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

Abstract

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

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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, Accepted Manuscript
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    Advantages of aptamers as ligands upon protein detection by AFM-based fishing

    T. Pleshakova, A. Kaysheva, J. Bayzyanova, A. Anashkina, V. Uchaikin, I. Shumov, V. Ziborov, V. Konev, A. Archakov and Y. Ivanov, Anal. Methods, 2017, Accepted Manuscript , DOI: 10.1039/C7AY01216K

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