Cell culture process for making a glycoprotein

Abstract

The instant application provides a method for screening batches of soy hydrolysate for a desired amount of a component thereof, such as ornithine or putrescine, and selecting only those batches of soy hydrolysate that have a desired amount of such component. The present disclosure also sets forth methods for culturing cells in media supplemented with selected batches of soy to produce more consistent, high quality lots of a protein of interest. Further, the present disclosure provides a plurality of protein preparations that have each been produced by culturing cells in media supplemented with separate batches of soy hydrolysate containing a desired amount of ornithine or putrescine, whereby each batch of protein produced exhibits improved quality of the protein of interest or amount of quality protein produced.

What is claimed is: 
     
         1 . A method of producing sarilumab comprising culturing a population of cells expressing sarilumab in a cell culture medium comprising soy hydrolysate, wherein the cell culture medium further comprises ornithine, putrescine, or a combination thereof, and wherein the concentration of ornithine, putrescine or the combination thereof in the cell culture medium is between 3.0 mg/L to 5.0 mg/L, thereby producing sarilumab. 
     
     
         2 . The method of  claim 1 , wherein the population of cells is obtained by clonal expansion of a cell comprising a recombinant polynucleotide encoding sarilumab. 
     
     
         3 . The method of  claim 1 , wherein the cell culture medium is chemically defined. 
     
     
         4 . The method of  claim 1 , wherein the cell culture medium comprises the twenty basic amino acids and cysteine. 
     
     
         5 . The method of  claim 1 , wherein the cell culture medium comprises lipids or free fatty acids. 
     
     
         6 . The method of  claim 1 , wherein the cells are a primary cells, BSC cells, HeLa cells, HepG2 cells, LLC-MK cells, CV-1 cells, COS cells, VERO cells, MDBK cells, MDCK cells, CRFK cells, RAF cells, RK cells, TCMK-1 cells, LLCPK cells, PK15 cells, LLC-RK cells, MDOK cells, BHK cells, BHK-21 cells, CHO cells, NS-1 cells, MRC-5 cells, WI-38 cells, BHK cells, 3T3 cells, 293 cells, RK cells, Per.C6 cells or chicken embryo cells. 
     
     
         7 . The method of  claim 1 , wherein the cells are CHO cells, 293 cells, or BHK cells. 
     
     
         8 . The method of  claim 7 , wherein the CHO cells are CHO-K1 cells. 
     
     
         9 . The method of  claim 1 , further comprising supplementing the cell culture medium with one or more point-of-use additions. 
     
     
         10 . The method of  claim 9 , wherein the one or more point-of-use additions comprise a growth factor, buffer, energy source, salt, amino acid, metal, chelator or combination thereof. 
     
     
         11 . The method of  claim 9 , wherein the one or more point-of-use additions comprises insulin. 
     
     
         12 . The method of  claim 11 , wherein the amount of insulin in the cell culture medium after addition is from about 0.1 μM to 10 μM. 
     
     
         13 . The method of  claim 10 , wherein the buffer comprises NaHCO 3  or HEPES. 
     
     
         14 . The method of  claim 10 , wherein the energy source comprises glucose. 
     
     
         15 . The method of  claim 14 , wherein the concentration of glucose in the cell culture medium after addition is about 1 mM to about 20 mM. 
     
     
         16 . The method of  claim 10 , wherein the metal comprises a metal salt. 
     
     
         17 . The method of  claim 16 , wherein the metal salt comprises copper sulfate, zinc sulfate, ferric chloride, nickel sulfate or a combination thereof. 
     
     
         18 . The method of  claim 10 , wherein the chelator comprises Tetrasodium EDTA dehydrate or citrate. 
     
     
         19 . The method of  claim 1 , further comprising purifying the sarilumab. 
     
     
         20 . A method of producing sarilumab comprising culturing a population of cells expressing sarilumab in a cell culture medium comprising soy hydrolysate, wherein the cell culture medium further comprises ornithine, putrescine, or a combination thereof, and wherein the concentration of ornithine, putrescine or the combination thereof in the cell culture medium is less than or equal to 5.0 mg/L, thereby producing sarilumab. 
     
     
         21 . The method of  claim 20 , wherein the concentration of ornithine, putrescine, or the combination thereof is between 0.5 mg/L to 2.0 mg/L, between 0.5 mg/L to 2.5 mg/L, between 0.5 mg/L to 3.0 mg/L, between 1.5 mg/L to 5 mg/L, or between 2.0 mg/L to 5.0 mg/L.