Identification of IgG1 Aggregation Initiation Region by Hydrogen Deuterium Mass Spectrometry
| Autor: | Tetsuo Torisu, Susumu Uchiyama, Kentaro Ishii, Takashi Tadokoro, Hiroaki Oyama, Katsumi Maenaka, Masanori Noda, Mika Yamauchi |
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| Rok vydání: | 2019 |
| Předmět: |
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Molecular Hydrogen Protein Conformation Pharmaceutical Science chemistry.chemical_element 02 engineering and technology Mass spectrometry 030226 pharmacology & pharmacy Mass Spectrometry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Humans Amino Acid Sequence Peptide sequence chemistry.chemical_classification Antibodies Monoclonal Deuterium Exchange Measurement Deuterium 021001 nanoscience & nanotechnology Fragment crystallizable region Amino acid Monomer chemistry Immunoglobulin G Biophysics Hydrogen–deuterium exchange 0210 nano-technology |
| Zdroj: | Journal of Pharmaceutical Sciences. 108:2323-2333 |
| ISSN: | 0022-3549 |
| DOI: | 10.1016/j.xphs.2019.02.023 |
| Popis: | Antibody aggregates are a potential risk for immunogenicity; therefore, rational approaches to improve associated aggregation properties need to be developed. Here, we report the amino acid region responsible for aggregation initiation. Two types of therapeutic IgG1 antibody monomer samples were prepared: IgG1 mAb40-3M stored at 40°C for 3 months, which existed in monodisperse state, and the monomer mAb65-5m, which was dissociated from small soluble aggregates by heating at 65°C for 5 min. Hydrogen deuterium exchange mass spectrometry of mAb40-3M identified 2 sites in the Fc region (site 1, F239-M256; site 2, S428-G450) with increased exchange rates. Site 1 includes a region reported as being susceptible to structural change induced by stress. Exposure of site 1 was undetected after 2 months of storage at 40°C but was subsequently detectable after 3 months. As site 2 is spatially close to site 1, the structural change of site 1 could propagate site 2. Besides these 2 regions, hydrogen deuterium exchange mass spectrometry of mAb65-5m identified an exposure of I257-W281 in Fc (site 3), within which a peptide sequence with high aggregation tendency was discovered. We thus concluded that exposure of site 3 is a trigger for the association of a partially denatured antibody. |
| Databáze: | OpenAIRE |
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