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Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics
A method for effective development of solid-phase immunoassays on a glass surface and for optimization of related protocols by highly sensitive quantitative monitoring of each assay step has been proposed and experimentally implemented. The method is based on the spectral correlation interferometry...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
A.I. Gordeyev
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3999470/ https://www.ncbi.nlm.nih.gov/pubmed/24772331 |
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author | Orlov, A. V. Burenin, A. G. Shipunova, V. O. Lizunova, A. A. Gorshkov, B. G. Nikitin, P. I. |
author_facet | Orlov, A. V. Burenin, A. G. Shipunova, V. O. Lizunova, A. A. Gorshkov, B. G. Nikitin, P. I. |
author_sort | Orlov, A. V. |
collection | PubMed |
description | A method for effective development of solid-phase immunoassays on a glass surface and for optimization of related protocols by highly sensitive quantitative monitoring of each assay step has been proposed and experimentally implemented. The method is based on the spectral correlation interferometry (SCI) that allows real-time measuring of the thickness of a biomolecular layer bound to the recognition molecular receptors on the sensor chip surface. The method is realized with compact 3-channel SCI-biosensors that employ as the sensor chips standard cover glass slips without deposition of any additional films. Different schemes for antibody immobilization on a glass surface have been experimentally compared and optimized toward a higher sorption capacity of the sensor chips. Comparative characterization of the kinetics of each immunoassay stage has been implemented with the optimized protocols: i) covalent immobilization of antibody on an epoxylated surface and ii) biotinylated antibody sorption on a biotinylated surface via a high-affinity biotin-streptavidin bond. We have shown that magnetic nanoparticles employed as labels with model detection of cardiac troponin I further amplify the SCI signal, resulting in 100-fold improvement of the detection limit. The developed protocols can also be used with the alternative immunoassay platforms, including the label methods based on registration of only the final assay result, which is the quantity of bound labels. |
format | Online Article Text |
id | pubmed-3999470 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | A.I. Gordeyev |
record_format | MEDLINE/PubMed |
spelling | pubmed-39994702014-04-25 Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics Orlov, A. V. Burenin, A. G. Shipunova, V. O. Lizunova, A. A. Gorshkov, B. G. Nikitin, P. I. Acta Naturae Research Article A method for effective development of solid-phase immunoassays on a glass surface and for optimization of related protocols by highly sensitive quantitative monitoring of each assay step has been proposed and experimentally implemented. The method is based on the spectral correlation interferometry (SCI) that allows real-time measuring of the thickness of a biomolecular layer bound to the recognition molecular receptors on the sensor chip surface. The method is realized with compact 3-channel SCI-biosensors that employ as the sensor chips standard cover glass slips without deposition of any additional films. Different schemes for antibody immobilization on a glass surface have been experimentally compared and optimized toward a higher sorption capacity of the sensor chips. Comparative characterization of the kinetics of each immunoassay stage has been implemented with the optimized protocols: i) covalent immobilization of antibody on an epoxylated surface and ii) biotinylated antibody sorption on a biotinylated surface via a high-affinity biotin-streptavidin bond. We have shown that magnetic nanoparticles employed as labels with model detection of cardiac troponin I further amplify the SCI signal, resulting in 100-fold improvement of the detection limit. The developed protocols can also be used with the alternative immunoassay platforms, including the label methods based on registration of only the final assay result, which is the quantity of bound labels. A.I. Gordeyev 2014 /pmc/articles/PMC3999470/ /pubmed/24772331 Text en Copyright ® 2014 Park-media Ltd. http://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Orlov, A. V. Burenin, A. G. Shipunova, V. O. Lizunova, A. A. Gorshkov, B. G. Nikitin, P. I. Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics |
title | Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics |
title_full | Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics |
title_fullStr | Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics |
title_full_unstemmed | Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics |
title_short | Development of Immunoassays Using Interferometric Real-Time Registration of Their Kinetics |
title_sort | development of immunoassays using interferometric real-time registration of their kinetics |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3999470/ https://www.ncbi.nlm.nih.gov/pubmed/24772331 |
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