Due to the rise of adeno-associated viruses (AAVs) as gene therapy delivery vectors, boundary sedimentation velocity analytical ultracentrifugation (boundary SV-AUC) has been developed into a widely used quality control assay even for release analytics. It can be considered as the “gold standard” for the determination of the loading status of empty, partially filled, and full capsids especially when conducted in multiwavelength (MWL) mode. It can be considered to provide the most accurate determination of the loading status, and it also provides information on the capsid titer, aggregates, and potential contaminants such as free DNA. MWL boundary SV-AUC can be regarded as a multi-attribute (MAM) method for the characterization of AAVs. One major drawback of the method is the high sample consumption both in terms of concentration and volume. Here, we compare two alternative AUC techniques, band SV-AUC and analytical CsCl density gradient sedimentation equilibrium AUC (CsCl SE-AUC) with the boundary SV-AUC and the MWL-SV-AUC experiment. Our data show a high consistency of the determined full/empty ratios between these techniques if the appropriate wavelengths and extinction coefficients are used.

由于腺相关病毒 (AAV) 作为基因治疗递送载体的兴起，边界沉降速度分析超速离心（边界 SV-AUC）已发展成为广泛使用的质量控制测定，甚至用于释放分析。它可以被认为是确定空、部分填充和完整衣壳加载状态的“金标准”，特别是在多波长 (MWL) 模式下进行时。它可以被认为提供了最准确的加载状态测定，并且还提供了有关衣壳滴度、聚集体和潜在污染物（例如游离 DNA）的信息。 MWL边界SV-AUC可以被视为一种用于表征AAV的多属性（MAM）方法。该方法的一个主要缺点是浓度和体积方面的样品消耗量较高。在这里，我们比较了两种替代 AUC 技术，带 SV-AUC 和分析 CsCl 密度梯度沉降平衡 AUC (CsCl SE-AUC) 与边界 SV-AUC 和 MWL-SV-AUC 实验。我们的数据显示，如果使用适当的波长和消光系数，这些技术之间确定的满/空比具有高度一致性。