Atomic force microscopy fishing and mass spectrometry identification of gp120 on immobilized aptamers

原子力显微镜钓取和质谱鉴定固定化适体上的 gp120

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作者:Yuri D Ivanov, Natalia S Bukharina, Tatyana O Pleshakova, Pavel A Frantsuzov, Elena Yu Andreeva, Anna L Kaysheva, Victor G Zgoda, Alexander A Izotov, Tatyana I Pavlova, Vadim S Ziborov, Sergey P Radko, Sergei A Moshkovskii, Alexander I Archakov

Abstract

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.

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