Glycosylated antibody

Abstract

The invention relates to a CHO cell-line capable of producing antibody, the cell-line having been co-transfected with a vector capable of expressing the light chain of the antibody and a vector capable of expressing the heavy chain of the antibody wherein the vectors contain independently selectable markers; also included is a CHO cell-line capable of producing a human antibody or an altered antibody, the cell-line having been transfected with a vector capable of expressing the light chain of the antibody and the heavy chain of the antibody; process for the production of antibody using a CHO cell-line and antibody having CHO glycosylation.

1 . An immunotherapy method of treating a human suffering from a disease or disorder, which method comprises the steps of: 
 (i) constructing a first recombinant expression vector encoding a light chain of a therapeutically effective antibody and constructing a second expression vector encoding a heavy chain of said therapeutically effective antibody;    (ii) introducing said vectors of step (i) into a Chinese hamster ovary (CHO) cell;    (iii) culturing said CHO cell in a culture medium so that said light and heavy chains are produced and a CHO glycosylated therapeutically effective recombinant antibody is thereby produced;    (iv) recovering said therapeutically effective recombinant antibody of step (iii);    (v) administering the recombinant antibody of step (iv) in a therapeutically effective amount to said human.    
     
     
         2 . The method of  claim 1  wherein the recombinant antibody is a human, chimeric, CDR-grafted or bi-specific antibody.  
     
     
         3 . The method of  claim 1  wherein the human is afflicted with a T-cell disorder.  
     
     
         4 . The method of  claim 3  wherein the T-cell disorder is severe vasculitis, rheumatoid arthritis or systemic lupus.  
     
     
         5 . The method of  claim 1  wherein the human is afflicted with an autoimmune disease.  
     
     
         6 . The method of  claim 5  wherein the autoimmune disease is multiple sclerosis, graft vs host disease, psoriasis, Juvenile onset diabetes, Sjogrens disease, thyroid disease, myasthenia gravis, transplant rejection or asthma.  
     
     
         7 . The method of  claim 1  wherein the human is afflicted with cancer.  
     
     
         8 . The method of  claim 7  wherein the cancer is non-Hodgkins lymphoma or multiple myeloma.  
     
     
         9 . The method of  claim 1  wherein the human is afflicted with an infectious disease.  
     
     
         10 . The method of  claim 9  wherein the infectious disease is HIV or herpes.  
     
     
         11 . An immunotherapy method of treating a human suffering from a disease or disorder, which method comprises the steps of: 
 (i) transforming a Chinese hamster ovary (CHO) cell with a recombinant expression vector such that said cell can express a recombinant antibody;    (ii) culturing said CHO cell in serum-free medium so that a CHO glycosylated therapeutically effective recombinant antibody is thereby produced;    (iii) recovering said therapeutically effective recombinant antibody of step (ii);    (iv) administering the recombinant antibody of step (iii) in a therapeutically effective amount to said human.    
     
     
         12 . The method of  claim 11  wherein the recombinant antibody is a human, chimaeric, CDR-grafted or bi-specific antibody.  
     
     
         13 . The method of  claim 11  wherein the human is afflicted with a T-cell disorder.  
     
     
         14 . The method of  claim 13 , wherein the T-cell disorder is severe vasculitis, rheumatoid arthritis or systemic lupus.  
     
     
         15 . The method of  claim 11  wherein the human is afflicted with an autoimmune disease.  
     
     
         16 . The method of  claim 15  wherein the autoimmune disease is multiple sclerosis, graft vs host disease, psoriasis, Juvenile onset diabetes, Sjogrens disease, thyroid disease, myasthenia gravis, transplant rejection or asthma.  
     
     
         17 . The method of  claim 11  wherein the human is afflicted with cancer.  
     
     
         18 . The method of  claim 17  wherein the cancer is non-Hodgkins lymphoma or multiple myeloma.  
     
     
         19 . The method of  claim 11  wherein the human is afflicted with an infectious disease.  
     
     
         20 . The method of  claim 19  wherein the infectious disease is HIV or herpes.  
     
     
         21 . The method of  claim 11  wherein the cell is cultured in said serum-free medium for greater than two months.  
     
     
         22 . The method of  claim 21  wherein the cell is cultured in said serum free medium for greater than five months.  
     
     
         23 . The method of  claim 11  wherein the culture undergoes multiple passage.  
     
     
         24 . The method of  claim 11  wherein the serum free medium comprises water, an osmolarity regulator, a buffer, an energy source, L-glutamine and at least one additional amino acid, an inorganic iron source and a recombinant growth factor wherein each component of said medium is obtained from a source other than directly from an animal source.  
     
     
         25 . The medium of  claim 11  wherein the medium is devoid of bovine serum albumin, pure human transferrin and soyabean lecithin.  
     
     
         26 . The method of  claim 11  wherein the growth factor is recombinant insulin.  
     
     
         27 . The method of  claim 11  wherein the basal medium component of said serum-free medium is an Iscove modification of DMEM.  
     
     
         28 . The method of  claim 11  wherein the serum-free medium consists essentially of the medium of WCM4.  
     
     
         29 . The method of  claim 11  wherein the serum-free medium consists essentially of the medium of WCM 5.  
     
     
         30 . The method of  claim 11  wherein the serium-free medium consists of the medium WCM 4.  
     
     
         31 . The method of  claim 11  wherein the serum-free medium consists of the medium WCM 5.  
     
     
         32 . An immunotherapy method of treating a human suffering from a disease or disorder, which method comprises the steps of: 
 administering to a human suffering from a disease or disorder, a therapeutically effective amount of a therapeutically effective recombinant human antibody or recombinant altered antibody, said recombinant antibody comprising two light chains and two heavy chains and having an Fc region and a Fab region, wherein said recombinant antibody is expressed in a Chinese hamster ovary (CHO) cell and glycosylated in said Fc region by said CHO cell.    
     
     
         33 . The method of  claim 32 , wherein said antibody has complement lysis activity in an in vitro assay.  
     
     
         34 . The method of  claim 32 , wherein when said antibody is bound to a target cell in said human, the cell to which it is bound is lysed.  
     
     
         35 . The method of  claim 32 , wherein said antibody contains human constant domains.  
     
     
         36 . The method of  claim 8 , wherein the cancer is non-Hodgkins lymphoma.  
     
     
         37 . The method of  claim 18 , wherein the cancer is non-Hodgkins lymphoma.  
     
     
         38 . The method of  claim 32 , wherein the cancer is non-Hodgkins lymphoma.