論文 - 詳細
| RRC ID | 65133 |
|---|---|
| 著者 | Senga Y, Imamura H, Miyafusa T, Watanabe H, Honda S. |
| タイトル | AlphaScreen-based homogeneous assay using a pair of 25-residue artificial proteins for high-throughput analysis of non-native IgG. |
| ジャーナル | Sci Rep |
| Abstract |
Therapeutic IgG becomes unstable under various stresses in the manufacturing process. The resulting non-native IgG molecules tend to associate with each other and form aggregates. Because such aggregates not only decrease the pharmacological effect but also become a potential risk factor for immunogenicity, rapid analysis of aggregation is required for quality control of therapeutic IgG. In this study, we developed a homogeneous assay using AlphaScreen and AF.2A1. AF.2A1 is a 25-residue artificial protein that binds specifically to non-native IgG generated under chemical and physical stresses. This assay is performed in a short period of time. Our results show that AF.2A1-AlphaScreen may be used to evaluate the various types of IgG, as AF.2A1 recognizes the non-native structure in the constant region (Fc region) of IgG. The assay was effective for detection of non-native IgG, with particle size up to ca. 500 nm, generated under acid, heat, and stirring conditions. In addition, this technique is suitable for analyzing non-native IgG in CHO cell culture supernatant and mixed with large amounts of native IgG. These results indicate the potential of AF.2A1-AlphaScreen to be used as a high-throughput evaluation method for process monitoring as well as quality testing in the manufacturing of therapeutic IgG. |
| 巻・号 | 7(1) |
| ページ | 12466 |
| 公開日 | 2017-9-29 |
| DOI | 10.1038/s41598-017-12693-w |
| PII | 10.1038/s41598-017-12693-w |
| PMID | 28963557 |
| PMC | PMC5622108 |
| MeSH | Animals CHO Cells Cricetulus Drug Stability High-Throughput Screening Assays* Hot Temperature Humans Hydrogen-Ion Concentration Immunoglobulin Fc Fragments / chemistry* Immunoglobulin G / chemistry* Immunoglobulins, Intravenous / chemistry* Kinetics Peptides / chemical synthesis Peptides / chemistry* Protein Aggregates Protein Binding Quality Control Solutions |
| IF | 3.998 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 6 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | 293T(RCB2202) |