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Open AccessArticle

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

1
Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
2
Scientific and Technological Centres (CCiT-UB), Molecular Interactions, University of Barcelona, 08028 Barcelona, Spain
3
Mass Spectrometry and Proteomics Core Facility, Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain
4
Department of Inorganic and Organic Chemistry, University of Barcelona, 08028 Barcelona, Spain
*
Author to whom correspondence should be addressed.
Pharmaceuticals 2020, 13(6), 134; https://doi.org/10.3390/ph13060134
Received: 2 June 2020 / Revised: 22 June 2020 / Accepted: 24 June 2020 / Published: 26 June 2020
(This article belongs to the Section Pharmaceutical Technology)
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. View Full-Text
Keywords: nanobodies; SPR; biophysics; thermodynamics; molecular recognition; EGF; antibody–antigen (Ag–Ab) interactions nanobodies; SPR; biophysics; thermodynamics; molecular recognition; EGF; antibody–antigen (Ag–Ab) interactions
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MDPI and ACS Style

Guardiola, S.; Varese, M.; Taulés, M.; Díaz-Lobo, M.; García, J.; Giralt, E. Probing the Kinetic and Thermodynamic Fingerprints of Anti-EGF Nanobodies by Surface Plasmon Resonance. Pharmaceuticals 2020, 13, 134.

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