研发用于肿瘤凝集素标志物筛选的三维微阵列芯片方法

In this proposal, a kind of novel three-dimensional lectin microarray-based assay is developed and employed for studying the interactions of lectins with cellular glycans because the interactions of proteins with glycans on living cell surface play an important role in the biological system. The three-dimensional lectin microarray is fabricated by photopolymerization reaction of acrylamide. Combination of a home-made detection system, the assay enables to evaluate glycan expression difference of living cells, the affinities of lectins with living cells and screen high specific lectin of tumor（i.e.,lectin biomakers of tumor）. Furthermore, specific lectin conjugated rare earth ions doped upconverting nanoparticles are also synthesized and used as contrast agents for in vivo upconversion luminescence and magnetic resonance imaging of specific tumors (i.e.,colorectal cancer), resulting to develop nanoparticle-based contrast agents for colorectal cancer diagnosis. The research results of the proposal provide a new method for glycomics studies of living systems and the early diagnosis of tumors.

本项目拟以在生命体系中起重要作用的糖基化合物与蛋白质间相互作用为主要研究对象，通过丙烯酰胺的光聚合反应构建三维凝集素微阵列芯片并研制相应的检测系统应用于活细胞表面糖基化合物和凝集素的相互作用研究。通过凝集素与活细胞亲和能力的差异获得不同细胞表面糖基化合物的表达差异；筛选与肿瘤细胞具有高亲和性和特异性的凝集素（即肿瘤凝集素标志物）。进一步通过共价修饰法合成凝集素标志物修饰的稀土掺杂上转换纳米粒子应用于特定肿瘤（结直肠癌）的活体上转换荧光和磁共振成像，考察所筛选凝集素标志物的肿瘤靶向性并获得能用于结直肠癌诊断的纳米造影剂。项目的研究成果能够为生命体的糖组学研究和肿瘤的早期诊断提供新方法。

应用光聚合反应构建了基于聚丙烯酰胺水凝胶、聚合物分子刷等的三维凝集素微阵列芯片（3D Lectin Microarray）并研制相应的检测装置；并以此为基础发展高通量方法应用于筛选对肿瘤具有高特异性的凝集素。以结直肠癌为研究对象，筛选特定结直肠癌细胞（SW480）的凝集素标志物。进一步构建凝集素修饰的稀土掺杂上转换纳米粒子（L-UCNP）应用于荷结直肠癌SW480裸鼠的活体上转换荧光（UCL）和磁共振（MR）双模态成像，获得能用于结直肠癌诊断的纳米造影剂。.本研究工作共在SCI等期刊发表学术论文29篇，申请中国发明专利3项，其中授权发明专利1项。