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An easy pipeline for one-step purification of SARS-CoV-2 nucleocapsid protein from insect cell suspension culture
The COVID-19 pandemic has demanded a range of biotechnological products for detection of SARS-CoV-2 variants and evaluation of human seroconversion after infection or vaccination. In this work, we describe an easy pipeline for expression of SARS-CoV-2 nucleocapsid (N) protein in insect cells followe...
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Published in: | Journal of virological methods 2022-01, Vol.299, p.114341-114341, Article 114341 |
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description | The COVID-19 pandemic has demanded a range of biotechnological products for detection of SARS-CoV-2 variants and evaluation of human seroconversion after infection or vaccination. In this work, we describe an easy pipeline for expression of SARS-CoV-2 nucleocapsid (N) protein in insect cells followed by its purification via affinity chromatography. The N gene was cloned into the genome of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) via transposition and the resulting recombinant baculovirus was used for infection of lepidopteran Sf9 cells adapted to high-density suspension. Using Tris−HCl pH 8.0 buffer as mobile phase and eluting bound proteins with 175 mM imidazole as part of a three-step gradient, an average of 1 mg N protein could be purified from each 50 mg of total protein from clarified supernatant. Such protein amount allows the manufacturing of serological tests and the development of basic studies on cellular responses to SARS-CoV-2. |
doi_str_mv | 10.1016/j.jviromet.2021.114341 |
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Such protein amount allows the manufacturing of serological tests and the development of basic studies on cellular responses to SARS-CoV-2.</description><subject>Affinity chromatography</subject><subject>Animals</subject><subject>Baculovirus</subject><subject>COVID-19</subject><subject>Humans</subject><subject>Insecta</subject><subject>Nucleocapsid</subject><subject>Nucleocapsid protein</subject><subject>Nucleocapsid Proteins - genetics</subject><subject>One-step purification</subject><subject>Pandemics</subject><subject>SARS-CoV-2</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNqFkUtv1DAUhS0EokPhL1RessngZ5xsEKNReUiVkCiwtRznGjzK2K6djNR_j6NpK1ixsiWfc-7x_RC6omRLCW3fHbaHk8_xCPOWEUa3lAou6DO0oZ3qG9J34jnaVGFb71xcoFelHAghUnH-El1w0fa9Uu0G3e0CBlPucfIJJh8Au5hxDNCUGRJOS_bOWzP7GHB0-Hb37bbZx58Nw2GxE0RrUvEjTjnO4AN2tRL2oYCdsYVpwmUpCUJZ7XaZ5iXDa_TCmanAm4fzEv34eP19_7m5-frpy35301ih6NwwI6hyiriO21EoR5kEaftanA6ctX0HtlXcDhKUAmI6JgflGO8p4eMwcMIv0ftzblqGI4wWwpzNpFP2R5PvdTRe__sS_G_9K550J3kvZVsD3j4E5Hi3QJn10Zf1UyZAXIpmslOECM5XaXuW2hxLyeCexlCiV176oB956ZWXPvOqxqu_Sz7ZHgFVwYezAOqqTh6yLtZDsDD6XJesx-j_N-MPmoislg</recordid><startdate>20220101</startdate><enddate>20220101</enddate><creator>de Camargo, Brenda R.</creator><creator>da Silva, Leonardo A.</creator><creator>de Oliveira, Athos S.</creator><creator>Ribeiro, Bergmann M.</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-1061-196X</orcidid></search><sort><creationdate>20220101</creationdate><title>An easy pipeline for one-step purification of SARS-CoV-2 nucleocapsid protein from insect cell suspension culture</title><author>de Camargo, Brenda R. ; da Silva, Leonardo A. ; de Oliveira, Athos S. ; Ribeiro, Bergmann M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c471t-2a417f70f83cd47f125e5c94691b32698ec673cb5e77e0a825b7f239103dbb303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Affinity chromatography</topic><topic>Animals</topic><topic>Baculovirus</topic><topic>COVID-19</topic><topic>Humans</topic><topic>Insecta</topic><topic>Nucleocapsid</topic><topic>Nucleocapsid protein</topic><topic>Nucleocapsid Proteins - genetics</topic><topic>One-step purification</topic><topic>Pandemics</topic><topic>SARS-CoV-2</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>de Camargo, Brenda R.</creatorcontrib><creatorcontrib>da Silva, Leonardo A.</creatorcontrib><creatorcontrib>de Oliveira, Athos S.</creatorcontrib><creatorcontrib>Ribeiro, Bergmann M.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>de Camargo, Brenda R.</au><au>da Silva, Leonardo A.</au><au>de Oliveira, Athos S.</au><au>Ribeiro, Bergmann M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An easy pipeline for one-step purification of SARS-CoV-2 nucleocapsid protein from insect cell suspension culture</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2022-01-01</date><risdate>2022</risdate><volume>299</volume><spage>114341</spage><epage>114341</epage><pages>114341-114341</pages><artnum>114341</artnum><issn>0166-0934</issn><eissn>1879-0984</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><notes>These authors contributed equally to this article.</notes><abstract>The COVID-19 pandemic has demanded a range of biotechnological products for detection of SARS-CoV-2 variants and evaluation of human seroconversion after infection or vaccination. 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source | ScienceDirect Journals |
subjects | Affinity chromatography Animals Baculovirus COVID-19 Humans Insecta Nucleocapsid Nucleocapsid protein Nucleocapsid Proteins - genetics One-step purification Pandemics SARS-CoV-2 |
title | An easy pipeline for one-step purification of SARS-CoV-2 nucleocapsid protein from insect cell suspension culture |
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