Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange support
A method suitable for the isolation of monoclonal antibodies (Mabs) is described. The protocol utilizes a zirconia based column modified with ethylenediamine- N, N′-tetra(methylenephosphonic) acid to create a novel cation-exchange chromatographic support. Initial experiments using a linear salt grad...
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| Veröffentlicht in: | Journal of Chromatography A 1999-01, Vol.831 (1), p.63-72 |
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| container_title | Journal of Chromatography A |
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| creator | Clausen, Andrew M Subramanian, Anuradha Carr, Peter W |
| description | A method suitable for the isolation of monoclonal antibodies (Mabs) is described. The protocol utilizes a zirconia based column modified with ethylenediamine-
N,
N′-tetra(methylenephosphonic) acid to create a novel cation-exchange chromatographic support. Initial experiments using a linear salt gradient demonstrate the ability of this support to efficiently separate Mab from transferrin and bovine serum albumin in a model matrix. Results of the purification of Mab from an actual cell culture supernatant over a range in protein concentrations are also shown. Analyses by enzyme-linked immunosorbent assay and gel electrophoresis demonstrate that Mabs can be recovered from a cell culture supernatant at high yield (92–98%) and high purity (>95%) in a single chromatographic step. |
| doi_str_mv | 10.1016/S0021-9673(98)00999-6 |
| format | Article |
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N,
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N,
N′-tetra(methylenephosphonic) acid to create a novel cation-exchange chromatographic support. Initial experiments using a linear salt gradient demonstrate the ability of this support to efficiently separate Mab from transferrin and bovine serum albumin in a model matrix. Results of the purification of Mab from an actual cell culture supernatant over a range in protein concentrations are also shown. Analyses by enzyme-linked immunosorbent assay and gel electrophoresis demonstrate that Mabs can be recovered from a cell culture supernatant at high yield (92–98%) and high purity (>95%) in a single chromatographic step.</description><subject>Adsorption</subject><subject>Antibodies, Monoclonal - isolation & purification</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cations</subject><subject>Cells, Cultured</subject><subject>Chelating Agents - pharmacology</subject><subject>Chromatography, Ion Exchange - methods</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Health. Pharmaceutical industry</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Monoclonal antibodies</subject><subject>Organophosphorus Compounds - pharmacology</subject><subject>Production of active biomolecules</subject><subject>Serum Albumin, Bovine - immunology</subject><subject>Transferrin - immunology</subject><subject>Zirconium</subject><issn>0021-9673</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1vFSEUhllobG39CRoWxuhiKjAzMKxM0_iVNNFEXZMzcKiYGbjCjLHGHy_3zo1254oAz_seeAh5zNkFZ1y-_MSY4I2Wqn2uhxeMaa0beY-c_j0-IQ9L-cYYV0yJB-SE15tBDPyU_P645uCDhSWkSJOnc4rJTinCRCEuYUwuYKE-p5lanCZq12lZM9Ky7jBHWCpU6FpCvKFQw66WoaO_QrYpBqAjlLrd6hv8ab9CvDmEdykv5-S-h6ngo-N6Rr68ef356l1z_eHt-6vL68Z2vVwar5mwfnC6BRix61zLVDuqTmsxOCeFbK3r-ag4dz1K5jrh2aikBw_glYX2jDzbenc5fV-xLGYOZf8biJjWYqTuNRuEqGC_gTanUjJ6s8thhnxrODN71eag2uydGj2Yg2oja-7JccA6zujupDbPFXh6BKBYmHyGaEP5x6la3vUVe7VhWG38CJhNsQGjRRcy2sW4FP7zkj94lqCe</recordid><startdate>19990122</startdate><enddate>19990122</enddate><creator>Clausen, Andrew M</creator><creator>Subramanian, Anuradha</creator><creator>Carr, Peter W</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>7X8</scope></search><sort><creationdate>19990122</creationdate><title>Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange support</title><author>Clausen, Andrew M ; Subramanian, Anuradha ; Carr, Peter W</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c456t-f902cf8d93aabe44d3073b749928dd6263cd51b711d5e60d42f0b76fafaaf7ca3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Adsorption</topic><topic>Antibodies, Monoclonal - isolation & purification</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cations</topic><topic>Cells, Cultured</topic><topic>Chelating Agents - pharmacology</topic><topic>Chromatography, Ion Exchange - methods</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Health. Pharmaceutical industry</topic><topic>Industrial applications and implications. Economical aspects</topic><topic>Monoclonal antibodies</topic><topic>Organophosphorus Compounds - pharmacology</topic><topic>Production of active biomolecules</topic><topic>Serum Albumin, Bovine - immunology</topic><topic>Transferrin - immunology</topic><topic>Zirconium</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Clausen, Andrew M</creatorcontrib><creatorcontrib>Subramanian, Anuradha</creatorcontrib><creatorcontrib>Carr, Peter W</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>MEDLINE - Academic</collection><jtitle>Journal of Chromatography A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Clausen, Andrew M</au><au>Subramanian, Anuradha</au><au>Carr, Peter W</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange support</atitle><jtitle>Journal of Chromatography A</jtitle><addtitle>J Chromatogr A</addtitle><date>1999-01-22</date><risdate>1999</risdate><volume>831</volume><issue>1</issue><spage>63</spage><epage>72</epage><pages>63-72</pages><issn>0021-9673</issn><coden>JOCRAM</coden><abstract>A method suitable for the isolation of monoclonal antibodies (Mabs) is described. The protocol utilizes a zirconia based column modified with ethylenediamine-
N,
N′-tetra(methylenephosphonic) acid to create a novel cation-exchange chromatographic support. Initial experiments using a linear salt gradient demonstrate the ability of this support to efficiently separate Mab from transferrin and bovine serum albumin in a model matrix. Results of the purification of Mab from an actual cell culture supernatant over a range in protein concentrations are also shown. Analyses by enzyme-linked immunosorbent assay and gel electrophoresis demonstrate that Mabs can be recovered from a cell culture supernatant at high yield (92–98%) and high purity (>95%) in a single chromatographic step.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>10028281</pmid><doi>10.1016/S0021-9673(98)00999-6</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Journal of Chromatography A, 1999-01, Vol.831 (1), p.63-72 |
| issn | 0021-9673 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Adsorption Antibodies, Monoclonal - isolation & purification Biological and medical sciences Biotechnology Cations Cells, Cultured Chelating Agents - pharmacology Chromatography, Ion Exchange - methods Electrophoresis, Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Fundamental and applied biological sciences. Psychology Health. Pharmaceutical industry Industrial applications and implications. Economical aspects Monoclonal antibodies Organophosphorus Compounds - pharmacology Production of active biomolecules Serum Albumin, Bovine - immunology Transferrin - immunology Zirconium |
| title | Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange support |
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