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Fluorescence Bioanalysis of Bevacizumab Using Pre-Column and Post-Column Derivatization – Liquid Chromatography After Immunoaffinity Magnetic Purification
https://gifu-pu.repo.nii.ac.jp/records/14308
https://gifu-pu.repo.nii.ac.jp/records/143081238c7f1-3165-4b8e-ab5d-ea26ad4ee423
Item type | 研究室原著論文(1) | |||||
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公開日 | 2021-03-08 | |||||
タイトル | ||||||
タイトル | Fluorescence Bioanalysis of Bevacizumab Using Pre-Column and Post-Column Derivatization – Liquid Chromatography After Immunoaffinity Magnetic Purification | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Fluorescence Bioanalysis of Bevacizumab Using Pre-Column and Post-Column Derivatization – Liquid Chromatography After Immunoaffinity Magnetic Purification | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Bevacizumab | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Fluorescence derivatization | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | High-temperature reversed-phase liquid chromatography | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Immunoaffinity purification | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Bioanalysis | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Bevacizumab | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Fluorescence derivatization | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | High-temperature reversed-phase liquid chromatography | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Immunoaffinity purification | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | Bioanalysis | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
アクセス権 | ||||||
アクセス権 | metadata only access | |||||
アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
抄録 | ||||||
値 | This report presents two fluorescence labeling methods for therapeutic monoclonal antibody, bevacizumab, to increase its detection sensitivity for fluorescence detection. One method is high-temperature reversed-phase LC (HT-RPLC) following post-column fluorogenic derivatization using o-phthalaldehyde with thiol. Another method is pre-column derivatization using Zenon Alexa Fluor 488 protein-tag following size-exclusion chromatography (SEC). The calibration curves of bevacizumab were 1–50 μg/mL (post-column method) and 0.1–10 μg/mL (pre-column method). Both methods showed good correlation coefficients (r2 > 0.991). The LOD and the LOQ of bevacizumab were, respectively, 0.13 and 0.43 μg/mL (post-column method) and 0.03 and 0.1 μg/mL (pre-column method). The sensitivities were about 2 and 10 times higher than that of native fluorescence detection. The proposed methods were applied to bevacizumab spiked human plasma samples. The bevacizumab in plasma samples was purified selectively with immunoaffinity beads and detected as a single peak using HT-RPLC or SEC with fluorescence detection. | |||||
書誌情報 |
CHROMATOGRAPHY en : CHROMATOGRAPHY 巻 41, 号 3, p. 115-122, 発行日 2020 |