Recombinant protein production in a human cell

Abstract

Methods and compositions for the production of recombinant proteins in a human cell line. The methods and compositions are particularly useful for generating stable expression of human recombinant proteins of interest that are modified post-translationally, for example, by glycosylation. Such proteins may have advantageous properties in comparison with their counterparts produced in non-human systems such as Chinese hamster ovary cells.

1 .- 57 . (canceled) 
     
     
         58 . A process for producing a protein of interest, comprising:
 providing an immortalized human embryonic retina cell that expresses an adenoviral E1 protein, said cell comprising a nucleotide sequence encoding the protein of interest;   culturing the cell in a serum-free culture medium to produce the protein of interest; and   harvesting the protein of interest from said culture medium,   wherein the cell does not produce structural adenoviral proteins.   
     
     
         59 . The process of  claim 58 , wherein the cell is a PER.C6® cell as deposited under ECACC number 96022940, modified by introduction therein of the nucleotide sequence encoding the protein of interest. 
     
     
         60 . The process of  claim 58 , wherein the nucleic acid encoding the protein of interest is integrated into the cell's genome. 
     
     
         61 . The process of  claim 58 , wherein the cell is in suspension during said culturing. 
     
     
         62 . The process of  claim 58 , wherein the nucleotide sequence encoding the protein of interest is under control of a heterologous promoter. 
     
     
         63 . The process of  claim 58 , wherein the protein of interest is a glycoprotein. 
     
     
         64 . The process of  claim 58 , wherein the protein of interest is a human protein. 
     
     
         65 . The process of  claim 58 , wherein the protein of interest comprises a variable domain of an immunoglobulin. 
     
     
         66 . The process of  claim 58 , wherein the protein of interest is an antibody. 
     
     
         67 . The process of  claim 58 , wherein the cell further expresses from a heterologous promoter a cDNA encoding an enzyme that is involved in post-translational modification of proteins. 
     
     
         68 . The process of  claim 67 , wherein the cDNA has been integrated into the cell's genome.