Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells
In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (G E ) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel ch...
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| Veröffentlicht in: | 3 Biotech 2019-11, Vol.9 (11), p.1-11, Article 385 |
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| creator | Targovnik, Alexandra Marisa Ferrari, Alejandro Mc Callum, Gregorio Juan Arregui, Mariana Bernadett Smith, Ignacio Bracco, Lautaro Fidel Alfonso, Victoria López, María Gabriela Martínez-Solís, María Herrero, Salvador Miranda, María Victoria |
| description | In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (G
E
) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (
polh
-
pSeL
), the expression level reached a yield of 4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard
polh
promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, G
E
was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein. |
| doi_str_mv | 10.1007/s13205-019-1920-4 |
| format | Article |
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E
) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (
polh
-
pSeL
), the expression level reached a yield of 4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard
polh
promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, G
E
was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein.</description><identifier>ISSN: 2190-572X</identifier><identifier>EISSN: 2190-5738</identifier><identifier>DOI: 10.1007/s13205-019-1920-4</identifier><identifier>PMID: 31656723</identifier><language>eng</language><publisher>Cham: Springer International Publishing</publisher><subject>Affinity chromatography ; Agriculture ; Antibodies ; Antigens ; Baculovirus ; Bioinformatics ; Biomaterials ; Biotechnology ; Cancer Research ; Cell culture ; Chemistry ; Chemistry and Materials Science ; Glycoprotein G ; Glycoproteins ; Insect cells ; Insects ; Lysates ; Lyssavirus ; Metal ions ; Original ; Original Article ; Rabies ; Stem Cells ; Vaccines ; Viruses</subject><ispartof>3 Biotech, 2019-11, Vol.9 (11), p.1-11, Article 385</ispartof><rights>King Abdulaziz City for Science and Technology 2019</rights><rights>Copyright Springer Nature B.V. 2019</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c495t-b151695ec39bfb5df66969a201cdc73206df8f7d86e8a137b51f08af039957be3</citedby><cites>FETCH-LOGICAL-c495t-b151695ec39bfb5df66969a201cdc73206df8f7d86e8a137b51f08af039957be3</cites><orcidid>0000-0003-4480-031X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6778167/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6778167/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,27926,27927,41490,42559,51321,53793,53795</link.rule.ids></links><search><creatorcontrib>Targovnik, Alexandra Marisa</creatorcontrib><creatorcontrib>Ferrari, Alejandro</creatorcontrib><creatorcontrib>Mc Callum, Gregorio Juan</creatorcontrib><creatorcontrib>Arregui, Mariana Bernadett</creatorcontrib><creatorcontrib>Smith, Ignacio</creatorcontrib><creatorcontrib>Bracco, Lautaro Fidel</creatorcontrib><creatorcontrib>Alfonso, Victoria</creatorcontrib><creatorcontrib>López, María Gabriela</creatorcontrib><creatorcontrib>Martínez-Solís, María</creatorcontrib><creatorcontrib>Herrero, Salvador</creatorcontrib><creatorcontrib>Miranda, María Victoria</creatorcontrib><title>Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells</title><title>3 Biotech</title><addtitle>3 Biotech</addtitle><description>In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (G
E
) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (
polh
-
pSeL
), the expression level reached a yield of 4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard
polh
promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, G
E
was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein.</description><subject>Affinity chromatography</subject><subject>Agriculture</subject><subject>Antibodies</subject><subject>Antigens</subject><subject>Baculovirus</subject><subject>Bioinformatics</subject><subject>Biomaterials</subject><subject>Biotechnology</subject><subject>Cancer Research</subject><subject>Cell culture</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Glycoprotein G</subject><subject>Glycoproteins</subject><subject>Insect cells</subject><subject>Insects</subject><subject>Lysates</subject><subject>Lyssavirus</subject><subject>Metal ions</subject><subject>Original</subject><subject>Original Article</subject><subject>Rabies</subject><subject>Stem Cells</subject><subject>Vaccines</subject><subject>Viruses</subject><issn>2190-572X</issn><issn>2190-5738</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNp1kUFrGzEQhUVpaILjH9CboJdcttWsLGnnEiihcQqBHppAexJarWQrrFeutBvwv6-MjUMCEYKRNN889HiEfAb2FRhT3zLwmomKAVaANasWH8hFDcgqoXjz8XSu_5yTec5PrCwBAoF9IuccpJCq5hfk711Yrfsddd4HG9ww0m2K3WTHEAcaPU2mDS7T55CmTFf9zsbSH10Y6JI6O8Yubky5lP3bYym5PFLr-j5fkjNv-uzmxzojj7c_Hm7uqvtfy5833-8ru0AxVi0IkCic5dj6VnReSpRoaga2s6pYlJ1vvOoa6RoDXLUCPGuMZxxRqNbxGbk-6G6nduM6Wzwk0-ttChuTdjqaoF93hrDWq_ispVINSFUEro4CKf6bXB71JuS9BTO4OGVdc4YNcFRY0C9v0Kc4paHY21MMpUDZFAoOlE0x5-T86TPA9D47fchOl-z0Pju9KDP1YSYXdli59KL8_tB_a7ubQw</recordid><startdate>20191101</startdate><enddate>20191101</enddate><creator>Targovnik, Alexandra Marisa</creator><creator>Ferrari, Alejandro</creator><creator>Mc Callum, Gregorio Juan</creator><creator>Arregui, Mariana Bernadett</creator><creator>Smith, Ignacio</creator><creator>Bracco, Lautaro Fidel</creator><creator>Alfonso, Victoria</creator><creator>López, María Gabriela</creator><creator>Martínez-Solís, María</creator><creator>Herrero, Salvador</creator><creator>Miranda, María Victoria</creator><general>Springer International Publishing</general><general>Springer Nature B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-4480-031X</orcidid></search><sort><creationdate>20191101</creationdate><title>Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells</title><author>Targovnik, Alexandra Marisa ; Ferrari, Alejandro ; Mc Callum, Gregorio Juan ; Arregui, Mariana Bernadett ; Smith, Ignacio ; Bracco, Lautaro Fidel ; Alfonso, Victoria ; López, María Gabriela ; Martínez-Solís, María ; Herrero, Salvador ; Miranda, María Victoria</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c495t-b151695ec39bfb5df66969a201cdc73206df8f7d86e8a137b51f08af039957be3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Affinity chromatography</topic><topic>Agriculture</topic><topic>Antibodies</topic><topic>Antigens</topic><topic>Baculovirus</topic><topic>Bioinformatics</topic><topic>Biomaterials</topic><topic>Biotechnology</topic><topic>Cancer Research</topic><topic>Cell culture</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Glycoprotein G</topic><topic>Glycoproteins</topic><topic>Insect cells</topic><topic>Insects</topic><topic>Lysates</topic><topic>Lyssavirus</topic><topic>Metal ions</topic><topic>Original</topic><topic>Original Article</topic><topic>Rabies</topic><topic>Stem Cells</topic><topic>Vaccines</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Targovnik, Alexandra Marisa</creatorcontrib><creatorcontrib>Ferrari, Alejandro</creatorcontrib><creatorcontrib>Mc Callum, Gregorio Juan</creatorcontrib><creatorcontrib>Arregui, Mariana Bernadett</creatorcontrib><creatorcontrib>Smith, Ignacio</creatorcontrib><creatorcontrib>Bracco, Lautaro Fidel</creatorcontrib><creatorcontrib>Alfonso, Victoria</creatorcontrib><creatorcontrib>López, María Gabriela</creatorcontrib><creatorcontrib>Martínez-Solís, María</creatorcontrib><creatorcontrib>Herrero, Salvador</creatorcontrib><creatorcontrib>Miranda, María Victoria</creatorcontrib><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>3 Biotech</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Targovnik, Alexandra Marisa</au><au>Ferrari, Alejandro</au><au>Mc Callum, Gregorio Juan</au><au>Arregui, Mariana Bernadett</au><au>Smith, Ignacio</au><au>Bracco, Lautaro Fidel</au><au>Alfonso, Victoria</au><au>López, María Gabriela</au><au>Martínez-Solís, María</au><au>Herrero, Salvador</au><au>Miranda, María Victoria</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells</atitle><jtitle>3 Biotech</jtitle><stitle>3 Biotech</stitle><date>2019-11-01</date><risdate>2019</risdate><volume>9</volume><issue>11</issue><spage>1</spage><epage>11</epage><pages>1-11</pages><artnum>385</artnum><issn>2190-572X</issn><eissn>2190-5738</eissn><abstract>In the present study, we developed a complete process to produce in insect cells a high amount of the ectodomain of rabies virus glycoprotein G (G
E
) as suitable antigen for detecting anti-rabies antibodies. Using the baculovirus expression vector system in Sf9 insect cells combined with a novel chimeric promoter (
polh
-
pSeL
), the expression level reached a yield of 4.1 ± 0.3 mg/L culture, which was significantly higher than that achieved with the standard
polh
promoter alone. The protein was recovered from the cell lysates and easily purified in only one step by metal ion affinity chromatography, with a yield of 95% and a purity of 87%. Finally, G
E
was successfully used in an assay to detect specific antibodies in serum samples derived from rabies-vaccinated animals. The efficient strategy developed in this work is an interesting method to produce high amounts of this glycoprotein.</abstract><cop>Cham</cop><pub>Springer International Publishing</pub><pmid>31656723</pmid><doi>10.1007/s13205-019-1920-4</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0003-4480-031X</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Affinity chromatography Agriculture Antibodies Antigens Baculovirus Bioinformatics Biomaterials Biotechnology Cancer Research Cell culture Chemistry Chemistry and Materials Science Glycoprotein G Glycoproteins Insect cells Insects Lysates Lyssavirus Metal ions Original Original Article Rabies Stem Cells Vaccines Viruses |
| title | Highly efficient production of rabies virus glycoprotein G ectodomain in Sf9 insect cells |
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