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原子力显微镜(AFM)的发明为测量分子间特异性相互作用力提供了新的技术手段.利用AFM单分子力谱(SMFS)技术分别测量了提纯的CD20,淋巴瘤Raji细胞表面的CD20和淋巴瘤病人B细胞表面的CD20与Rituximab(抗CD20单克隆抗体)之间的相互作用力.通过探针功能化技术,将Rituximab连接到AFM针尖;通过基底功能化技术,将提纯的CD20分子吸附到云母表面,对CD20分子进行了AFM成像,并测量了CD20与Rituximab之间的相互作用力;通过静电吸附和化学固定,将淋巴瘤Raji细胞和淋巴瘤病人细胞固定到载玻片表面,对Raji细胞和病人细胞进行了AFM成像,并分别测量了Raji细胞表面的CD20和病人B细胞表面的CD20与Rituximab之间的相互作用力.比较并分析了在提纯的CD20分子表面、Raji细胞表面和病人B细胞表面测量CD20-Rituximab相互作用力的差异,实验结果表明Raji细胞表面的CD20与Rituximab之间的相互作用力明显小于提纯的CD20以及淋巴瘤病人B细胞表面的CD20与Rituximab之间的相互作用力,为深入研究造成Rituximab耐药性差异的分子机理提供了技术思路和实验方法.
The invention of atomic force microscope (AFM) has provided a new technical measure for measuring the intermolecular specific interaction force.It has been used to measure the purified CD20, CD20 and lymphoma on the surface of Raji cells by AFM single molecule force spectrum (SMFS) The interaction between CD20 and Rituximab (anti-CD20 mAb) on the patient’s B cell surface Rituximab is attached to the AFM tip by probe functionalization technology; the purified CD20 molecule is adsorbed to the mica by a substrate functionalization technique Surface, AFM imaging of CD20 molecules was performed and the interaction between CD20 and Rituximab was measured. Raji cells and lymphoma cells were immobilized on the surface of the slide by electrostatic adsorption and chemical fixation. Raji cells And the patient’s cell AFM imaging and measured the Raji cell surface CD20 and the patient’s B cell surface between the CD20 and Rituximab interaction between the comparison and analysis of purified CD20 molecule surface Raji cell surface and the patient B The difference in CD20-Rituximab interaction forces was measured on the cell surface. The experimental results show that the interaction between CD20 and Rituximab on the surface of Raji cells is significantly less than that of purified CD20 and the interaction between CD20 and Rituximab on the surface of B lymphocytes in patients with lymphoma provide technical ideas and experimental methods for further research on the molecular mechanism of the differences in Rituximab resistance.