采用 UltraScan 多波长分析超速离心技术对腺相关病毒衣壳的装载状态进行表征和定量分析。
Characterization and quantification of adeno-associated virus capsid-loading states by multi-wavelength analytical ultracentrifugation with UltraScan.
机构信息
Department of Chemistry & Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
Division of Biology & Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
出版信息
Nanomedicine (Lond). 2023 Sep;18(22):1519-1534. doi: 10.2217/nnm-2023-0156. Epub 2023 Oct 25.
We present multi-wavelength (MW) analytical ultracentrifugation (AUC) methods offering superior accuracy for adeno-associated virus characterization and quantification. Experimental design guidelines are presented for MW sedimentation velocity and analytical buoyant density equilibrium AUC. Our results were compared with dual-wavelength AUC, transmission electron microscopy and mass photometry. In contrast to dual-wavelength AUC, MW-AUC correctly quantifies adeno-associated virus capsid ratios and identifies contaminants. In contrast to transmission electron microscopy, partially filled capsids can also be detected and quantified. In contrast to mass photometry, first-principle results are obtained. Our study demonstrates the improved information provided by MW-AUC, highlighting the utility of several recently integrated UltraScan programs, and reinforces AUC as the gold-standard analysis for viral vectors.
我们提出了多波长（MW）分析超速离心（AUC）方法，为腺相关病毒的表征和定量提供了更高的准确性。 为 MW 沉降速度和分析浮力平衡 AUC 提出了实验设计指南。 我们的结果与双波长 AUC、透射电子显微镜和质谱法进行了比较。 与双波长 AUC 相比，MW-AUC 可正确定量腺相关病毒衣壳比并识别污染物。 与透射电子显微镜相比，也可以检测和定量部分填充的衣壳。 与质光法相比，获得了第一性原理结果。 我们的研究表明，MW-AUC 提供了改进的信息，突出了最近集成的几个 UltraScan 程序的实用性，并强化了 AUC 作为病毒载体的金标准分析。
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