Combination of the 2A/furin technology with an animal component free cell line development platform process
The recently described 2A/furin technology combines both chains of the antibody in a single open reading frame. Upon translation and secretion, the peptide is processed by the cell to generate native fully functional IgG antibodies. Here, we describe the results of an evaluation study of this techno...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2010-07, Vol.87 (4), p.1517-1524 |
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| creator | Jostock, Thomas Dragic, Zorica Fang, Jianmin Jooss, Karin Wilms, Burkhard Knopf, Hans-Peter |
| description | The recently described 2A/furin technology combines both chains of the antibody in a single open reading frame. Upon translation and secretion, the peptide is processed by the cell to generate native fully functional IgG antibodies. Here, we describe the results of an evaluation study of this technology for an industrial CHO cell line development process. The 2A/furin expression cassette setup was combined with a Novartis vector system. A transfection, selection, and cloning procedure in chemically defined media was established at Novartis and applied for a monoclonal test antibody. The productivity of 2A/furin-vector-derived clones in non-optimized generic shake flask fed-batch models was in a comparable range with clones derived from the reference control vector. Higher clonal production stability was seen for the majority of clones generated with the 2A/furin technology compared to the clones generated with the reference control vector. Product quality was analyzed by SDS-PAGE and no significant difference was detected between the two systems. Thus, it was shown that the 2A/furin technology can be successfully combined with a Novartis CHO expression system and platform. Due to the single ORF setup, the 2A/furin technology may therefore offer a suitable approach to reduce vector size and complexity. |
| doi_str_mv | 10.1007/s00253-010-2625-0 |
| format | Article |
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Upon translation and secretion, the peptide is processed by the cell to generate native fully functional IgG antibodies. Here, we describe the results of an evaluation study of this technology for an industrial CHO cell line development process. The 2A/furin expression cassette setup was combined with a Novartis vector system. A transfection, selection, and cloning procedure in chemically defined media was established at Novartis and applied for a monoclonal test antibody. The productivity of 2A/furin-vector-derived clones in non-optimized generic shake flask fed-batch models was in a comparable range with clones derived from the reference control vector. Higher clonal production stability was seen for the majority of clones generated with the 2A/furin technology compared to the clones generated with the reference control vector. Product quality was analyzed by SDS-PAGE and no significant difference was detected between the two systems. Thus, it was shown that the 2A/furin technology can be successfully combined with a Novartis CHO expression system and platform. Due to the single ORF setup, the 2A/furin technology may therefore offer a suitable approach to reduce vector size and complexity.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-010-2625-0</identifier><identifier>PMID: 20461511</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>2A/furin ; Animals ; Antibodies ; Biological and medical sciences ; Biomedical and Life Sciences ; Biotechnology ; Biotechnology - methods ; Cell culture ; CHO ; CHO Cells ; Cloning ; Cricetinae ; Cricetulus ; Fundamental and applied biological sciences. Psychology ; Furin - genetics ; Furin - metabolism ; Gene amplification ; Gene Expression ; Genetic Vectors - genetics ; Genetic Vectors - metabolism ; Genomes ; Life Sciences ; Light ; Methods and Protocols ; Microbial Genetics and Genomics ; Microbiology ; Plasmids ; Productivity ; Recombinant antibody production ; Studies</subject><ispartof>Applied microbiology and biotechnology, 2010-07, Vol.87 (4), p.1517-1524</ispartof><rights>Springer-Verlag 2010</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c456t-3acbbdd93e2c9da9d1ec907727a9ce8c7b3c45dc52fbaa692467abcecf169d133</citedby><cites>FETCH-LOGICAL-c456t-3acbbdd93e2c9da9d1ec907727a9ce8c7b3c45dc52fbaa692467abcecf169d133</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-010-2625-0$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-010-2625-0$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23005201$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20461511$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jostock, Thomas</creatorcontrib><creatorcontrib>Dragic, Zorica</creatorcontrib><creatorcontrib>Fang, Jianmin</creatorcontrib><creatorcontrib>Jooss, Karin</creatorcontrib><creatorcontrib>Wilms, Burkhard</creatorcontrib><creatorcontrib>Knopf, Hans-Peter</creatorcontrib><title>Combination of the 2A/furin technology with an animal component free cell line development platform process</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>The recently described 2A/furin technology combines both chains of the antibody in a single open reading frame. Upon translation and secretion, the peptide is processed by the cell to generate native fully functional IgG antibodies. Here, we describe the results of an evaluation study of this technology for an industrial CHO cell line development process. The 2A/furin expression cassette setup was combined with a Novartis vector system. A transfection, selection, and cloning procedure in chemically defined media was established at Novartis and applied for a monoclonal test antibody. The productivity of 2A/furin-vector-derived clones in non-optimized generic shake flask fed-batch models was in a comparable range with clones derived from the reference control vector. Higher clonal production stability was seen for the majority of clones generated with the 2A/furin technology compared to the clones generated with the reference control vector. Product quality was analyzed by SDS-PAGE and no significant difference was detected between the two systems. Thus, it was shown that the 2A/furin technology can be successfully combined with a Novartis CHO expression system and platform. Due to the single ORF setup, the 2A/furin technology may therefore offer a suitable approach to reduce vector size and complexity.</description><subject>2A/furin</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Biotechnology - methods</subject><subject>Cell culture</subject><subject>CHO</subject><subject>CHO Cells</subject><subject>Cloning</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Furin - genetics</subject><subject>Furin - metabolism</subject><subject>Gene amplification</subject><subject>Gene Expression</subject><subject>Genetic Vectors - genetics</subject><subject>Genetic Vectors - metabolism</subject><subject>Genomes</subject><subject>Life Sciences</subject><subject>Light</subject><subject>Methods and Protocols</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Plasmids</subject><subject>Productivity</subject><subject>Recombinant antibody production</subject><subject>Studies</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkU-L1TAUxYMoznP0A7jRIAyu6tz8b5fDw1FhwIXOOqRp8l7HNqlJq8y3N6VPB1woBLK4v3POTQ5CLwm8IwDqMgNQwSogUFFJRQWP0I5wRiuQhD9GOyBKVEo09Rl6lvMdAKG1lE_RGQUuiSBkh77t49j2wcx9DDh6PB8dpleXfkl9wLOzxxCHeLjHP_v5iE0opx_NgG0cpxhcmLFPzmHrhgEPfXC4cz_cEKdxHU2DmX1MI55StC7n5-iJN0N2L073Obq9fv91_7G6-fzh0_7qprJcyLlixrZt1zXMUdt0pumIsw0oRZVprKutalkBOyuob42RDeVSmdY664ksMGPn6O3mW3K_Ly7PeuzzuqIJLi5ZK85l09Ba_J9kjIuaNSv55i_yLi4plGesEBDG6zWYbJBNMefkvJ5S-a50rwnotTG9NaZLY3ptTEPRvDoZL-3ouj-K3xUV4OIEmGzN4JMJts8PHAMQFFaOblwuo3Bw6WHDf6W_3kTeRG0OqRjfflndgNSScqHYL2Xqt9g</recordid><startdate>20100701</startdate><enddate>20100701</enddate><creator>Jostock, Thomas</creator><creator>Dragic, Zorica</creator><creator>Fang, Jianmin</creator><creator>Jooss, Karin</creator><creator>Wilms, Burkhard</creator><creator>Knopf, Hans-Peter</creator><general>Berlin/Heidelberg : Springer-Verlag</general><general>Springer-Verlag</general><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>IQODW</scope><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>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>7QO</scope><scope>7T5</scope><scope>H94</scope></search><sort><creationdate>20100701</creationdate><title>Combination of the 2A/furin technology with an animal component free cell line development platform process</title><author>Jostock, Thomas ; Dragic, Zorica ; Fang, Jianmin ; Jooss, Karin ; Wilms, Burkhard ; Knopf, Hans-Peter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c456t-3acbbdd93e2c9da9d1ec907727a9ce8c7b3c45dc52fbaa692467abcecf169d133</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>2A/furin</topic><topic>Animals</topic><topic>Antibodies</topic><topic>Biological and medical sciences</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Biotechnology - methods</topic><topic>Cell culture</topic><topic>CHO</topic><topic>CHO Cells</topic><topic>Cloning</topic><topic>Cricetinae</topic><topic>Cricetulus</topic><topic>Fundamental and applied biological sciences. 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| subjects | 2A/furin Animals Antibodies Biological and medical sciences Biomedical and Life Sciences Biotechnology Biotechnology - methods Cell culture CHO CHO Cells Cloning Cricetinae Cricetulus Fundamental and applied biological sciences. Psychology Furin - genetics Furin - metabolism Gene amplification Gene Expression Genetic Vectors - genetics Genetic Vectors - metabolism Genomes Life Sciences Light Methods and Protocols Microbial Genetics and Genomics Microbiology Plasmids Productivity Recombinant antibody production Studies |
| title | Combination of the 2A/furin technology with an animal component free cell line development platform process |
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