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Counter-pressure filtration of proteins

A protein and filter technology, applied in the direction of peptide/protein components, filtration separation, fixed filter element filter, etc., can solve problems such as damage, reduction of processing yield, and protein destruction

Active Publication Date: 2012-11-21
TAKEDA PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, when the filtration device is operated at a pressure sufficient to ideally produce the desired processing flow rate, the increased flow rate (and the concomitant increase in shear) tends to reduce the processing yield, such as by damaging or destroying the protein , and / or by reducing the filter rate over time

Method used

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  • Counter-pressure filtration of proteins
  • Counter-pressure filtration of proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-8

[0033] The aqueous mixture of shear-sensitive, coagulation cascade protein vWF is sterilized and filtered to determine the effect of pressure (and back pressure) and other process variables (such as filter size and type) on the effectiveness of filtration, such as based on the filtrate The activity of vWF being recovered, the average flow rate of the filtrate and the ability to scale up the processing.

[0034] The aqueous mixture of recombinant vWF ("rVWF:Rco") has an activity of 125IU / ml and the concentration is 1159μg / ml (the bicinchoninic acid ("BCA") experiment). This mixture was prepared for the following Example 1- 8 in each of the examples. A low molecular weight salt is added to adjust the pH of the mixture to 7.3 and the osmolality to a value of about 400 mOsmol / l (milligrams osmolality / liter), respectively. Since Examples 1-8 are used to isolate the hydrodynamic effects (such as shearing) on ​​the protein near the filter, no other dispersed contaminants or materials (...

Embodiment 9-20

[0055] Aqueous solutions containing factor VIII in a SARTOCLEAN pre-filter The filter is tested to determine the effect of back pressure on the effectiveness of the filter, for example based on the required filter surface area. In Examples 9-13, no back pressure filtration was performed; the initial applied pressure was 100 mbar to 500 mbar. In Examples 14-16, the pressure difference was 200 mbar, and in Examples 17-20, the pressure difference was reduced to less than 150 mbar. The surface area of ​​each filter is 1.2m 2 . Table 5 contains the Factor VIII activity before and after filtration in each experiment.

[0056] Table 5. Recovery of FVIII in the filtrate

[0057]

[0058] Previous data proves that when back pressure filtration is used for the same amount of active material, the necessary filtration area is much less. Back pressure filtration stabilizes the filtration, and when the pressure difference is optimized, the average activity yield is improved.

Embodiment 21-25

[0060] The solution containing factor XIII is The N66 polyamide filter was tested to detect the effect of back pressure on the effectiveness of filtration, for example based on the recovered activity and protein concentration of factor XIII in the filtrate. The area of ​​each filter is 0.82m 2 . In Examples 21-23, no back pressure filtration was performed; the applied pressure used in these examples was 600 mbar. In Examples 24-26, the pressure difference is approximately 100 mbar. Table 6 contains the protein concentration, activity and yield before and after filtration for each experiment.

[0061] Table 6. Recovery of FXIII in the filtrate

[0062]

[0063] As shown in Table 6, by using back pressure, the activity yield and protein yield during filtration have been considerably improved.

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Abstract

A method is disclosed for filtering a protein in a liquid mixture in a manner that does not substantially damage or otherwise limit the recovery of the protein in the filtration filtrate. The method generally includes passing a liquid mixture containing a protein (e.g., an aqueous vWF mixture) through a filter while applying a counter pressure to the liquid mixture filtrate to accurately reduce and control the pressure differential across the filter. The disclosed method has the advantage that relatively high filtration flow rates can be achieved at relatively low pressure differentials, in contrast to high pressure differentials, which actually reduce the filtration flow rate of protein liquid mixtures. Further, the method can recover substantially all of the protein that is initially present in the liquid mixture.

Description

[0001] Cross references to related applications [0002] This application enjoys the priority of U.S. Provisional Patent Application No. 61 / 017,418 filed on December 28, 2007 according to 35 U.S.C. §119(e), which is incorporated herein by reference. Technical field [0003] The present disclosure generally relates to filtration methods for protein purification. More specifically, this application relates to, for example, low-shear, back-pressure sterilization filtration of proteins that are susceptible to shear damage when being transported in a liquid (such as shear-sensitive proteins and proteins in the coagulation cascade). Background technique [0004] The purified protein mixture can be administered to patients for various treatments. The purified protein mixture prepared for infusion into the patient must be sterilized before use. A suitable sterilization process for some proteins includes membrane filtration of the purified protein mixture. When the protein can pass throug...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/08A61K47/12A61K47/18A61K47/26A61K38/00B01D29/00
CPCA61K47/183A61K47/12A61K47/26A61K9/08B01D61/00
Inventor 奈博伊沙·尼科利奇米谢拉·弗雷沃夫冈·格雷博梅耶托马斯·简西克马蒂亚斯·弗里德克劳斯·蔡彻蔡科维斯基科特·施内克巴巴拉·瑞格勒阿尔玛·卡萨波维奇
Owner TAKEDA PHARMA CO LTD
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