用于复杂生物样品体系分离与识别的分子印迹技术最新进展

Recent advances of molecular imprinting technology for the separation and recognition of complex biological sample systems

由生物样品中复杂组分所导致的基质效应会严重影响分离分析技术的准确性、灵敏度与可靠性。免疫亲和技术作为降低或消除基质效应的方法已被广泛应用于诊断分析和蛋白纯化等领域,但该技术仍存在明显的缺点,如成本高昂、制备流程繁琐、保存条件苛刻以及配体浸出等问题。目前,如何通过有效降低或消除复杂生物样品中的基质效应来实现痕量目标分析物的分离及识别仍是一个具有挑战性的问题。分子印迹技术(molecular imprinting technology,MIT)一直被广泛应用于固相萃取与色谱分离等领域,随着MIT的发展,各种新型印迹策略被提出;其中,分子印迹聚合物(molecularly imprinted polymer,MIP)作为一种能够模拟抗原-抗体间相互作用的高分子聚合物,可以从各种复杂生物样品中提取出目标分析物,从而有效消除基质效应的影响。MIP不仅拥有高特异性与高亲和力的优点,而且与抗体和适配体等生物大分子相比,MIP还具有稳定性高、成本低廉以及制备简便等优势。近年来一些基于MIT的传统分离技术得到了深入发展,其中包括色谱固定相以及固相萃取吸附剂等。此外,结合了MIT与高灵敏检测技术的分析方法在疾病诊断和生物成像等领域也受到了广泛关注。本文着重介绍了近年来发展的新型印迹策略,并介绍了基于MIP的分离分析方法在各领域中的应用以及现阶段存在的不足,最后对MIT的未来发展方向做出了展望。

Given continuous improvements in industrial production and living standards,the analysis and detection of complex biological sample systems has become increasingly important.Common complex biological samples include blood,serum,saliva,and urine.At present,the main methods used to separate and recognize target analytes in complex biological systems are electrophoresis,spectroscopy,and chromatography.However,because biological samples consist of complex components,they suffer from the matrix effect,which seriously affects the accuracy,sensitivity,and reliability of the selected separation analysis technique.In addition to the matrix effect,the detection of trace components is challenging because the content of the analyte in the sample is usually very low.Moreover,reasonable strategies for sample enrichment and signal amplification for easy analysis are lacking.In response to the various issues described above,researchers have focused their attention on immuno-affinity technology with the aim of achieving efficient sample separation based on the specific recognition effect between antigens and antibodies.Following a long period of development,this technology is now widely used in fields such as disease diagnosis,bioimaging,food testing,and recombinant protein purification.Common immuno-affinity technologies include solid-phase extraction(SPE)magnetic beads,affinity chromatography columns,and enzyme linked immunosorbent assay(ELISA)kits.Immuno-affinity techniques can successfully reduce or eliminate the matrix effect;however,their applications are limited by a number of disadvantages,such as high costs,tedious fabrication procedures,harsh operating conditions,and ligand leakage.Thus,developing an effective and reliable method that can address the matrix effect remains a challenging endeavor.Similar to the interactions between antigens and antibodies as well as enzymes and substrates,biomimetic molecularly imprinted polymers(MIPs)exhibit high specificity and affinity.Furthermore,compared with many other biomacromolecules such as antigens and aptamers,MIPs demonstrate higher stability,lower cost,and easier fabrication strategies,all of which are advantageous to their application.Therefore,molecular imprinting technology(MIT)is frequently used in SPE,chromatographic separation,and many other fields.With the development of MIT,researchers have engineered different types of imprinting strategies that can specifically extract the target analyte in complex biological samples while simultaneously avoiding the matrix effect.Some traditional separation technologies based on MIP technology have also been studied in depth;the most common of these technologies include stationary phases used for chromatography and adsorbents for SPE.Analytical methods that combine MIT with highly sensitive detection technologies have received wide interest in fields such as disease diagnosis and bioimaging.In this review,we highlight the new MIP strategies developed in recent years,and describe the applications of MIT-based separation analysis methods in fields including chromatographic separation,SPE,diagnosis,bioimaging,and proteomics.The drawbacks of these techniques as well as their future development prospects are also discussed.