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Impact of assay temperature on antibody binding characteristics in living cells: A case study
Ridgeview Instruments AB Uppsala.ORCID iD: 0000-0002-4509-4106
Charles Univ. Prague Czech Republic.
Karlstad University, Faculty of Health, Science and Technology (starting 2013), Department of Engineering and Chemical Sciences (from 2013).ORCID iD: 0000-0002-7123-2066
Ridgeview Instruments AB Uppsala; Uppsala universitet.
2017 (English)In: BIiomedical reports, ISSN 2049-9434, Vol. 7, no 5, p. 400-406Article in journal (Refereed) Published
Abstract [en]

Kinetic and thermodynamic studies of ligand-receptor interactions are essential for increasing the understanding of receptor activation mechanisms and drug behavior. The characterization of molecular interactions on living cells in real-time goes beyond most current binding assays, and provides valuable information about the dynamics and underlying mechanism of the molecules in a living system. The effect of temperature on interactions in cell-based assays is, however, rarely discussed. In the present study, the effect of temperature on binding of monoclonal antibodies, cetuximab and pertuzumab to specific receptors on living cancer cells was evaluated, and the affinity and kinetics of the interactions were estimated at selected key temperatures. Changes in the behavior of the interactions, particularly in the on- and off-rates were observed, leading to greatly extended time to reach the equilibrium at 21 degrees C compared with at 37 degrees C. However, the observed changes in kinetic characteristics were less than a factor of 10. It was concluded that it is possible to conduct real-time measurements with living cells at different temperatures, and demonstrated that influences of the ambient temperature on the interaction behavior are likely to be less than one order of magnitude.

Place, publisher, year, edition, pages
2017. Vol. 7, no 5, p. 400-406
National Category
Pharmaceutical Sciences
Identifiers
URN: urn:nbn:se:kau:diva-65945DOI: 10.3892/br.2017.982ISI: 000417416000002PubMedID: 29181152OAI: oai:DiVA.org:kau-65945DiVA, id: diva2:1177654
Available from: 2018-01-25 Created: 2018-01-25 Last updated: 2018-07-02Bibliographically approved

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Fornstedt, Torgny

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Encarnacao, Joao CrispimFornstedt, Torgny
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CiteExportLink to record
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