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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Imaging recognition events between human IgG and rat anti-human IgG by atomic force microscopy
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Imaging recognition events between human IgG and rat anti-human IgG by atomic force microscopy

机译：通过原子力显微镜对人IgG和大鼠抗人IgG之间的成像识别事件进行成像

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摘要

Chemically immobilized rat anti-human immunoglobulin (IgG) monolayers on thiols modified gold substrates were fabricated using self-assembled monolayer (SAM) method. The antibody monolayers were imaged before and after free human IgG treated, whilst recognition events between antigen and antibody were monitored by contact mode atomic force microscopy (CM-AFM) and tapping mode AFM (TM-AFM), with topographic and/or phase images being recorded. The obtained images with different surface compositions show distinct nanostructures, indicating occurrence of recognition and binding events of antigen-antibody. The size of the observed surface structures of the antibody monolayer, when tip broaden effect had been taken into account, was very close to the actual size of the antibody molecule. Thus, these results suggest CM-AFM is capable of, and proven satisfactory in detecting protein-protein interactions (PPIs), providing the sample was prepared appropriately and the scanning parameters were set adequately. Moreover, phase imaging can serve as a real time contrast enhancement technique to TM-AFM in terms of highlighting edges and clearly observing fine features.
机译：使用自组装单层（SAM）方法在硫醇修饰的金底物上化学固定的大鼠抗人免疫球蛋白（IgG）单层。在游离人IgG处理之前和之后对抗体单层成像，同时通过接触模式原子力显微镜（CM-AFM）和敲击模式AFM（TM-AFM）监视抗原和抗体之间的识别事件，并显示地形图和/或相图被记录。获得的具有不同表面组成的图像显示出不同的纳米结构，表明发生了抗原-抗体的识别和结合事件。当考虑到尖端加宽效应时，观察到的抗体单层表面结构的尺寸非常接近抗体分子的实际尺寸。因此，这些结果表明，只要适当地准备了样品并且适当地设置了扫描参数，CM-AFM就能够并且证明在检测蛋白质-蛋白质相互作用（PPI）方面令人满意。此外，就突出边缘和清晰观察精细特征而言，相位成像可以作为TM-AFM的实时对比度增强技术。

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