分子印迹技术研究进展

分子印迹是制备具有分子特异识别功能聚合物的一种技术。本文从分子印迹聚合物的识别机理、分子印迹聚合制备条件和制备技术三个方面综述了分子印迹的研究进展,最后展望了分子印迹发展前景。引用文献66篇。     

水相识别分子印迹技术

在各种基于超分子方法的仿生识别体系中,分子印迹聚合物已经证明是一种有潜力的合成受体,受到了广泛的关注。传统的分子印迹技术通常是在有机溶剂中制备对小分子具有选择性的印迹聚合物,而在水相中制备及识别生物大分子的研究仍具有相当的挑战性。从小分子到生物大分子、从有机相到水相,反映了分子印迹技术的发展趋势。本文对最近几年分子印迹在水相制备与识别方面的最新进展进行了总结与评述,探讨了水相识别印迹聚合物的设计策略与制备方法;着重介绍了水相识别技术在固相萃取、色谱固定相、药物控释、中药有效成份提取以及生物分子识别等方面的应用;指出了提高水相识别选择性的途径并对其将来的发展进行了建议与展望。     

分子印迹-仿生传感器的研究进展

分子印迹技术是制备具有选择性分子识别能力聚合物(分子印迹聚合物)的新兴化学合成技术。分子印迹聚合物的一个重要应用是在生物传感器中取代生物分子作为识别元件,研制耐受性强、低成本的分子印迹仿生传感器。综述了分子印迹技术的基本原理及其在仿生传感器方面的应用研究现状,并对分子印迹仿生传感器的发展前景进行了评述。引用文献24篇。     

分子印迹聚合物微球的制备及应用研究进展

球形分子印迹聚合物具有制备简单、使用方便；分子识别效率高且便于功能设计等优点，近年来成为分子印迹技术领域研究的热点之一。对球形分子印迹聚合物微球的制备及其应用研究进展作了较为详细的介绍。     

溶菌酶分子印迹聚合物膜的制备及其吸附性能

以溶菌酶为模板蛋白质，结合分子印迹技术在硅烷化的基质玻片上制备了溶菌酶分子印迹聚合物膜。实验优化了溶菌酶聚合物膜的印迹体系，考察了溶菌酶分子印迹聚合物膜的吸附平衡时间、最大吸附量、特异识别能力、重复使用性以及对实际样品中溶菌酶的分离情况。结果表明，在最优条件下，制备的分子印迹聚合物膜对溶菌酶具有特异吸附能力，印迹因子为3.0，吸附平衡时间为5 min，吸附行为符合Langmuir吸附模型，理论最大吸附量为42.5 mg/g，实际样品中的吸附量为30 mg/g。且此印迹聚合物膜在重复使用5次后，最大吸附量仅下降了5%，具有良好的重复使用性。该方法为复杂生物样品中目标蛋白质的分离富集提供了一种快速、高效的手段。     

腈菌唑分子印迹聚合物的制备及识别性能研究

以腈菌唑为模板分子，采用原位分子印迹技术，制备具有特定识别性能的连续棒状分子印迹聚合物。考察了流动相中酸量对分离的影响，研究了几种结构类似物在所得分子印迹柱上的保留特性。结果表明，这种棒状分子印迹聚合物比相应的空白聚合物有高的识别性能和选择性。     

分子印迹技术研究进展

分子印迹技术是制备对特定目标分子具有特异性识别能力的高分子材料的技术,所制备的高分子材料被称为分子印迹聚合物.分子印迹聚合物因具有预定性、识别性和实用性三大优点已广泛应用于分离、模拟抗体与受体、催化剂以及仿生传感器等方面和领域,显示出了广泛的应用前景.作者对分子印迹技术的发展历史、基本原理、分类、应用现状以及一些新的研究热点进行了综述.     

分子印迹技术在毛细管电色谱中的应用

分子印迹技术是制备具有分子识别功能聚合物，即分子印迹聚合物（MIPs)的一种新技术；毛细管电色谱（CEC）是一个具有发展前途的色谱新技术。将分子印迹技术和毛细管电色谱两种新技术相结合，优势互补，具有极大的发展潜力。本文对分子印迹技术在毛细管电色谱中的应用，以及各类MIPs-CEC毛细管柱的制备方法进行了较为全面的综述，引用文献52篇。     

纳米结构蛋白质分子印迹聚合物的研究进展

在生物技术和生命科学等领域常依赖抗体的特异识别作用来实现目标蛋白质的分离、分析和检测。以蛋白质为模板的分子印迹聚合物有望取代这些昂贵的抗体,但交联聚合物网络中大分子的传质限制是蛋白质分子印迹研究中面临的一个重要问题。纳米结构的蛋白质印迹材料具有较高的表面积与体积比及较浅的印迹位点,很大程度上可克服这一问题。本文详细介绍各种纳米蛋白质印迹聚合物的制备方法,分析各自的优势和不足,并简要介绍其应用。     

分子印迹技术在蛋白质分离分析中的研究进展

蛋白质结构复杂,种类多样,与各种生命活动密切相关。大部分蛋白质在生物体中含量极低,对其分析检测带来极大困难。因此实现复杂生物样品中蛋白质的选择性识别与分离,对实现蛋白质的分离分析意义重大。通过分子印迹技术制备的分子印迹聚合物含有与模板分子大小、形状一致,官能团相互匹配的三维印迹空穴,在蛋白质的选择性识别与分离领域显示出了巨大的发展潜力。但是,由于蛋白质具有尺寸较大、构型易变、结构复杂等特点,分子印迹技术在蛋白质印迹中面临着巨大挑战。该文在介绍几种新型分子印迹技术包括表面印迹、抗原决定基印迹和金属螯合物印迹的基础上,综述了近3年分子印迹技术在蛋白质分离分析方面的应用,并对其发展进行了总结与展望。     

蛋白质分子印迹技术载体形式的研究进展

现代生物技术产品分离成本很高,分子印迹技术以其优良的操作稳定性为蛋白质分离提纯提供了一种新的方法,合成蛋白质分子印迹聚合物具有巨大的应用价值,又极具挑战性,已成为各国科学工作者们研究的热点。本文对蛋白质分子印迹过程中使用的载体形式进行了综述,对不同形式载体的使用特点进行了总结,详细叙述了常见的载体形式如硅胶、合成树脂球、高分子膜、云母、凝胶以及一些新型的载体类似形式如环糊精和壳聚糖等,并探讨了目前蛋白质分子印迹技术存在的问题及其应用前景。     

Silica nanoparticle supported molecularly imprinted polymer layers with varied degrees of crosslinking for lysozyme recognition

Surface imprinting over nanosized support materials is particularly suitable for protein templates, considering the problems with mass transfer limitation and low binding capacity. Previously we have demonstrated a strategy for surface protein imprinting over vinyl-modified silica nanopartiles with lysozyme as a model template by polymerization in high-dilution monomer solution to prevent macrogelation. Herein, the synthesis process was further studied toward enhancement of the imprinting performance by examining the effect of several synthesis conditions. Interestingly, the feed crosslinking degree was found to have a great impact on the thickness of the formed imprinting polymer layers and the recognition properties of the resulting imprinted materials. The imprinted particles with a crosslinking degree up to 50% showed the best imprinting effect. The imprinting factor achieved 2.89 and the specific binding reached 23.3 mg g⁻¹, which are greatly increased compared to those of the lowly crosslinked imprinted materials reported previously. Moreover, the relatively high crosslinking degree led to no significant retarding of the binding kinetics to the imprinted particles, and the saturated adsorption was reached within 10 min. Therefore, this may be a promising method for protein imprinting.     

二维蛋白质分子印迹聚合物*

本文对二维蛋白质分子印迹进行了综述：介绍了二维蛋白分子印迹的基本概念;阐述了常见二维蛋白质分子印迹方法的基本原理,包括表位印迹法、溶胶-凝胶法、射频光电等离子沉积法和Langmuir单层法等;根据二维蛋白分子印迹材料的不同形式,详细叙述了二维蛋白质分子印迹薄膜、核-壳微球、纳米线、Langmuir脂质体单层的制备过程、结合能力、选择识别性能;分析了目前二维蛋白质分子印迹技术存在的一些不足和进一步研究的方向。     

基于银增强金标记物的阳极溶出伏安免疫分析用于补体第三成分的测定

分子印迹是制备对特定分子具有专一性结合能力的聚合物的技术,所制备的聚合物被称为分子印迹聚合物（Molecularly imprinted polymers,MIPs）,此类聚合物在分离提纯、模拟酶和传感器等方面均显示出广阔的应用前景,迄今,小分子化合物的印迹技术已经十分成熟。     

表面分子印迹聚合物纳米线用于蛋白质的特异性识别

手性配体交换色谱是拆分手性化合物,特别是氨基酸和羟基酸对映体的一种有效方法,通常以光活性氨基酸或其衍生物为手性选择子,可通过键合及涂渍制备手性固定相,也可作为流动相添加剂来实现手性配体交换色谱分离分析,配体交换键合固定相需要完成载体和手性选择子之间的偶联,键合量因受到载体和制备条件的影响而较难控制,且柱效较低。     

Advances in the development of molecularly imprinted polymers for the separation and analysis of proteins with liquid chromatography

This review documents recent advances in the design, synthesis, characterization, and application of molecularly imprinted polymers in the form of monoliths and particles/beads for the use in the separation and analysis of proteins with solid‐phase extraction or liquid chromatography. The merits of three‐dimensional molecular imprinting, whereby the molecular template is randomly embedded in the polymer, and two‐dimensional imprinting, in which the template is confined to the surface, are described. Target protein binding can be achieved by either using the entire protein as a template or by using a protein substructure as template, that is, a peptide, as in the "epitope" approach. The intended approach and strategy then determine the choice of polymerization method. A synopsis has been provided on methods used for the physical, chemical, and functional characterizations and associated performance evaluations of molecularly imprinted and nonimprinted control polymers, involving a diverse range of analytical techniques commonly used for low and high molecular mass analytes. Examples of recent applications demonstrate that, due to the versatility of imprinting methods, molecularly imprinted monoliths or particles/beads can be adapted to protein extraction/depletion and separation procedures relevant to, for example, protein biomarker detection and quantification in biomedical diagnostics and targeted proteomics.     

The effect of protein structural conformation on nanoparticle molecular imprinting of ribonuclease A using miniemulsion polymerization

One of the major difficulties faced in the molecular imprinting of proteins is the inherently fragile and flexible nature of the protein template which makes it incompatible with most polymerization systems. Miniemulsion polymerization is a possible approach for preparing molecularly imprinted nanoparticles, and in this study, the method of initiation, the high-shear homogenization, and the surfactant used for the polymerization reaction had been considered as possible factors that can denature the template protein, ribonuclease A (RNase A). The conformation of the protein in a miniemulsion was studied using circular dichroism (CD). It was found that redox initiation was more suitable for protein imprinting and that the addition of poly(vinyl alcohol) (PVA) as a co-surfactant had proved to be effective in preserving the template protein structural integrity. On the basis of the results of the study, polymeric nanoparticles imprinted with RNase A were prepared via miniemulsion polymerization using methyl methacrylate (MMA) and ethylene glycol dimethacrylate (EGDMA) as the functional and cross-linker monomers, respectively, with the conditions of the polymerization system optimized to best preserve the integrity of the protein template. In the subsequent investigation for the recognition properties of the prepared nanoparticles through batch and competitive rebinding tests, the imprinted nanoparticles prepared through the conventional (nonoptimized) miniemulsion polymerization lacked the target specificity as displayed by those prepared under the optimized conditions. This illustrated the importance of protein structural integrity in protein imprinting.     

分子印迹技术用于蛋白质的识别*

分子印迹技术是一种新型的高效分离技术。合成含识别蛋白质的分子印迹聚合物具有极大的应用价值,又极具挑战性,已成为许多分子识别领域工作者的研究热点。本文总结了近10年来该领域的研究进展,讨论了已有不同方法的发展状况以及相对优缺点,阐明了其可能发展的方向和前景。     

Bovine serum albumin-imprinted polyacrylamide gel beads prepared via inverse-phase seed suspension polymerization

Synthetic materials capable of specifically recognition proteins are important in bioseparation and biosensors. In this study, bovine serum albumin-imprinted polyacrylamide gel beads were synthesized via inverse-phase seed suspension polymerization, using high-density crosslinked gel beads as core, low-density crosslinked polyacrylamide gel as imprinting shell. The surface of gel bead had a large quantity of well-distributed macropores, which were suited to let the proteins pass in and out. The selectivity test showed that imprinting gel beads exhibited good recognition for template proteins, as compared to the control protein. We consider the formation of multiple hydrogen bonds and complementary shape between the imprinting cavities and the template proteins are the two factors that lead to the imprinting effect. The imprinting beads had quick adsorption rate and possessed improved regeneration property in comparison with those prepared directly via inverse-phase suspension polymerization.     

The effectively specific recognition of bovine serum albumin imprinted silica nanoparticles by utilizing a macromolecularly functional monomer to stabilize and imprint template

Structural stability of the template is one of the most important considerations during the preparation of protein imprinting technology. To address this limitation, we propose a novel and versatile strategy of utilizing macromolecularly functional monomers to imprint biomacromolecules. Results from circular dichroism and synchronous fluorescence experiments reflect the macromolecularly functional monomers tendency to interact with the protein surface instead of permeating it and destroying the hydrogen bonds that maintain the protein’s structural stability, therefore stabilizing the template protein structure during the preparation of imprinted polymers. The imprinted polymers composed of macromolecularly functional monomers or their equivalent micromolecularly functional monomers over silica nanoparticles were characterized and carried out in batch rebinding test and competitive adsorption experiments. In batch rebinding test, the imprinted particles prepared with macromolecularly functional monomers exhibited an imprinting factor of 5.8 compared to those prepared by micromolecularly functional monomers with the imprinting factor of 3.4. The selective and competitive adsorption experiments also demonstrated the imprinted particles made by macromolecularly functional monomers possessed much better selectivity and specific recognition ability for template protein. Therefore, using macromolecularly functional monomers to imprint may overcome the mutability of biomacromolecule typically observed during the preparation of imprinted polymers, and thus promote the further development of imprinting technology.