化学发光功能化金属有机骨架探针在免疫分析中的应用

Development of detection protocols for extrinsic harmful contaminants in traditional Chinese medicines is of great significance for the insurance of drug safety. The reported protocols for the detection of these contaminants often suffer from low testing efficiency and unsatisfied selectivity. Various functional metal-organic frameworks (MOFs) have attracted increasing interest due to their unique physical and chemical properties, easy modification, exceptional tunability of structures and inner porosity. In the present project, MOFs will be utilized to conduct several chemiluminescent (CL) immunoassay approaches for the detection of harmful contaminants in traditional Chinese medicines. Firstly, some MOFs possessing high enzyme-like activity, such as peroxidase mimetics, hydrolase mimetics and reductase mimetics, will be adopted as CL probes for the development of enzyme-mimetic MOFs-based CL immunoassay. Secondly, by utilizing the encapsulating of CL reagents such as acridinium ester and luminol, MOFs with enormous pores will be used as probe carriers for the development of highly sensitive MOFs-based CL immunoassays. Thirdly, due to the increased CL signals by Fenton-like MOFs, MOFs will be utilized as CL enhancers in Fenton-like MOFs-based CL immunoassays. The project can achieve stable and reliable MOFs-based CL probes for the development of highly sensitive and selective immunoassay platform for the detection of extrinsic harmful contaminants in traditional Chinese medicines.

外源性有害残留物的检测对中药质量评价具有重要的意义，有助于严格限制中药中污染物的残留量，进而确保病人的身体健康。为了解决已有检测方法在检测效率和特异性上的缺陷，本项目基于组分多样、结构可控以及易修饰的MOFs纳米材料，研究针对外源性有害残留物的新型化学发光免疫分析方法。具体内容包括：①将具有高效类酶活性（例如氧化酶、水解酶、还原酶）的MOFs作为信号探针，催化化学发光反应，构建MOFs纳米酶型免疫分析技术。②将高负载量的MOFs作为载体，高效载入化学发光体系的催化剂、发光剂（例如吖啶酯、鲁米诺），提高发光体系的发光信号，构建化学发光试剂-MOFs复合物型免疫分析方法。③将具备高氧化活性的MOFs作为增敏剂，显著地增敏化学发光体系信号，构建MOFs类芬顿试剂型免疫分析平台。本研究有望制备稳定性良好、发光性能优异的MOFs化学发光探针，并构建用于外源性有害残留物检测的免疫分析平台。

外源性有害残留物对中药安全的危害是不容忽视的，因而它是中药质量评价的重要一环。目前针对中药外源性有害残留物的检测方法，主要是基于大型仪器的检测技术，这些方法的检测效率和特异性有待提高。本项目基于组分多样、结构可控以及易修饰的MOFs纳米材料，构建高灵敏、高选择性的新型化学发光免疫分析方法，用于外源性有害残留物的快速筛查和现场分析。代表性研究成果如下：1）基于Pearson的MOFs“硬酸-硬碱/软酸-软碱配对”理论，以Fe MIL-53为模板成功合成了Co单原子催化剂，可将鲁米诺体系化学发光信号增敏1349倍。将其用作信号探针，采用夹心免疫分析模式，即可完成肌钙蛋白I的化学发光免疫分析。检测范围为0.01-50 ng/mL,其检出限为3.3 pg/mL。2）基于Pearson的MOFs“硬酸-硬碱/软酸-软碱配对”理论，以UiO-66为模板成功合成了Co单原子催化剂，可将鲁米诺体系化学发光信号增敏3775倍。Ti3C2 MXenes对Co单原子催化剂-鲁米诺化学发光体系信号猝灭率高达94.6%，将其用作化学信号猝灭型探针，构建侧向层析免疫夹心分析方法用于肌钙蛋白I的特异性检测。该检测范围为1.0-100 pg/mL，检出限为0.33 pg/mL。3）以yolk-shell型ZIF-8@SiO2纳米球为载体，采用原位碱金属刻蚀技术，成功制备了掺杂型Co 单原子催化剂，其可对鲁米诺化学发光体系增强3133.7倍。将其作为化学发光探针标记黄曲霉毒素B1抗体，初步建立了该待检物的高灵敏度免疫分析方法。其检测范围为10-1000 pg/mL，其检出限为 0.44 pg/mL。4）采用低温NaBH4还原技术，以C3N4为模板制备了双位点Co 单原子催化剂，其可对鲁米诺化学发光体系增强22829倍。将其作为化学发光探针标记黄曲霉毒素B1抗体，初步建立了该待检物的高灵敏度免疫分析方法。其检测范围为1.0-500 pg/mL，其检出限为0.33 pg/mL。在项目经费的资助下，以第一作者以及通讯作者在Analytical Chemistry、Chemistry-A European Journal和ACS Applied Materials & Interfaces刊物上发表SCI论文3篇；以第一完成人身份申请发明专利1项；以第三完成人身份获得2018年度“重庆市自然科学奖”二等奖。