基于分子印迹与表面增强拉曼散射的蛋白质疾病标志物检测研究进展

doi:10.6023/A20080364

化学学报 ›› 2021, Vol. 79 ›› Issue (1): 45-57.DOI: 10.6023/A20080364 上一篇下一篇

综述

基于分子印迹与表面增强拉曼散射的蛋白质疾病标志物检测研究进展

贺晖^a, 周玲俐^a, 刘震^a^,^*()

a 南京大学化学化工学院生命分析化学国家重点实验室南京 210023

投稿日期:2020-08-14 发布日期:2020-11-27
通讯作者: 刘震

作者简介:

贺晖, 2012年本科毕业于西北农林科技大学, 2015年硕士毕业于西北农林科技大学, 2019年博士毕业于南京大学化学化工学院, 取得博士学位后留校工作任副研究员. 主要研究方向为等离激元增强效应研究、生物分子质谱分析及单细胞分析.

周玲俐, 2017年本科毕业于郑州大学化学与分子工程学院, 2020年硕士毕业于南京大学化学化工学院, 主要从事基于分子印迹聚合物和表面增强拉曼光谱的蛋白质检测方向的研究.

刘震, 1998年于中国科学院大连化学物理研究所获博士学位. 2000~2002年, 日本兵库大学日本学术振兴会(JSPS)特别研究员; 2002~2005年, 加拿大滑铁卢大学博士后. 2005年获聘南京大学教授, 2006年起任博士生导师, 2011~2014年任加拿大滑铁卢大学工程学院兼职教授, 2014年获国家杰出青年科学基金. 主要研究方向: 以仿生分子识别为核心, 结合纳米技术、先进材料、分离科学、生物质谱和拉曼光谱等手段, 发展生物分离、组学分析、疾病诊断分析、单细胞分析、癌症靶向及免疫治疗新方法和新材料.

* E-mail: zhenliu@nju.edu.cn

基金资助:
国家自然科学基金重大科研仪器研制项目(21627810); 国家自然科学基金重点项目(21834003)

Advances in Protein Biomarker Assay via the Combination of Molecular Imprinting and Surface-enhanced Raman Scattering

Hui He^a, Lingli Zhou^a, Zhen Liu^a^,^*()

a State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China

Received:2020-08-14 Published:2020-11-27
Contact: Zhen Liu
Supported by:
the Key Scientific Instrument and Equipment Development Project(21627810); the Key Grant of the National Natural Science Foundation of China.(21834003)

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摘要/Abstract

异常的蛋白质表达与疾病的发生与发展密切相关, 因此蛋白质已作为疾病标志物广泛应用于疾病的早期诊断、治疗监测和预后评估. 然而, 临床样本中的蛋白质疾病标志物通常含量极低, 并存在高丰度的基质干扰, 对检测方法的特异性和灵敏度提出挑战. 目前, 蛋白质疾病标志物的检测方法主要是免疫分析. 但是, 免疫分析主要依赖抗体进行特异性识别, 而抗体具有不易制备、稳定性较差和成本高等缺点. 同时, 免疫分析常通过荧光和化学发光等技术实现高灵敏检测, 但存在操作繁琐、光漂白、光谱宽等不足. 分子印迹聚合物已发展成为在特异性和亲和力方面可媲美抗体的仿生识别材料, 且具有容易制备、稳定性好和成本低等优势. 表面增强拉曼散射技术具有超高灵敏度、光谱窄、快速、无损检测等优势而广泛应用于化学和生物分析. 近年来, 分子印迹技术和表面增强拉曼散射技术的结合产生了系列先进的蛋白质检测方法, 展现了独特的优势, 受到了广泛的关注. 本综述旨在介绍该联用分析技术的主要进展, 在分别介绍分子印迹和表面增强拉曼散射及其在蛋白质检测中单独应用的基础上, 着重介绍基于两种技术的蛋白质疾病标志物的检测方法的研究进展. 最后, 对该联用技术的未来发展做了展望.

关键词: 分子印迹聚合物, 表面增强拉曼散射, 蛋白质, 检测, 疾病标志物

Abnormal protein expression closely correlates with the occurrence and development of diseases. Thus, proteins have been widely used as disease markers for early diagnosis, treatment monitoring and prognosis evaluation of diseases. However, protein biomarkers in clinical samples are usually present in extremely low concentration while the detection often suffers from severe interference by high-abundance sample matrix, which challenges the specificity and sensitivity of protein biomarker assay methods. Currently, the main detection method of protein biomarkers is immunoassay. Nevertheless, immunoassay mainly relies on antibodies for specific recognition, and antibodies are associated with disadvantages such as difficulty in preparation, poor stability, and high-cost. Meanwhile, immunoassay is mainly based on fluorescence and chemiluminescence to achieve high-sensitivity detection, but they have inherent defects such as cumbersome operation, photobleaching and broad spectrum. Molecularly imprinted polymers (MIP) have developed as biomimetic recognition materials with antibody-comparable specificity and affinity, while offering advantages such as ease in preparation, good stability and low cost. On the other hand, surface-enhanced Raman scattering (SERS) has been widely used in chemical or biological assays due to its merits including ultra-high sensitivity, narrow spectrum, speed, non-destructivity, and so on. In recent years, the combination of molecular imprinting and SERS has generated a series of advanced protein detection methods that exhibited unique strengths, which has gained wide attention. This review aims to survey the main advances of this hybrid analytical technique. After introduction of MIP and SERS as well as their separate applications in the detection of protein biomarkers, we mainly focus on the combination of MIP and SERS as well as its application in protein biomarker detection. We finally briefly sketch the future development of this hybrid analytical method.

Key words: molecularly imprinted polymer, surface-enhanced Raman scattering, protein, detection, disease biomarker

引用此文

贺晖, 周玲俐, 刘震. 基于分子印迹与表面增强拉曼散射的蛋白质疾病标志物检测研究进展[J]. 化学学报, 2021, 79(1): 45-57.

Hui He, Lingli Zhou, Zhen Liu. Advances in Protein Biomarker Assay via the Combination of Molecular Imprinting and Surface-enhanced Raman Scattering[J]. Acta Chimica Sinica, 2021, 79(1): 45-57.

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图/表 12

图1. 分子印迹的原理示意图[47]

Figure 1. Schematic of the principle of molecular imprinting[47]

图2. 光刻硼亲和分子印迹法原理示意图[34]

Figure 2. Schematic of the principle of photolithographic boronate affinity molecular imprinting[34]

图3. 硼亲和可控定向表面印迹法原理示意图[72]

Figure 3. Schematic of the principle of boronate affinity controllable- oriented surface imprinting[72]

图4. 硼亲和锚定的可控定向表位印迹法原理示意图[100]

Figure 4. Schematic of the principle of controllable oriented surface imprinting of boronate affinity-anchored epitopes[100]

图5. 基于MIP荧光传感器应用于Cyt C的检测原理示意图[105]

Figure 5. Overview of MIP-based fluorescence sensor applied to Cyt C detection[105]

图6. SERS和SERS的电磁增强机理[117]

Figure 6. SERS and its electromagnetic enhancement mechanism[117]

图7. 蛋白质无标记SERS检测的原理示意图[124]

Figure 7. Overview of label-free SERS detection of protein[124]

图8. 基于SERS的肝癌抗原多重检测过程[131]

Figure 8. Schematic representation of the process of SERS-based multiplex detection of liver cancer antigens[131]

图9. PISA法用于(A)单细胞[140]和(B)复杂样品[141]中蛋白质的检测原理示意图

Figure 9. Schematic of PISA for the detection of proteins in single cells[140] and complex samples[141]

图10. 基于(A) Au纳米星[148]和(B) Au@Ag核壳结构的MIP和抗体联用的SERS检测原理示意图[149]

Figure 10. Schematic of the combination of MIP and antibodies modified (A) Au nanostar[148] and (B) Au@Ag nanostructure[149] for SERS detection

图11. 基于双重表位MIP识别的蛋白质间接SERS检测原理示意图[102]

Figure 11. Schematic of the duMIP-based indirect SERS detection of target protein[102]

图12. (A)用于选择性SERS检测转铁蛋白的分子印迹等离子体纳米传感器[151]和(B)覆有印迹层的金纳米棒SERS传感器[152]

Figure 12. Overview of (A) molecularly imprinted plasmonic nanosensor for selective SERS detection of transferrin[151] and (B) gold nanorods coated with imprinted layer (MIP@AuNR) SERS sensor[152]

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基于分子印迹与表面增强拉曼散射的蛋白质疾病标志物检测研究进展

Advances in Protein Biomarker Assay via the Combination of Molecular Imprinting and Surface-enhanced Raman Scattering

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