反蛋白石结构分子印迹膜的制备及其在肿瘤标志物检测中的应用研究

Tumors have become the main cause of human death. Rapid detection of tumor markers in serum has significant biological importance and clinical values. Glycoproteins are a large family of carbohydrate-conjugated proteins that have been employed as tumor markers. Boronate affinity has been an important means for detection of glycoproteins. However, traditional boronate affinity method suffers from some disadvantages, such as the glycoproteins should be bound under alkaline conditions, large instrument was needed to transfer the identification process to readable signal. Herein, molecular imprinted polymer, photonic crystal and boronate affinity were combined together to fabricate imprinted membrane for naked-eye detection of tumor markers. The selectivity of the material would be improved by the high selectivity of the imprinted material to the target and the specific recognition of the boric acid to the glycoprotein. The mass transfer rate would be improved by the porous structure of the inverse opal skeleton. In addition, by designing a synthesized boric acid monomer with lower pKa value, the material could combined to the target molecule at physiological pH. Meanwhile, owing to the inverse opal structure of the imprinted membrane, it can be used for naked eye detection of the tumor markers. This project will expand the application field of boric acid affinity materials, and provide new ideas for the detection of tumor markers. It is expected to acquire new technology for tumor markers detection with independent intellectual property rights, which has important theoretical significance and application value. Therefore, this approach has good application prospects in many areas.

恶性肿瘤已成为导致人类死亡的主要因素。建立快速检测血清中肿瘤标志物的方法对肿瘤的早期诊断具有重要的临床价值。糖蛋白是非常重要的一类肿瘤标志物，传统硼酸亲和材料须在碱性条件下结合糖蛋白且需借助大型仪器将分子识别过程转换成可读信号。针对此问题，本项目拟结合硼酸亲和技术、表面印迹技术和光子晶体技术，制备具有信号自表达性能的印迹膜，构建快速检测糖蛋白类肿瘤标志物的方法。旨在通过印迹材料对目标物的高选择性及硼酸对糖蛋白特有的识别性，提高材料对糖蛋白的选择性。借助反蛋白石贯穿的孔道结构，提高传质速率。通过设计合成较低pKa值的硼酸单体，实现生理pH下的检测。同时，光子晶体完美的光学调控性能，赋予材料信号自表达性，可实现肿瘤标志物的裸眼检测。本项目的研究将拓展硼酸亲和材料的应用领域，为肿瘤标志物的检测提供新思路，有望获得具有自主知识产权的肿瘤标志物检测新技术，具有重要的理论意义和应用价值。

恶性肿瘤已成为导致人类死亡的主要因素。建立快速检测血清中肿瘤标志物的方法对肿瘤的早期诊断具有重要的临床价值。糖蛋白是非常重要的一类肿瘤标志物，传统硼酸亲和材料须在碱性条件下结合糖蛋白且需借助大型仪器将分子识别过程转换成可读信号。本研究将硼酸亲和配体、表面印迹技术和光子晶体技术结合，制备了具有信号自表达性能的硼酸基印迹光子晶体传感器，实现了糖蛋白类肿瘤标志物的快速、裸眼检测。首先以二氧化硅纳米球为基底，辣根过氧化物酶（HRP）为印迹模板，丙烯酰胺为功能单体，采用表面印迹技术，制备了对HRP具有较高吸附性能的印迹聚合物（SiO2@DFFPBA/MIPs）。印迹过程中，采用（2,4-二氟-3-甲酰基苯基硼酸，DFFPBA）将模板糖蛋白固定于二氧化硅纳米球表面，显著的提高了SiO2@DFFPBA/MIPs的吸附容量。考察了交联剂用量，对目标糖蛋白吸附量的影响。然后，采用蒸发诱导自组装法将SiO2@DFFPBA/MIPs组装成高度有序的三维光子晶体阵列（CPICA）。考察了离子强度和溶液pH对其响应性的影响。所制备的CPICA衍射峰随糖蛋白浓度的增加逐渐红移，并伴随着明显的颜色变化，最大红移量为87 nm，检出限为3.0 × 10−13 mol/mL。同时，由于DFFPBA具有较低的pKa值，实现了糖蛋白的高选择性、生理pH检测。总之，该研究中制备了具有信号自表达性能的印迹光子晶体传感器，探讨了其检测机理，建立了低成本，快速、裸眼检测糖蛋白类肿瘤标志物的新方法。本项目的研究为发展高灵敏度、低成本肿瘤标志物检测方法提供了新的思路，为光子晶体在生化传感器的应用拓展和检测机理提供了理论基础。

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