Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins

Currently, there is great interest in the development of highly sensitive bioanalytical systems for diagnosing diseases at an early stage, when pathological biomarkers are present in biological fluids at low concentrations and there are no clinical manifestations. A promising direction is the use of...

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Main Authors: Maria O. Ershova, Amir Taldaev, Petr V. Konarev, Georgy S. Peters, Anastasia A. Valueva, Irina A. Ivanova, Sergey V. Kraevsky, Andrey F. Kozlov, Vadim S. Ziborov, Yuri D. Ivanov, Alexander I. Archakov, Tatyana O. Pleshakova
Format: Article
Language:English
Published: MDPI AG 2023-12-01
Series:Biomolecules
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Online Access:https://www.mdpi.com/2218-273X/13/12/1776
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author Maria O. Ershova
Amir Taldaev
Petr V. Konarev
Georgy S. Peters
Anastasia A. Valueva
Irina A. Ivanova
Sergey V. Kraevsky
Andrey F. Kozlov
Vadim S. Ziborov
Yuri D. Ivanov
Alexander I. Archakov
Tatyana O. Pleshakova
author_facet Maria O. Ershova
Amir Taldaev
Petr V. Konarev
Georgy S. Peters
Anastasia A. Valueva
Irina A. Ivanova
Sergey V. Kraevsky
Andrey F. Kozlov
Vadim S. Ziborov
Yuri D. Ivanov
Alexander I. Archakov
Tatyana O. Pleshakova
author_sort Maria O. Ershova
collection DOAJ
description Currently, there is great interest in the development of highly sensitive bioanalytical systems for diagnosing diseases at an early stage, when pathological biomarkers are present in biological fluids at low concentrations and there are no clinical manifestations. A promising direction is the use of molecular detectors―highly sensitive devices that detect signals from single biomacromolecules. A typical detector in this class is the atomic force microscope (AFM). The high sensitivity of an AFM-based bioanalysis system is determined by the size of the sensing element of an atomic force microscope―the cantilever―the radius of the curvature of which is comparable to that of a biomolecule. Biospecific molecular probe–target interactions are used to ensure detection system specificity. Antibodies, aptamers, synthetic antibodies, and peptides can be used as molecular probes. This study has demonstrated the possibility of using aptamers as molecular probes for AFM-based detection of the ovarian cancer biomarker CA125. Antigen detection in a nanomolar solution was carried out using AFM chips with immobilized aptamers, commercially available or synthesized based on sequences from open sources. Both aptamer types can be used for antigen detection, but the availability of sequence information enables additional modeling of the aptamer structure with allowance for modifications necessary for immobilization of the aptamer on an AFM chip surface. Information on the structure and oligomeric composition of aptamers in the solution was acquired by combining small-angle X-ray scattering and molecular modeling. Modeling enabled pre-selection, before the experimental stage, of aptamers for use as surface-immobilized molecular probes.
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spelling doaj.art-db7e3b81c4924f898331b5ad0410d92b2023-12-22T13:56:04ZengMDPI AGBiomolecules2218-273X2023-12-011312177610.3390/biom13121776Selection of Aptamers for Use as Molecular Probes in AFM Detection of ProteinsMaria O. Ershova0Amir Taldaev1Petr V. Konarev2Georgy S. Peters3Anastasia A. Valueva4Irina A. Ivanova5Sergey V. Kraevsky6Andrey F. Kozlov7Vadim S. Ziborov8Yuri D. Ivanov9Alexander I. Archakov10Tatyana O. Pleshakova11Institute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaA.V. Shubnikov Institute of Crystallography of Federal Scientific Research Centre “Crystallography and Photonics” of Russian Academy of Sciences, Leninsky Ave. 59, 119333 Moscow, RussiaNational Research Centre “Kurchatov Institute”, Akademika Kurchatova Square 1, 123182 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaInstitute of Biomedical Chemistry, Pogodinskaya Str. 10/8, 119121 Moscow, RussiaCurrently, there is great interest in the development of highly sensitive bioanalytical systems for diagnosing diseases at an early stage, when pathological biomarkers are present in biological fluids at low concentrations and there are no clinical manifestations. A promising direction is the use of molecular detectors―highly sensitive devices that detect signals from single biomacromolecules. A typical detector in this class is the atomic force microscope (AFM). The high sensitivity of an AFM-based bioanalysis system is determined by the size of the sensing element of an atomic force microscope―the cantilever―the radius of the curvature of which is comparable to that of a biomolecule. Biospecific molecular probe–target interactions are used to ensure detection system specificity. Antibodies, aptamers, synthetic antibodies, and peptides can be used as molecular probes. This study has demonstrated the possibility of using aptamers as molecular probes for AFM-based detection of the ovarian cancer biomarker CA125. Antigen detection in a nanomolar solution was carried out using AFM chips with immobilized aptamers, commercially available or synthesized based on sequences from open sources. Both aptamer types can be used for antigen detection, but the availability of sequence information enables additional modeling of the aptamer structure with allowance for modifications necessary for immobilization of the aptamer on an AFM chip surface. Information on the structure and oligomeric composition of aptamers in the solution was acquired by combining small-angle X-ray scattering and molecular modeling. Modeling enabled pre-selection, before the experimental stage, of aptamers for use as surface-immobilized molecular probes.https://www.mdpi.com/2218-273X/13/12/1776AFMaptamersprotein detectionCA125bio specific interactionSAXS
spellingShingle Maria O. Ershova
Amir Taldaev
Petr V. Konarev
Georgy S. Peters
Anastasia A. Valueva
Irina A. Ivanova
Sergey V. Kraevsky
Andrey F. Kozlov
Vadim S. Ziborov
Yuri D. Ivanov
Alexander I. Archakov
Tatyana O. Pleshakova
Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins
Biomolecules
AFM
aptamers
protein detection
CA125
bio specific interaction
SAXS
title Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins
title_full Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins
title_fullStr Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins
title_full_unstemmed Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins
title_short Selection of Aptamers for Use as Molecular Probes in AFM Detection of Proteins
title_sort selection of aptamers for use as molecular probes in afm detection of proteins
topic AFM
aptamers
protein detection
CA125
bio specific interaction
SAXS
url https://www.mdpi.com/2218-273X/13/12/1776
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