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目的合成能特异性识别肿瘤细胞的聚丁二炔生物传感器。方法超声乳化-共价修饰法制备聚丁二炔/磷脂(polydiacetylene/Phospholipid,PDA/PC)纳米囊泡,通过共价修饰法将鼠抗人细胞角蛋白抗体CK19共价修饰固定在PDA/PC纳米囊泡表面,制备可变色的PDA/PC生物传感器,用于检测肿瘤细胞。透射电镜负染技术,激光散射粒径测定仪、紫外-可见分光光度计扫描,计算比色响应(colorimetric response,CR)等对合成的PC/PDA生物传感器进行表征,以乳腺癌细胞株MCF-7为模型,模拟考察PDA/PC生物传感器识别鉴定外周血循环肿瘤细胞(circulating tumorcells,CTCs)可行性。结果 TEM检查结果显示制备的PDA/PC生物传感器呈球形或类球形、粒径均匀,平均粒径约为(223.4±23.6)nm;激光散射粒径分析仪显示生物传感器的强均粒径和多分散系数为298.4 nm和0.184;特异性抗原加入后,纳米粒发生团聚,颜色出现由蓝至红的变化,且CR值随着抗原浓度增加而增加,与高表达的乳腺癌细胞MCF-7发生特异性结合,充分混匀后,溶液颜色产生由蓝至红的变化。结论成功合成由单克隆抗体抗CK19修饰的PDA/PC生物传感器,通过免疫化学显色技术,可快速、有效、灵敏的检测溶液中微量乳腺癌MCF-7细胞,为PDA/PC生物传感器技术在CTC识别上的应用奠定基础。
Objective To synthesize polybutadienyne biosensors that can specifically recognize tumor cells. METHODS: Phacoemulsification-covalent modification was used to prepare polyydiacetylene / phospholipid (PDA / PC) nanovesicles. The covalent modification of mouse anti-human cytokeratin antibody CK19 was immobilized on PDA / PC Nanofibrillar surfaces were prepared for a variable color PDA / PC biosensor for detection of tumor cells. The PC / PDA biosensor was characterized by transmission electron microscopy (TEM) staining, laser scattering particle size analyzer, UV-visible spectrophotometer and colorimetric response (CR) 7 as a model to simulate the feasibility of identifying peripheral blood circulating tumor cells (CTCs) by PDA / PC biosensor. Results The results of TEM examination showed that the prepared PDA / PC biosensor was spherical or spheroidal with average particle diameter of (223.4 ± 23.6) nm. The laser scattering particle size analyzer showed that the biosensor had a strong average particle diameter The dispersion coefficients were 298.4 nm and 0.184, respectively. After the specific antigen was added, the nanoparticles agglomerated and the color changed from blue to red, and the CR value increased with the increase of antigen concentration, which coincided with the high expression of MCF-7 in breast cancer cells Specific binding, the full mix, the solution color changes from blue to red. Conclusion The PDA / PC biosensor modified by the monoclonal antibody against CK19 was successfully synthesized. The immunoprecipitation of MCF-7 cells in solution could be rapidly, effectively and sensitively detected by immunochemical staining. CTC identification based on the application laid the foundation.