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

对生物药物中病毒和DNA的浓缩和纯化进行了研究。具体澄清如下：1.组蛋白-DNA和染色质在平均孔径为15 nm的再生中空纤维中的渗透率显著低于游离DNA的渗透率。当γ-球蛋白溶液通过平均孔径为35 nm和15 nm的脱毒膜时，通量急剧增加，饲料溶液中的DNA量减少，通常以ppb量级的污染物存在于蛋白质溶液中。动态光散射法在γ-球蛋白溶液中检测到粒径为135-225 nm的γ-DNA-球蛋白复合物。在γ-球蛋白溶液中加入0.4mol/L的氯化钠可破坏γ-DNA-球蛋白复合体。γ-球蛋白溶液通过膜的通量随NaC l浓度的增加而急剧增加，达到0.1m o l/L，这是由于存在NaC l的γ-球蛋白溶液中γ-球蛋白复合体的减少所致。通量对氯化钠浓度的影响取决于膜和膜材料的孔径大小和结构。DNA-γ-球蛋白复合体也被γ-球蛋白溶液中的DNA酶破坏。γ-球蛋白溶液经脱氧核糖核酸酶处理后，膜通量急剧增加。这是因为在含有γ-球蛋白溶液的DNA-γ-球蛋白复合体中，DNA-γ-球蛋白复合体的DNA片段减少了。结果表明，DNA分子和病毒可以通过变形成线状的形式扩散到再生纤维素固体中比分子大小更小的孔隙中。

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.