Cell culture media and methods

Patent metadata

Field	Value
Publication number	US-9410118-B2
Application number	US-201214367413-A
Country	US
Kind code	B2
Filing date	Dec 21, 2012
Priority date	Dec 22, 2011
Publication date	Aug 9, 2016
Grant date	Aug 9, 2016

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

What is claimed is:

1. A method of making a eukaryotic cell culture medium, feed or supplement composition comprising a protected and stable labile component, the method comprising:
(i) adding a minimal volume of an aqueous solution to a dry powder format of a medium, feed or supplement comprising a labile component 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; and
(v) drying the bead to form the composition containing said protected and stable labile component.

2. The method of claim 1 , wherein the bead is added aseptically into a bioreactor.

3. The method of claim 1 , wherein the cell culture medium, feed or supplement composition is protein free.

4. (Examiner Amendment) The method of claim 1 , wherein the powder of step (i) used for preparing the microsuspension is in an AGT (advanced granulation technology cell culture medium) format.

5. A medium, feed or supplement composition comprising a protected and stable labile component and prepared as a microsuspension by:
(i) adding a minimal volume of an aqueous solution to a dry powder format of a medium, feed or supplement comprising a labile component to make a paste;
(ii) mixing the paste vigorously to prepare the microsuspension;
(iii) encapsulating the microsuspension in a capsular material to form a bead; and
(iv) drying the bead to form the composition containing said protected and stable labile component.

6. The medium, feed or supplement composition of claim 5 , further comprising an anti-oxidant within the bead.

7. The medium, feed or supplement composition of claim 6 , wherein the bead is made of a 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.

8. The medium, feed or supplement composition of claim 7 , wherein the bead is coated with a coating solution.

9. The medium, feed or supplement composition of claim 8 , wherein the coating solution is selected from the group consisting of poly-glycolic acid, PLGA (poly-lactic-co-glycolic acid), collagen, polyhydroxyalkanoates (PHA), poly-e-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.

10. The medium, feed or supplement composition of claim 8 , further comprising a reactive species that is chelated using a chelator.

11. The medium, feed or supplement composition of claim 10 , wherein the reactive species is a cation, a metal ion or a trace element.

12. The medium, feed or supplement composition of claim 10 , wherein the chelator is selected from the group consisting of EDTA, citrate, succinate, cyclodextrin, clatharates, dendrimers and amino acids.

13. The medium, feed or supplement composition of claim 5 , wherein the medium, feed or supplement composition is irradiated.

14. The medium, feed or supplement composition of claim 13 , wherein the irradiation is with gamma-rays.

15. The medium, feed or supplement composition of claim 14 , wherein the gamma-rays are 25-100 kGy.

16. The medium, feed or supplement composition of claim 14 , wherein the gamma-rays are 30-50 kGy.

17. The medium, feed or supplement composition of claim 16 , wherein the gamma-ray is 30 kGy.

18. A process for increasing the shelf life of a medium, feed or supplement composition comprising a protected and stable labile component, the process comprising:
(i) adding a minimal volume of an aqueous solution to a dry powder format of a medium, feed or supplement comprising a labile component 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; and
(v) drying the bead to form the composition containing said protected and stable labile component to provide for the medium, feed or supplement composition at an ambient temperature.

19. The process of claim 18 , wherein the bead further comprises a molecule selected from the group consisting of vitamins, amino acids, peptides, macromolecules, hormones and growth factors.

20. The process of claim 19 , wherein the microsuspension of step (iv) concentrates the molecule within the bead.

Assignees

• Life Technologies Corp

Inventors

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

Classifications

• B01J13/22

  Coating · CPC title

• B29L2009/00

  Layered products · CPC title

• C12N2533/70

  Polysaccharides · CPC title

• B01J13/06

  by phase separation · CPC title

• C12N2533/74

  Alginate · CPC title