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Optimization of separation and digestion conditions in immune complexome analysis
http://hdl.handle.net/10069/33939
http://hdl.handle.net/10069/33939730a1603-9081-4ac5-9d3c-0712fee69374
名前 / ファイル | ライセンス | アクション |
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Table S1.pdf (59.5 kB)
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AnaBio443_181.pdf (1.3 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2013-11-05 | |||||
タイトル | ||||||
タイトル | Optimization of separation and digestion conditions in immune complexome analysis | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Gradient elution time | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Immune complexome analysis | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Microwave-assisted digestion | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Nano-liquid chromatography–tandem mass spectrometry | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Tryptic digestion | |||||
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 | |||||
著者 |
Baba, Miyako
× Baba, Miyako× Ohyama, Kaname× Kishikawa, Naoya× Kuroda, Naotaka |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Immune complexome analysis is a method for identifying and profiling of antigens in circulating immune complexes (CICs); it involves separation of immune complexes from serum, direct tryptic digestion of these complexes, and protein analysis via nano-liquid chromatography–tandem mass spectrometry (nano-LC–MS/MS). To improve this method, we initially investigated the effects of two factors—the gradient elution program and nano-LC column type (C18-packed, C8-packed, or packed spray capillary column)—on the numbers of peptides and proteins identified. Longer gradient elution times resulted in higher identification capability throughout the range of 25–400 min. Moreover, the packed spray capillary column supported identification of more peptides and proteins than did any other column. In addition, microwave-assisted digestion was compared with conventional digestion, which involved incubation overnight at 37 °C. Microwave-assisted digestion produced more partially digested peptides than did conventional digestion. However, the percentages of miscleaved peptides in all of the identified peptides in microwave-assisted digestion of immune complexes (a protein mixture) were lower than those in the physical stimulation-assisted digestion of a model protein. Microwave-assisted digestion is slightly inferior to, or as effective as, conventional digestion, but it drastically reduces the digestion time. | |||||
書誌情報 |
Analytical Biochemistry 巻 443, 号 2, p. 181-186, 発行日 2013 |
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出版者 | ||||||
出版者 | Elsevier Inc. | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 00032697 | |||||
DOI | ||||||
関連タイプ | isVersionOf | |||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1016/j.ab.2013.08.026 | |||||
権利 | ||||||
権利情報 | © 2013 Elsevier Inc. All rights reserved. | |||||
著者版フラグ | ||||||
出版タイプ | AM | |||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | Analytical Biochemistry, 443(2), pp.181-186; 2013 |