Microparticles for cell disruption and/or biomolecule recovery

Patent metadata

Field	Value
Publication number	US-2016200760-A1
Application number	US-201414913337-A
Country	US
Kind code	A1
Filing date	Aug 25, 2014
Priority date	Aug 23, 2013
Publication date	Jul 14, 2016
Grant date	—

Abstract

The present invention provides novel methods of cell disruption and release of biomolecules from a cell. The invention comprises the use of positively and/or negatively charged microparticles comprising ground resin. It is particularly useful for purification of biomolecules from cell culture.

First claim

1 .- 18 . (canceled)

19 . A method for recovering biomolecules from a biological fluid, the method comprising
a) adding to the biological fluid positively charged microparticles comprising a ground polymeric anion-exchange resin or negatively charged microparticles comprising a ground polymeric cation exchange resin, and b) recovering the biomolecules from the biological fluid.

20 . The method of claim 19 , wherein the biomolecule is a polypeptide or polynucleotide.

21 . The method of claim 19 , wherein the microparticles form flocs.

22 . The method of claim 19 , wherein the biological fluid comprises cells and the microparticles cause a disruption of the cells or extraction of the biomolecules from the cells following mixing.

23 . The method of claim 19 , wherein the anion-exchange resin and the cation exchange resin are polystyrene-based, Hydroxyethyl methacrylate (HEMA)-based, dimethylamino ethylmethacrylate (DMAEMA)-based, dimethylamino ethylmethacrylate (pDMAEMA), polyacrylamide based, or methacrylic acid (MAA)-based.

24 . The method of claim 19 , wherein the cation exchange resin and anion-exchange resin are polystyrene cross-linked with divinylbenzene.

25 . The method of claim 19 , wherein the microparticles have an average particle size of less than about 5 μm.

26 . The method of claim 19 , wherein the positively charged microparticles or negatively charged microparticles are obtained by grinding a polymeric anion-exchange or cation-exchange resin, respectively.

27 . The method of claim 19 , wherein the anion-exchange resin is AMBERLITE® IRA-400, AMBERLITE®IRA-485, DOWEX® 1X2-100, DOWEX® 1-8-100, DOWEX® MARATHON® A2 or DIAION® SA 20A.

28 . The method of claim 19 , wherein the cation exchange resin is AMBERLITE® IRC-748, DOWEX® 50 WX2-100, DOWEX® 50 WX8-100, DOWEX® MARATHON® MSC or DIAION® SK 110.

29 . The method of claim 22 , wherein the cells are eukaryotic or prokaryotic cells.

30 . The method of claim 19 , further comprising allowing the microparticles to form flocs before recovering the biomolecules from the biological fluids.

31 . The method of claim 30 , wherein recovering the biomolecules from biological fluids comprises removing the flocs from the biological fluids and desorbing the biomolecules from the flocs.

32 . A method of disrupting cells in a cell suspension or extracting biomolecules from the cells comprising adding positively charged or negatively charged microparticles to the cell suspension.

33 . The method of claim 32 , wherein the biomolecules are released from the cells upon disruption or extraction of the biomolecules from the cells.

34 . The method of claim 33 , wherein the biomolecules are polypeptides or polynucleotides.

35 . A biological fluid comprising positively charged microparticles comprising a ground polymeric anion-exchange resin or negatively charged microparticles comprising a ground polymeric cation exchange resin.

36 . The biological fluid of claim 35 further comprising flocs.

Assignees

• Boehringer Ingelheim Rcv Gmbh
• Sandoz Ag

Inventors

• Hahn Rainer
• Jungbauer Alois
• Trefilov Alexandru

Classifications

• B01J39/26

  Cation exchangers for chromatographic processes · CPC title

• C12Y115/01001

  Superoxide dismutase (1.15.1.1) · CPC title

• B01J39/04 Primary

  Processes using organic exchangers · CPC title

• B01J41/20

  Anion exchangers for chromatographic processes · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title