Understanding viral partitioning in two-phase aqueous nonionic micellar systems: 2. Effect of entrained micelle-poor domains.
Understanding viral partitioning in two-phase aqueous nonionic micellar systems: 2. Effect of entrained micelle-poor domains.
复制标题
了解两相水性非离子胶束系统中的病毒分配：2. 夹带的胶束贫乏结构域的影响。
DOI：
10.1002/bit.10194
复制
发表时间：
2002
影响因子：
3.8
通讯作者：
Blankschtein,Daniel
中科院分区：
文献类型：
--
作者：
Kamei,DanielT;King,JonathanA;Wang,DanielIC;Blankschtein,Daniel
与亲水性、水溶性蛋白质的分配行为不同，病毒在两相非离子癸基四（环氧乙烷）（C10 E4）胶束体系中的分配行为不能用我们小组最近开发的排除体积理论完全解释。因此，排除体积理论的核心假设-宏观相分离平衡的实现-在实验和理论上受到挑战。拍摄了不同体积比的两相水性C10 E4胶束系统的照片，以证明宏观、顶部、胶束富集相中的胶束贫乏（病毒富集）域的夹带随着体积比的降低而降低。然后用模型病毒噬菌体P22和模型蛋白质细胞色素在不同的操作温度和不同的体积比下进行分配实验。对于噬菌体P22，当体积比从10降低到0.1时，在每个温度下测得的病毒分配系数降低约一个数量级，这清楚地表明夹带是影响病毒分配的重要因素。对于cytochromec，测得的蛋白质分配系数没有变化，这表明这种夹带效应对蛋白质分配的影响可以忽略不计。还开发了一种新的分配理论描述，将排除体积理论与夹带效应相结合。在该理论中，一个拟合参数-在宏观、顶部、胶束富集相中夹带的胶束贫乏域的体积分数-用于解释夹带。为了拟合该参数，对于给定的体积比，仅需要单个分配实验，而与分配溶质无关。分配的新的理论描述产生了很好的定量预测的病毒分配系数。因此，可以得出结论，控制两相水性C10 E4胶束系统中病毒分配的主要机制是宏观、顶部、胶束富集相中胶束贫乏（病毒富集）域的夹带以及病毒和胶束之间的排斥体积相互作用。© 2002 Wiley Periodicals，Inc. Biotechnol Bioeng 78：203-216，2002; DOI 10.1002/bit.10194
Unlike the partitioning behavior of hydrophilic, water‐soluble proteins, the partitioning behavior of viruses in the two‐phase aqueous nonionicn‐decyl tetra(ethylene oxide) (C10E4) micellar system cannot be fully explained using the excluded‐volume theory developed recently by our group. A central assumption underlying the excluded‐volume theory—that macroscopic phase separation equilibrium is attained—was therefore challenged experimentally and theoretically. Photographs of the two‐phase aqueous C10E4micellar system were taken for different volume ratios to demonstrate that the entrainment of micelle‐poor (virus‐rich) domains in the macroscopic, top, micelle‐rich phase decreases with a decrease in the volume ratio. Partitioning experiments were then conducted with the model virus bacteriophage P22 and the model protein cytochromecat different operating temperatures for different volume ratios. For bacteriophage P22, the measured viral partition coefficient at each temperature decreased by about an order of magnitude when the volume ratio was decreased from 10 to 0.1, which clearly indicated that entrainment is an important factor influencing viral partitioning. For cytochromec, the measured protein partition coefficient did not change, which demonstrated that this entrainment effect negligibly influences protein partitioning. A new theoretical description of partitioning was also developed that combines the excluded‐volume theory with this entrainment effect. In this theory, one fitted parameter—the volume fraction of entrained micelle‐poor domains in the macroscopic, top, micelle‐rich phase—is used to account for the entrainment. To fit this parameter, only a single partitioning experiment is required for a given volume ratio, irrespectively of the partitioning solute. The new theoretical description of partitioning yielded very good quantitative predictions of the viral partition coefficients. Accordingly, it can be concluded that the primary mechanisms governing viral partitioning in the two‐phase aqueous C10E4micellar system are the entrainment of micelle‐poor (virus‐rich) domains in the macroscopic, top, micelle‐rich phaseandthe excluded‐volume interactions that operate between the viruses and the micelles. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 203–216, 2002; DOI 10.1002/bit.10194