Identifying and modulating disulfide formation in the biopharmaceutical production of a recombinant protein vaccine candidate
Structural conversion of the serotype A recombinant botulinum neurotoxin heavy chain fragment (rBoNTA(H c)) produced intracellularly in Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the origina...
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| Veröffentlicht in: | Journal of biotechnology 2003-08, Vol.103 (3), p.257-271 |
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| creator | Bouvier, Anne Chapline, Jaymi Boerner, Renee Jeyarajah, Shanthini Cook, Susan Acharya, Prathima S Henderson, Ian Schrimsher, Jeffrey L Shepard, Scot R |
| description | Structural conversion of the serotype A recombinant botulinum neurotoxin heavy chain fragment (rBoNTA(H
c)) produced intracellularly in
Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the original recovered species into its derived counterpart and a number of analytical tools such as peptide mapping by LC/MS confirmed the formation of a disulfide bond, especially in samples of neutral to basic pH. A cation exchange chromatographic method proved useful in following the incidence of the reaction in various rBoNTA(H
c) samples. The disulfide formation kinetics were characterized using a one-quarter quadratic factorial design, following the investigation and development of controlled oxidation conditions using cysteine and cystamine as the redox pair. Temperature, pH and concentration of the redox pair had a significant effect on the yield and rate of the disulfide formation. This controlled reaction was eventually introduced as a functional unit operation in the purification process. The summation of preliminary scale-up and potency data showed scalability and robustness in the production of an active disulfide-bonded form of a recombinant botulism vaccine candidate. The presence of the disulfide bond did not effect the vaccine potency and it enhanced the molecule's thermal stability. |
| doi_str_mv | 10.1016/S0168-1656(03)00106-8 |
| format | Article |
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c)) produced intracellularly in
Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the original recovered species into its derived counterpart and a number of analytical tools such as peptide mapping by LC/MS confirmed the formation of a disulfide bond, especially in samples of neutral to basic pH. A cation exchange chromatographic method proved useful in following the incidence of the reaction in various rBoNTA(H
c) samples. The disulfide formation kinetics were characterized using a one-quarter quadratic factorial design, following the investigation and development of controlled oxidation conditions using cysteine and cystamine as the redox pair. Temperature, pH and concentration of the redox pair had a significant effect on the yield and rate of the disulfide formation. This controlled reaction was eventually introduced as a functional unit operation in the purification process. The summation of preliminary scale-up and potency data showed scalability and robustness in the production of an active disulfide-bonded form of a recombinant botulism vaccine candidate. The presence of the disulfide bond did not effect the vaccine potency and it enhanced the molecule's thermal stability.</description><identifier>ISSN: 0168-1656</identifier><identifier>EISSN: 1873-4863</identifier><identifier>DOI: 10.1016/S0168-1656(03)00106-8</identifier><identifier>PMID: 12890612</identifier><identifier>CODEN: JBITD4</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Amino Acid Sequence ; Animals ; Biological and medical sciences ; Biopharmaceutical production ; Biopharmaceutics - methods ; Botulinum neurotoxins ; Botulinum Toxins, Type A - chemical synthesis ; Botulinum Toxins, Type A - chemistry ; Botulinum Toxins, Type A - metabolism ; Botulinum Toxins, Type A - therapeutic use ; Botulism - prevention & control ; Disulfide formation ; Disulfides - chemistry ; Drug Design ; Drug Stability ; Fundamental and applied biological sciences. Psychology ; Hydrogen-Ion Concentration ; Kinetics ; Mice ; Molecular Sequence Data ; Oxidation ; Oxidation-Reduction ; Pichia - chemistry ; Pichia - metabolism ; Protein Engineering - methods ; Recombinant protein folding ; Recombinant Proteins - chemistry ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - metabolism ; Recombinant Proteins - therapeutic use ; Structure-Activity Relationship ; Temperature ; Vaccines, Synthetic - chemistry ; Vaccines, Synthetic - isolation & purification ; Vaccines, Synthetic - metabolism ; Vaccines, Synthetic - therapeutic use</subject><ispartof>Journal of biotechnology, 2003-08, Vol.103 (3), p.257-271</ispartof><rights>2003 Elsevier B.V.</rights><rights>2003 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c459t-30cc9b03b67a4f15454eedb2fc9d1d44d52741b168af815a89a9aaa39bb4a8bc3</citedby><cites>FETCH-LOGICAL-c459t-30cc9b03b67a4f15454eedb2fc9d1d44d52741b168af815a89a9aaa39bb4a8bc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0168-1656(03)00106-8$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,782,786,3552,27931,27932,46002</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14990264$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12890612$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bouvier, Anne</creatorcontrib><creatorcontrib>Chapline, Jaymi</creatorcontrib><creatorcontrib>Boerner, Renee</creatorcontrib><creatorcontrib>Jeyarajah, Shanthini</creatorcontrib><creatorcontrib>Cook, Susan</creatorcontrib><creatorcontrib>Acharya, Prathima S</creatorcontrib><creatorcontrib>Henderson, Ian</creatorcontrib><creatorcontrib>Schrimsher, Jeffrey L</creatorcontrib><creatorcontrib>Shepard, Scot R</creatorcontrib><title>Identifying and modulating disulfide formation in the biopharmaceutical production of a recombinant protein vaccine candidate</title><title>Journal of biotechnology</title><addtitle>J Biotechnol</addtitle><description>Structural conversion of the serotype A recombinant botulinum neurotoxin heavy chain fragment (rBoNTA(H
c)) produced intracellularly in
Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the original recovered species into its derived counterpart and a number of analytical tools such as peptide mapping by LC/MS confirmed the formation of a disulfide bond, especially in samples of neutral to basic pH. A cation exchange chromatographic method proved useful in following the incidence of the reaction in various rBoNTA(H
c) samples. The disulfide formation kinetics were characterized using a one-quarter quadratic factorial design, following the investigation and development of controlled oxidation conditions using cysteine and cystamine as the redox pair. Temperature, pH and concentration of the redox pair had a significant effect on the yield and rate of the disulfide formation. This controlled reaction was eventually introduced as a functional unit operation in the purification process. The summation of preliminary scale-up and potency data showed scalability and robustness in the production of an active disulfide-bonded form of a recombinant botulism vaccine candidate. The presence of the disulfide bond did not effect the vaccine potency and it enhanced the molecule's thermal stability.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biopharmaceutical production</subject><subject>Biopharmaceutics - methods</subject><subject>Botulinum neurotoxins</subject><subject>Botulinum Toxins, Type A - chemical synthesis</subject><subject>Botulinum Toxins, Type A - chemistry</subject><subject>Botulinum Toxins, Type A - metabolism</subject><subject>Botulinum Toxins, Type A - therapeutic use</subject><subject>Botulism - prevention & control</subject><subject>Disulfide formation</subject><subject>Disulfides - chemistry</subject><subject>Drug Design</subject><subject>Drug Stability</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hydrogen-Ion Concentration</subject><subject>Kinetics</subject><subject>Mice</subject><subject>Molecular Sequence Data</subject><subject>Oxidation</subject><subject>Oxidation-Reduction</subject><subject>Pichia - chemistry</subject><subject>Pichia - metabolism</subject><subject>Protein Engineering - methods</subject><subject>Recombinant protein folding</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - metabolism</subject><subject>Recombinant Proteins - therapeutic use</subject><subject>Structure-Activity Relationship</subject><subject>Temperature</subject><subject>Vaccines, Synthetic - chemistry</subject><subject>Vaccines, Synthetic - isolation & purification</subject><subject>Vaccines, Synthetic - metabolism</subject><subject>Vaccines, Synthetic - therapeutic use</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEFvFCEUx4nR2G31I2i42OhhFGYYFk7GNNY2aeJBPZMHPCxmhlkHpkkPfvcyuxt77AXC4_fnPX6EvOHsI2dcfvpRF9Vw2cv3rPvAGGeyUc_Ihqtt1wglu-dk8x85Iac5_2GMCd3zl-SEt0ozydsN-XftMZUY7mP6TSF5Ok5-GaCsRx_zMoTokYZpHmttSjQmWm6R2jjtbqEWHS4lOhjobq5Bt2emQIHO6KbRxgSprHcFa_IOnIsJqauNooeCr8iLAEPG18f9jPy6_Prz4qq5-f7t-uLLTeNEr0vTMee0ZZ2VWxCB96IXiN62wWnPvRC-b7eC2_pbCIr3oDRoAOi0tQKUdd0ZOT-8Wyf5u2AuZozZ4TBAwmnJhivFlJCqgv0BdPOU84zB7OY4wnxvODOrd7P3blaphnVm792subfHBosd0T-mjqIr8O4IQK66wgzJxfzICa1ZK0XlPh84rDruIs4mu4jJoY_VaDF-ik-M8gB4OaK8</recordid><startdate>20030815</startdate><enddate>20030815</enddate><creator>Bouvier, Anne</creator><creator>Chapline, Jaymi</creator><creator>Boerner, Renee</creator><creator>Jeyarajah, Shanthini</creator><creator>Cook, Susan</creator><creator>Acharya, Prathima S</creator><creator>Henderson, Ian</creator><creator>Schrimsher, Jeffrey L</creator><creator>Shepard, Scot R</creator><general>Elsevier B.V</general><general>Elsevier</general><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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20030815</creationdate><title>Identifying and modulating disulfide formation in the biopharmaceutical production of a recombinant protein vaccine candidate</title><author>Bouvier, Anne ; Chapline, Jaymi ; Boerner, Renee ; Jeyarajah, Shanthini ; Cook, Susan ; Acharya, Prathima S ; Henderson, Ian ; Schrimsher, Jeffrey L ; Shepard, Scot R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c459t-30cc9b03b67a4f15454eedb2fc9d1d44d52741b168af815a89a9aaa39bb4a8bc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biopharmaceutical production</topic><topic>Biopharmaceutics - methods</topic><topic>Botulinum neurotoxins</topic><topic>Botulinum Toxins, Type A - chemical synthesis</topic><topic>Botulinum Toxins, Type A - chemistry</topic><topic>Botulinum Toxins, Type A - metabolism</topic><topic>Botulinum Toxins, Type A - therapeutic use</topic><topic>Botulism - prevention & control</topic><topic>Disulfide formation</topic><topic>Disulfides - chemistry</topic><topic>Drug Design</topic><topic>Drug Stability</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hydrogen-Ion Concentration</topic><topic>Kinetics</topic><topic>Mice</topic><topic>Molecular Sequence Data</topic><topic>Oxidation</topic><topic>Oxidation-Reduction</topic><topic>Pichia - chemistry</topic><topic>Pichia - metabolism</topic><topic>Protein Engineering - methods</topic><topic>Recombinant protein folding</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - metabolism</topic><topic>Recombinant Proteins - therapeutic use</topic><topic>Structure-Activity Relationship</topic><topic>Temperature</topic><topic>Vaccines, Synthetic - chemistry</topic><topic>Vaccines, Synthetic - isolation & purification</topic><topic>Vaccines, Synthetic - metabolism</topic><topic>Vaccines, Synthetic - therapeutic use</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bouvier, Anne</creatorcontrib><creatorcontrib>Chapline, Jaymi</creatorcontrib><creatorcontrib>Boerner, Renee</creatorcontrib><creatorcontrib>Jeyarajah, Shanthini</creatorcontrib><creatorcontrib>Cook, Susan</creatorcontrib><creatorcontrib>Acharya, Prathima S</creatorcontrib><creatorcontrib>Henderson, Ian</creatorcontrib><creatorcontrib>Schrimsher, Jeffrey L</creatorcontrib><creatorcontrib>Shepard, Scot R</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bouvier, Anne</au><au>Chapline, Jaymi</au><au>Boerner, Renee</au><au>Jeyarajah, Shanthini</au><au>Cook, Susan</au><au>Acharya, Prathima S</au><au>Henderson, Ian</au><au>Schrimsher, Jeffrey L</au><au>Shepard, Scot R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identifying and modulating disulfide formation in the biopharmaceutical production of a recombinant protein vaccine candidate</atitle><jtitle>Journal of biotechnology</jtitle><addtitle>J Biotechnol</addtitle><date>2003-08-15</date><risdate>2003</risdate><volume>103</volume><issue>3</issue><spage>257</spage><epage>271</epage><pages>257-271</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><coden>JBITD4</coden><abstract>Structural conversion of the serotype A recombinant botulinum neurotoxin heavy chain fragment (rBoNTA(H
c)) produced intracellularly in
Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the original recovered species into its derived counterpart and a number of analytical tools such as peptide mapping by LC/MS confirmed the formation of a disulfide bond, especially in samples of neutral to basic pH. A cation exchange chromatographic method proved useful in following the incidence of the reaction in various rBoNTA(H
c) samples. The disulfide formation kinetics were characterized using a one-quarter quadratic factorial design, following the investigation and development of controlled oxidation conditions using cysteine and cystamine as the redox pair. Temperature, pH and concentration of the redox pair had a significant effect on the yield and rate of the disulfide formation. This controlled reaction was eventually introduced as a functional unit operation in the purification process. The summation of preliminary scale-up and potency data showed scalability and robustness in the production of an active disulfide-bonded form of a recombinant botulism vaccine candidate. The presence of the disulfide bond did not effect the vaccine potency and it enhanced the molecule's thermal stability.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>12890612</pmid><doi>10.1016/S0168-1656(03)00106-8</doi><tpages>15</tpages></addata></record> |
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| ispartof | Journal of biotechnology, 2003-08, Vol.103 (3), p.257-271 |
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| subjects | Amino Acid Sequence Animals Biological and medical sciences Biopharmaceutical production Biopharmaceutics - methods Botulinum neurotoxins Botulinum Toxins, Type A - chemical synthesis Botulinum Toxins, Type A - chemistry Botulinum Toxins, Type A - metabolism Botulinum Toxins, Type A - therapeutic use Botulism - prevention & control Disulfide formation Disulfides - chemistry Drug Design Drug Stability Fundamental and applied biological sciences. Psychology Hydrogen-Ion Concentration Kinetics Mice Molecular Sequence Data Oxidation Oxidation-Reduction Pichia - chemistry Pichia - metabolism Protein Engineering - methods Recombinant protein folding Recombinant Proteins - chemistry Recombinant Proteins - isolation & purification Recombinant Proteins - metabolism Recombinant Proteins - therapeutic use Structure-Activity Relationship Temperature Vaccines, Synthetic - chemistry Vaccines, Synthetic - isolation & purification Vaccines, Synthetic - metabolism Vaccines, Synthetic - therapeutic use |
| title | Identifying and modulating disulfide formation in the biopharmaceutical production of a recombinant protein vaccine candidate |
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