Design of Temperature-Responsive Surfaces and Its Application for Aqueous Chromatography

温度响应表面的设计及其在水色谱中的应用

• 批准号: 07408030
• 负责人: SAKURAI Yasuhisa
• 金额: $ 17.47万
• 依托单位: Tokyo Women's Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07408030/
• 关键词: Thermo-responsive surface; Aqueous chromatography; Poly (N-isopropylacrylamide); Hydrophilic; Hydrophobic; Hydrophobic interaction; Steroid hormones; Polypeptides; クロマトグラフィー; 親・疎水性

This project aimed to develop a novel aqueous chromatography system in which the polarity of stationary phases is controlled by external temperature changes to separate and purify solutes without changing their structure and functions. This system is quite different from reversed phase chromatography in which the polarity of stationary phases is altered by the addition of organic solvents in water with various mixing ration. In the proposed system, the surface of the aminated silica stationary phase was modified with temperature-responsive poly (N-isopropylacrylamide) (PIPAAm) with a carboxyl end. The successful modification of the surfaces with polymers were confirmed by means of titration of residual surface functional groups, X-ray photoelectron spectroscopy, and atmic force microscope. These surfaces showed temperature-responsive wettability changes, being hydrophilic at lower temperature below polymer transition temperature, and become hydrophobic above the transition temperature. … More The elution profiles of five steroids were then investigated by changing temperature using high-performance liquid chromatography column packed with PIPAAm-grafted silica using aqueous mobile phase. Retention times of hydrophobic steroids increased with increasing temperature without any change in the eluent composition. Retarded elution time of hydrophobic steroids resulted from the stronger interaction of these compounds with the PIPAAm-grafted surfaces in which the surface property becomes hydrophobic with temperature increase. As a consequence, gradient elution-like effect is achieved with a single mobile phase by controlling external temperature. We further investigated the possibility of this temperature-responsive chromatography for polypeptide separation. Polypeptides with similar number of amino acid residues were successfully separated at higher temperature where hydrophobic interaction of solutes with stationary phases are primarily influenced the separation efficacy. In conclusion, elution of target substances is controlled only by a small change in column temperature without any further modification of the aqueous mobile phase in the proposed chromatography. Less

本项目旨在开发一种新型的水色谱系统，该系统通过外部温度的变化来控制固定相极性，从而在不改变溶质结构和功能的情况下分离和纯化溶质。该体系与反相色谱不同，反相色谱通过在不同混合比的水中加入有机溶剂来改变固定相极性。在所提出的体系中，胺化硅胶固定相的表面用带有羧基的温度响应性聚(N-异丙基丙烯酰胺)(PIPAAm)修饰。通过表面剩余官能团滴定、X-射线光电子能谱、原子力显微镜等手段证实了聚合物成功地对表面进行了改性。这些表面表现出对温度敏感的润湿性变化，在低于聚合物转变温度的较低温度下是亲水的，在转变温度以上变得疏水。…在此基础上，采用PIPAAm接枝硅胶的高效液相色谱柱，采用水相流动相，通过变温的方法研究了5种甾体化合物的洗脱曲线。疏水类固醇的保留时间随着温度的升高而增加，洗脱液的组成没有任何变化。疏水类固醇洗脱时间的延迟是由于这些化合物与PIPAAm接枝表面的相互作用较强，表面性质随着温度的升高而变得疏水。因此，通过控制外部温度，利用单一流动相实现了类似于梯度洗脱的效果。我们进一步研究了这种温度响应层析用于多肽分离的可能性。在较高的温度下，氨基酸残基数目相近的多肽被成功分离，其中溶质与固定相的疏水相互作用主要影响分离效果。总之，目标物质的洗脱仅通过柱温的微小变化来控制，而不需要对所提出的层析中的水相流动相进行任何进一步的修改。较少

项目成果

Y.G.Takei,T.Aoki,K.Sanui,N.Ogata,Y.Sakurai,T.Okano: "Temperature-modulated platelet and lymphocyte interactions with poly (N-isopropylacrylamide)-grafted surfaces" Biomaterials. 16. 667-673 (1995)

Y.G.Takei、T.Aoki、K.Sanui、N.Ogata、Y.Sakurai、T.Okano：“温度调节血小板和淋巴细胞与聚（N-异丙基丙烯酰胺）接枝表面的相互作用”生物材料。

薬師寺大二: "温度応答性高分子をグラフトした表面を用いた新しい温度制御型タンパク質クロマトグラフィー" 治療と診断のシステム 化学工学シンポジウムシリーズ55 化学工学会治療システム研究会編. 67-70 (1996)

Daiji Yakushiji：“使用接枝有温度响应聚合物的表面的新型温度控制蛋白质色谱”治疗和诊断系统化学工程研讨会系列 55 由化学工程师学会处理系统研究组编辑 67-70 (1996)。

H.Kanazawa: "Analysis of peptides and proteins temperature-responsive chromatographic system using N-isopropylacrylamide polymer-modified columns" J.Pharamaceu.Biomed.Anal.(印刷中). (1997)

H. Kanazawa：“使用 N-异丙基丙烯酰胺聚合物改性柱分析肽和蛋白质温度响应色谱系统”J.Pharamaceu.Biomed.Anal.（出版中）。

T.Okano,N.Yamada,M Okuhara,H.Sakai,Y.Sakurai: "Mechanism of cell detachment from temperature-modulated hydrophilic-hydrophobic polymer surfaces" Biomaterials. 16. 297-303 (1995)

T.Okano，N.Yamada，M Okuhara，H.Sakai，Y.Sakurai：“细胞从温度调节的亲水-疏水聚合物表面脱离的机制”生物材料。

T.Okano,A.Kikuchi,Y.Sakurai,Y.Takei,N.Ogata: "Temperature-responsive poly (N-isopropylacrylamide) as a modulator for alteration of hydrophilic/hydrophobic surface properties to control activation/inactivation of platelets" Journal of Controlled Release. 3

T.Okano、A.Kikuchi、Y.Sakurai、Y.Takei、N.Ogata：“温度响应型聚（N-异丙基丙烯酰胺）作为改变亲水/疏水表面特性以控制血小板活化/失活的调节剂”期刊