A comprehensive LC-MS based study of factors influencing biotinylation of critical reagents.

IF 1.9 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2024-09-04 DOI:10.1080/17576180.2024.2394317

Wanlu Qu, Jim Glick, Paola Dessanti, Jennifer Cunliffe

引用次数: 0

Abstract

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.

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基于 LC-MS 的关键试剂生物素化影响因素综合研究。

目的：关键试剂（CR）应用于配体结合测定（LBA），而生物素化是一种广泛用于关键试剂的连接方法。然而，表征不充分和生物素化不一致会导致 LBA 失败，需要进行大量的故障排除工作。本论文开发了生物素化 CR 的检测方法，并评估了生物素化条件的效率，以确保试剂的可靠性和在 LBA 中实施时的准确性：应用完整质量分析鉴定具有复杂糖基化和生物素化模式的CR。结果：生物素酰化程度和位点均发生了变化：结果：生物素化程度和位点清晰可见：结论：成功鉴定了 A CR 及其生物素化。结论：成功鉴定了一种 CR 及其生物素化特征，明确了生物素化效率与标记条件之间的关系。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.