Development of polycation-immobilized cellulose particles for selective DNA adsorption based its multipoint attachment

开发基于多点附着的用于选择性 DNA 吸附的聚阳离子固定化纤维素颗粒

• 批准号: 18550198
• 负责人: SAKATA Masayo
• 金额: $ 2.68万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18550198/
• 关键词: DNA adsorption; polvcation; dimethvlaminopropylacrylamide; ligand; polvethvleneimine; chromatographic separation; pore size; ionic strength; セルロース粒子; イオン相互作用; イオン強度

We have focused a method for the selective removal of DNA from various cellular products using columns packed with polycation-immobilized cellulose beads. Polyethyleneimine (PEI), poly-N,N-dimethylaminopropylacrylamide (poly (DAPA)) and poly (ε-lysine) (PεL), all of which have cationic properties, were used as the ligands on the beads. Cellufine-GC15[○!R] and -CPC^[○!R] were used as cellulose matrices. Adsorption of DNA by the beads was determined using a batchwise method or a column method. Each bead type showed high DNA-adsorbing activity at pH 7.0 and ionic strengths of μ= 0.05-0.8. The larger the pore size of the beads, the larger the DNA-adsorbing activity. The DNA- adsorbing capacities per wet-mL of PEI-, poly(DAPA)- and PεL-immobilized Cellufine-CPC with large pore sizes, were 3.7, 3.2 and 1.8 mg, respectively. When a protein, such as bovine serum albumin (BSA) or γ-globulin, was present in solution with the DNA under physiological conditions (pH 7.0,μ= 0.2), the DNA selectivity of the PEI-immobilized Cellufine-CPC columns was unsatisfactory, because both the DNA and the protein were adsorbed onto the column. In contrast, the poly (DAPA) -immobilized Cellufine-CPC column selectively removed DNA from each protein solution contaminated with DNA under similar conditions : the DNA concentration in each treated protein solution was below 10 ng /mL, and high recovery of each protein (>92%) was obtained.

我们重点研究了一种使用聚阳离子固定化纤维素珠填充的柱子选择性地从各种细胞产品中去除DNA的方法。用具有阳离子性质的聚乙烯亚胺、聚N，N-二甲氨基丙基丙烯酰胺和聚ε-赖氨酸(P-εL)作为微球的配体。纤维素基质为-CellufineGC15[0！R]和-CPC^[0！R]。用间歇法或柱法测定小球对DNA的吸附。当pH值为7.0时，每种类型的微珠都表现出较高的DNA吸附活性，离子强度μ=0.0 5~0.8。小球的孔径越大，其对DNA的吸附活性越大。大孔PEI、Poly(DAPA)和P-εL固定化细胞微球对DNA的单位湿量吸附容量分别为3.7、3.2和1.8 mg。当蛋白质，如牛血清白蛋白或γ球蛋白，在生理条件下(pH 7.0，μ=0.2)与DNA存在时，由于DNA和蛋白质都被吸附在柱上，因此固定化的CELUFINE-CPC柱的DNA选择性不理想。相反，聚(DAPA)固定化Cellufinin-CPC柱在相似的条件下选择性地从每个被DNA污染的蛋白质溶液中去除DNA：每个处理蛋白质溶液中的DNA浓度都在10 ng/mL以下，并且每个蛋白质的回收率都很高(~gt；92%)。

项目成果

Preparation of nearly monodisphered polystyrene particles formed by polymerization in a surfactant-free emulsion with silica particles

二氧化硅颗粒在无表面活性剂乳液中聚合制备近单二球聚苯乙烯颗粒

• 发表时间: 2007
• 期刊: Chemistry Letters 36(6)
• 作者: S.;Kawano;S.;Nishi;Masayo;Sakata;M.;Kunitake
• 通讯作者: Kunitake

Preparation of polycation-immobilized cellulose beads and their application to selective removal of endotoxin

聚阳离子固定纤维素珠的制备及其选择性去除内毒素的应用

• 发表时间: 2007
• 期刊: Cellulose Communications 14 (1)
• 作者: M.;Sakata;M.;Todokoro
• 通讯作者: Todokoro

Selective removal of DNA from bioproducts by polycationimmobilized cellulose beads

通过聚阳离子固定化纤维素珠选择性去除生物制品中的 DNA

• 发表时间: 2006
• 期刊: Journal of Liquid Chromatography & Related Technologies 29
• 作者: M.;Sakata;M.;Nakayama;K.;Yanagi;M.;Sasaki;M.;Kunitake;C.;Hirayama
• 通讯作者: Hirayama

Formation of nearly monodispersed polystyrene particles by spontaneous emulsion polymerization into silica particle solutions without surfactants

通过自发乳液聚合形成几乎单分散的聚苯乙烯颗粒到不含表面活性剂的二氧化硅颗粒溶液中

• 发表时间: 2007
• 作者: S.;Kawano;S.;Nishi;M.;Sakata;M.;Kunitake
• 通讯作者: Kunitake

Selective removal of DNA from bioproducts by polycation-immobilized cellulose beads

通过聚阳离子固定纤维素珠选择性去除生物产品中的 DNA

• 发表时间: 2006
• 期刊: Journal of Liquid Chromatography&Related Technology 29
• 作者: S.;Kawano;S.;Nishi;Masayo;Sakata;M.;Kunitake;Sakata M.(第一著者名);坂田 眞砂代(第一著者名);Kawano S.(第一著者名);Sakata M.
• 通讯作者: Sakata M.