【作者】 王立新；

【导师】 屠一锋；

【作者基本信息】 苏州大学 ， 化学， 2023， 硕士

【摘要】 冠心病(CHD)是威胁人类健康的主要疾病,其发病机制是冠状动脉中粥样斑块的形成,致使血管狭窄甚至堵塞。与其它生物标志物相比,脂蛋白相关的磷脂酶A2(Lp-PLA2)参与了动脉粥样硬化的多个过程,有大量研究表明,人血液中Lp-PLA2水平升高是冠心病事件的一个高风险因素,并且独立于其它的心血管危险因素和多种炎症生物标志物,这一发现有利于对临床患者进行风险评估,以此帮助预警与动脉粥样硬化相关的冠心病风险。据目前的商业化检测方法和之前报道的文献,Lp-PLA2的检测方法主要有质谱法、分光光度法、乳胶增强比浊法、放射免疫分析法、酶联免疫法和光电化学免疫法等。然而,这些方法多成本昂贵、操作复杂和费时费力,不利于临床或居家的即时应用。因此,迫切需求设计开发一种快速、便捷、准确的Lp-PLA2检测方法。本研究以钴铁普鲁士蓝类似物(CoFe PBA)和金纳米粒子(AuNPs)构建的多功能纳米复合材料(AuNPs@CoFe PBA)作为传感基底,开发了一种高灵敏的用于检测Lp-PLA2的电化学发光(ECL)免疫传感器。得益于CoFe PBA和AuNPs的协同作用,纳米复合材料具有优异的类过氧化物酶样活性,能够催化鲁米诺ECL反应,极大的增强了 ECL信号约29倍,为传感响应提供了充分的基础水准。同时,纳米复合材料的比表面积增加和大量的AuNPs为固定更多的抗体提供了条件,从而提升了该ECL免疫传感器的灵敏度和响应范围。当目标物Lp-PLA2被免疫传感器捕获时,基于免疫复合物的形成导致了传质阻力和电子转移阻力的增加,免疫传感器的ECL信号被抑制。在优化的条件下,构建的ECL免疫传感器拥有从1 ng/mL到2200 ng/mL的宽线性范围,检测限低至0.21 ng/mL。此外,该传感器还具有良好的特异性、稳定性和重现性。该研究为CHD的预防和诊断提供了一种新的策略,并进一步扩大了普鲁士蓝类似物在ECL传感器领域中的应用。更多还原

【Abstract】 Coronary heart disease(CHD)has become a major threat to human health,which the pathogenesis is the formation of atheromatous plaques in the coronary ventricles.Compared to other biomarkers,Lipoprotein-associated phospholipase A2(Lp-PLA2)is involved in multiple processes of atherosclerosis.Numerous studies have demonstrated that higher levels of Lp-PLA2 in human blood are a significant risk factor for coronary events.This risk factor is independent of other cardiovascular risk factors and a number of inflammatory biomarkers.This finding is useful for risk assessment of clinical patients as an early warning of the risk of coronary heart disease associated with atherosclerosis.From the current commercial assays and previously reported literature,the detection methods for Lp-PLA2 include mass spectrometry,spectrophotometry,latexenhanced turbidimetry,radioimmunoassay,enzyme-linked immunoassay and photoelectrochemical immunoassay.However,these methods are expensive,complex and time-consuming to operate,which are not conducive to timely patient detection.Therefore,there is an urgent need to design and develop a method that can detect LpPLA2 quickly,conveniently and accurately.Using a multifunctional nanocomposite(AuNPs@CoFe PBA)containing CoFe Prussian Blue Analogue(CoFe PBA)and AuNPs as sensing platform,an electrochemiluminescent immunosensor for the highly sensitive detection of LpPLA2 was developed.Benefitting the synergistic effect of CoFe PBA and AuNPs,nanocomposite had excellent peroxidase-like activity,and was able to catalyze the luminol ECL reaction,amplifying the ECL signal by～29-fold.Meanwhile,the increased specific surface area of the nanocomposite and the abundant AuNPs provided conditions for immobilization of more antibodies,which increased sensitivity of this ECL immunosensor.When the target Lp-PLA2 was captured,the ECL immunosensor responded with a reduced ECL signal,which was attributed to an increase in mass transfer resistance due to the formation of immune complexes.Under optimized conditions,the constructed ECL immunosensor displayed a broad linear range from 1 ng/mL to 2200 ng/mL,and the low detection limit was 0.21 ng/mL.Additionally,it offered high specificity,stability,and repeatability.This work provides a new approach to diagnosing CHD and broads the application of PBA in the field of ECL biosensors.更多还原