Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology

Risa Yamashiro,1 Akikazu Sakudo,1 Masaaki Nagatsu2 1Laboratory of Biometabolic Chemistry, School of Health Sciences, University of the Ryukyus, Nishihara, Okinawa 903-0215, Japan; 2Research Institute of Electronics, Shizuoka University, Hamamatsu, Shizuoka 432-8561, Japan Background: Rota...

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Autores principales:	Yamashiro R, Sakudo A, Nagatsu M
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	rotavirus antibody-integrated magnetic beads nanobeads nanoparticle Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/374f33e18b5e41a3aeae2e52681938b3
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spelling	oai:doaj.org-article:374f33e18b5e41a3aeae2e52681938b32021-12-02T11:34:40ZEfficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology1178-2013https://doaj.org/article/374f33e18b5e41a3aeae2e52681938b32019-03-01T00:00:00Zhttps://www.dovepress.com/efficient-recovery-and-enrichment-of-infectious-rotavirus-using-separa-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Risa Yamashiro,1 Akikazu Sakudo,1 Masaaki Nagatsu2 1Laboratory of Biometabolic Chemistry, School of Health Sciences, University of the Ryukyus, Nishihara, Okinawa 903-0215, Japan; 2Research Institute of Electronics, Shizuoka University, Hamamatsu, Shizuoka 432-8561, Japan Background: Rotavirus is the representative cause of severe acute gastroenteritis in young children. A characteristic feature of rotavirus is low infectious dose and robustness of the virion, suggesting sanitation and hygiene will have little impact. Thus, development of a vaccine should be given priority. Efficient capture of infectious viruses is an important step in generating a vaccine. Previously, antibody-integrated magnetic nanobeads (MNBs) have been developed for virus capture. This study examines the applicability of this method for infectious rotavirus recovery and enrichment.Materials and methods: Graphite-encapsulated MNBs were treated with radio frequency (RF) excited Ar/NH3 gas mixture plasma to introduce amino groups onto their surfaces. Rotavirus viral protein 7 (VP7) antibody was attached to the surface of MNBs via these amino groups using a coupling agent, N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP). The antibody-integrated MNBs were incubated with rotavirus-infected cell lysate and then separated from the supernatant by applying a magnetic field. After thorough washing, rotavirus was recovered and enrichment analysis done by polymerase chain reaction (PCR), immunochromatography, and an infection analysis using MA104 cells.Results and discussion: Immunochromatography and PCR indicate that anti-rotavirus antibody-integrated MNPs efficiently capture rotavirus with the capsid protein and viral RNA. The estimated recovery rate was 80.2% by PCR and 90.0% by infection analysis, while the concentrating factor was 7.9-fold by PCR and 6.7-fold by infection analysis. In addition, the absence of non-specific binding to the antibody-integrated MNPs was confirmed using anti-dengue virus antibody-integrated MNBs as a negative control.Conclusion: These results suggest that this capture procedure is a useful tool for recovery and enrichment of infectious rotavirus. Moreover, when combined with a suitable virus assay this capture procedure can increase the sensitivity of rotavirus detection. Therefore, this capture method is a valuable tool for generating vaccines as well as for developing sensitive detection systems for viruses. Keywords: rotavirus, antibody-integrated, magnetic beads, nanobeads, nanoparticleYamashiro RSakudo ANagatsu MDove Medical Pressarticlerotavirusantibody-integratedmagnetic beadsnanobeadsnanoparticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 14, Pp 1865-1876 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	rotavirus antibody-integrated magnetic beads nanobeads nanoparticle Medicine (General) R5-920
spellingShingle	rotavirus antibody-integrated magnetic beads nanobeads nanoparticle Medicine (General) R5-920 Yamashiro R Sakudo A Nagatsu M Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
description	Risa Yamashiro,1 Akikazu Sakudo,1 Masaaki Nagatsu2 1Laboratory of Biometabolic Chemistry, School of Health Sciences, University of the Ryukyus, Nishihara, Okinawa 903-0215, Japan; 2Research Institute of Electronics, Shizuoka University, Hamamatsu, Shizuoka 432-8561, Japan Background: Rotavirus is the representative cause of severe acute gastroenteritis in young children. A characteristic feature of rotavirus is low infectious dose and robustness of the virion, suggesting sanitation and hygiene will have little impact. Thus, development of a vaccine should be given priority. Efficient capture of infectious viruses is an important step in generating a vaccine. Previously, antibody-integrated magnetic nanobeads (MNBs) have been developed for virus capture. This study examines the applicability of this method for infectious rotavirus recovery and enrichment.Materials and methods: Graphite-encapsulated MNBs were treated with radio frequency (RF) excited Ar/NH3 gas mixture plasma to introduce amino groups onto their surfaces. Rotavirus viral protein 7 (VP7) antibody was attached to the surface of MNBs via these amino groups using a coupling agent, N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP). The antibody-integrated MNBs were incubated with rotavirus-infected cell lysate and then separated from the supernatant by applying a magnetic field. After thorough washing, rotavirus was recovered and enrichment analysis done by polymerase chain reaction (PCR), immunochromatography, and an infection analysis using MA104 cells.Results and discussion: Immunochromatography and PCR indicate that anti-rotavirus antibody-integrated MNPs efficiently capture rotavirus with the capsid protein and viral RNA. The estimated recovery rate was 80.2% by PCR and 90.0% by infection analysis, while the concentrating factor was 7.9-fold by PCR and 6.7-fold by infection analysis. In addition, the absence of non-specific binding to the antibody-integrated MNPs was confirmed using anti-dengue virus antibody-integrated MNBs as a negative control.Conclusion: These results suggest that this capture procedure is a useful tool for recovery and enrichment of infectious rotavirus. Moreover, when combined with a suitable virus assay this capture procedure can increase the sensitivity of rotavirus detection. Therefore, this capture method is a valuable tool for generating vaccines as well as for developing sensitive detection systems for viruses. Keywords: rotavirus, antibody-integrated, magnetic beads, nanobeads, nanoparticle
format	article
author	Yamashiro R Sakudo A Nagatsu M
author_facet	Yamashiro R Sakudo A Nagatsu M
author_sort	Yamashiro R
title	Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
title_short	Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
title_full	Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
title_fullStr	Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
title_full_unstemmed	Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
title_sort	efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/374f33e18b5e41a3aeae2e52681938b3
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Efficient recovery and enrichment of infectious rotavirus using separation with antibody-integrated graphite-encapsulated magnetic nanobeads produced by argon/ammonia gas plasma technology

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