Method of concentrating shear-sensitive biopolymers using hollow fibre membranes

A biopolymer, anti-shear technology, applied in the field of biopolymers, can solve the problems of reducing shear stress, not providing alternative methods, reducing and other problems, and achieving the effect of avoiding protein precipitation

Active Publication Date: 2014-07-09
TAKEDA PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In general, the prior art does not teach or suggest to one of ordinary skill in the art a cost-effective method of concentrating shear-sensitive biopolymers without bioaggregation due to protein precipitation, membrane clogging, and membrane surface adsorption massive loss of property
Similarly, reducing the shear stress in the device by reducing the flow rate of the biopolymer-containing mixture does not provide an effective alternative, as it still requires a certain minimum flow rate to avoid membrane clogging and sedimentation adsorption

Method used

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  • Method of concentrating shear-sensitive biopolymers using hollow fibre membranes
  • Method of concentrating shear-sensitive biopolymers using hollow fibre membranes
  • Method of concentrating shear-sensitive biopolymers using hollow fibre membranes

Examples

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Effect test

Embodiment 1

[0046] Four experiments were performed using vWF as a shear sensitive biopolymer and hollow fiber dialysis components. The hollow fiber dialysis module has 3000cm 2 Membrane surface area, 30 μm thick membrane, 100 mm fiber length and 200 μm fiber inner diameter. The membrane material is polyethersulfone. The concentrations of the feed streams were 0.7 grams of vWF protein per liter (gvWF / L), 0.56 gvWF / L, 0.39 g vWF / L, 0.27 g vWF / L. The concentrations of the retentate after concentration were 2.52 g vWF / L, 4.59 g vWF / L, 2.23 g vWF / L and 1.26 g vWF / L, respectively. The experiment took approximately 2 to 4 hours to complete.

[0047] The solution buffer in the feed stream was 20 mM (millimolar) HEPES and 150 mM NaCl buffer, pH 7.4 at room temperature. The dialysis buffer was 20 mM citrate and 15 mM glycine buffer, pH 7.3 at room temperature. The HEPES concentration shrunk from greater than 15 mM in the feed to less than 1 mM in the retentate.

[0048] Table 2: Data for Tria...

Embodiment 2

[0051] Two experiments were performed using vWF as a shear sensitive biopolymer and a hollow fiber dialysis module. The hollow fiber dialysis module has 7000cm 2 Membrane surface area, 30 μm thick membrane, 100 mm fiber length and 200 μm fiber inner diameter. The membrane material is polyethersulfone. Experiments were performed at a feed flow rate of 300 ml / min, an initial volume reduction rate of 2 L / hr and a dialysis rate of 5 L / hr. The shear rate applied by the feed flow rate is approximately 571 seconds -1. Feedstream concentrations were about 0.18 g vWF / L and about 0.22 g vWF / L. The retentate concentrations were about 0.88 g vWF / L and about 0.95 g vWF / L, respectively.

[0052] The solution buffer in the feed stream was 20 mM (millimolar) HEPES and 150 mM NaCl buffer, pH 7.4 at room temperature. The dialysis buffer was a salt-free, 20 mM citrate and 15 mM glycine buffer, pH 7.3 at room temperature. The HEPES concentration shrunk from greater than 15 mM in the feed t...

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Abstract

The present invention generally relates to a method of concentrating a mixture comprising a shear-sensitive biopolymer such as von Willebrand factor. Conventional methods of concentrating biopolymers impose too much shear stress, which can cause degradation of shear-sensitive biopolymers. The methods disclosed herein reduce shear stress while maintaining high filtrate flux rates. Disclosed herein is a method of concentrating a shear-sensitive biopolymer comprising flowing a mixture containing the shear-sensitive biopolymer into a hollow fiber dialysis module to form a retentate in which the The concentration of the shear-sensitive biopolymer is higher than the concentration of the shear-sensitive biopolymer in the mixture. Hollow fiber dialysis modules have high filtrate throughput and low shear rates at low flow rates. This ensures high product yields and minimal loss of shear-sensitive biopolymers.

Description

technical field [0001] The disclosure of the present application generally relates to methods of concentrating shear-sensitive biopolymers such as von Willebrand Factor (vWF). Background technique [0002] Known methods of concentrating (and diafiltration) biopolymers include tangential flow ultrafiltration (and diafiltration) in flat plate and hollow fiber devices. These devices operate at sufficiently high flow rates and transmembrane pressures to ensure a filtrate flux suitable for cost-effective operation. However, these operating conditions result in high shear rates. Additionally, these devices may include screens to further increase filtrate throughput. These sieves also increase the shear stress applied to the biopolymer. Such shear stress is particularly undesirable when attempting to concentrate shear-sensitive biopolymers such as proteins or virions, since shear stress can damage, denature or inactivate the biopolymer. [0003] There are various known methods ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D61/24A61M1/16
CPCB01D2325/04B01D61/243B01D2311/165A61P7/08A61M1/16B01D63/02B01D71/06
Inventor A·米特雷尔M·哈斯拉赫尔C·迈尔
Owner TAKEDA PHARMA CO LTD
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