Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags
Express and highly sensitive immunoassays for the quantitative registration of cardiac troponin I (cTnI) are in high demand for early point-of-care differential diagnosis of acute myocardial infarction. The selection of antibodies that feature rapid and tight binding with antigens is crucial for imm...
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| Format: | Article |
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MDPI AG
2022-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/9/4474 |
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| author | Alexey V. Orlov Juri A. Malkerov Denis O. Novichikhin Sergey L. Znoyko Petr I. Nikitin |
| author_facet | Alexey V. Orlov Juri A. Malkerov Denis O. Novichikhin Sergey L. Znoyko Petr I. Nikitin |
| author_sort | Alexey V. Orlov |
| collection | DOAJ |
| description | Express and highly sensitive immunoassays for the quantitative registration of cardiac troponin I (cTnI) are in high demand for early point-of-care differential diagnosis of acute myocardial infarction. The selection of antibodies that feature rapid and tight binding with antigens is crucial for immunoassay rate and sensitivity. A method is presented for the selection of the most promising clones for advanced immunoassays via simultaneous characterization of interaction kinetics of different monoclonal antibodies (mAb) using a direct label-free method of multiplex spectral correlation interferometry. mAb-cTnI interactions were real-time registered on an epoxy-modified microarray glass sensor chip that did not require activation. The covalent immobilization of mAb microdots on its surface provided versatility, convenience, and virtually unlimited multiplexing potential. The kinetics of tracer antibody interaction with the “cTnI—capture antibody” complex was characterized. Algorithms are shown for excluding mutual competition of the tracer/capture antibodies and selecting the optimal pairs for different assay formats. Using the selected mAbs, a lateral flow assay was developed for rapid quantitative cTnI determination based on electronic detection of functionalized magnetic nanoparticles applied as labels (detection limit—0.08 ng/mL, dynamic range > 3 orders). The method can be extended to other molecular biomarkers for high-throughput screening of mAbs and rational development of immunoassays. |
| first_indexed | 2024-03-10T04:08:40Z |
| format | Article |
| id | doaj.art-e8d624799c5c4a32a6a06431ca9cb1a3 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:08:40Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e8d624799c5c4a32a6a06431ca9cb1a32023-11-23T08:17:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-04-01239447410.3390/ijms23094474Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic NanotagsAlexey V. Orlov0Juri A. Malkerov1Denis O. Novichikhin2Sergey L. Znoyko3Petr I. Nikitin4Prokhorov General Physics Institute of the Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow, RussiaProkhorov General Physics Institute of the Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow, RussiaProkhorov General Physics Institute of the Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow, RussiaProkhorov General Physics Institute of the Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow, RussiaProkhorov General Physics Institute of the Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow, RussiaExpress and highly sensitive immunoassays for the quantitative registration of cardiac troponin I (cTnI) are in high demand for early point-of-care differential diagnosis of acute myocardial infarction. The selection of antibodies that feature rapid and tight binding with antigens is crucial for immunoassay rate and sensitivity. A method is presented for the selection of the most promising clones for advanced immunoassays via simultaneous characterization of interaction kinetics of different monoclonal antibodies (mAb) using a direct label-free method of multiplex spectral correlation interferometry. mAb-cTnI interactions were real-time registered on an epoxy-modified microarray glass sensor chip that did not require activation. The covalent immobilization of mAb microdots on its surface provided versatility, convenience, and virtually unlimited multiplexing potential. The kinetics of tracer antibody interaction with the “cTnI—capture antibody” complex was characterized. Algorithms are shown for excluding mutual competition of the tracer/capture antibodies and selecting the optimal pairs for different assay formats. Using the selected mAbs, a lateral flow assay was developed for rapid quantitative cTnI determination based on electronic detection of functionalized magnetic nanoparticles applied as labels (detection limit—0.08 ng/mL, dynamic range > 3 orders). The method can be extended to other molecular biomarkers for high-throughput screening of mAbs and rational development of immunoassays.https://www.mdpi.com/1422-0067/23/9/4474molecular biomarkersdetection of biomoleculessuperparamagnetic iron oxide nanoparticleshigh-throughput sensingcardiac markersreal-time optical biosensors |
| spellingShingle | Alexey V. Orlov Juri A. Malkerov Denis O. Novichikhin Sergey L. Znoyko Petr I. Nikitin Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags International Journal of Molecular Sciences molecular biomarkers detection of biomolecules superparamagnetic iron oxide nanoparticles high-throughput sensing cardiac markers real-time optical biosensors |
| title | Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags |
| title_full | Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags |
| title_fullStr | Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags |
| title_full_unstemmed | Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags |
| title_short | Multiplex Label-Free Kinetic Characterization of Antibodies for Rapid Sensitive Cardiac Troponin I Detection Based on Functionalized Magnetic Nanotags |
| title_sort | multiplex label free kinetic characterization of antibodies for rapid sensitive cardiac troponin i detection based on functionalized magnetic nanotags |
| topic | molecular biomarkers detection of biomolecules superparamagnetic iron oxide nanoparticles high-throughput sensing cardiac markers real-time optical biosensors |
| url | https://www.mdpi.com/1422-0067/23/9/4474 |
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