Cell culture media and methods

Patent metadata

Field	Value
Publication number	US-2016376548-A1
Application number	US-201615198237-A
Country	US
Kind code	A1
Filing date	Jun 30, 2016
Priority date	Dec 22, 2011
Publication date	Dec 29, 2016
Grant date	—

Abstract

Compositions and methods are described for preparing media, feeds, and supplements. Such methods and medias may display increased stability of labile components and may use, for example, microsuspension and/or encapsulation technologies, chelation, and optionally, coating and/or mixing the labile compounds with anti-oxidants. The compositions may withstand thermal and/or irradiation treatment and have reduced virus number. These techniques may result in product with extended shelf-life, extended release of their internal components into culture, or in product that can be added aseptically into a bioreactor using minimal volumes. The compositions and methods may optimize the bioproduction workflow and increase efficiency.

First claim

1 - 21 . (canceled)

22 . A method of making a recombinant polypeptide or virus, the method comprising:
a) preparing a cell culture medium, feed or supplement comprising a microsuspension by:
(i) adding a minimal volume of an aqueous solution to a dry powder medium, feed or supplement to make a paste;
(ii) mixing the paste vigorously to prepare a microsuspension;
(iii) optionally, adding an effective amount of an anti-oxidant to the microsuspension to form a mixture;
(iv) encapsulating the microsuspension of (ii), or the mixture of (iii), in a capsular material to form a bead;
(v) drying the bead to form the cell culture medium, feed or supplement,
b) dissolving the cell culture medium, feed or supplement of (v) in a solvent to form a liquid cell culture medium, c) contacting said liquid cell culture medium with a cell expressing the recombinant polypeptide or virus, d) culturing said cell in said liquid cell culture medium under conditions favoring the expression of the recombinant polypeptide or virus.

23 . The method of claim 22 , wherein the cell culture medium comprises a labile substance.

24 . The method of claim 22 , wherein the capsular material that is selected from the group consisting of alginate, poly-L-lactic acid, chitosan, agarose, gelatin, hyaluronic acid, chondroitin sulfate, dextran, dextran sulfate, heparin, heparin sulfate, heparan sulfate, gellan gum, xanthan gum, guar gum, water soluble cellulose derivatives and carrageenan.

25 . The method of claim 22 , wherein the bead is coated with a coating solution.

26 . The method of claim 25 , wherein the coating solution is selected form the group consisting of poly-glycolic acid, PLGA (poly-lactic-co-glycolic acid), collagen, polyhydroxyalkanoates (PHA), poly-c-caprolactone, poly-ortho esters, poly-anhydrides, poly-phosphazenes, poly-amino acids, polydimethylsiloxane,polyurethranes, poly-tetrafluoroethylene, polyethylene, polysulphone, poly-methyl methacrylate, poly-2-hydroxyethylmethacrylate, polyamides, polypropylene, poly-vinyl chloride, polystyrene, poly-vinyl pyrrolidone, poly-L-lysine and polyornithine.

27 . The method of claim 25 , wherein the bead further comprises a reactive species that is chelated using a chelator.

28 . The method of claim 27 , wherein the reactive species is a cation, a metal ion or a trace element.

29 . The method of claim 27 , wherein the chelating moieties are selected from the group consisting of EDTA, citrate, succinate, cyclodextrin, clatharates, dendrimers and amino acids.

30 . The method of claim 22 , wherein the cell culture medium, feed or supplement is irradiated.

31 . The method of claim 30 wherein the irradiation is with gamma-rays.

32 . The method of claim 31 , wherein the gamma-rays are 25-100 kGy.

33 . The method of claim 22 wherein the cell is a mammalian cell.

34 . The method of claim 22 , wherein the cell culture medium, feed or supplement is protein free.

35 . The method of claim 22 , wherein the powdered cell culture medium used for the microsuspension is AGT (advanced granulation technology cell culture medium).

36 . A method of culturing a cell comprising:
a) preparing a medium, feed or supplement comprising a microsuspension by:
(i) adding a minimal volume of an aqueous solution to a dry powder medium, feed or supplement to make a paste;
(ii) mixing the paste vigorously to prepare a microsuspension;
(iii) optionally, adding an effective amount of an anti-oxidant to the microsuspension to form a mixture;
(iv) encapsulating the microsuspension of (ii), or the mixture of (iii), in a capsular material to form a bead;
(v) drying the bead to form the medium, feed or supplement comprising the microsuspension,
b) dissolving the medium, feed or supplement of (v) in a solvent to form a liquid cell culture medium, c) contacting said liquid cell culture medium with said cell to be cultured.

37 . The method of culturing a cell of claim 36 wherein, the medium, feed or supplement comprising the microsuspension is stored at ambient temperatures.

38 . The method of culturing a cell of claim 36 wherein the cell is a mammalian cell.

39 . The method of culturing a cell of claim 38 wherein the mammalian cell is selected from the group consisting of Chinese Hamster Ovary (CHO) cells, 293. BHK, Vero, PerC6, MDBK and MDCK cells.

40 . The method of culturing a cell of claim 39 wherein the mammalian cell is a CHO cell.

41 . The method of culturing a cell of claim 36 wherein the medium, feed or supplement is irradiated.

Assignees

• Life Technologies Corp

Inventors

• Fike Richard
• Barrett Shawn
• Jiang Zhou
• Judd David

Classifications

• B29L2009/00

  Layered products · CPC title

• C12N2533/40

  Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers · CPC title

• B29C67/24

  characterised by the choice of material · CPC title

• C12N2533/74

  Alginate · CPC title

• C12N2511/00

  Cells for large scale production · CPC title