Compounded media powder formulation and method of preparation of liquid medium for cell culture

Patent metadata

Field	Value
Publication number	US-10227559-B2
Application number	US-201515302488-A
Country	US
Kind code	B2
Filing date	Apr 7, 2015
Priority date	Apr 10, 2014
Publication date	Mar 12, 2019
Grant date	Mar 12, 2019

Abstract

The present invention provides a compounded cell culture medium powder formulation comprising: a basal medium powder and a cell culture media supplement, wherein the cell culture media supplement comprises and one or more salts; one or more growth factors; one or more inorganic ions; an amino acid supplement comprising one or more of asparagine, glutamine, histidine, and serine; one or more buffers; and one or more anti-foaming agents. The invention further provides methods of making a compounded cell culture medium powder formulation methods of making a cell culture medium for growing mammalian cells and methods of producing a protein of interest by culturing cells in the cell culture medium and isolating the protein of interest.

First claim

What is claimed is:

1. A method of making a compounded cell culture medium powder formulation for growing mammalian cells, comprising:
combining a basal medium powder having the following components at the following concentrations upon hydration:
i) 0.0003-0.003 g/L biotin;
ii) 0.035-0.33 g/L calcium chloride;
iii) 0.003-0.03 g/L choline chloride;
iv) 0.0002-0.002 g/L cyanocobalamin (B 12);
v) 1-10 g/L D-mannose;
vi) 0.001-0.01 g/L D-calcium pantothenate;
vii) 0.3-3.0 g/L dextrose (anhydrous);
viii) 0.00003-0.0003 g/L DL-alpha-lipoic acid;
ix) 0.00001-0.00015 g/L ferric nitrate;
x) 0.00005-0.0015 g/L ferrous sulfate;
xi) 0.001-0.01 g/L folic acid;
xii) 0.007-0.20 g/L glycine;
xiii) 0.001-0.01 g/L hypoxanthine 2Na;
xiv) 0.005-0.05 g/LI-inositol;
xv) 0.003-0.03 g/L L-alanine;
xvi) 0.08-1.4 /L L-arginine;
xvii) 0.006-0.16 g/L L-asparagine;
xviii) 0.005-0.10 g/L L-aspartic acid;
xix) 0.005-0.05 g/L L-cysteine HCl;
xx) 0.02-0.2 g/L L-cystine 2HCl;
xxi) 0.005-0.15 g/L L-glutamic acid (anhydrous);
xxii) 0.02-1.5 g/L L-glutamine;
xxiii) 0.0003-0.003 g/L L-glutathione;
xxiv) 0.02-0.2 g/L L-histidine HCl;
xxv) 0.03-0.9 g/L L-isoleucine;
xxvi) 0.03-0.9 g/L L-leucine;
xxvii) 0.05-1.5 g/L L-lysine;
xxviii) 0.01-0.3 g/L L-methionine;
xxix) 0.02-0.6 g/L L-phenylalanine;
xxx) 0.008-0.25 g/L L-proline;
xxxi) 0.009-0.25 g/L L-serine;
xxxii) 0.03-0.9 g/L L-threonine;
xxxiii) 0.006-0.16 g/L L-tryptophan;
xxxiv) 0.02-0.9 g/L L-tyrosine 2Na;
xxxv) 0.03-0.9 g/L L-valine;
xxxvi) 0.01-0.18 g/L magnesium chloride;
xxxvii) 0.02-0.12 g/L magnesium sulfate anhydrous;
xxxviii) 0.001-0.01 g/L niacinamide;
xxxix) 0.0005-0.005 g/L O-phoshphoryl-ethanolamine;
xl) 0.1-1.0 g/L potassium chloride;
xli) 0.00002-0.0002 g/L putrescine 2HCl;
xlii) 0.001-0.01 g/L pyridoxal HCl;
xliii) 0.00001-0.0001 g/L pyridoxine HCl;
xliv) 0.0001-0.001 g/L riboflavin;
xlv) 2.0-15 g/L sodium chloride;
xlvi) 0.01-0.2 g/L sodium phosphate monobasic;
xlvii) 0.02-0.2 g/L sodium phosphate dibasic anhydrous;
xlviii) 0.015-0.15 g/L sodium pyruvate;
xlix) 0.001-0.01 g/L thiamine HCl;
l) 0.0001-0.001 g/L thymidine;
li) 0.00006-0.0015 g/L zinc sulfate;
lii) 0.0000003-0.000006 g/L cupric sulfate;
liii) 0.0000005-0.000008 g/L selenium dioxide;
liv) 0.00001-0.0001 g/L linoleic acid;
lv) 0.0001-0.001 g/L beta-mercaptoethanol;
lvi) 0.0003-0.005 g/L ethanolamine;

with basal media supplement powder having the following components at the following concentrations upon hydration:
a) 0.5-5 g/L magnesium chloride;
b) 0.5-15 mg/mL insulin;
c) one or more additional trace metals selected from the group consisting of:
0.0005-0.01 mg/L ammonium molybdate tetrahydrate,
0.0001-0.01 mg/L chromium potassium sulfate dodecahydrate,
0.001-0.125 mg/L cupric sulfate pentahydrate,
0.001-0.1 mg/L of litium chloride (anhydrous)
0.00004-0.004 mg/L manganese sulfate hydrate, and
0.04-4.2 mg/L of sodium metasilicate nonahydrate;

d) one or more additional amino acid selected from the group consisting of 0.007-0.07 g/L asparagine hydrate, 0.25-2.5 g/L glutamine, 0.5-5 g/L histidine free base, and 0.01-0.1 g/L serine;
e) one or more buffers; and
f) one or more anti-foaming agents,

to make a cell culture medium powder, wherein the cell culture media when hydrated completely dissolves the media components.

Assignees

• Bayer Healthcare Llc

Inventors

• Shimoni Yuval
• Möhrle Volker

Classifications

• C12N2500/42

  Organic phosphate, e.g. beta glycerophosphate · CPC title

• C12N5/0018 Primary

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12N2500/32

  Amino acids · CPC title

• C12N2500/34

  Sugars · CPC title

• C12N2500/44

  Thiols, e.g. mercaptoethanol · CPC title