Cold Spring Harb Protoc. 2020 Aug 3;2020(8):095604. doi: 10.1101/pdb.prot095604.
Centrifugation to equilibrium in cesium chloride gradients has been used for more than 40 yr to purify viruses. The application of high G-forces for a long period of time to a solution of CsCl generates a density gradient that allows separation of empty, partially packaged, and fully packaged viral particles from cellular debris, proteins, and nucleic acids in the crude viral lysate on the basis of their buoyant densities. This protocol describes the use of CsCl gradients to purify AAV vectors from crude viral lysates.
离心平衡氯化铯梯度法已被用于病毒纯化 40 余年。在 CsCl 溶液中长时间施加高离心力会产生密度梯度,根据其漂浮密度,可将空的、部分包装的和完全包装的病毒颗粒与粗病毒裂解液中的细胞碎片、蛋白质和核酸分离。本方案描述了使用 CsCl 梯度从粗病毒裂解液中纯化 AAV 载体。