Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium
The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 me...
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| Veröffentlicht in: | Biologicals 2009-04, Vol.37 (2), p.108-118 |
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| creator | Batista, Fabiana R.X. Moraes, Ângela M. Büntemeyer, Heino Noll, Thomas |
| description | The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected
Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO
2 were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO
2 of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98
g/10
7 cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery. |
| doi_str_mv | 10.1016/j.biologicals.2008.11.001 |
| format | Article |
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Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO
2 were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO
2 of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98
g/10
7 cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery.</description><identifier>ISSN: 1045-1056</identifier><identifier>EISSN: 1095-8320</identifier><identifier>DOI: 10.1016/j.biologicals.2008.11.001</identifier><identifier>PMID: 19059791</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Aeration ; Amino Acids - metabolism ; Animals ; Carbohydrate Metabolism - drug effects ; Cell Culture Techniques ; Cell Line - metabolism ; Cell Line - physiology ; Cell Proliferation - drug effects ; Culture Media, Serum-Free - pharmacology ; Drosophila melanogaster ; Fed-batch culture ; Growth kinetics ; Hydrogen-Ion Concentration ; Lactic Acid - metabolism ; Oxygen Consumption - drug effects ; Rabies virus ; Rabies virus - genetics ; Rabies virus - metabolism ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; RVGP ; S2 cells ; Time Factors ; Viral Fusion Proteins - isolation & purification ; Viral Fusion Proteins - metabolism</subject><ispartof>Biologicals, 2009-04, Vol.37 (2), p.108-118</ispartof><rights>2008 The International Association for Biologicals</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c406t-5c380bbdf31dd1feee3b300339c4178159e9c45c62aea1f869af0eeb98fe65aa3</citedby><cites>FETCH-LOGICAL-c406t-5c380bbdf31dd1feee3b300339c4178159e9c45c62aea1f869af0eeb98fe65aa3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.biologicals.2008.11.001$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19059791$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Batista, Fabiana R.X.</creatorcontrib><creatorcontrib>Moraes, Ângela M.</creatorcontrib><creatorcontrib>Büntemeyer, Heino</creatorcontrib><creatorcontrib>Noll, Thomas</creatorcontrib><title>Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium</title><title>Biologicals</title><addtitle>Biologicals</addtitle><description>The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected
Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO
2 were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO
2 of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98
g/10
7 cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery.</description><subject>Aeration</subject><subject>Amino Acids - metabolism</subject><subject>Animals</subject><subject>Carbohydrate Metabolism - drug effects</subject><subject>Cell Culture Techniques</subject><subject>Cell Line - metabolism</subject><subject>Cell Line - physiology</subject><subject>Cell Proliferation - drug effects</subject><subject>Culture Media, Serum-Free - pharmacology</subject><subject>Drosophila melanogaster</subject><subject>Fed-batch culture</subject><subject>Growth kinetics</subject><subject>Hydrogen-Ion Concentration</subject><subject>Lactic Acid - metabolism</subject><subject>Oxygen Consumption - drug effects</subject><subject>Rabies virus</subject><subject>Rabies virus - genetics</subject><subject>Rabies virus - metabolism</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>RVGP</subject><subject>S2 cells</subject><subject>Time Factors</subject><subject>Viral Fusion Proteins - isolation & purification</subject><subject>Viral Fusion Proteins - metabolism</subject><issn>1045-1056</issn><issn>1095-8320</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkctuFDEQRS0EIiHwC8hs2HXj6reXaHhFisQCWFtud3nwyG0PfoyU7-CHcTMjkR1ZuSzfquu6h5A3wGpgMLw71LPx1u-NkjbWDWNTDVAzBk_INTDeV1PbsKdb3fUVsH64Ii9iPBQBdGP3nFwBZz0fOVyT37dO24xOIfWaqmxTDkiVd4tJxrtIvaMBlV9n46RL9EPw0R9_GivpilY6v5cxYaDfGqrQ2kiPwS9ZGbenQc4GIz2ZkCPd23vly1tC4_7amJNMuNByixjyWumAWEYuJq8vyTNdFsNXl_OG_Pj08fvuS3X39fPt7v1dpTo2pKpX7cTmedEtLAtoRGznlrG25aqDcYKeY6l6NTQSJehp4FIzxJlPGodeyvaGvD3PLf_6lTEmsZq4bSEd-hzFMJa8eD_8V9iwbhzGritCfhaqElMMqMUxmFWGewFMbOjEQTxAJzZ0AkAUMqX39cUkzyWHf50XVkWwOwuwZHIyGERUZiO3mEIoicWbR9j8AWSgtXg</recordid><startdate>20090401</startdate><enddate>20090401</enddate><creator>Batista, Fabiana R.X.</creator><creator>Moraes, Ângela M.</creator><creator>Büntemeyer, Heino</creator><creator>Noll, Thomas</creator><general>Elsevier Ltd</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>7SS</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20090401</creationdate><title>Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium</title><author>Batista, Fabiana R.X. ; Moraes, Ângela M. ; Büntemeyer, Heino ; Noll, Thomas</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c406t-5c380bbdf31dd1feee3b300339c4178159e9c45c62aea1f869af0eeb98fe65aa3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Aeration</topic><topic>Amino Acids - metabolism</topic><topic>Animals</topic><topic>Carbohydrate Metabolism - drug effects</topic><topic>Cell Culture Techniques</topic><topic>Cell Line - metabolism</topic><topic>Cell Line - physiology</topic><topic>Cell Proliferation - drug effects</topic><topic>Culture Media, Serum-Free - pharmacology</topic><topic>Drosophila melanogaster</topic><topic>Fed-batch culture</topic><topic>Growth kinetics</topic><topic>Hydrogen-Ion Concentration</topic><topic>Lactic Acid - metabolism</topic><topic>Oxygen Consumption - drug effects</topic><topic>Rabies virus</topic><topic>Rabies virus - genetics</topic><topic>Rabies virus - metabolism</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>RVGP</topic><topic>S2 cells</topic><topic>Time Factors</topic><topic>Viral Fusion Proteins - isolation & purification</topic><topic>Viral Fusion Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Batista, Fabiana R.X.</creatorcontrib><creatorcontrib>Moraes, Ângela M.</creatorcontrib><creatorcontrib>Büntemeyer, Heino</creatorcontrib><creatorcontrib>Noll, Thomas</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Biologicals</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Batista, Fabiana R.X.</au><au>Moraes, Ângela M.</au><au>Büntemeyer, Heino</au><au>Noll, Thomas</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium</atitle><jtitle>Biologicals</jtitle><addtitle>Biologicals</addtitle><date>2009-04-01</date><risdate>2009</risdate><volume>37</volume><issue>2</issue><spage>108</spage><epage>118</epage><pages>108-118</pages><issn>1045-1056</issn><eissn>1095-8320</eissn><abstract>The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected
Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO
2 were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO
2 of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98
g/10
7 cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>19059791</pmid><doi>10.1016/j.biologicals.2008.11.001</doi><tpages>11</tpages></addata></record> |
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| ispartof | Biologicals, 2009-04, Vol.37 (2), p.108-118 |
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| subjects | Aeration Amino Acids - metabolism Animals Carbohydrate Metabolism - drug effects Cell Culture Techniques Cell Line - metabolism Cell Line - physiology Cell Proliferation - drug effects Culture Media, Serum-Free - pharmacology Drosophila melanogaster Fed-batch culture Growth kinetics Hydrogen-Ion Concentration Lactic Acid - metabolism Oxygen Consumption - drug effects Rabies virus Rabies virus - genetics Rabies virus - metabolism Recombinant Proteins - genetics Recombinant Proteins - metabolism RVGP S2 cells Time Factors Viral Fusion Proteins - isolation & purification Viral Fusion Proteins - metabolism |
| title | Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium |
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