met protein have not been assessed. Accordingly, in the current study we aim to evaluate the kinetic and thermodynamic properties of c-Met interaction with these scFvs as anticancer agents by means of surface plasmon resonance (SPR) technique. Phage-scFvs were immobilized on the 11-mercaptoundecanoic acid gold chips after carboxylic groups activation by N-ethyl-N-(3-diethylaminopropyl) carbodiimide/N-hydroxysuccinimide and, then the c-Met binding to each scFvs (ES1, ES2, and ES3) at different concentrations (ranging from 20 to 665 μM) was explored. Kinetic studies revealed that ES1 has the highest affinity (KD = 3.36 × 10-8) toward its target at 25°C. Calculation of thermodynamic parameters also showed positive values for enthalpy and entropy changes, which was representative of hydrophobic forces between c-Met and ES1. Furthermore, the positive value of Gibbs free energy indicated that c-Met binding to ES1 was enthalpy-driven. Taken together, we concluded that produced ES1 can be applied as promising scFv-based therapy for diagnosis or targeting of c-Met in various cancers.
Insights
The study evaluated the molecular interaction of c-Met specific single chain fragment variables (scFvs) as cancer therapies. ES1 demonstrated the highest affinity for c-Met, indicating its potential for cancer diagnosis and treatment.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- The mesenchymal epithelial transition factor (c-Met) is a key target in cancer therapy.
- Previous research identified c-Met-specific single chain fragment variables (scFvs) as potential cancer treatments.
- The molecular interactions of these scFvs with c-Met required detailed assessment.
Purpose of the Study:
- To evaluate the kinetic and thermodynamic properties of c-Met interaction with scFvs.
- To assess the potential of scFvs as anticancer agents through molecular interaction analysis.
- To determine the binding characteristics of specific scFvs (ES1, ES2, ES3) to c-Met.
Main Methods:
- Surface Plasmon Resonance (SPR) technique was employed to analyze molecular interactions.
- Phage-scFvs were immobilized on gold chips for binding assays.
- Binding of varying concentrations of c-Met to immobilized scFvs was measured.
Main Results:
- The scFv ES1 exhibited the highest binding affinity to c-Met, with a KD of 3.36 × 10-8 at 25°C.
- Thermodynamic analysis indicated that the binding of c-Met to ES1 is driven by hydrophobic forces and is enthalpy-driven.
- Positive enthalpy and entropy changes were observed, characteristic of hydrophobic interactions.
Conclusions:
- The scFv ES1 shows significant potential as an anticancer therapeutic agent.
- ES1 can be utilized for the diagnosis or targeted therapy of c-Met in various cancers.
- The characterized molecular interactions provide a basis for further development of scFv-based cancer treatments.
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