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Pilot-scale process for magnetic bead purification of antibodies directly from non-clarified CHO cell culture
AdBIOPRO, VINNOVA Competence Ctr Adv Bioprod Continuous Pro, Stockholm, Sweden;KTH Royal Inst Technol, Sch Engn Sci Chem Biotechnol & Hlth, Dept Ind Biotechnol, Cell Technol Grp CETEG, Stockholm, Sweden.
PE Bioproc Consulting AB, Strangnas, Sweden.
AdBIOPRO, VINNOVA Competence Ctr Adv Bioprod Continuous Pro, Stockholm, Sweden;Lab On A Bead AB, Uppsala, Sweden.
Stockholm Univ, Dept Organ Chem, Stockholm, Sweden.
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2019 (English)In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 35, no 3, article id e2775Article in journal (Refereed) Published
Abstract [en]

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 x 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. (c) 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2775, 2019.

Place, publisher, year, edition, pages
John Wiley & Sons, 2019. Vol. 35, no 3, article id e2775
Keywords [en]
magnetic beads, purification, monoclonal antibody, pilot-scale, downstream-bioprocess
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-389831DOI: 10.1002/btpr.2775ISI: 000471314600026PubMedID: 30629859OAI: oai:DiVA.org:uu-389831DiVA, id: diva2:1339355
Funder
Vinnova, 2016-04152Vinnova, 2016-05181Available from: 2019-07-29 Created: 2019-07-29 Last updated: 2019-07-29Bibliographically approved

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