https://scholars.lib.ntu.edu.tw/handle/123456789/636828
標題: | Real-time bottom-up characterization of protein mixtures enabled by online microdroplet-assisted enzymatic digestion (MAED) | 作者: | Ma, Cheng-Hua Chen, Chih-Lin CHENG-CHIH HSU |
公開日期: | 19-十月-2023 | 卷: | 59 | 期: | 84 | 起(迄)頁: | 12585 | 來源出版物: | Chemical communications (Cambridge, England) | 摘要: | Enzymatic digestion remains one of the "rate-determining steps" in the bottom-up analysis of proteins. However, by performing digestion in microdroplets generated from electrosonic spray, the reaction could be accelerated to a timescale lower than milliseconds. Here, we describe a simple and rapid online digestion platform named online microdroplet-assisted enzymatic digestion (MAED). It involves the integration of intact protein separation with enzymatic digestion in microdroplets. Via online MAED, various protein standards, including an antibody standard, were characterized in a bottom-up manner without prior digestion, and high sequence coverages were obtained. We further extended the application of online MAED to a more complex sample, mouse brain extract, where protein identifications were successfully yielded. Compared with the conventional bottom-up approach, a more comprehensive characterization could be obtained particularly for low molecular weight proteins. In short, we provide a rapid and alternative bottom-up analysis in a top-down fashion as well as a new possibility for microdroplet chemistry. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/636828 | ISSN: | 13597345 | DOI: | 10.1039/d3cc03509c |
顯示於: | 化學系 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。