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Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture
High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoclonal antibodies (mAbs) from non‐clarified CHO cell broth using a pilot‐scale magnetic separator. The LOABeads had a maximum binding capacity of 65 mg/mL and...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley & Sons, Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6617771/ https://www.ncbi.nlm.nih.gov/pubmed/30629859 http://dx.doi.org/10.1002/btpr.2775 |
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author | Brechmann, Nils A. Eriksson, Per‐Olov Eriksson, Kristofer Oscarsson, Sven Buijs, Jos Shokri, Atefeh Hjälm, Göran Chotteau, Véronique |
author_facet | Brechmann, Nils A. Eriksson, Per‐Olov Eriksson, Kristofer Oscarsson, Sven Buijs, Jos Shokri, Atefeh Hjälm, Göran Chotteau, Véronique |
author_sort | Brechmann, Nils A. |
collection | PubMed |
description | High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoclonal antibodies (mAbs) from non‐clarified CHO cell broth using a pilot‐scale magnetic separator. The LOABeads had a maximum binding capacity of 65 mg/mL and an adsorption capacity of 25–42 mg IgG/mL bead in suspension for an IgG concentration of 1 to 8 g/L. Pilot‐scale separation was initially tested in a mAb capture step from 26 L clarified harvest. Small‐scale experiments showed that similar mAb adsorptions were obtained in cell broth containing 40 × 10(6) cells/mL as in clarified supernatant. Two pilot‐scale purification runs were then performed on non‐clarified cell broth from fed‐batch runs of 16 L, where a rapid mAb adsorption ≥96.6% was observed after 1 h. This process using 1 L of magnetic beads had an overall mAb yield of 86% and 16 times concentration factor. After this single protein A capture step, the mAb purity was similar to the one obtained by column chromatography, while the host cell protein content was very low, <10 ppm. Our results showed that this magnetic bead mAb purification process, using a dedicated pilot‐scale separation device, was a highly efficient single step, which directly connected the culture to the downstream process without cell clarification. Purification of mAb directly from non‐clarified cell broth without cell separation can provide significant savings in terms of resources, operation time, and equipment, compared to legacy procedure of cell separation followed by column chromatography step. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2775, 2019. |
format | Online Article Text |
id | pubmed-6617771 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | John Wiley & Sons, Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-66177712019-07-22 Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture Brechmann, Nils A. Eriksson, Per‐Olov Eriksson, Kristofer Oscarsson, Sven Buijs, Jos Shokri, Atefeh Hjälm, Göran Chotteau, Véronique Biotechnol Prog RESEARCH ARTICLES High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoclonal antibodies (mAbs) from non‐clarified CHO cell broth using a pilot‐scale magnetic separator. The LOABeads had a maximum binding capacity of 65 mg/mL and an adsorption capacity of 25–42 mg IgG/mL bead in suspension for an IgG concentration of 1 to 8 g/L. Pilot‐scale separation was initially tested in a mAb capture step from 26 L clarified harvest. Small‐scale experiments showed that similar mAb adsorptions were obtained in cell broth containing 40 × 10(6) cells/mL as in clarified supernatant. Two pilot‐scale purification runs were then performed on non‐clarified cell broth from fed‐batch runs of 16 L, where a rapid mAb adsorption ≥96.6% was observed after 1 h. This process using 1 L of magnetic beads had an overall mAb yield of 86% and 16 times concentration factor. After this single protein A capture step, the mAb purity was similar to the one obtained by column chromatography, while the host cell protein content was very low, <10 ppm. Our results showed that this magnetic bead mAb purification process, using a dedicated pilot‐scale separation device, was a highly efficient single step, which directly connected the culture to the downstream process without cell clarification. Purification of mAb directly from non‐clarified cell broth without cell separation can provide significant savings in terms of resources, operation time, and equipment, compared to legacy procedure of cell separation followed by column chromatography step. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2775, 2019. John Wiley & Sons, Inc. 2019-01-30 2019 /pmc/articles/PMC6617771/ /pubmed/30629859 http://dx.doi.org/10.1002/btpr.2775 Text en © 2019 The Authors. Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | RESEARCH ARTICLES Brechmann, Nils A. Eriksson, Per‐Olov Eriksson, Kristofer Oscarsson, Sven Buijs, Jos Shokri, Atefeh Hjälm, Göran Chotteau, Véronique Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture |
title | Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture |
title_full | Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture |
title_fullStr | Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture |
title_full_unstemmed | Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture |
title_short | Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture |
title_sort | pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified cho cell culture |
topic | RESEARCH ARTICLES |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6617771/ https://www.ncbi.nlm.nih.gov/pubmed/30629859 http://dx.doi.org/10.1002/btpr.2775 |
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