Phoenix Laboratory of Gene Therapy and Cell Therapy, Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, MD11, Basement 1, 10, Medical Drive, Singapore 117597, Singapore.
Infectious Diseases Translational Research Programme, Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117545, Singapore.
Int J Mol Sci. 2024 Jul 30;25(15):8342. doi: 10.3390/ijms25158342.
Adeno-associated viruses (AAVs) have emerged as promising tools for gene therapy due to their safety and efficacy in delivering therapeutic genes or gene editing sequences to various tissues and organs. AAV serotype 9 (AAV9), among AAV serotypes, stands out for its ability to efficiently target multiple tissues, thus holding significant potential for clinical applications. However, existing methods for purifying AAVs are cumbersome, expensive, and often yield inconsistent results. In this study, we explore a novel purification strategy utilizing Dynabeads™ CaptureSelect™ magnetic beads. The AAV9 magnetic beads capture AAV9 with high specificity and recovery between 70 and 90%, whereas the AAVX magnetic beads did not bind to the AAV9. Through continuous interaction with AAVs in solution, these beads offer enhanced clearance of genomic DNA and plasmids even in the absence of endonuclease. The beads could be regenerated at least eight times, and the used beads could be stored for up to six months and reused without a significant reduction in recovery. The potency of the AAV9-purified vectors in vivo was comparable to that of iodixanol purified vectors.
腺相关病毒 (AAV) 因其在将治疗性基因或基因编辑序列递送至各种组织和器官方面的安全性和有效性，已成为基因治疗的有前途的工具。在 AAV 血清型中，血清型 9 (AAV9) 因其能够有效地靶向多种组织而脱颖而出，因此具有重要的临床应用潜力。然而，现有的 AAV 纯化方法繁琐、昂贵，且常常产生不一致的结果。在这项研究中，我们探索了一种利用 Dynabeads™CaptureSelect™磁珠的新型纯化策略。AAV9 磁珠对 AAV9 具有高特异性和 70%至 90%的回收率，而 AAVX 磁珠则不与 AAV9 结合。通过与溶液中的 AAV 持续相互作用，这些珠子即使在没有内切酶的情况下，也能增强对基因组 DNA 和质粒的清除。这些珠子至少可以再生 8 次，并且使用过的珠子可以储存长达 6 个月，并且在不显著降低回收率的情况下可以重复使用。体内纯化的 AAV9 载体的效价与碘克沙醇纯化的载体相当。
Zotero 插件
