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Mixed Bed Ion Exchange Adsorber

Inactive Publication Date: 2017-10-19
MILLIPORE CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent aims to develop new technologies to simplify and reduce the costs of downstream processes by using less expensive and disposable adsorbent media. These technologies would eliminate the need for costly cleaning and storage of columns after use, and also increase performance by targeting a wider variety of impurities.

Problems solved by technology

However, it is often less effective in removing positively charged impurities, such as basic HCP, product aggregates and fragments.
The process of screening hybridoma libraries for candidate mABs is both time consuming and labor intensive.
However, the higher cost of these sophisticated ligands on resins precludes their employment for single use or in disposable processes.
Consequently, the separation process is inherently slow since the rate of mass transport is typically controlled by pore diffusion.
However, the use of small diameter beads comes at the price of increased column pressure drop.
Chromatography media typically has a very high cost (>$1000 / L) and significant quantities are required for large scale production columns.
As a result, biopharmaceutical manufacturers recycle chromatography resins hundreds of times. Each of these regeneration cycles consumes substantial quantities of media, and each step incurs additional costs associated with the validation of each cleaning, sterilization, and column packing operation.
However, in such constructions, poor uniformity in gel location and mass generally leads to poor efficiencies (shallow breakthrough and elution fronts).
In addition, resistance to flow can be high, even for short bed depths, a problem often aggravated by gel compression under modest pressure loads.
The current downstream process (DSP) is complex and expensive.
This is challenging to accomplish with resin- and membrane-based adsorbent media.

Method used

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  • Mixed Bed Ion Exchange Adsorber
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  • Mixed Bed Ion Exchange Adsorber

Examples

Experimental program
Comparison scheme
Effect test

example 1

Graft Polymerization of Un-Modified Nylon Fibers

[0207]10 g Allasso nylon fibers and water (466 ml) are added into a 500 ml bottle. 14 ml 1M HNO3 (14.4 mmol) are added to the reaction mixture, followed by the addition of 1.2 ml of a 0.4 M ammonium cerium(IV) nitrate solution in 1M HNO3 (0,480 mmol). The reaction mixture is agitated for 15 minutes. 3.39 g Glycidyl methacrylate (GMA, 24 mmol) are added. Now the agitated reaction mixture is heated to 35° C. for 1 hour. After cooling down to room temperature, the solids are washed with DI water (3×300 ml) and the damp material is used immediately in the following step.

example 2

Q-Functionalization of Epoxy-Functionalized Fibers (AEX Fiber Media)

[0208]The damp GMA functionalized fibers from example 1 are added into a 2 L bottle together with water (500 ml) and a solution of 50 wt % trimethylamine (aq.) in methanol (500 ml). The mixture is agitated for 18 hours at room temperature. Then the fiber solids are subsequently washed with a solution of 0.2 M ascorbic acid in 0.5 M sulphuric acid (3×400 ml), DI water (3×400 ml), 1M sodium hydroxide solution (3×400 ml), DI water (3×400 ml) and ethanol (1×400 ml). Subsequently, the material is placed in an oven to dry at 40° C. for 48 hours.

Yield: 11.74 g of a white fibrous solid

example 3

Graft Polymerization of Un-Modified Nylon Fibers (CEX Fiber Media)

[0209]10 g Allasso nylon fibers and water (460 ml) are added into a 1000 ml bottle. 29.8 ml 1M HNO3 solution (30 mmol) are added to the reaction mixture, followed by the addition of a solution 7.46 ml of a 0.4 M ammonium cerium(IV) nitrate solution in 1M HNO3 (3.00 mmol). The reaction mixture is agitated for 15 minutes. Then 61.5 g 3-sulfopropylmethacrylate potassium salt (3-SPMA, 250 mmol) are added and the resulting agitated reaction mixture is heated to 35° C. for 18 hours. After cooling to room temperature, the fiber solids from each bottle are washed with DI water (3×300 ml), 0.2 M ascorbic acid in 0.5 M sulphuric acid (3×300 ml), DI water (3×300 ml), 1M sodium hydroxide solution (3×300 ml), DI water 3×300 ml) and ethanol (1×300 ml). The prepared material is then placed in an oven to dry at 40° C.

Yield: 11.38 g of a white fibrous solid

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Abstract

The present invention refers to new species of an ion exchange adsorber which is suitable for the separation of host cell proteins (HCPs), antibody fragments and low molecular weight substances from solutions containing antibodies. The invention especially refers to a process for purifying biological samples by separating biomolecules of interest and impurities, comprising steps of contacting a sample with said chromatography media consisting of fibers, said fibers having imparted thereon functionality enabling ion exchange chromatography and / or hydrophobic interaction.

Description

[0001]The present invention refers to new species of an ion exchange adsorber which is suitable for the separation of host cell proteins (HCPs), antibody fragments and low molecular weight substances from solutions containing antibodies. The invention especially refers to a process for purifying biological samples by separating biomolecules of interest and impurities, comprising steps of contacting a sample with said chromatography media consisting of fibers, said fibers having imparted thereon functionality enabling ion exchange chromatography and / or hydrophobic interaction.BACKGROUNDPurification of Monoclonal Antibodies[0002]Since monoclonal antibodies (mAbs) are used for pharmaceutical applications, they are required in exceptionally high purities [A. Jungbauer, G. Carta, in: Protein Chromatography, Process Development and Scale-Up; WILEY-VCH Verlag, Weinheim (Germany) 2010].[0003]In general mammalian cell cultures are employed to manufacture the majority of therapeutic monoclona...

Claims

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Application Information

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IPC IPC(8): C07K1/20B01D63/02B01D15/38C07K1/36C07K1/18
CPCC07K1/20C07K1/18B01D2311/2626B01D63/02B01D15/3804C07K1/36
Inventor STONE, MATTHEW T.AMARA, JOHN P.
Owner MILLIPORE CORP
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