LC-MS/MS-BASED METHODS FOR CHARACTERIZING PROTEINS
收藏国家林业和草原科学数据中心2023-02-12 更新2024-03-07 收录
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The embodiments herein provide methods to analyze the in vitro stability of recalcitrant or membrane-bound proteins in simulated gastric fluid (SGF) comprising the proteolytic enzyme， pepsin， and in combination with a novel pepsin-trypsin assay employing state-of-the-art mass spectrometric approaches， such as LC-MS/MS， to monitor the precise degradation products. The extent of protein digestion can be evaluated by the appearance of peptic products and the disappearance of tryptic peptide products (as a proxy for intact protein). The embodiments herein also provide methods for protein quantitation using high-sensitivity LC-MRM-MS quantification. The methods embodied herein are particularly useful in charactering proteins produced in transgenic plants， such as canola genetically engineered to produce long chain omega-3 polyunsaturated fatty acids. The embodiments herein provide methods to analyze the in vitro stability of recalcitrant or membrane-bound proteins in simulated gastric fluid (SGF) comprising the proteolytic enzyme， pepsin， and in combination with a novel pepsin-trypsin assay employing state-of-the-art mass spectrometric approaches， such as LC-MS/MS， to monitor the precise degradation products. The extent of protein digestion can be evaluated by the appearance of peptic products and the disappearance of tryptic peptide products (as a proxy for intact protein). The embodiments herein also provide methods for protein quantitation using high-sensitivity LC-MRM-MS quantification. The methods embodied herein are particularly useful in charactering proteins produced in transgenic plants， such as canola genetically engineered to produce long chain omega-3 polyunsaturated fatty acids.
提供机构：
国家林业和草原科学数据中心
创建时间：
2023-02-12
