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Impact of assay tem...
Impact of assay temperature on antibody binding characteristics in living cells : A case study
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- Encarnacao, Joao Crispim (author)
- Ridgeview Instruments AB, Skillsta 4, S-74020 Uppsala, Sweden.
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- Barta, Pavel (author)
- Charles Univ Prague, Fac Pharm, Dept Biophys & Phys Chem, Hradec Kralove 50005, Czech Republic.
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- Fornstedt, Torgny, 1957- (author)
- Karlstads universitet,Institutionen för ingenjörs- och kemivetenskaper (from 2013)
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- Andersson, Karl, 1972- (author)
- Uppsala universitet,Medicinsk strålningsvetenskap,Ridgeview Instruments AB, Skillsta 4, S-74020 Uppsala, Sweden,Ridgeview Instruments AB Uppsala; Uppsala universitet
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Ridgeview Instruments AB, Skillsta 4, S-74020 Uppsala, Sweden Charles Univ Prague, Fac Pharm, Dept Biophys & Phys Chem, Hradec Kralove 50005, Czech Republic. (creator_code:org_t)
- 2017-09-14
- 2017
- English.
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In: BIOMEDICAL REPORTS. - : Spandidos Publications. - 2049-9434 .- 2049-9442. ; 7:5, s. 400-406
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Abstract
Subject headings
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- 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.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Farmaceutiska vetenskaper (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Pharmaceutical Sciences (hsv//eng)
Keyword
- drug kinetics
- thermodynamics
- real-time interactions
- clinical monoclonal antibodies
- growth factor receptors
- Kemi
Publication and Content Type
- ref (subject category)
- art (subject category)
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