Wanlu Qu, Jim Glick, Paola Dessanti, Jennifer Cunliffe
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A comprehensive LC-MS based study of factors influencing biotinylation of critical reagents.
Aim: Critical reagents (CR) are applied in ligand binding assays (LBA) and biotinylation is a widely conjugation method used for critical reagents. However, insufficient characterization and inconsistent biotinylation can lead to LBA failures and necessitate extensive troubleshooting. This publication developed the detection of biotinylated CR and evaluates efficiency of biotinylation conditions to ensure the reliability of reagents and accuracy when implemented in LBA.Materials & methods: Intact mass analysis was applied to characterize a CR with complex glycosylation and biotinylation patterns. Peptide mapping was developed to identify the biotinylation sites.Results: Biotinylation degrees and sites were clearly illustrated.Conclusion: A CR and its biotinylation were successfully characterized. The relationship between biotinylation efficiency and labeling conditions was clearly illustrated.
期刊介绍：
Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance.
Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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