Probing the Kinetic and Thermodynamic Fingerprints of Anti-EGF Nanobodies by Surface Plasmon Resonance

Autor: Salvador Guardiola, Monica Varese, Marta Taulés, Mireia Díaz-Lobo, Jesús García, Ernest Giralt
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: Pharmaceuticals, Vol 13, Iss 6, p 134 (2020)
Druh dokumentu: article
ISSN: 1424-8247
DOI: 10.3390/ph13060134
Popis: Despite the widespread use of antibodies in clinical applications, the precise molecular mechanisms underlying antibody–antigen (Ab–Ag) interactions are often poorly understood. In this study, we exploit the technical features of a typical surface plasmon resonance (SPR) biosensor to dissect the kinetic and thermodynamic components that govern the binding of single-domain Ab or nanobodies to their target antigen, epidermal growth factor (EGF), a key oncogenic protein that is involved in tumour progression. By carefully tuning the experimental conditions and transforming the kinetic data into equilibrium constants, we reveal the complete picture of binding thermodynamics, including the energetics of the complex-formation transition state. This approach, performed using an experimentally simple and high-throughput setup, is expected to facilitate mechanistic studies of Ab-based therapies and, importantly, promote the rational development of new biological drugs with suitable properties.
Databáze: Directory of Open Access Journals
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