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On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen
Sensitive quantification of protein biomarkers is highly desired for clinical diagnosis and treatment. Yet, unlike DNA/RNA which can be greatly amplified by PCR/RT-PCR, the amplification and detection of trace amount of proteins remain a great challenge. Here, we combined allosteric probe (AP) with...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Vienna
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9342938/ https://www.ncbi.nlm.nih.gov/pubmed/35915288 http://dx.doi.org/10.1007/s00604-022-05404-4 |
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author | Ling, Min Luo, Na Cui, Lanyu Cao, Yongqiang Ning, Xueping Sun, Jian Xu, Xiaoping He, Shengbin |
author_facet | Ling, Min Luo, Na Cui, Lanyu Cao, Yongqiang Ning, Xueping Sun, Jian Xu, Xiaoping He, Shengbin |
author_sort | Ling, Min |
collection | PubMed |
description | Sensitive quantification of protein biomarkers is highly desired for clinical diagnosis and treatment. Yet, unlike DNA/RNA which can be greatly amplified by PCR/RT-PCR, the amplification and detection of trace amount of proteins remain a great challenge. Here, we combined allosteric probe (AP) with magnetic bead (MB) for assembling an on-bead DNA synthesis system (named as APMB) to amplify protein signals. The AP is designed and conjugated onto the MB, enabling the protein biomarker to be separated and enriched. Once recognizing the biomarker, the AP alters its conformation to initiate DNA synthesis on beads for primary signal amplification. During the DNA synthesis, biotin-dATPs are incorporated into the newly synthesized DNA strands. Then, the biotin-labeled DNA specifically captures streptavidin (STR)–conjugated horseradish peroxidase (HRP), which is used to catalyze a colorimetric reaction for secondary signal amplification. By using carcinoembryonic antigen (CEA) as a protein model, the APMB can quantify protein biomarkers of as low as 0.01 ng/mL. The response values measured by APMB are linearly related to the protein concentrations in the range 0.05 to 20 ng/mL. Clinical examination demonstrated good practicability of the APMB in quantifying serum protein biomarker. The on-bead DNA synthesis could be exploited to improve protein signal amplification, thus facilitating protein biomarker detection of low abundance for early diagnosis. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00604-022-05404-4. |
format | Online Article Text |
id | pubmed-9342938 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Springer Vienna |
record_format | MEDLINE/PubMed |
spelling | pubmed-93429382022-08-02 On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen Ling, Min Luo, Na Cui, Lanyu Cao, Yongqiang Ning, Xueping Sun, Jian Xu, Xiaoping He, Shengbin Mikrochim Acta Original Paper Sensitive quantification of protein biomarkers is highly desired for clinical diagnosis and treatment. Yet, unlike DNA/RNA which can be greatly amplified by PCR/RT-PCR, the amplification and detection of trace amount of proteins remain a great challenge. Here, we combined allosteric probe (AP) with magnetic bead (MB) for assembling an on-bead DNA synthesis system (named as APMB) to amplify protein signals. The AP is designed and conjugated onto the MB, enabling the protein biomarker to be separated and enriched. Once recognizing the biomarker, the AP alters its conformation to initiate DNA synthesis on beads for primary signal amplification. During the DNA synthesis, biotin-dATPs are incorporated into the newly synthesized DNA strands. Then, the biotin-labeled DNA specifically captures streptavidin (STR)–conjugated horseradish peroxidase (HRP), which is used to catalyze a colorimetric reaction for secondary signal amplification. By using carcinoembryonic antigen (CEA) as a protein model, the APMB can quantify protein biomarkers of as low as 0.01 ng/mL. The response values measured by APMB are linearly related to the protein concentrations in the range 0.05 to 20 ng/mL. Clinical examination demonstrated good practicability of the APMB in quantifying serum protein biomarker. The on-bead DNA synthesis could be exploited to improve protein signal amplification, thus facilitating protein biomarker detection of low abundance for early diagnosis. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00604-022-05404-4. Springer Vienna 2022-08-01 2022 /pmc/articles/PMC9342938/ /pubmed/35915288 http://dx.doi.org/10.1007/s00604-022-05404-4 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Original Paper Ling, Min Luo, Na Cui, Lanyu Cao, Yongqiang Ning, Xueping Sun, Jian Xu, Xiaoping He, Shengbin On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
title | On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
title_full | On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
title_fullStr | On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
title_full_unstemmed | On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
title_short | On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
title_sort | on-bead dna synthesis triggered by allosteric probe for detection of carcinoembryonic antigen |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9342938/ https://www.ncbi.nlm.nih.gov/pubmed/35915288 http://dx.doi.org/10.1007/s00604-022-05404-4 |
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