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- Publisher Website: 10.1038/s41598-022-06817-0
- Scopus: eid_2-s2.0-85125155086
- PMID: 35194086
- WOS: WOS:000759999200046
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Article: A multiparametric fluorescence assay for screening aptamer–protein interactions based on microbeads
Title | A multiparametric fluorescence assay for screening aptamer–protein interactions based on microbeads |
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Authors | |
Issue Date | 2022 |
Citation | Scientific Reports, 2022, v. 12, n. 1, article no. 2961 How to Cite? |
Abstract | For improving aptamer-ligand binding we have developed a screening system that defines optimal binding buffer composition. Using multiplex assays, one buffer system is needed which guarantees the specific binding of all aptamers. We investigated nine peer-reviewed DNA aptamers. Non-specific binding of aptamers is an obstacle. To address this, we investigated 16 proteins as specificity controls bound covalently to encoded microbeads in a multiplex assay. Increasing the NaCl concentration decreased the binding for all aptamers. Changing pH values by one unit higher or lower did not influence the aptamer binding significantly. However, pH < 5 led to non-specific binding for all aptamers. The PfLDH-aptamer selected in the absence of divalent cations exhibited doubling of its binding signal by the addition of Ca2+ and Mg2+. We confirmed Ca2+ and Mg2+ dependency of the aptamers for streptavidin and thrombin by observing a 90% and 50% binding decrease, respectively. We also achieved a doubling of binding for the streptavidin aptamer when replacing Ca2+ and Mg2+ by Mn2+. A buffer suitable for all aptamers can have considerable variations in pH or ionic strength, but divalent cations (Ca2+, Mg2+, Mn2+) are essential. |
Persistent Identifier | http://hdl.handle.net/10722/334810 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Schmidt, Carsten | - |
dc.contributor.author | Kammel, Anne | - |
dc.contributor.author | Tanner, Julian A. | - |
dc.contributor.author | Kinghorn, Andrew B. | - |
dc.contributor.author | Khan, Muhammad Moman | - |
dc.contributor.author | Lehmann, Werner | - |
dc.contributor.author | Menger, Marcus | - |
dc.contributor.author | Schedler, Uwe | - |
dc.contributor.author | Schierack, Peter | - |
dc.contributor.author | Rödiger, Stefan | - |
dc.date.accessioned | 2023-10-20T06:50:54Z | - |
dc.date.available | 2023-10-20T06:50:54Z | - |
dc.date.issued | 2022 | - |
dc.identifier.citation | Scientific Reports, 2022, v. 12, n. 1, article no. 2961 | - |
dc.identifier.uri | http://hdl.handle.net/10722/334810 | - |
dc.description.abstract | For improving aptamer-ligand binding we have developed a screening system that defines optimal binding buffer composition. Using multiplex assays, one buffer system is needed which guarantees the specific binding of all aptamers. We investigated nine peer-reviewed DNA aptamers. Non-specific binding of aptamers is an obstacle. To address this, we investigated 16 proteins as specificity controls bound covalently to encoded microbeads in a multiplex assay. Increasing the NaCl concentration decreased the binding for all aptamers. Changing pH values by one unit higher or lower did not influence the aptamer binding significantly. However, pH < 5 led to non-specific binding for all aptamers. The PfLDH-aptamer selected in the absence of divalent cations exhibited doubling of its binding signal by the addition of Ca2+ and Mg2+. We confirmed Ca2+ and Mg2+ dependency of the aptamers for streptavidin and thrombin by observing a 90% and 50% binding decrease, respectively. We also achieved a doubling of binding for the streptavidin aptamer when replacing Ca2+ and Mg2+ by Mn2+. A buffer suitable for all aptamers can have considerable variations in pH or ionic strength, but divalent cations (Ca2+, Mg2+, Mn2+) are essential. | - |
dc.language | eng | - |
dc.relation.ispartof | Scientific Reports | - |
dc.title | A multiparametric fluorescence assay for screening aptamer–protein interactions based on microbeads | - |
dc.type | Article | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1038/s41598-022-06817-0 | - |
dc.identifier.pmid | 35194086 | - |
dc.identifier.scopus | eid_2-s2.0-85125155086 | - |
dc.identifier.volume | 12 | - |
dc.identifier.issue | 1 | - |
dc.identifier.spage | article no. 2961 | - |
dc.identifier.epage | article no. 2961 | - |
dc.identifier.eissn | 2045-2322 | - |
dc.identifier.isi | WOS:000759999200046 | - |