期刊
ANALYTICAL METHODS
卷 9, 期 42, 页码 6049-6060出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c7ay01216k
关键词
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资金
- Russian Science Foundation [14-25-00132]
- Ministry of Education and Science of the Russian Federation [RFMEFI62117X0017, 14.621.21.0017]
- Russian Federation President scholarship [SP-4280.2016.4]
- Russian Science Foundation [17-25-00016] Funding Source: Russian Science Foundation
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.
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