Highly galactosylated anti-tnf-alpha antibodies and uses thereof

Abstract

In one aspect, the disclosure relates to highly galactosylated anti-TNF-alpha antibodies and compositions thereof. In one aspect, the disclosure relates to populations of anti-TNF-alpha antibodies with a high level of galactosylation, and compositions thereof. In one aspect, the disclosure relates to methods of production and use of highly galactosylated anti-TNF-alpha antibodies and populations of anti-TNF-alpha antibodies with a high level of galactosylation. In some embodiments, the anti-TNF-alpha antibody is adalimumab.

What is claimed is: 
     
         1 - 77 . (canceled) 
     
     
         78 . A transgenic non-human mammal comprising mammary gland epithelial cells, wherein the mammary gland epithelial cells of the transgenic non-human mammal comprise cells that are genetically engineered to express a nucleic acid sequence encoding the heavy chain of the anti-TNF-alpha antibody and a nucleic acid sequence encoding the light chain of an anti-TNF-alpha antibody. 
     
     
         79 . The transgenic non-human mammal of  claim 78 , wherein the nucleic acid sequence encoding the heavy chain of the anti-TNF-alpha antibody comprises a nucleic acid sequence of SEQ ID NO: 3, and wherein the nucleic acid sequence encoding the light chain of the anti-TNF-alpha antibody comprises a nucleic acid sequence of SEQ ID NO: 4. 
     
     
         80 . The transgenic non-human mammal of  claim 79 , wherein the nucleic acid sequence encoding the heavy chain of the anti-TNF-alpha antibody and the nucleic acid sequence encoding the light chain of the anti-TNF-alpha antibody are physically linked. 
     
     
         81 . The transgenic non-human mammal of  claim 79 , wherein the nucleic acid sequence encoding the heavy chain of the anti-TNF-alpha antibody and the nucleic acid sequence encoding the light chain of the anti-TNF-alpha antibody are operably linked to a milk specific promoter. 
     
     
         82 . The transgenic non-human mammal of  claim 81 , wherein the milk specific promoter is a casein promoter, a b-casein promoter, a r-casien promoter, a K-casein promoter, a whey acidic protein promoter, or a α-lactalbumin promoter. 
     
     
         83 . The transgenic non-human mammal of  claim 79 , wherein the non-human mammal is a goat, a sheep, a bison, a camel, a cow, a pig, a rabbit, a buffalo, a horse, a rat, a mouse or a llama. 
     
     
         84 . The transgenic non-human mammal of  claim 83 , wherein the non-human mammal is a goat. 
     
     
         85 . The transgenic non-human mammal of  claim 83 , wherein the mammary gland epithelial cells expresses a population of anti-TNF-alpha antibodies. 
     
     
         86 . The transgenic non-human mammal of  claim 85 , wherein the level of galactosylation of the population of anti-TNF-alpha antibodies is at least 70%. 
     
     
         87 . The transgenic non-human mammal of  claim 85 , wherein the population of anti-TNF-alpha antibodies comprises mono-galactosylated N-glycans and/or bi-galactosylated N-glycans. 
     
     
         88 . The transgenic non-human mammal of  claim 86 , wherein the level of fucosylation of the population of anti-TNF-alpha antibodies is at least 50%. 
     
     
         89 . The transgenic non-human mammal of  claim 88 , wherein the ratio of the level of galactosylation to the level of fucosylation of the population of anti-TNF-alpha antibodies is between 1.0 and 1.4. 
     
     
         90 . The transgenic non-human mammal of  claim 89 , wherein at least 35% of the antibodies in the population of anti-TNF-alpha antibody comprise bi-galactosylated N-glycans and at least 25% of the antibodies in the population of anti-TNF-alpha antibodies comprises mono-galactosylated N-glycans. 
     
     
         91 . The transgenic non-human mammal of  claim 85 , wherein the population of anti-TNF-alpha antibodies has an increased level of complement dependent cytotoxicity (CDC) activity when compared to a population of antibodies not produced in mammary gland epithelial cells. 
     
     
         92 . The transgenic non-human mammal of  claim 85 , wherein the population of anti-TNF-alpha antibodies has an increased level of antibody-dependent cellular cytotoxicity (ADCC) activity when compared to a population of antibodies not produced in mammary gland epithelial cells. 
     
     
         93 . The transgenic non-human mammal of  claim 79 , wherein the anti-TNF-alpha antibody comprises a heave chain, which comprises an amino acid sequence of SEQ ID NO: 1, and a light chain, which comprises an amino acid sequence of SEQ ID NO: 2. 
     
     
         94 . The transgenic non-human mammal of  claim 85 , wherein at least 30% of the antibodies in the population of antibodies contain at least one oligomannose; and wherein the level of fucosylation of the antibodies in the population of antibodies is at least 30%. 
     
     
         95 . The transgenic non-human mammal of 94, wherein the antibodies in the population of antibodies exhibit a high mannose glycosylation pattern. 
     
     
         96 . The transgenic non-human mammal of  claim 95 , wherein at least one chain of the antibodies in the population of antibodies contains an oligomannose and is non-fucosylated. 
     
     
         97 . The transgenic non-human mammal of  claim 94 , wherein the major carbohydrate of the antibodies in the population of antibodies is non-fucosylated.