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Pilot-scale process for magnetic bead purification of antibodies directly from non-clarified CHO cell culture
Stockholms universitet, Naturvetenskapliga fakulteten, Institutionen för organisk kemi.
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Antal upphovsmän: 82019 (Engelska)Ingår i: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 35, nr 3, artikel-id e2775Artikel i tidskrift (Refereegranskat) 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.

Ort, förlag, år, upplaga, sidor
2019. Vol. 35, nr 3, artikel-id e2775
Nyckelord [en]
magnetic beads, purification, monoclonal antibody, pilot-scale, downstream-bioprocess
Nationell ämneskategori
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
Identifikatorer
URN: urn:nbn:se:su:diva-171178DOI: 10.1002/btpr.2775ISI: 000471314600026PubMedID: 30629859OAI: oai:DiVA.org:su-171178DiVA, id: diva2:1340249
Tillgänglig från: 2019-08-02 Skapad: 2019-08-02 Senast uppdaterad: 2019-08-02Bibliografiskt granskad

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Brechmann, Nils A.
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Institutionen för organisk kemi
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Biotechnology progress (Print)
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

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