生物质谱技术在腺相关病毒(AAV)载体制剂质量控制中的应用  

Application of biological mass spectrometry in quality control of adeno-asso⁃ciated virus carrier preparations

作　　者：李孟效 李惠琳 LI Mengxiao;LI Huilin(School of Pharmaceutical Science,Sun Yat-Sen University,Guangzhou 510006,China)

机构地区：[1]中山大学药学院,广州510006

出　　处：《中国药科大学学报》2023年第6期682-694,共13页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81872836,No.91953102);广东省自然科学基金资助项目(No.2019A1515011265)。

摘　　要：腺相关病毒(adeno-associated virus,AAV)是一种基因治疗中常用的病毒载体。由于其安全性较高且能够靶向多种细胞,在临床前和临床研究中得到了较多的应用。不过在设计和生产的过程中,AAV载体有着诸多会影响其安全性和疗效的关键质量属性。生物质谱技术的发展和应用为生物大分子的研究提供了一个便捷的平台,尤其是在蛋白质序列、结构和相互作用方面。对于AAV载体而言,质谱技术可以实现衣壳蛋白比率、翻译后修饰、血清型、空衣壳比率的测定或表征,从而协助对AAV载体的质量控制。与现有方法相比,质谱技术具有样品需求量少、分析快速灵敏、适用于完整AAV载体的分析和质量分辨率高的优点,并且可以区分空衣壳、满衣壳和部分包封的衣壳。未来,通过将更加高效的蛋白质分离技术与质谱技术联用、开发新的信息处理软件平台和新的质谱检测方法,质谱技术有望在AAV载体的设计和生产中发挥更加重要的作用。Adeno-associated virus(AAV)is a common viral vector used in gene therapy.Because of its high safety and its ability to target a variety of cells,it has been widely used in preclinical and clinical studies.However,during the design and production,AAV vectors have many key quality attributes that affect their safety and efficacy.The development and application of biological mass spectrometry technology provides a convenient platform for the research on biological macromolecules,especially in the aspects of protein sequence,structure and interaction.For AAV vectors,mass spectrometry can facilitate the determination or characterization of capsid protein ratio,post-translational modification,serotype,and empty capsid ratio,thus assisting in the quality control of AAV vectors.Compared with the existing methods,mass spectrometry has the advantages of smaller amount of sample size,faster and more sensitive analysis,being more suitable for the analysis of complete AAV vectors with higher mass resolution,and can distinguish empty capsids,full capsids and partial capsids.In the future,mass spectrometry technology is expected to play a more important role in the design and production of AAV vectors through the coupling of more efficient protein separation technology with mass spectrometry,the development of new information processing software platforms and new mass spectrometry detection techniques.

关 键 词：腺相关病毒 质谱 质量控制 非变性质谱 电荷检测质谱 

分 类 号：R392[医药卫生—免疫学]