Cell culture media and methods of antibody production

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.

1 . A method of producing bevacizumab, or a fragment thereof, comprising the step of culturing a mammalian cell comprising a nucleic acid encoding bevacizumab or fragment thereof in a cell culture medium, wherein the cell culture medium comprises two or more components selected from the group consisting of copper, insulin, and cystine, and wherein the cell produces bevacizumab, or a fragment thereof. 
     
     
         2 - 6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the cell culture medium comprises insulin at a concentration of from about 1.0 mg/L to about 100.0 mg/L, about 10.0 mg/L to about 100.0 mg/L, about 10.0 mg/L to about 50.0 mg/L, about 10.0 mg/L to about 35.0 mg/L, about 10.0 mg/L to about 25.0 mg/L, or about 25 mg/L. 
     
     
         8 - 12 . (canceled) 
     
     
         13 . The method of  claim 1 , wherein the cell culture medium comprises copper at a concentration of from about 69 nM to about 1,000 nM, about 325 nM to about 375 nM, about 325 nM to about 350 nM, or about any one of 330 nM, 335 nM, 339 nM, 340 nM, 345 nM or 350 nM. 
     
     
         14 - 30 . (canceled) 
     
     
         31 . The method of  claim 1 , wherein the cell culture medium comprises insulin and the method further comprises the step of adding an additional amount of insulin to the cell culture medium. 
     
     
         32 - 40 . (canceled) 
     
     
         41 . The method of  claim 1 , wherein the method further comprises the step of adding cysteine to the cell culture medium. 
     
     
         42 - 43 . (canceled) 
     
     
         44 . The method of  claim 1 , wherein the method further comprises the step of adding cystine to the cell culture medium. 
     
     
         45 - 48 . (canceled) 
     
     
         49 . The method of  claim 1 , 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. 
     
     
         50 - 51 . (canceled) 
     
     
         52 . Bevacizumab, or fragment thereof, produced by the method of  claim 1 . 
     
     
         53 . A composition comprising: (i) bevacizumab, or a fragment thereof, produced by the method of  claim 1  and (ii) a pharmaceutically acceptable carrier. 
     
     
         54 . A method of culturing a mammalian cell comprising a nucleic acid encoding bevacizumab, or a fragment thereof, the method comprising the step of contacting the mammalian cell with a cell culture medium comprising two or more components selected from the group consisting of copper, insulin and cystine. 
     
     
         55 - 59 . (canceled) 
     
     
         60 . The method of  claim 54 , wherein the cell culture medium comprises insulin at a concentration of from about 1.0 mg/L to about 100.0 mg/L, about 10.0 mg/L to about 100.0 mg/L, about 10.0 mg/L to about 50.0 mg/L, about 10.0 mg/L to about 35.0 mg/L, about 10.0 mg/L to about 25.0 mg/L, or about 25 mg/L. 
     
     
         61 - 65 . (canceled) 
     
     
         66 . The method of  claim 54 , wherein the cell culture medium comprises copper at a concentration of from about 69 nM to about 1,000 nM about 325 nM to about 375 nM, about 325 nM to about 350 nM, or about any one of 330 nM, 335 nM, 339 nM, 340 nM, 345 nM or 350 nM. 
     
     
         67 - 83 . (canceled) 
     
     
         84 . The method of  claim 54 , wherein the cell culture medium comprises insulin and the method further comprises the step of adding an additional amount of insulin to the cell culture medium. 
     
     
         85 - 93 . (canceled) 
     
     
         94 . The method of  claim 54 , wherein the method further comprises the step of adding cysteine to the cell culture medium. 
     
     
         95 - 96 . (canceled) 
     
     
         97 . The method of  claim 54 , wherein the method further comprises the step of adding cystine to the cell culture medium. 
     
     
         98 - 102 . (canceled) 
     
     
         103 . The method of  claim 54 , 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. 
     
     
         104 - 106 . (canceled) 
     
     
         107 . A kit for supplementing a cell culture medium for use in culturing a mammalian cell comprising a nucleic acid encoding bevacizumab, or a fragment thereof, the kit comprising at least two of components (i)-(iii): (i) insulin in an amount to provide from about 1.0 mg/L to about 100.0 mg/L insulin in the cell culture medium; (ii) cystine in an amount to provide from about 0.7 mM to about 2.0 mM cystine in the cell culture medium; (iii) and copper in an amount to provide from about 69.0 nM to about 1,000.0 nM copper in the cell culture medium. 
     
     
         108 - 111 . (canceled) 
     
     
         112 . A cell culture medium for use in culturing a mammalian cell comprising a nucleic acid encoding bevacizumab, or a fragment thereof, the cell culture medium comprising at least two of components (i)-(iii): (i) from about 1.0 mg/L to about 100.0 mg/L insulin; (ii) from about 69.0 nM to about 1,000.0 nM copper; and (iii) from about 0.7 mM to about 2.0 mM cystine. 
     
     
         113 - 120 . (canceled) 
     
     
         121 . A composition comprising (a) a mammalian cell comprising a nucleic acid encoding bevacizumab, or a fragment thereof; and (b) a cell culture medium according to  claim 112 . 
     
     
         122 . A composition comprising: (a) bevacizumab, or a fragment thereof; and (b) a cell culture medium according to  claim 112 . 
     
     
         123 - 225 . (canceled)