Photodisruption of the Structurally Conserved Cys-Cys-Trp Triads Leads to Reduction-Resistant Scrambled Intrachain Disulfides in an IgG1 Monoclonal Antibody
| Autor: | Paula Lee Tao, Aaron T. Wecksler, Alavattam Sreedhara, Galahad Deperalta, Jian Yin, Bruce Kabakoff |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
medicine.drug_class Stereochemistry Size-exclusion chromatography Population Acrylic Resins Pharmaceutical Science Monoclonal antibody 030226 pharmacology & pharmacy Mass Spectrometry 03 medical and health sciences Electron transfer 0302 clinical medicine Drug Discovery medicine Amino Acid Sequence Cysteine Disulfides Photodegradation education education.field_of_study Photolysis Chemistry Disulfide bond Tryptophan Antibodies Monoclonal Dipeptides 030104 developmental biology Immunoglobulin G Molecular Medicine Photochemical degradation Oxidation-Reduction |
| Zdroj: | Molecular pharmaceutics. 15(4) |
| ISSN: | 1543-8392 |
| Popis: | Photostability conditions as prescribed by ICH guidelines induced highly reduction-resistant scrambled disulfides that contribute to the population of apparent nonreducible aggregates in an IgG1 mAb. Photoinduced cross-linked species were isolated under reducing conditions using an organic phase size exclusion chromatography (OP-SEC) method, followed by O18-labeling tryptic mapping to identify cross-linked peptides. Disulfide scrambling was observed within the IgG1 structurally conserved-intrachain cysteine-cysteine-tryptophan triads (Cys-Cys-Trp), and correlated with Trp-to-kynurenine (Kyn) photodegradation within these triads. We hypothesize that intrachain disulfides protect the proximal Trp within the Cys-Cys-Trp triads from photodegradation by enabling dissipation of Trp-absorbed UV energy via electron transfer to the disulfide bond. Finally, we propose three distinct mechanisms of photochemical degradation of monoclonal antibodies mediated by Trp residues. |
| Databáze: | OpenAIRE |
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