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Use of depth filtration in series with continuous centrifugation to clarify mammalian cell cultures

A mammalian, deep filtration technology, applied in the direction of separation of microorganisms, microorganisms, microorganisms, etc., can solve the problem of reducing the ability of filtration steps

Inactive Publication Date: 2007-08-08
BIOGEN IDEC MA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, mammalian supernatants produced by methods taught by the prior art contain substantial debris below 1.5 μm, which reduces the capacity of any subsequent filtration steps thereafter

Method used

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  • Use of depth filtration in series with continuous centrifugation to clarify mammalian cell cultures
  • Use of depth filtration in series with continuous centrifugation to clarify mammalian cell cultures
  • Use of depth filtration in series with continuous centrifugation to clarify mammalian cell cultures

Examples

Experimental program
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Embodiment 1

[0057] Clarification of CHO Cell Suspensions Using a Continuously Operated Disk-Stack Centrifuge

[0058] The initial feasibility study was carried out using a rented disc stack centrifuge (SC-6 separator from Westfalia). The machine is dimensioned to operate within a volumetric flow rate of 60-1000 L / h, well suited for clarification of suspensions produced by 200 L bioreactors. A total of three experimental processes (operations, run) with the following main objectives were carried out:

[0059] (1) To evaluate the overall compatibility of large-scale cell suspensions with a continuous disk stack centrifuge, expressed as particle removal (rate) and cell disruption;

[0060] (2) To determine the required filtration step downstream of centrifugation before applying the supernatant to the first chromatographic column;

[0061] (3) To find the operating window for estimating the size of the centrifuge needed to clarify cell suspensions at production scale and for determining ...

Embodiment 2

[0178] Clarification of NSO cell cultures for purification of secreted antibodies

[0179] Clarification of NSO cell cultures was achieved using a centrifuge in conjunction with a depth filter. The cells of the cell culture secrete the antibody of interest so that the clarification process can be used as an initial step in antibody purification. Clarification involves the use of a centrifuge followed by a series of depth filters, pre-filters, and membrane filters to remove insoluble material (including cells, cell debris, and all insoluble material in cell culture), on the antibody-containing liquid The clear liquid is basically clear. This clarity is usually described as 0.2 μm clarity, which is referenced to the pore size rating of the last filter in the series assembly.

[0180] The clarification method was developed to be suitable for the clarification of NSO cell cultures with approximately 1% (wet weight / volume) solid matter and a viability between 3% and 100%. Incl...

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Abstract

Methods for clarification of cell samples using centrifugation in combination with depth filtration.

Description

[0001] related application [0002] This application claims priority to US Patent Application Serial No. 10 / 871,261, filed June 21, 2004, which is hereby incorporated in its entirety. technical field [0003] The present invention generally relates to solid-liquid separation methods, eg, to separate cells and cell debris from biological samples to prepare clarified cell-free samples. This method is used as a purification step when isolating proteins secreted by cells in culture, such as secreted therapeutic proteins. Background technique [0004] The centrifugal separation method is a perfect continuous operation solid-liquid separation method. Although centrifugation is used for the separation of bacterial and yeast suspensions in industrial production, the use of continuous industrial-scale centrifugation has only recently been applied to mammalian cell cultures. [0005] Filtration equipment for industrial centrifugation (eg, samples over 2200 L) is bulky and represents...

Claims

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Application Information

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IPC IPC(8): B01D61/00C12N1/00C12N1/02
CPCC07K1/30C07K1/36C07K1/34C12M47/02C12N1/02Y10S435/803C12N1/00B01D61/00
Inventor 克里斯蒂娜·Y·彭约尔格·特梅斯
Owner BIOGEN IDEC MA INC