Cold Spring Harb Protoc. 2020 Aug 3;2020(8):095679. doi: 10.1101/pdb.prot095679.
AAV virions are built from three major capsid proteins, VP1, VP2, and VP3, at a ratio of 1:1:18. On a silver-stained SDS-polyacrylamide gel, VP1, VP2, and VP3 should be the only visible bands in a highly purified recombinant adeno-associated virus (rAAV) preparation, migrating at approximately 87, 73, and 62 kDa, respectively. This protocol describes how SDS-PAGE and silver staining can be used to determine the purity of an rAAV preparation. In addition, using a highly purified rAAV preparation whose particle titer is known, this assay can be used to derive a semiquantitative estimate of the particle concentration of a test vector.
AAV 病毒粒子由三种主要衣壳蛋白 VP1、VP2 和 VP3 以 1:1:18 的比例构建而成。在银染 SDS-聚丙烯酰胺凝胶上，高度纯化的重组腺相关病毒（rAAV）制剂中仅应可见 VP1、VP2 和 VP3 三条带，分别迁移至约 87、73 和 62 kDa。本方案介绍了 SDS-PAGE 和银染如何用于确定 rAAV 制剂的纯度。此外，还可以使用已知颗粒滴度的高度纯化 rAAV 制剂，通过该测定法可以对半定量估计测试载体的颗粒浓度。
