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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...

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Detalles Bibliográficos
Autores principales: Ling, Min, Luo, Na, Cui, Lanyu, Cao, Yongqiang, Ning, Xueping, Sun, Jian, Xu, Xiaoping, He, Shengbin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Vienna 2022
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.
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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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