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Continuous production of recombinant proteins

A technology of recombinant protein and bioreactor, applied in the direction of immunoglobulin, specific-purpose bioreactor/fermenter, biochemical instrument, etc., can solve problems such as limited experience

Pending Publication Date: 2021-12-21
GENZYME CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Limited experience with continuous downstream operations as technical requirements for upstream and downstream development vary considerably

Method used

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  • Continuous production of recombinant proteins
  • Continuous production of recombinant proteins
  • Continuous production of recombinant proteins

Examples

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Embodiment 1

[0063] Embodiment 1: Continuous production of recombinant protein

[0064] A 100-L booster perfusion bioreactor coupled with dual ATF was used to produce recombinant monoclonal antibodies (mAbs) over a 70-day time course. Cell-free mAb-containing medium (harvest) was continuously pumped to a 100-L harvest buffer vessel for further processing.

[0065] Capture using a 2X 1-L (8x 20cm) prepacked and irradiated Protein A affinity column operated in bind and elute MCC mode on a pilot-scale steam-in-place PCC slide Medium containing antibodies. The capture step was performed such that one of the two columns was always loaded from the harvest buffer vessel and the average inlet volumetric flow rate to the capture step matched the average outlet volumetric flow rate from the perfusion bioreactor. The loading on the Protein A column was controlled by ΔUV to approximately 3% breakthrough. Prior to virus inactivation, separate trap column eluates were collected in a 5-L mixing vessel...

Embodiment 2

[0071] Embodiment 2: Continuous production of recombinant protein

[0072] A 100-L booster perfusion bioreactor coupled with dual ATF was used to produce recombinant monoclonal antibodies (mAbs) over a 27 day time course. Cell-free mAb-containing medium (harvest) was continuously pumped to a 100-L harvest buffer vessel for further processing.

[0073] Using a 2X 1-L (8x20cm) prepacked and irradiated Protein A affinity column operated in bind and elute MCC mode on a pilot-scale steam-in-place PCC slide to capture the protein A affinity column containing Antibody medium. The capture step was performed such that one of the two columns was always loaded from the harvest buffer vessel and the average inlet volumetric flow rate to the capture step matched the average outlet volumetric flow rate from the perfusion bioreactor. The loading on the Protein A column was controlled by ΔUV to approximately 3% breakthrough. Prior to virus inactivation, separate trap column eluates were co...

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Abstract

The present disclosure relates to methods and systems for the continuous production of recombinant proteins. In particular embodiments, the disclosure relates to methods and systems using capture chromatography, post-capture chromatography, virus filtration, and ultrafiltration / diafiltration for the continuous production of recombinant proteins.

Description

[0001] This application claims the benefit of U.S. Provisional Application No. 62 / 828,755, filed April 3, 2019, the disclosure of which is expressly incorporated herein by reference. technical field [0002] The present disclosure relates to methods and systems for the continuous production of recombinant proteins. In specific embodiments, the present disclosure relates to methods and systems for the continuous production of recombinant proteins using capture chromatography, post-capture chromatography, viral filtration, and ultrafiltration / diafiltration. Background technique [0003] Continuous manufacturing is a routine mode of production used in industries such as petrochemical and food production. The benefits of continuous processes include steady state operation, reduced equipment size, high volumetric productivity, streamlined process flow, low cycle time, and reduced capital costs. Konstantinov et al., J. Pharm. Sci. 104(3):813-20 (2015). [0004] Continuous manufa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00B01D15/18C07K1/36C07K16/00C12M1/00
CPCC12P21/005C07K16/00C07K1/36B01D15/1864B01D15/1807C12M47/10C07K1/22C07K1/34B01D61/14B01D61/145B01D2311/2688B01D2315/16B01D2315/08B01D61/58B01D2311/2697C12M33/14C12M37/02
Inventor M·科尔堡T·维特尔C·瓦尔纳K·布劳尔
Owner GENZYME CORP