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Issue 42, 2017
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Advantages of aptamers as ligands upon protein detection by AFM-based fishing

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Abstract

A combined AFM/MS method was employed for protein registration in solution. This method is based on reversible specific capturing of a target protein from a large volume of analyzed solution onto a small sensor area of a chip with immobilized aptamer ligands. Fishing of the core antigen of hepatitis C virus (HCVcoreAg) from 10−12 M solution of this protein in buffer was carried out. It has been shown that the AFM/MS method allows the detection of HCVcoreAg in the form of a protein conjugate as well as in the presence of protein matrix (components of human serum). At the first stage, aptamer/antigen complexes were AFM-registered on the chip surface after its incubation in 10−12 M solution of antigen in buffer. A technique for the analysis was developed, and criteria for the evaluation of AFM analysis data based on the counting of the aptamer/antigen complexes were proposed. At the second stage, the mass spectrometric identification of protein objects on the chip surface was accomplished; this allowed the reliable identification of the target protein (the core antigen of hepatitis C virus, HCVcoreAg) and confirmed the AFM analysis data.

Graphical abstract: Advantages of aptamers as ligands upon protein detection by AFM-based fishing

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Publication details

The article was received on 12 May 2017, accepted on 21 Sep 2017 and first published on 28 Sep 2017


Article type: Paper
DOI: 10.1039/C7AY01216K
Citation: Anal. Methods, 2017,9, 6049-6060
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    Advantages of aptamers as ligands upon protein detection by AFM-based fishing

    T. O. Pleshakova, A. L. Kaysheva, J. M. Bayzyanova, A. S. Anashkina, V. F. Uchaikin, I. D. Shumov, V. S. Ziborov, V. A. Konev, A. I. Archakov and Yu. D. Ivanov, Anal. Methods, 2017, 9, 6049
    DOI: 10.1039/C7AY01216K

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