Pore-Size Controlled and Aminated Poly(γ-methyl L-glutamate) Particles for Selective Removal of Nucleic Acids
Pore-Size Controlled and Aminated Poly(γ-methyl L-glutamate) Particles for Selective Removal of Nucleic Acids
复制标题
用于选择性去除核酸的孔径控制和胺化聚（γ-L-谷氨酸甲酯）颗粒
DOI：
10.1295/polymj.30.616
复制
发表时间：
1998
期刊：
影响因子：
2.8
通讯作者：
K. Ohkuma
中科院分区：
文献类型：
--
作者：
C. Hirayama;M. Sakata;K. Moriguchi;H. Mizokami;H. Ihara;K. Ohkuma
本报告描述了一种从各种蛋白质溶液中选择性去除核酸的方法，使用胺化聚（γ-甲基L-谷氨酸酯）（PMLG-NH 2）颗粒。核酸（从鲑鱼精巢纯化的DNA）的吸附活性增加的氨基含量或孔径（分子量排阻的多糖，Mlim）的颗粒的增加。在pH 7.0和离子强度μ=0.05的条件下，随着孔径从2×103增加到2×105，DNA和颗粒（氨基含量：3.5 meq g−1）之间的表观解离常数从8.2× 10 −13 mol l−1（M）降低到2.0×10−13 M。在另一方面，牛血清白蛋白的吸附活性增加，增加Mlim的颗粒，但随着缓冲液的离子强度的增加而下降。结果，当Mlim为2×103，氨基含量为3.5 meq g−1时，PMLG-NH 2颗粒在pH 7.0和离子强度（μ）为0.2的条件下从各种含DNA的样品（如蛋白质溶液）中去除DNA。颗粒还从源自支气管败血波氏杆菌和多杀性巴氏杆菌的粗抗原溶液中去除DNA。在去除DNA后，每种样品溶液均获得了保护性抗原的高回收率（100%），其中DNA的浓度降至10 ng ml−1以下。
This report describes a method for selective removal of nucleic acids from various protein solutions, using aminated poly(γ-methyl L-glutamate) (PMLG-NH2) particles. The adsorbing activity for nucleic acids (purified DNA from salmon spermary) increased with increase in either the amino-group content or pore-size (molecular mass exclusion of polysaccharide, Mlim) of the particles. The apparent dissociation constant between the DNA and the particles (amino-group content: 3.5 meq g−1) decreased from 8.2×10−13 mol l−1 (M) to 2.0×10−13 M with increase in pore size from 2×103 to 2×105, at pH 7.0 and an ionic strength of μ=0.05. On the other hand, the adsorbing activity of bovine serum albumin increased with increasing Mlim of the particles, but decreased with increasing ionic strength of the buffer. As a result, when Mlim was 2×103 and amino-group content was 3.5 meq g−1, PMLG-NH2 particles removed DNA from various DNA-containing samples, such as protein solutions, at pH 7.0 and ionic strength (μ) of 0.2. The particles also removed DNA from crude antigen solutions originating from Bordetella bronchiseptica and Pasteurella multocida. A high recovery of protective antigen (100 %) was obtained with each sample solution after removal of the DNA, and the concentration of DNA in it decreased to below 10 ng ml−1.