Cell culture media and methods of antibody production

Patent metadata

Field	Value
Publication number	US-2016102141-A1
Application number	US-201514852382-A
Country	US
Kind code	A1
Filing date	Sep 11, 2015
Priority date	Mar 15, 2013
Publication date	Apr 14, 2016
Grant date	—

Abstract

Cell culture media are provided herein as are methods of using the media for cell culture and antibody production from cells. Compositions comprising antibodies and fragments thereof, produced by the methods herein are also provided.

First claim

1 - 225 . (canceled)

226 . A method of producing bevacizumab or a fragment thereof, comprising a step of culturing a mammalian cell comprising a nucleic acid encoding bevacizumab or a fragment thereof in a cell culture medium, wherein initial cell culture medium in a cell culture cycle comprises two or more components selected from the group consisting of copper at a concentration of from about 69 nM to about 1,000 nM, insulin at a concentration of from about 1.0 mg/L to about 100.0 mg/L, and cystine at a concentration of from about 0.7 mM to about 2.0 mM, and wherein the cell produces bevacizumab or the fragment.

227 . The method of claim 226 , wherein the initial cell culture medium comprises (1) copper and insulin; (2) copper and cystine; (3) insulin and cystine; or (4) copper, insulin, and cystine.

228 . The method of claim 226 , wherein the initial cell culture medium comprises insulin at a concentration of from about 10.0 mg/L to about 50.0 mg/L.

229 . The method of claim 228 , wherein the initial cell culture medium comprises insulin at a concentration of from about 10.0 mg/L to about 20.0 mg/L.

230 . The method of claim 228 , wherein the initial cell culture medium comprises insulin at a concentration of about any one of 10.0 mg/L, 15 mg/L, 20 mg/L, and 25 mg/L.

231 . The method of claim 226 , wherein the initial cell culture medium comprises copper at a concentration of from about 325 nM to about 375 nM.

232 . The method of claim 226 , wherein the initial cell culture medium comprises copper at a concentration of from about 325 nM to about 350 nM.

233 . The method of claim 226 , wherein the initial cell culture medium comprises copper at a concentration of about any one of 330 nM, 335 nM, 339 nM, 340 nM, 345 nM and 350 nM.

234 . The method of claim 226 , wherein the initial cell culture medium comprises cystine at a concentration of from about 0.7 mM to about 2.0 mM.

235 . The method of claim 226 , wherein the initial cell culture medium comprises cystine at a concentration of from about 1.0 mM to about 1.6 mM.

236 . The method of claim 226 , wherein the initial cell culture medium comprises cystine at a concentration of about any one of 1.0 mM, 1.2 mM, 1.3 mM, 1.4 mM, 1.5 mM and 1.6 mM.

237 . The method of claim 226 , wherein the initial cell culture medium comprises an animal-derived hydrolysate and/or a plant-derived hydrolysate.

238 . The method of claim 226 , wherein the initial cell culture medium comprises an animal-derived hydrolysate at a concentration of from about 6.0 g/L to about 20.0 g/L.

239 . The method of claim 226 , wherein the initial cell culture medium comprises an animal-derived hydrolysate at a concentration of from about 8.0 g/L to about 12.0 g/L.

240 . The method of claim 226 , wherein the initial cell culture medium comprises an animal-derived hydrolysate at a concentration of from about 9.0 g/L to about 11.0 g/L.

241 . The method of claim 226 , wherein the initial cell culture medium comprises an animal-derived hydrolysate at a concentration of about 13 g/L.

242 . The method of claim 226 , wherein the initial cell culture medium comprises a plant-derived hydrolysate at a concentration of from about 1.0 g/L to about 10.0 g/L.

243 . The method of claim 226 , wherein the cell culture medium comprises a plant-derived hydrolysate at a concentration of from about 2.0 g/L to about 3.0 g/L.

244 . The method of claim 226 , wherein the cell culture medium comprises a plant-derived hydrolysate at a concentration of from about 2.25 g/L to about 2.75 g/L.

245 . The method of claim 226 , wherein the initial cell culture medium comprises a plant-derived hydrolysate at a concentration of about 2.5 g/L.

246 . The method of claim 226 , wherein the cell culture medium comprises both an animal-derived hydrolysate and a plant-derived hydrolysate, and wherein the animal-derived hydrolysate is present in a greater amount than the plant-derived hydrolysate.

247 . The method of claim 226 , wherein the initial cell culture medium comprises insulin and the method further comprises a step of adding an additional amount of insulin to the cell culture medium during the cell culture cycle.

248 . The method of claim 247 , wherein the additional amount of insulin is added to the cell culture medium at least once, at least twice, at least three times, at least four times, at least five times, or at least six times during the cell culture cycle.

249 . The method of claim 248 , wherein the insulin added each time is from about 5 mg/L to about 25 mg/L.

250 . The method of claim 248 , wherein the insulin added each time is about any one of 5 mg/L, 10 mg/L, 15 mg/L, 20 mg/L, and 25 mg/L.

251 . The method of claim 248 , wherein the cumulative amount of insulin added during the cell culture cycle is from about 20 mg/L to about 100 mg/L.

252 . The method of claim 251 , wherein the cumulative amount of insulin added during the cell culture cycle is about any one of 20 mg/L, 25 mg/L, 30 mg/L, 35 mg/L, 40 mg/L, 45 mg/L, 50 mg/L, 55 mg/L, 60 mg/L, 65 mg/L, 70 mg/L, 75 mg/L, 80 mg/L and 85 mg/L.

253 . The method of claim 226 , wherein the initial cell culture medium comprises cystine and the method further comprises a step of adding an additional amount of cystine to the cell culture medium during the cell culture cycle.

254 . The method of claim 253 , wherein cystine is added in an amount to provide from about 0.1 to about 1.5 mM additional cystine in the cell culture medium.

255 . The method of claim 254 , wherein cystine is added in an amount to provide about 0.4 to about 0.7 mM additional cystine in the cell culture medium.

256 . The method of claim 253 , wherein cystine is added in a batch feed during the cell culture cycle.

257 . The method of claim 226 , wherein the method further comprises at least one batch feed during the cell culture cycle.

258 . The method of claim 257 , wherein the method comprises two, three, or four batch feeds during the cell culture cycle.

259 . The method of claim 257 , wherein the batch feed medium comprises an animal-derived hydrolysate and/or a plant-derived hydrolysate.

260 . The method of claim 226 , wherein during the cell culture cycle, the temperature of the medium is reduced by at least about 2, at least about 3, at least about 4, or at least about 5 degrees C. relative to the temperature at the beginning of the culturing.

261 . The method of claim 260 , wherein the temperature of the medium is reduced at least once or at least twice during the cell culture cycle.

262 . The method of claim 261 , wherein the temperature is reduced on day 8 and day 10 after the beginning of the culturing.

263 . The method of claim 226 , wherein the cell is cultured at a temperature ranging from about 31° C. to about 35° C.

264 . The method of claim 263 , wherein the cell is cultured at a first temperature of about 35° C. for a first period of time, is cultured at a second temperature of about 33° C. for a second period of time, and is cultured at a third temperature of about 31° C. for a third period of time.

265 . The method of claim

Assignees

• Genentech Inc

Inventors

• Carvalhal Veronica
• Vijayasankaran Natarajan
• Brown Lauren
• Dirocco Thomas
• Mcknight Nathan

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C12N2510/02

  Cells for production · CPC title

• C07K16/00

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

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C12N5/0682

  Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells (oocytes C12N5/0609) · CPC title