{"id":{"repo_id":"aachen","oai_identifier":"oai:publications.rwth-aachen.de:51729"},"canonical_url":"https://search.dev.ndltd.org/etd/aachen/oai:publications.rwth-aachen.de:51729","repository":{"repo_id":"aachen","name":"RWTH Aachen University","base_url":"https://publications.rwth-aachen.de/oai2d"},"display":{"title":"Pilot-scale process development for the purification of the recombinant antibody 2G12 from transgenic tobacco","abstract":"The production of recombinant therapeutic proteins is becoming increasingly important as demand rises and the industry reaches its production capacity. Transgenic plants offer an alternative production system to the established platforms, but this requires the development of downstream processing schemes tailored for both the platform and the product. The aim of this thesis, within the EU-funded project Pharma-Planta, was to develop and optimize downstream processing strategies for the purification of the recombinant anti-HIV antibody 2G12. Different techniques were successfully combined into a process strategy to fulfill this objective. Clinical-grade recombinant antibody was purified using by the following process steps and industrially available equipment: manual harvest, dispersion and extraction, pH shift in the crude extract, fiber removal by filtration, clarification by filtration, capture by affinity Protein A membrane chromatography, intermediate purification by ceramic hydroxyapatite chromatography, polishing by virus filtration, concentration by ultrafiltration, diafiltration by ultrafiltration, final concentration and formulation. The process recovery of 60% resulted in a final yield of 3.7 g of clinical-grade tobacco-derived antibody purified from 216 kg leaf material, equal to 880 l crude extract. The antibody was polished to a purity >99%, consisting of 99.9% monomeric Ab with an in vitro binding activity of 93% and an in vitro neutralizing activity of 103% relative to the CHO2G12. This thesis established the basis for reproducible and predictable processing of plant material at the hundred-kilogram scale, and shows how further upscaling could be achieved.","abstract_html":"The production of recombinant therapeutic proteins is becoming increasingly important as demand rises and the industry reaches its production capacity. Transgenic plants offer an alternative production system to the established platforms, but this requires the development of downstream processing schemes tailored for both the platform and the product. The aim of this thesis, within the EU-funded project Pharma-Planta, was to develop and optimize downstream processing strategies for the purification of the recombinant anti-HIV antibody 2G12. Different techniques were successfully combined into a process strategy to fulfill this objective. Clinical-grade recombinant antibody was purified using by the following process steps and industrially available equipment: manual harvest, dispersion and extraction, pH shift in the crude extract, fiber removal by filtration, clarification by filtration, capture by affinity Protein A membrane chromatography, intermediate purification by ceramic hydroxyapatite chromatography, polishing by virus filtration, concentration by ultrafiltration, diafiltration by ultrafiltration, final concentration and formulation. The process recovery of 60% resulted in a final yield of 3.7 g of clinical-grade tobacco-derived antibody purified from 216 kg leaf material, equal to 880 l crude extract. The antibody was polished to a purity >99%, consisting of 99.9% monomeric Ab with an in vitro binding activity of 93% and an in vitro neutralizing activity of 103% relative to the CHO2G12. 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Transgenic plants offer an alternative production system to the established platforms, but this requires the development of downstream processing schemes tailored for both the platform and the product. The aim of this thesis, within the EU-funded project Pharma-Planta, was to develop and optimize downstream processing strategies for the purification of the recombinant anti-HIV antibody 2G12. Different techniques were successfully combined into a process strategy to fulfill this objective. Clinical-grade recombinant antibody was purified using by the following process steps and industrially available equipment: manual harvest, dispersion and extraction, pH shift in the crude extract, fiber removal by filtration, clarification by filtration, capture by affinity Protein A membrane chromatography, intermediate purification by ceramic hydroxyapatite chromatography, polishing by virus filtration, concentration by ultrafiltration, diafiltration by ultrafiltration, final concentration and formulation. The process recovery of 60% resulted in a final yield of 3.7 g of clinical-grade tobacco-derived antibody purified from 216 kg leaf material, equal to 880 l crude extract. The antibody was polished to a purity >99%, consisting of 99.9% monomeric Ab with an in vitro binding activity of 93% and an in vitro neutralizing activity of 103% relative to the CHO2G12. This thesis established the basis for reproducible and predictable processing of plant material at the hundred-kilogram scale, and shows how further upscaling could be achieved."]},{"key":"dc:source","label":"Dc Source","values":["Aachen : Publikationsserver der RWTH Aachen University III, 104 S. : Ill., graph. Darst. (2009). = Aachen, Techn. Hochsch., Diss., 2009"]},{"key":"dc:title","label":"Title","values":["Pilot-scale process development for the purification of the recombinant antibody 2G12 from transgenic tobacco"]}]}],"canonical_facts":{"dc:contributor":["Fischer, Rainer"],"dc:coverage":["DE"],"dc:creator":["Lobedann, Martin"],"dc:date":["2009"],"dc:description":["The production of recombinant therapeutic proteins is becoming increasingly important as demand rises and the industry reaches its production capacity. 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Clinical-grade recombinant antibody was purified using by the following process steps and industrially available equipment: manual harvest, dispersion and extraction, pH shift in the crude extract, fiber removal by filtration, clarification by filtration, capture by affinity Protein A membrane chromatography, intermediate purification by ceramic hydroxyapatite chromatography, polishing by virus filtration, concentration by ultrafiltration, diafiltration by ultrafiltration, final concentration and formulation. The process recovery of 60% resulted in a final yield of 3.7 g of clinical-grade tobacco-derived antibody purified from 216 kg leaf material, equal to 880 l crude extract. The antibody was polished to a purity >99%, consisting of 99.9% monomeric Ab with an in vitro binding activity of 93% and an in vitro neutralizing activity of 103% relative to the CHO2G12. 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