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Method for chromatography reuse

a chromatography and reuse technology, applied in the field of chromatography, can solve the problems of protein carryover, protein cost increase, and protein cost increas

Inactive Publication Date: 2015-04-02
GENENTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention provides methods to clean and regenerate chromatography materials for reuse. The methods involve passing different buffer solutions through the material to remove impurities and prepare it for use with different products. The cleaning process involves multiple steps, including equilibration, elution, and regeneration steps. The methods can be used with various types of chromatography materials, such as resins and affinity materials, and can be used for large-scale production of polypeptides. Overall, the invention provides a reliable and effective way to clean and regenerate chromatography materials for reuse.

Problems solved by technology

However, Protein A resin costs are significant, comprising a substantial portion of the raw material costs in MAb manufacturing (Fahrner, R. L., et al., Biotechnol. Appl. Biochem. 1999, 30, 121-128; Kelley, B., Biotechnol. Prog. 2007, 23:995-1008).
This expense is further exacerbated by resin underuse, such that a single packed Protein A column is used only 10% of its potential lifetime (in the pilot plant and during clinical production).
Protein A resin reuse for multiple products is not a common practice as reuse can result in protein carryover, not only from previous runs, but also from previously purified products.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Protein Carryover

[0301]This example describes an effort to quantify protein carryover from sample to sample a pre-cleaning test run was performed on lab scale using a standard Protein A affinity material (0.66×20 cm). This run was called a “mock run” as the process was run according to a standard purification procedure except the load cycle was simulated with the equilibration buffer. The elution pool was collected, as per a typical Protein A process, and analyzed to determine the presence of protein. The analysis revealed that 20-30 ppm of protein carryover was present in the “mock elution” in the absence of any additional column cleaning. The result was confirmed with a second run.

[0302]In order to determine safe carryover levels, a risk assessment was conducted to determine acceptable immunoglobulins (IgG) and protein carryover levels in mAbs, and a substance-specific Acceptable Daily Exposure (ADE) for IgG was calculated. A comparison of the ADE to EDI, resulted in a “worst case...

example 2

Evaluation of Ion Exchange Columns for Multi-Product Use

[0333]Studies were performed to determine if a similar cleaning process could be developed for ion exchange chromatographies. MAbA and MAbB from ProA pools were loaded onto cation exchange columns (POROS) or anion exchange columns (QSFF). Following normal elution, the columns were then subject to a “mock elution”. Fractions were analyzed for the presence of MAbA and / or MabB using a MabSelectSure assay, limit of detection was 0.82 ng / mL. As seen in FIG. 13, mock elution results indicated the need for additional cleaning of the columns

[0334]The following clean-in-place (CIP) procedure was tested.

I.3 CV Equilibration BufferII.2 CV 0.5N NaOH10 minute static holdIII.1 CV 0.5N NaOH10 minute static holdIV.1 CV 0.5N NaOHV.Post-cleaning mock run

[0335]Samples were conditioned with low concentration of detergent (0.1% polysorbate 20, 0.05% sodium azide) to prevent sample from sticking to the wall of the container. Samples were adjusted to...

example 3

Evaluation of ProSep a Column for Multi-Product Use

[0339]Different cleaning solutions were evaluated at small scale in order to assess which solutions were most effective at reducing product carryover. Several different categories of solutions were tested, including acids, chaotropes, salts, and organic solvents. This study was designed to follow the standard protein A antibody process in order to best mimic generic process conditions, although actual processing conditions may differ depending on the specific product used.

[0340]Flow was directed in a downflow direction through the column for all processes with the exception of the cleaning cycle. Throughout the cleaning cycle, flow was directed upwards through the column in hopes of creating a best case cleaning scenario. Since feedstock is directed through the column in a downward direction during the loading cycle, the top of the Protein A column would theoretically foul the most. By directing flow of the cleaning cycle upwards, t...

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Abstract

The present invention provides methods for cleaning or regenerating a chromatography materiel for reuse. The methods of the invention can be used for cleaning or regenerating chromatography columns for reuse in the large-scale manufacture of multiple polypeptide products.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]This application claims the benefit of priority to U.S. Provisional Ser. No. 61 / 874,305, filed on Sep. 5, 2013, entitled METHOD FOR CHROMATOGRAPHY REUSE, which is hereby incorporated by reference in its entirety for all purposes.FIELD OF THE INVENTION[0002]The present invention provides methods for chromatography.BACKGROUND OF THE INVENTION[0003]Recombinant monoclonal antibodies (mAbs) are used in medicine and diagnostics (Albrecht, H., et al., Drugs Today 2009, 45:199-211; Takimoto, C. H.; Principles of Oncologic Pharmacotherapy Calvo, E. In Cancer Management: A Multidisciplinary Approach Medical, Surgical & Radiation Oncolog, 11th ed.; Pazdur, R.; Wagman, L. D.; Camphausen, K. A.; Hoskins, W. J., Eds. CMP Healthcare Media LLC: Lawrence, Kans., USA, 2008). Industrially, recombinant mAbs are bioproduced in living cells, such as Chinese Hamster Ovary (CHO) cells (Fahrner, R. L., et al., Biotechnol. Genet. Eng. Rev. 2001, 18:301-327). Once p...

Claims

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Application Information

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IPC IPC(8): B01J49/00B01D15/20B01D15/38C07K1/18C07K1/22
CPCB01J49/0078C07K1/18B01D15/203B01D15/3809C07K1/22B01J49/60
Inventor MAHAJAN, EKTAKOTHARY, KAPILSO, JOANNAWERBER, JAY
Owner GENENTECH INC
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