Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection
Two subunits of the ternary troponin complex, I and C, have cardiac muscle specific isoforms, and hence could be applied as highly-selective markers of acute coronary syndrome. We aimed at paving the way for the development of a robust cardiac troponin I-detecting sandwich assay by replacing antibod...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-07-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/14/4963 |
| _version_ | 1797562522042105856 |
|---|---|
| author | Zoltán János Tolnai Judit András Zsuzsanna Szeitner Krisztina Percze László Ferenc Simon Róbert E. Gyurcsányi Tamás Mészáros |
| author_facet | Zoltán János Tolnai Judit András Zsuzsanna Szeitner Krisztina Percze László Ferenc Simon Róbert E. Gyurcsányi Tamás Mészáros |
| author_sort | Zoltán János Tolnai |
| collection | DOAJ |
| description | Two subunits of the ternary troponin complex, I and C, have cardiac muscle specific isoforms, and hence could be applied as highly-selective markers of acute coronary syndrome. We aimed at paving the way for the development of a robust cardiac troponin I-detecting sandwich assay by replacing antibodies with nuclease resistant aptamer analogues, so-called spiegelmers. To complement the previously generated spiegelmers that were specific for the N-terminus of cTnI, spiegelmers were selected for an amino acid stretch in the proximity of the C-terminal part of the protein by using a D-amino acid composed peptide. Following the selection, the oligonucleotides were screened by filter binding assay, and surface plasmon resonance analysis of the most auspicious candidates demonstrated that this approach could provide spiegelmers with subnanomolar dissociation constant. To demonstrate if the selected spiegelmers are functional and suitable for cTnI detection in a sandwich type arrangement, AlphaLisa technology was leveraged and the obtained results demonstrated that spiegelmers with different epitope selectivity are suitable for specific detection of cTnI protein even in human plasma containing samples. These results suggest that spiegelmers could be considered in the development of the next generation cTnI monitoring assays. |
| first_indexed | 2024-03-10T18:29:28Z |
| format | Article |
| id | doaj.art-3dd25e0752d6401bb73361191228e3a4 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T18:29:28Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-3dd25e0752d6401bb73361191228e3a42023-11-20T06:41:36ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-07-012114496310.3390/ijms21144963Spiegelmer-Based Sandwich Assay for Cardiac Troponin I DetectionZoltán János Tolnai0Judit András1Zsuzsanna Szeitner2Krisztina Percze3László Ferenc Simon4Róbert E. Gyurcsányi5Tamás Mészáros6Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó u. 37-47., H-1094 Budapest, HungaryDepartment of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó u. 37-47., H-1094 Budapest, HungaryDepartment of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó u. 37-47., H-1094 Budapest, HungaryDepartment of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó u. 37-47., H-1094 Budapest, HungaryBME “Lendület” Chemical Nanosensors Research Group, Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szt. Gellért tér 4, H-1111 Budapest, HungaryBME “Lendület” Chemical Nanosensors Research Group, Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szt. Gellért tér 4, H-1111 Budapest, HungaryDepartment of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó u. 37-47., H-1094 Budapest, HungaryTwo subunits of the ternary troponin complex, I and C, have cardiac muscle specific isoforms, and hence could be applied as highly-selective markers of acute coronary syndrome. We aimed at paving the way for the development of a robust cardiac troponin I-detecting sandwich assay by replacing antibodies with nuclease resistant aptamer analogues, so-called spiegelmers. To complement the previously generated spiegelmers that were specific for the N-terminus of cTnI, spiegelmers were selected for an amino acid stretch in the proximity of the C-terminal part of the protein by using a D-amino acid composed peptide. Following the selection, the oligonucleotides were screened by filter binding assay, and surface plasmon resonance analysis of the most auspicious candidates demonstrated that this approach could provide spiegelmers with subnanomolar dissociation constant. To demonstrate if the selected spiegelmers are functional and suitable for cTnI detection in a sandwich type arrangement, AlphaLisa technology was leveraged and the obtained results demonstrated that spiegelmers with different epitope selectivity are suitable for specific detection of cTnI protein even in human plasma containing samples. These results suggest that spiegelmers could be considered in the development of the next generation cTnI monitoring assays.https://www.mdpi.com/1422-0067/21/14/4963spiegelmertroponinIsandwich assay |
| spellingShingle | Zoltán János Tolnai Judit András Zsuzsanna Szeitner Krisztina Percze László Ferenc Simon Róbert E. Gyurcsányi Tamás Mészáros Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection International Journal of Molecular Sciences spiegelmer troponinI sandwich assay |
| title | Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection |
| title_full | Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection |
| title_fullStr | Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection |
| title_full_unstemmed | Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection |
| title_short | Spiegelmer-Based Sandwich Assay for Cardiac Troponin I Detection |
| title_sort | spiegelmer based sandwich assay for cardiac troponin i detection |
| topic | spiegelmer troponinI sandwich assay |
| url | https://www.mdpi.com/1422-0067/21/14/4963 |
| work_keys_str_mv | AT zoltanjanostolnai spiegelmerbasedsandwichassayforcardiactroponinidetection AT juditandras spiegelmerbasedsandwichassayforcardiactroponinidetection AT zsuzsannaszeitner spiegelmerbasedsandwichassayforcardiactroponinidetection AT krisztinapercze spiegelmerbasedsandwichassayforcardiactroponinidetection AT laszloferencsimon spiegelmerbasedsandwichassayforcardiactroponinidetection AT robertegyurcsanyi spiegelmerbasedsandwichassayforcardiactroponinidetection AT tamasmeszaros spiegelmerbasedsandwichassayforcardiactroponinidetection |