Thermoresponsive chromatography with regulated electrostatic interactions for effective separation of nucleic acids

具有受控静电相互作用的热响应色谱可有效分离核酸

基本信息

批准号：
13680955
负责人：
KIKUCHI Akihiko
金额：
$ 1.22万
依托单位：
Tokyo Women's Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

This research focused on the design and characterization of novel thermoresponsive chromatography system in which the surface characteristics can change hydrophilic/hydrophobic properties and, at the same time, surface charge densities by temperature as a sole variant in aqueous milieu. Such surfaces was prepared for separation of bioactive nucleic acids efficiently without use of several chromatography columns working with different separation modes. We have already elucidated the surface property alterations of the cationic thermoresponsive poly(N-isopropylacrylamide) (PIPAAm) modified surfaces with temperature in terms of hydrophobicity changes and surface potential changes. Furthermore, separation of adenosine nucleotides, AMP, ADP, and ATP was successfully achieved with this column matrix. In this fiscal year, separation of adenosine oligonucleotides and thymidine oligonucleotides with repeating number of 2-5, were investigated using cationic, thermoresponsive column. Thermorespon … More sive column matrices were prepared by radical polymerization of IPAAm monomer and 3,3-dimethylaminopropylacrylamide (DMAPAA) on the initiator immobilized silica beads. The sample mixture was injected into polymer-modified silica packed column (4.6mmφ, 150mm length), and elution behavior of these nucleotides were monitored. For nonionic PIPAAm homopolymer column, retention times of the oligonucleotides are almost same regardless of the number of repeating units. In contrast, retention of oligonucleotides is increased with increasing number of repeating units on cationic, thermoresponsive surfaces. Polarity of the nucleotide bases is significantly important to alter oligonucleotide retention on cationic thermoresponsive column matrices. Thus the retention times of the thymidine oligonucleotides are slightly longer than those of the adenosine nucleotides with the same amount of the phosphate groups. The interaction of oligonucleotides and cationic surfaces is multi-point interaction of polyelectrolytes, and thus the retention increased with increasing the number of repeating units. In conclusion, the obtained results suggest that the prepared cationic, thermoresponsive surfaces can be used to separate oligonucleotides through the recognition of charge densities and hydrophobicity of sample molecules. Less

这项研究的重点是新型热响应色谱系统的设计和表征，在该系统中，表面特性可以改变疏水/疏水性能，同时，与水环中的唯一变体一样，表面电荷密度是温度的。准备这样的表面以有效地分离生物活性核酸，而无需使用几个具有不同分离模式的色谱柱。我们已经阐明了阳离子热响应聚（N-异丙基丙烯酰胺）（PIPAAM）的表面特性变化，并在疏水性变化和表面潜在的变化方面随温度而变化。此外，使用该柱矩阵成功实现了腺苷核苷酸，AMP，ADP和ATP的分离。在这个财政年度，使用阳离子，热响应柱研究了腺苷寡核苷酸和胸苷寡核苷酸的分离。热响应柱…通过在固定二氧化硅珠的引发剂上对IPAAM单体和3,3-二甲基氨基丙烯酰胺（DMAPAA）的根本性聚合制备了更多的柱。将样品混合物注入聚合物修饰的二氧化硅堆积柱（4.6mmφ，150mm长），并监测这些核动肽的洗脱行为。对于非离子Pipaam均聚物柱，无论重复单位的数量多少，寡核苷酸的保留时间几乎相同。相比之下，随着阳离子，热响应表面上的重复单位数量的增加，寡核苷酸的保留量增加。核苷酸碱基的极性对于改变阳离子热响应柱材料的寡核苷酸保留非常重要。胸苷寡核苷酸的保留时间比磷酸基团量相同的腺苷核苷酸的保留时间略长。寡核苷酸和阳离子表面的相互作用是聚电解质的多点相互作用，因此保留量随着重复单元的数量增加而增加。总之，获得的结果表明，制备的阳离子阳离子表面可通过识别电荷密度和样品分子的疏水性来分离寡核苷酸。较少的

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Chikako SAKAMOTO, Yuji OKADA, Hideko KANAZAWA, Eri AYANO, T.NISHIMURA, M.ANDO, Akihiko KIKUCHI, Teruo OKANO: "Temperature-and pH-responsive aminopropyl silica ion-exchange columns grafted with copolymers of N-isopropylacrylamide"Journal of Chromatography

Chikako SAKAMOTO、Yuji OKADA、Hideko KANAZAWA、Eri AYANO、T.NISHIMURA、M.ANDO、Akihiko KIKUCHI、Teruo OKANO：“接枝有 N-异丙基丙烯酰胺共聚物的温度和 pH 响应型氨丙基二氧化硅离子交换柱”Journal of Chromatography