ANTIBODIES WITH IMPROVED STABILITY TO INTESTINAL DIGESTION, POLYNUCLEOTIDES THEREOF AND METHODS OF USE THEREOF TO TREAT DISEASE
The present invention provides for recombinant antibodies having the features of ruminant early colostrum antibodies that impart resistance to proteases and intestinal digestion. It is a feature of the present invention that when administered to animals including humans, pharmaceutical compositions comprising the novel recombinant antibodies of the present invention, advantageously exhibit resistance to proteases and intestinal digestion. Thus, pharmaceutical compositions of the recombinant antibodies of the invention may be used to deliver antibody therapeutics particularly by oral delivery to the gastrointestinal tract when oral delivery is advantageous.
1 . An isolated recombinant monoclonal antibody comprising (a) heavy and light chain CDRs from a non-bovine antibody and (b) a constant region comprising all or a portion of a bovine IgG1 constant region.
2 . The antibody of claim 1 , wherein the non-bovine antibody is a human antibody.
3 . The antibody of claim 1 , wherein the constant region comprises all or a portion of a bovine IgG1 hinge region.
4 . The antibody of claim 1 , wherein the constant region comprises at least one of
(i) all or a portion of a bovine IgG1 CH1 domain;
(ii) all or a portion of a bovine IgG1 CH2 domain; and
(iii) all or a portion of a bovine IgG1 CH3 domain.
5 . The antibody of claim 1 , wherein the antibody comprises one or more of the following features found in bovine IgG1:
a) a disulfide bond linking the N-terminus of the CH1 domain to the N-terminus of the hinge region,
b) a disulfide bond linking the N-terminus of the CH1 domain to the C-terminus of the light chain, and
c) a cluster of three disulfide bonds linking the hinge regions of two heavy chains together.
6 . The antibody of claim 1 , wherein the bovine IgG1 constant region is selected from any of SEQ ID NOs: 1-3.
7 . The antibody of claim 1 , wherein the constant region further comprises a portion of a human constant region.
8 . The antibody of claim 7 , wherein the human constant region is an IgG1 or IgG2.
9 . An isolated recombinant monoclonal human antibody, wherein the constant region of the antibody comprises one or more of the following features found in bovine IgG1:
a) a disulfide bond linking the N-terminus of the CH1 domain to the N-terminus of the hinge region,
b) a disulfide bond linking the N-terminus of the CH1 domain to the C-terminus of the light chain, and
c) a cluster of three disulfide bonds linking the hinge regions of two heavy chains together.
10 . The antibody of claim 9 , wherein the antibody binds to a biological antigen.
11 . The antibody of claim 10 , wherein the antigen is TNF-α.
12 . The antibody of claim 11 , wherein the antibody comprises the heavy and light chain variable region sequences of infliximab.
13 . The antibody of claim 1 , wherein the antibody retains antigen binding after protease digestion.
14 . The antibody of claim 1 , wherein the antibody comprises constant region comprising one or more of the following substitutions (Kabat numbering): threonine at position 252 is substituted with methionine; glycine at position 255 is substituted with arginine; glutamine at position 309 is substituted with leucine; threonine at position 314 is substituted with leucine; and glycine at position 315 is substituted with asparagine.
15 . A recombinant fully bovine monoclonal IgG1 antibody.
16 . A nucleic acid encoding the antibody of claim 1 .
17 . An expression vector comprising the nucleic acid of claim 16 .
18 . A host cell comprising the nucleic acid of claim 16 .
19 . A pharmaceutical composition comprising the antibody of claim 1 and a carrier and, optionally, a preservative, the composition being optionally formulated for oral administration.
20 . The composition of claim 19 which is lyophilized.
21 . A method of treating a disease of the digestive tract comprising orally administering to an individual in need thereof the antibody of claim 1 .
22 . The method of claim 21 , wherein the disease is selected from the group consisting of ulcerative colitis and Crohn's disease.
23 . A method of producing the antibody of claim 1 , comprising expressing the antibody in a host cell and isolating the antibody from the cell.