乳液界面各向异性生长辅助构建单孔中空MMIPs选择性分离纯化2′-脱氧腺苷的研究

It is of paramount scientific, economic and social significance to explore the new methods bearing independent intellectual property rights, for selective separation and purification of 2'-deoxyadenosine (dA), whilst improving the general yield and purity of dA. In this project, we intend to construct a hollow composite magnetic molecularly polymers (MMIPs) with single holes via anisotropic growth onto emulsion interface, which is subjected further to separate and purify dA molecules selectively and effectively. In this study, hydrophilic (or hydrophobic) silane coupling agents is to be introduced into the emulsion droplet interface as precursors, and catalytic hydrolysis of the precursor in the internal phase will contributes to the anisotropic growth of silane at the emulsion interface and form an a hollow silica nanoparticle with single holes; a imprinting polymer layer consisting of the functional derivative monomer from 2'-deoxythymidine and 2'-deoxyuridine iodide substitutes and dA as template molecule is designed, and we aim to embrace the obtained silica nanoparticles with the imprinted polymeric layer to introduce numerous molecular recognition sites; Moreover, under sonication, we can readily filled ultrasmall magnetic Fe3O4 into the single holes arising from silica nanoparticles, and finally facilitate the preparation of targeted hollow composite MMIPs with unique single holes, which also have the ability to recover their magnetic strengths reversibly. On this basis, we will carefully study the behavior of MMIPs towards the selective separation and purification of dA, and mainly concentrate on its equilibrium, kinetics and thermodynamics. Additionally, the behavior and mechanism of dA recognition will be elucidated further to establish and optimize the adsorption, separation and mass transfer models, with the aim to advance the separation and purification process synergistically. To this end, a new method for selective separation and purification of dA will established to achieve the ultimate goal of obtaining dA products with high yield and purity.

探索具有自主知识产权的2′-脱氧腺苷(dA)选择性分离纯化新方法、提高产物中dA得率和纯度，具有极其重要的科学、经济和社会意义。本课题拟开展乳液界面各向异性生长辅助构建单孔中空复合磁性印迹聚合物(MMIPs)选择性分离纯化dA分子的工作。研究中拟在乳液液滴界面引入亲水(油)功能性硅烷偶联剂作为前驱体，经内相催化前驱体的水解诱导其乳液界面各向异性生长并形成单孔中空硅骨架；以2′-脱氧胸苷和2′-脱氧尿苷碘取代物为原料设计合成功能单体，以dA为模板分子，在单孔中空硅骨架内外表面可控构筑印迹聚合层；在超声浴下经单孔填充小尺寸磁性Fe3O4，制备磁强度“可逆”恢复的单孔中空MMIPs；探索MMIPs选择性分离纯化dA分子过程中的平衡、动力学和热力学行为，研究其对dA的识别能力与机制，建立相应的分离与传质模型；优化分离纯化过程，建立适用于选择性分离纯化dA的新方法，获得高得率和纯度的dA产品。

2′-脱氧腺苷(2′-deoxyadenosine, dA)是一种天然的脱氧核苷，是脱氧核糖核酸DNA 的结构片段，是基因药物和基因工程研究的重要原材料，同时具有很好的生理活性，是很多抗病毒、抗肿瘤、抗艾滋病药物公认的良好中间体，因此在市场上有极其广泛需求。目前，常用的dA分离纯化方法有：结晶法和吸附法等。前者在应用过程中能耗高、辅助器材体积庞大，且不适于处理低浓度 dA 产品；吸附法是利用吸附剂来捕获dA，再将dA洗脱下来，达到dA与其它杂质分离的要求。吸附法对产品中微量dA具有提取率高且易连续操作的优点，但常见吸附剂的选择性、吸附容量和传质速率仍有待进一步提高。因此，根据dA分子结构的特点，从dA与吸附剂吸附位点的相互作用机理出发，开发新型吸附剂、调控吸附剂结构、强化吸附剂功能，建立和完善dA分离纯化的新方法，有效提高 dA 产品得率和纯度是目前急需解决的课题。.分子印迹聚合物(MIPs)，通常称为合成抗体模拟物，能够对靶分子进行特异性分子识别。与天然抗体相比，MIPs更稳定，易于满足广泛应用的要求而且价格低廉。表面印迹聚合物(SMIPs)是把分子印迹识别位点构筑在基质材料表面的聚合吸附剂，显示出高结合能力、快速传质和快速结合动力学。.本研究通过将Alg-Ca2+ Pickering乳液与Janus-MIPs纳米片和超顺磁性氧化铁纳米颗粒相结合，构造了马赛克型的海藻酸盐磁性复合MIPs吸附剂（J-SNs-MMIPs-Pickering），并将其用于选择性分离2'-脱氧腺苷（dA）。J-SNs-MMIPs-Pickering具有三个显着优点。首先，J-SNs-MIPs纳米片由于具有不对称的化学结构可作为Pickering乳液的稳定粒子，可以有效地解决由于纳米片堆叠而造成的位点掩蔽的难题。其次，J-SNs-MMIPs-Pickering具有优异的识别选择性，较大的结合能力和快速的结合速度，嘧啶碱基对dA的特异性匹配作用是分子印迹的识别的作用。第三，通过外部磁场可以快速将J-SNs-MMIPs-Pickering与悬浮液分离。因此，本实验为利用Janus纳米片和Pickering乳液反应器制备具有高选择性和高吸附能力的复合吸附剂提供了一种简便的方法，并提供了一种选择性分离和分析复杂生物样品中核苷和脱氧核苷化合物的方法。