Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests

Abstract Enzymes are the cornerstone of modern biotechnology. Achromopeptidase (ACP) is a well-known enzyme that hydrolyzes a number of proteins, notably proteins on the surface of Gram-positive bacteria. It is therefore used for sample preparation in nucleic acid tests. However, ACP inhibits DNA am...

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Autores principales:	Georgios Chondrogiannis, Shirin Khaliliazar, Anna Toldrà, Pedro Réu, Mahiar M. Hamedi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5683787a9cc1402793b2f139fb92178c
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spelling	oai:doaj.org-article:5683787a9cc1402793b2f139fb92178c2021-12-02T17:05:45ZNitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests10.1038/s41598-021-85481-22045-2322https://doaj.org/article/5683787a9cc1402793b2f139fb92178c2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85481-2https://doaj.org/toc/2045-2322Abstract Enzymes are the cornerstone of modern biotechnology. Achromopeptidase (ACP) is a well-known enzyme that hydrolyzes a number of proteins, notably proteins on the surface of Gram-positive bacteria. It is therefore used for sample preparation in nucleic acid tests. However, ACP inhibits DNA amplification which makes its integration difficult. Heat is commonly used to inactivate ACP, but it can be challenging to integrate heating into point-of-care devices. Here, we use recombinase polymerase amplification (RPA) together with ACP, and show that when ACP is immobilized on nitrocellulose paper, it retains its enzymatic function and can easily and rapidly be activated using agitation. The nitrocellulose-bound ACP does, however, not leak into the solution, preventing the need for deactivation through heat or by other means. Nitrocellulose-bound ACP thus opens new possibilities for paper-based Point-of-Care (POC) devices.Georgios ChondrogiannisShirin KhaliliazarAnna ToldràPedro RéuMahiar M. HamediNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Georgios Chondrogiannis Shirin Khaliliazar Anna Toldrà Pedro Réu Mahiar M. Hamedi Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
description	Abstract Enzymes are the cornerstone of modern biotechnology. Achromopeptidase (ACP) is a well-known enzyme that hydrolyzes a number of proteins, notably proteins on the surface of Gram-positive bacteria. It is therefore used for sample preparation in nucleic acid tests. However, ACP inhibits DNA amplification which makes its integration difficult. Heat is commonly used to inactivate ACP, but it can be challenging to integrate heating into point-of-care devices. Here, we use recombinase polymerase amplification (RPA) together with ACP, and show that when ACP is immobilized on nitrocellulose paper, it retains its enzymatic function and can easily and rapidly be activated using agitation. The nitrocellulose-bound ACP does, however, not leak into the solution, preventing the need for deactivation through heat or by other means. Nitrocellulose-bound ACP thus opens new possibilities for paper-based Point-of-Care (POC) devices.
format	article
author	Georgios Chondrogiannis Shirin Khaliliazar Anna Toldrà Pedro Réu Mahiar M. Hamedi
author_facet	Georgios Chondrogiannis Shirin Khaliliazar Anna Toldrà Pedro Réu Mahiar M. Hamedi
author_sort	Georgios Chondrogiannis
title	Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
title_short	Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
title_full	Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
title_fullStr	Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
title_full_unstemmed	Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
title_sort	nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/5683787a9cc1402793b2f139fb92178c
work_keys_str_mv	AT georgioschondrogiannis nitrocelluloseboundachromopeptidaseforpointofcarenucleicacidtests AT shirinkhaliliazar nitrocelluloseboundachromopeptidaseforpointofcarenucleicacidtests AT annatoldra nitrocelluloseboundachromopeptidaseforpointofcarenucleicacidtests AT pedroreu nitrocelluloseboundachromopeptidaseforpointofcarenucleicacidtests AT mahiarmhamedi nitrocelluloseboundachromopeptidaseforpointofcarenucleicacidtests
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Nitrocellulose-bound achromopeptidase for point-of-care nucleic acid tests

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