Flow field-flow fractionation: new method for separating, purifying, and characterizing the diffusivity of viruses
Flow field-flow fractionation: new method for separating, purifying, and characterizing the diffusivity of viruses
复制标题
流场流分级分离：分离、纯化和表征病毒扩散性的新方法
作者：
J. Giddings;F. Yang;M. N. Myers
描述了流场流分级分离的性质和理论，并讨论了其对病毒样颗粒的潜在适用性。不同的病毒类型被证明保留在不同的水平。保留可以通过实验参数的变化来控制，与理论非常一致。然而，表明了温和的吸附效应，需要开发测量扩散系数的替代策略。对于 Qbeta，我们的值与文献值的吻合度在 10% 以内；使用 Abeta 作为内标获得的其他病毒的值未经测试。最后，证明流场流分离可以干净地分离两种病毒，以及从白蛋白杂质中分离出两种病毒，可以分析大至几毫克的样品，并且该方法在病毒系统的定量和定性分析中具有潜在的实用性。
The nature and theory of flow field-flow fractionation is described, and its potential applicability to virus-like particles is discussed. Different virus types are shown to be retained at different levels. Retention can be controlled by variation of the experimental parameters, in good agreement with theory. However, a mild adsorption effect is indicated and requires the development of alternate strategies for measuring diffusion coefficients. For Qbeta, our value agrees well within 10% of literature values; the values obtained for other viruses, using Abeta as an internal standard, are untested. Finally, it is demonstrated that flow field-flow fractionation can cleanly fractionate two viruses from one another and from an albumin impurity, that samples as large as several milligrams in size can be analyzed, and that the method has potential utility in the quantitative and qualitative analysis of virus systems.