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Separation method using polymer multi phase systems

A multiphase system, polymer technology, applied in the field of separation of at least one target compound, can solve the problem of reducing the solubility of proteins in the system

Inactive Publication Date: 2010-03-24
GE HEALTHCARE BIO SCI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of the system are: (a) the polymer still includes expensive polysaccharides; (b) the polymer is then further chemically modified; (c) the mentioned high molecular weight (Mw2200000) and intrinsic phase viscosity; and (c) ) relatively high polymer concentration required for phase formation, which is expected to bind water molecules and reduce the solubility of system proteins

Method used

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  • Separation method using polymer multi phase systems
  • Separation method using polymer multi phase systems
  • Separation method using polymer multi phase systems

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 - Preparation and Phase Diagram of the Two-Phase System of the Invention

[0073] raw material

[0074] Polymer: Poly(ethylene glycol) 4000 (Merck), PEG 8000 (Sigma-Aldrich), sodium polyacrylate ex Aldrich, CAS No.: 9003-04-7, molecular weight 30000 (in 40 wt% aqueous solution), molecular weight 8000 (in 45 wt% aqueous solution). NaCl and Na 2 SO 4 Methanol, Ba(NO 3 ) 2 (from Merck and P.A. quality). Millipore filtered water was used in all solutions.

[0075] Determination of phase diagram

[0076] The phase boundaries of the stem (doublet) are determined by the well-known method titration [see Methods in Enzymology, Vol. 228, Aqueous Two-Phase Systems, Harry Walter and G. Johansson eds. Academic Press, New York, 1994]. In this case, a system was fabricated with a composition suspected to be in the two-phase region. If the system becomes turbid upon mixing, it indicates the presence of a two-phase system. The polymer of this system was diluted ...

Embodiment 2

[0095] Example 2 - Effect of Salt and pH on Two-Phase Systems of the Invention

[0096] Two-phase systems were prepared as described above. The effect of pH on the EOPO 3900 and NaPAA 15000 systems was investigated. The results are presented in Table 3 below, which provides an observation of the effect of pH on phase volume ratio and phase system formation in a biphasic system containing EOPO and NaPAA in 200 mM NaP buffer. At these polymer molecular weights, concentrations and salt conditions, two phases were formed at pH 6-8 instead of pH 5.

[0097] Table 3: Effect of Salt and pH on EOPO 3900 NaPAA 15000 Two-Phase System

[0098] EOPO3900

Embodiment 3

[0099] Embodiment 3-Protein purification method

[0100] In this example, it is explained how to use Figure 4 Outlined method to purify protein. More specifically, a two-phase polymer system is used to clarify the sample during the aforementioned affinity chromatography step.

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Abstract

The present invention relates to a process of isolating one or more target compounds, wherein the clarification of feed is performed using partitioning in a multiphase system comprising a first polymer, which is a synthetic poly(acid), a second synthetic polymer, which is a poly(ether), and at least one salt, which clarification is followed by at least one step of affinity chromatography. The molecular weight of the poly(acid) may be in the range of 1000-100,000 Da. The target compound is preferably a biomolecule, such as a monoclonal antibody.

Description

technical field [0001] The invention relates to a method for isolating at least one target compound from a liquid, the process comprising at least one separation step by means of a differential partitioning of said target compound between two aqueous phases in Formed naturally in the presence of certain polymers and added salts. The invention also includes a method for isolating monoclonal antibodies using such partitioning between two aqueous phases that naturally form in the presence of certain polymers and added salts; and involving Kits for making such assignments. Background of the invention [0002] Revolutions in biotechnology, including the development of modern biopharmaceuticals and the mapping of the human genome, were made possible by the development of separation methods such as chromatography and electrophoresis. Such methods can be used in small scale as well as in large scale, and are called flexible methods, which can be used for a variety of substances, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/20C07K16/00C08L33/02C08L71/02
CPCC08L33/02C07K1/145C08L71/00C08L33/08C08L33/06C07K1/20C07K1/22C08G2650/58C08L71/02C08L2666/14C08L2666/04
Inventor R·约特K·拉基E·马塞多G·马尔姆奎斯特J·沙纳加J·范阿尔施泰恩
Owner GE HEALTHCARE BIO SCI CORP