Yan Lu, Chao-Xuan Zhang, Patrick Rodrigues, Lei Yang, Aaron Shafer, Zoran Rankovic, Frank Fazio
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引用次数: 0
摘要
基于慢病毒载体的基因治疗在治疗各种疾病方面越来越受欢迎。圣犹达正在开发各种LV载体，用于在细胞基因治疗中转导细胞。一些LV载体是使用稳定的293T包装细胞系生产的，其中包括gag-pol-rev-tat和病毒糖蛋白。反式激活因子（transactuator of transcription[Tat]）是一种能显著提高慢病毒转录效率的调节蛋白。Tat的残留分析对基因载体的质量和安全性至关重要。在这项工作中，我们开发了一种高灵敏度的液相色谱-串联质谱法来分析慢病毒中残留的Tat，作为酶联免疫吸附测定的替代方法。左心室中残留的Tat可以高特异性准确定量，检测限为0.3 ng/mL。
Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for Sensitive Analysis of Residual Protein Tat Bh1-101 in Lentiviral Vectors for Gene Therapy.
Lentiviral (LV) vector-based gene therapy is gaining popularity for treating a wide range of diseases. Various LV vectors are being developed for transducing cells in cellular gene therapy at St. Jude. Some LV vectors are produced using stable 293T packaging cell lines, which includes gag-pol-rev-tat and virus-glycoprotein. Transactivating factor (transactivator of transcription [Tat]) is a regulatory protein that drastically increases the efficiency of lentiviral transcription. Residual analysis of Tat is critical for gene vector quality and safety. In this work, we developed a highly sensitive liquid chromatography-tandem mass spectrometry method for analysis of residual Tat in Lentivirus as an alternative to enzyme-linked immunosorbent assay. Residual Tat in LV can be accurately quantified with high specificity with a limit of detection of 0.3 ng/mL.
期刊介绍：
Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases.
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