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[固定化适配体上 gp120 的原子力显微镜捕获及其质谱鉴定]

[Atomic force microscopy fishing of gp120 on immobilized aptamer and its mass spectrometry identification].

作者信息

Bukharina N S, Ivanov Yu D, Pleshakova T O, Frantsuzov P A, Andreeva E Yu, Kaysheva A L, Izotov A A, Pavlova T I, Ziborov V S, Radko S P, Archakov A I

机构信息

Institute of Biomedical Chemistry, Moscow, Russia.

出版信息

Biomed Khim. 2015 May-Jun;61(3):363-72. doi: 10.18097/PBMC20156103363.

Abstract

A method of atomic force microscopy-based fishing (AFM fishing) has been developed for protein detection in the analyte solution using a chip with an immobilized aptamer. This method is based on the biospecific fishing of a target protein from a bulk solution onto the small AFM chip area with the immobilized aptamer to this protein used as the molecular probe. Such aptamer-based approach allows to increase an AFM image contrast compared to the antibody-based approach. Mass spectrometry analysis used after the biospecific fishing to identify the target protein on the AFM chip has proved complex formation. Use of the AFM chip with the immobilized aptamer avoids interference of the antibody and target protein peaks in a mass spectrum.

摘要

一种基于原子力显微镜的垂钓法(AFM垂钓法)已被开发出来,用于在分析物溶液中检测蛋白质,该方法使用固定有适配体的芯片。此方法基于将目标蛋白质从大量溶液中通过生物特异性方式垂钓到具有固定该蛋白质适配体的小AFM芯片区域上,该适配体用作分子探针。与基于抗体的方法相比,这种基于适配体的方法能够提高AFM图像对比度。在生物特异性垂钓后使用质谱分析来鉴定AFM芯片上的目标蛋白质,已证明存在复合物形成。使用固定有适配体的AFM芯片可避免质谱中抗体和目标蛋白质峰的干扰。

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