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Integrated Top-Down and Bottom-Up Mass Spectrometry for Characterization of Diselenide Bridging Patterns of Synthetic Selenoproteins
Analytical Chemistry ( IF 7.4 ) Pub Date : 2022-08-05 , DOI: 10.1021/acs.analchem.2c01433
Eleanor Watts 1 , Ross Thyer 2 , Andrew D Ellington 3 , Jennifer S Brodbelt 1
Affiliation  

With the rapid acceleration in the design and development of new biotherapeutics, ensuring consistent quality and understanding degradation pathways remain paramount, requiring an array of analytical methods including mass spectrometry. The incorporation of non-canonical amino acids, such as for synthetic selenoproteins, creates additional challenges. A comprehensive strategy to characterize selenoproteins should serve dual purposes of providing sequence confirmation and mapping of selenocysteine bridge locations and the identification of unanticipated side products. In the present study, a combined approach exploiting the benefits of both top-down and bottom-up mass spectrometry was developed. Both electron-transfer/higher-energy collision dissociation and 213 nm ultraviolet photodissociation were utilized to provide complementary information, allowing high quality characterization, localization of diselenide bridges for complex proteins, and the identification of previously unreported selenoprotein dimers.

中文翻译:

用于表征合成硒蛋白的二硒化物桥接模式的集成自上而下和自下而上质谱

随着新生物治疗药物设计和开发的快速加速,确保一致的质量和了解降解途径仍然至关重要,这需要包括质谱在内的一系列分析方法。非规范氨基酸的掺入,例如合成硒蛋白,带来了额外的挑战。表征硒蛋白的综合策略应具有双重目的,即提供序列确认和硒代半胱氨酸桥位置的定位以及意外副产物的鉴定。在本研究中,开发了一种利用自上而下和自下而上质谱法优点的组合方法。电子转移/高能碰撞解离和 213 nm 紫外光解离都被用来提供补充信息,
更新日期:2022-08-05
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