Ivanov Yuri D, Bukharina Natalia S, Pleshakova Tatyana O, Frantsuzov Pavel A, Andreeva Elena Yu, Kaysheva Anna L, Zgoda Victor G, Izotov Alexander A, Pavlova Tatyana I, Ziborov Vadim S, Radko Sergey P, Moshkovskii Sergei A, Archakov Alexander I
Department of Personalized Medicine, Orekhovich Institute of Biomedical Chemistry of the Russian Academy of Medical Sciences, Moscow, Russia.
Department of Personalized Medicine, Orekhovich Institute of Biomedical Chemistry of the Russian Academy of Medical Sciences, Moscow, Russia ; PostgenTech Ltd., Moscow, Russia.
Int J Nanomedicine. 2014 Oct 3;9:4659-70. doi: 10.2147/IJN.S66946. eCollection 2014.
Atomic force microscopy (AFM) was applied to carry out direct and label-free detection of gp120 human immunodeficiency virus type 1 envelope glycoprotein as a target protein. This approach was based on the AFM fishing of gp120 from the analyte solution using anti-gp120 aptamers immobilized on the AFM chip to count gp120/aptamer complexes that were formed on the chip surface. The comparison of image contrasts of fished gp120 against the background of immobilized aptamers and anti-gp120 antibodies on the AFM images was conducted. It was shown that an image contrast of the protein/aptamer complexes was two-fold higher than the contrast of the protein/antibody complexes. Mass spectrometry identification provided an additional confirmation of the target protein presence on the AFM chips after biospecific fishing to avoid any artifacts.
应用原子力显微镜（AFM）对作为靶蛋白的1型人类免疫缺陷病毒包膜糖蛋白gp120进行直接且无标记的检测。该方法基于使用固定在AFM芯片上的抗gp120适体从分析物溶液中对gp120进行AFM捕捉，以计数在芯片表面形成的gp120/适体复合物。在AFM图像上，对捕捉到的gp120与固定适体和抗gp120抗体背景的图像对比度进行了比较。结果表明，蛋白质/适体复合物的图像对比度比蛋白质/抗体复合物的对比度高两倍。质谱鉴定为生物特异性捕捉后AFM芯片上靶蛋白的存在提供了额外的确认，以避免任何假象。
