Antibody capture by mixed-mode chromatography: A comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins
► Mixed mode resins PPA HyperCel, MEP HyperCel, HEA HyperCel and Capto adhere are effective as a capture step for antibody purification from a crude culture supernatant of CHO. ► Optimal conditions of antibody purification were defined using a microplate approach and validated in column. ► Interacti...
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| Veröffentlicht in: | Journal of Chromatography A 2011-11, Vol.1218 (45), p.8197-8208 |
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| container_title | Journal of Chromatography A |
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| creator | Pezzini, Jerome Joucla, Gilles Gantier, René Toueille, Magali Lomenech, Anne-Marie Le Sénéchal, Caroline Garbay, Bertrand Santarelli, Xavier Cabanne, Charlotte |
| description | ► Mixed mode resins PPA HyperCel, MEP HyperCel, HEA HyperCel and Capto adhere are effective as a capture step for antibody purification from a crude culture supernatant of CHO. ► Optimal conditions of antibody purification were defined using a microplate approach and validated in column. ► Interactions between ligand and proteins in these mixed mode resins were identified. ► HCPs that are potentially coeluted with the antibody were identified by mass spectrometry.
We evaluated mixed mode chromatography for the capture of recombinant antibodies from CHO cell culture supernatants. We studied PPA HyperCel, HEA HyperCel, MEP HyperCel and Capto adhere resins, which all contain hydrophobic and cationic groups. A microplate approach combined with DoE modeling allowed the exploration of the complex behaviors of these mixed mode resins. Optimal conditions for antibody purification and host cell proteins (HCPs) elimination were determined and then directly up-scaled to laboratory columns. Then we used mass spectrometry to identify the major HCPs potentially coeluted with the antibody. Differences between the four resins in terms of amount, complexity and identity of the HCPs present in the elution fractions were investigated. |
| doi_str_mv | 10.1016/j.chroma.2011.09.036 |
| format | Article |
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We evaluated mixed mode chromatography for the capture of recombinant antibodies from CHO cell culture supernatants. We studied PPA HyperCel, HEA HyperCel, MEP HyperCel and Capto adhere resins, which all contain hydrophobic and cationic groups. A microplate approach combined with DoE modeling allowed the exploration of the complex behaviors of these mixed mode resins. Optimal conditions for antibody purification and host cell proteins (HCPs) elimination were determined and then directly up-scaled to laboratory columns. Then we used mass spectrometry to identify the major HCPs potentially coeluted with the antibody. Differences between the four resins in terms of amount, complexity and identity of the HCPs present in the elution fractions were investigated.</description><identifier>ISSN: 0021-9673</identifier><identifier>EISSN: 1873-3778</identifier><identifier>DOI: 10.1016/j.chroma.2011.09.036</identifier><identifier>PMID: 21982448</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Antibodies, Monoclonal - chemistry ; Antibodies, Monoclonal - isolation & purification ; Antibody ; cell culture ; CHO Cells ; chromatography ; Chromatography, Liquid - instrumentation ; Chromatography, Liquid - methods ; Cricetinae ; Cricetulus ; Design of experiment ; Electric Conductivity ; High-Throughput Screening Assays ; Host cell proteins ; Hydrogen-Ion Concentration ; Hydrophobic and Hydrophilic Interactions ; hydrophobicity ; Laboratory Chemicals ; Mass Spectrometry ; Mixed-mode chromatography ; Models, Chemical ; proteins ; Proteins - chemistry ; Proteins - classification ; recombinant antibodies ; Recombinant Proteins - chemistry ; Recombinant Proteins - isolation & purification ; Reproducibility of Results ; resins ; Screening</subject><ispartof>Journal of Chromatography A, 2011-11, Vol.1218 (45), p.8197-8208</ispartof><rights>2011 Elsevier B.V.</rights><rights>Copyright © 2011 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c333t-c182b8ca9b1d16d9e5b0520332c860f8e65022514d4d323b353251a82ab52a23</citedby><cites>FETCH-LOGICAL-c333t-c182b8ca9b1d16d9e5b0520332c860f8e65022514d4d323b353251a82ab52a23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0021967311013975$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21982448$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pezzini, Jerome</creatorcontrib><creatorcontrib>Joucla, Gilles</creatorcontrib><creatorcontrib>Gantier, René</creatorcontrib><creatorcontrib>Toueille, Magali</creatorcontrib><creatorcontrib>Lomenech, Anne-Marie</creatorcontrib><creatorcontrib>Le Sénéchal, Caroline</creatorcontrib><creatorcontrib>Garbay, Bertrand</creatorcontrib><creatorcontrib>Santarelli, Xavier</creatorcontrib><creatorcontrib>Cabanne, Charlotte</creatorcontrib><title>Antibody capture by mixed-mode chromatography: A comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins</title><title>Journal of Chromatography A</title><addtitle>J Chromatogr A</addtitle><description>► Mixed mode resins PPA HyperCel, MEP HyperCel, HEA HyperCel and Capto adhere are effective as a capture step for antibody purification from a crude culture supernatant of CHO. ► Optimal conditions of antibody purification were defined using a microplate approach and validated in column. ► Interactions between ligand and proteins in these mixed mode resins were identified. ► HCPs that are potentially coeluted with the antibody were identified by mass spectrometry.
We evaluated mixed mode chromatography for the capture of recombinant antibodies from CHO cell culture supernatants. We studied PPA HyperCel, HEA HyperCel, MEP HyperCel and Capto adhere resins, which all contain hydrophobic and cationic groups. A microplate approach combined with DoE modeling allowed the exploration of the complex behaviors of these mixed mode resins. Optimal conditions for antibody purification and host cell proteins (HCPs) elimination were determined and then directly up-scaled to laboratory columns. Then we used mass spectrometry to identify the major HCPs potentially coeluted with the antibody. Differences between the four resins in terms of amount, complexity and identity of the HCPs present in the elution fractions were investigated.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - chemistry</subject><subject>Antibodies, Monoclonal - isolation & purification</subject><subject>Antibody</subject><subject>cell culture</subject><subject>CHO Cells</subject><subject>chromatography</subject><subject>Chromatography, Liquid - instrumentation</subject><subject>Chromatography, Liquid - methods</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>Design of experiment</subject><subject>Electric Conductivity</subject><subject>High-Throughput Screening Assays</subject><subject>Host cell proteins</subject><subject>Hydrogen-Ion Concentration</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>hydrophobicity</subject><subject>Laboratory Chemicals</subject><subject>Mass Spectrometry</subject><subject>Mixed-mode chromatography</subject><subject>Models, Chemical</subject><subject>proteins</subject><subject>Proteins - chemistry</subject><subject>Proteins - classification</subject><subject>recombinant antibodies</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Reproducibility of Results</subject><subject>resins</subject><subject>Screening</subject><issn>0021-9673</issn><issn>1873-3778</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9Uctu1DAUtRAVnQ78AQIv2ST4kYfDAmlUQUGq1EXL2nLsmxmPJnawnYr5Kz6xjlK6ZGX73vO4vgeh95SUlNDm87HUh-BHVTJCaUm6kvDmFdpQ0fKCt614jTaEMFp0Tcsv0VWMR0JoS1r2Bl0y2glWVWKD_u5csr03Z6zVlOYAuD_j0f4BU4zeAF49kt8HNR3OX_AOaz9OAQ7gon0EHNOcuUMGYQMJwmidStY77Afsp2RHdcLTHOxg9VrX3hm73CJOHlsD2f-lmUm5n9Sq4vb44GPCGk5ZJPgE1sW36GJQpwjvns8tevj-7eH6R3F7d_PzendbaM55KjQVrBdadT01tDEd1D2pGeGcadGQQUBTE8ZqWpnKcMZ7XvP8UoKpvmaK8S36tMpm398zxCRHG5dBlAM_R0kbvggs296iaoXq4GMMMMgp5H-Hs6RELlHJo1zXKJeoJOlkjirTPjw7zP0I5oX0L5sM-LgCBuWl2gcb5a_7rFDnHAVnhGTE1xUBeRGPFoKM2oLTYGwAnaTx9v8zPAGd-LTE</recordid><startdate>20111111</startdate><enddate>20111111</enddate><creator>Pezzini, Jerome</creator><creator>Joucla, Gilles</creator><creator>Gantier, René</creator><creator>Toueille, Magali</creator><creator>Lomenech, Anne-Marie</creator><creator>Le Sénéchal, Caroline</creator><creator>Garbay, Bertrand</creator><creator>Santarelli, Xavier</creator><creator>Cabanne, Charlotte</creator><general>Elsevier B.V</general><scope>FBQ</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>7QH</scope><scope>7UA</scope><scope>C1K</scope><scope>F1W</scope><scope>H97</scope><scope>L.G</scope></search><sort><creationdate>20111111</creationdate><title>Antibody capture by mixed-mode chromatography: A comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins</title><author>Pezzini, Jerome ; Joucla, Gilles ; Gantier, René ; Toueille, Magali ; Lomenech, Anne-Marie ; Le Sénéchal, Caroline ; Garbay, Bertrand ; Santarelli, Xavier ; Cabanne, Charlotte</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c333t-c182b8ca9b1d16d9e5b0520332c860f8e65022514d4d323b353251a82ab52a23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - chemistry</topic><topic>Antibodies, Monoclonal - isolation & purification</topic><topic>Antibody</topic><topic>cell culture</topic><topic>CHO Cells</topic><topic>chromatography</topic><topic>Chromatography, Liquid - instrumentation</topic><topic>Chromatography, Liquid - methods</topic><topic>Cricetinae</topic><topic>Cricetulus</topic><topic>Design of experiment</topic><topic>Electric Conductivity</topic><topic>High-Throughput Screening Assays</topic><topic>Host cell proteins</topic><topic>Hydrogen-Ion Concentration</topic><topic>Hydrophobic and Hydrophilic Interactions</topic><topic>hydrophobicity</topic><topic>Laboratory Chemicals</topic><topic>Mass Spectrometry</topic><topic>Mixed-mode chromatography</topic><topic>Models, Chemical</topic><topic>proteins</topic><topic>Proteins - chemistry</topic><topic>Proteins - classification</topic><topic>recombinant antibodies</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Reproducibility of Results</topic><topic>resins</topic><topic>Screening</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pezzini, Jerome</creatorcontrib><creatorcontrib>Joucla, Gilles</creatorcontrib><creatorcontrib>Gantier, René</creatorcontrib><creatorcontrib>Toueille, Magali</creatorcontrib><creatorcontrib>Lomenech, Anne-Marie</creatorcontrib><creatorcontrib>Le Sénéchal, Caroline</creatorcontrib><creatorcontrib>Garbay, Bertrand</creatorcontrib><creatorcontrib>Santarelli, Xavier</creatorcontrib><creatorcontrib>Cabanne, Charlotte</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aqualine</collection><collection>Water Resources Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><jtitle>Journal of Chromatography A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pezzini, Jerome</au><au>Joucla, Gilles</au><au>Gantier, René</au><au>Toueille, Magali</au><au>Lomenech, Anne-Marie</au><au>Le Sénéchal, Caroline</au><au>Garbay, Bertrand</au><au>Santarelli, Xavier</au><au>Cabanne, Charlotte</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Antibody capture by mixed-mode chromatography: A comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins</atitle><jtitle>Journal of Chromatography A</jtitle><addtitle>J Chromatogr A</addtitle><date>2011-11-11</date><risdate>2011</risdate><volume>1218</volume><issue>45</issue><spage>8197</spage><epage>8208</epage><pages>8197-8208</pages><issn>0021-9673</issn><eissn>1873-3778</eissn><abstract>► Mixed mode resins PPA HyperCel, MEP HyperCel, HEA HyperCel and Capto adhere are effective as a capture step for antibody purification from a crude culture supernatant of CHO. ► Optimal conditions of antibody purification were defined using a microplate approach and validated in column. ► Interactions between ligand and proteins in these mixed mode resins were identified. ► HCPs that are potentially coeluted with the antibody were identified by mass spectrometry.
We evaluated mixed mode chromatography for the capture of recombinant antibodies from CHO cell culture supernatants. We studied PPA HyperCel, HEA HyperCel, MEP HyperCel and Capto adhere resins, which all contain hydrophobic and cationic groups. A microplate approach combined with DoE modeling allowed the exploration of the complex behaviors of these mixed mode resins. Optimal conditions for antibody purification and host cell proteins (HCPs) elimination were determined and then directly up-scaled to laboratory columns. Then we used mass spectrometry to identify the major HCPs potentially coeluted with the antibody. Differences between the four resins in terms of amount, complexity and identity of the HCPs present in the elution fractions were investigated.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>21982448</pmid><doi>10.1016/j.chroma.2011.09.036</doi><tpages>12</tpages></addata></record> |
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| ispartof | Journal of Chromatography A, 2011-11, Vol.1218 (45), p.8197-8208 |
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| subjects | Animals Antibodies, Monoclonal - chemistry Antibodies, Monoclonal - isolation & purification Antibody cell culture CHO Cells chromatography Chromatography, Liquid - instrumentation Chromatography, Liquid - methods Cricetinae Cricetulus Design of experiment Electric Conductivity High-Throughput Screening Assays Host cell proteins Hydrogen-Ion Concentration Hydrophobic and Hydrophilic Interactions hydrophobicity Laboratory Chemicals Mass Spectrometry Mixed-mode chromatography Models, Chemical proteins Proteins - chemistry Proteins - classification recombinant antibodies Recombinant Proteins - chemistry Recombinant Proteins - isolation & purification Reproducibility of Results resins Screening |
| title | Antibody capture by mixed-mode chromatography: A comprehensive study from determination of optimal purification conditions to identification of contaminating host cell proteins |
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