Establishment of Membrane Separation and Purification using Conformational Change of DNA and Virus

对生物制药中病毒和DNA的浓缩和纯化进行了研究。具体说明如下：1。组蛋白- dna和染色质通过平均孔径为15 nm的再生纤维素中空纤维的渗透率明显低于游离dna的渗透率2。当γ-球蛋白溶液通过平均孔径为35 nm和15 nm的病毒去除膜时，发现通量急剧增加，饲料溶液中的DNA减少，而蛋白质溶液中通常含有以ppb数量级的污染。用动态光散射法在γ-球蛋白溶液中检测到粒径为135 ~ 225 nm的DNA-γ-球蛋白复合物。在γ-球蛋白溶液中加入0.4 mol/l NaCl可破坏DNA-γ-球蛋白复合物。当NaCl浓度达到0.1 mol/l时，γ-球蛋白溶液通过膜的通量急剧增加。这可以解释为在γ-球蛋白溶液中NaCl存在时DNA-γ-球蛋白复合物的减少。发现通量对NaCl浓度的影响取决于膜和膜材料的孔径和结构。DNA-γ-球蛋白复合物也被DNA酶在γ-球蛋白溶液中的处理破坏。生物药物溶液经DNAase处理后，γ-球蛋白溶液通过膜的通量急剧增加。这可以解释为含有γ-球蛋白溶液的DNA-γ-球蛋白复合物中DNA的分割导致DNA-γ-球蛋白复合物减少。由此得出结论，DNA分子和病毒可以通过变形成线状扩散到比分子大小更小的再生纤维素固体孔隙中。

Concentration and purification of Virus and DNA from biopharmaceuticals was investigated. The followings are specifically clarified ;1. The permeation of histone-DNA and chromatin through regenerated cellulose hollow fibers having a mean pore diameter of 15 nm was substantially lower than the permeation of free DNA.2. When γ-globulin solution permeated through the virus removal membranes having a mean pore diameter of 35 nm and 15 nm, a drastic increase in flux was found with a decrease in DNA in the feed solution, which was normally contained in the protein solution as a contamination in the order of ppb. A DNA-γ-globulin complex having a particle size of 135-225 nm was detected by dynamic light scattering measurements in the γ-globulin solution.3. The DNA-γ-globulin complex was destroyed by the addition of 0.4 mol/l NaCl in the γ-globulin solution. The flux of the γ-globulin solution through the membranes drastically increased with the increase in NaCl concentration up to 0.1 mol/l. This is explained by a decrease in the DNA-γ-globulin complex in the presence of NaCl in the γ-globulin solution. The effect of flux on the concentration of NaCl was found to depend on the pore sizes and structure of the membranes and membrane materials.4. The DNA-γ-globulin complex was also destroyed by the treatment of DNAase in the γ-globulin solution. The flux of the γ-globulin solution through the membranes drastically increased when the biodrug solution was treated with DNAase. This is explained by a decrease in the DNA-γ-globulin complex from segmentation of DNA in the DNA-γ-globulin complex containing the γ-globulin solution.5. It is concluded that the DNA molecule and virus could diffuse into the smaller pores in the regenerated cellulose solid than the size of the molecule by the deformation of the shape into a thread-like shape.