| Autor: |
Xiaobin, Xu, Jessica Ann, O'Callaghan, Zachary, Guarnero, Haibo, Qiu, Ning, Li, Terra, Potocky, Douglas E, Kamen, Kenneth S, Graham, Mohammed, Shameem, Teng-Chieh, Yang |
| Rok vydání: |
2021 |
| Předmět: |
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| Zdroj: |
Biophysical journal. 121(6) |
| ISSN: |
1542-0086 |
| Popis: |
Protein glycation is a common, normally innocuous, post-translational modification in therapeutic monoclonal antibodies. However, when glycation occurs on complementarity-determining regions (CDRs) of a therapeutic monoclonal antibody, its biological activities (e.g., potency) may be impacted. Here, we present a comprehensive approach to understanding the mechanism of protein glycation using a bispecific antibody. Cation exchange chromatography and liquid chromatography-mass spectrometry were used to characterize glycation at a lysine residue within a heavy chain (HC) CDR (HC-CDR3-Lys98) of a bispecific antibody. Thermodynamic analysis revealed that this reaction is reversible and can occur under physiological conditions with an apparent affinity of 8-10 mM for a glucose binding to HC-CDR3-Lys98. Results from kinetic analysis demonstrated that this reaction follows Arrhenius behavior in the temperature range of 5°C-45°C and can be well predicted in vitro and in a non-human primate. In addition, this glycation reaction was found to be driven by an unusually low pK |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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