Qu Wanlu, Glick Jim, Dessanti Paola, Cunliffe Jennifer
Biomedical Research, Novartis, Cambridge, MA 02139, USA.
Novartis Pharmaceuticals Corporation, East Hanover, NJ 07936, USA.
Bioanalysis. 2024;16(16):895-904. doi: 10.1080/17576180.2024.2394317. Epub 2024 Sep 4.
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. Intact mass analysis was applied to characterize a CR with complex glycosylation and biotinylation patterns. Peptide mapping was developed to identify the biotinylation sites.: Biotinylation degrees and sites were clearly illustrated. A CR and its biotinylation were successfully characterized. The relationship between biotinylation efficiency and labeling conditions was clearly illustrated.
关键试剂（CR）应用于配体结合分析（LBA），生物素化是一种广泛用于关键试剂的共轭方法。然而，表征不足和生物素化不一致会导致LBA失败，并需要进行大量的故障排除。本出版物开发了生物素化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以及在LBA中实施时的准确性。采用完整质量分析来表征具有复杂糖基化和生物素化模式的CR。开发了肽图谱以识别生物素化位点。生物素化程度和位点得到了清晰的说明。成功表征了一种CR及其生物素化。清楚地说明了生物素化效率与标记条件之间的关系。
