Compositions and methods for generating a synthetic antibody
Disclosed herein is a composition including a recombinant nucleic acid sequence that encodes an antibody. Also disclosed herein is a method of generating a synthetic antibody in a subject by administering the composition to the subject. The disclosure also provides a method of preventing and/or treating disease in a subject using said composition and method of generation.
1. A method of generating a synthetic antibody in a subject, the method comprising administering to the subject a composition comprising a recombinant nucleic acid sequence encoding an antibody or an antigen binding fragment thereof, wherein the recombinant nucleic acid sequence is expressed in the subject to generate the synthetic antibody, wherein the nucleic acid sequence comprises a nucleic acid sequence encoding at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 41 and 43.
2. The method of claim 1 , wherein the antibody comprises a heavy chain polypeptide, or an antigen binding fragment thereof, and a light chain polypeptide, or an antigen binding fragment thereof.
3. The method of claim 2 , wherein the heavy chain polypeptide, or antigen binding fragment thereof, is encoded by a first nucleic acid sequence and the light chain polypeptide, or antigen binding fragment thereof, is encoded by a second nucleic acid sequence.
4. The method of claim 3 , wherein the recombinant nucleic acid sequence comprises the first nucleic acid sequence and the second nucleic acid sequence.
5. The method of claim 4 , wherein the recombinant nucleic acid sequence further comprises a promoter for expressing the first nucleic acid sequence and the second nucleic acid sequence as a single transcript in the subject.
6. The method of claim 5 , wherein the promoter is a cytomegalovirus (CMV) promoter.
7. The method of claim 5 , wherein the recombinant nucleic acid sequence further comprises a third nucleic acid sequence encoding a protease cleavage site, wherein the third nucleic acid sequence is located between the first nucleic acid sequence and second nucleic acid sequence.
8. The method of claim 7 , wherein a protease of subject recognizes and cleaves the protease cleavage site.
9. The method of claim 8 , wherein the recombinant nucleic acid sequence is expressed in the subject to generate an antibody polypeptide sequence, wherein the antibody polypeptide sequence comprises the heavy chain polypeptide, or antigen binding fragment thereof, the protease cleavage site, and the light chain polypeptide, or antigen binding fragment thereof, wherein the protease of the subject recognizes and cleaves the protease cleavage site of the antibody polypeptide sequence thereby generating a cleaved heavy chain polypeptide and a cleaved light chain polypeptide, wherein the synthetic antibody is generated by the cleaved heavy chain polypeptide and the cleaved light chain polypeptide.
10. The method of claim 4 , wherein the recombinant nucleic acid sequence comprises a first promoter for expressing the first nucleic acid sequence as a first transcript and a second promoter for expressing the second nucleic acid sequence as a second transcript, wherein the first transcript is translated to a first polypeptide and the second transcript is translated into a second polypeptide, wherein the synthetic antibody is generated by the first and second polypeptide.
11. The method of claim 10 , wherein the first promoter and the second promoter are the same.
12. The method of claim 11 , wherein the promoter is a cytomegalovirus (CMV) promoter.
13. The method of claim 2 , wherein the heavy chain polypeptide comprises a variable heavy region and a constant heavy region 1.
14. The method of claim 2 , wherein the heavy chain polypeptide comprises a variable heavy region, a constant heavy region 1, a hinge region, a constant heavy region 2 and a constant heavy region 3.
15. The method of claim 2 , wherein the light chain polypeptide comprises a variable light region and a constant light region.
16. The method of claim 1 , wherein the recombinant nucleic acid sequence further comprises a Kozak sequence.
17. The method of claim 1 , wherein the recombinant nucleic acid sequence further comprises an immunoglobulin (Ig) signal peptide.
18. The method of claim 17 , wherein the Ig signal peptide comprises an IgE or IgG signal peptide.
19. The method of claim 1 , wherein the recombinant nucleic acid sequence comprises at least one nucleic acid sequence selected from the group consisting of SEQ ID NOs: 40 and 42.
20. The method of claim 4 , wherein the first nucleic acid sequence is expressed in the subject to generate a first polypeptide and the second nucleic acid sequence is expressed in the subject to generate a second polypeptide, wherein the synthetic antibody is generated by the first and second polypeptides.
21. The method of claim 20 , wherein the first nucleic acid sequence further comprises a first promoter for expressing the first polypeptide in the subject and wherein the second nucleic acid sequence further comprises a second promoter for expressing the second polypeptide in the subject.
22. The method of claim 21 , wherein the first promoter and second promoter are the same.
23. The method of claim 22 , wherein the promoter is a cytomegalovirus (CMV) promoter.
24. The method of claim 20 , wherein the heavy chain polypeptide comprises a variable heavy region and a constant heavy region 1.
25. The method of claim 20 , wherein the heavy chain polypeptide comprises a variable heavy region, a constant heavy region 1, a hinge region, a constant heavy region 2 and a constant heavy region 3.
26. The method of claim 20 , wherein the light chain polypeptide comprises a variable light region and a constant light region.
27. The method of claim 20 , wherein the first nucleic acid sequence and the second nucleic acid sequence further comprise a Kozak sequence.
28. The method of claim 20 , wherein the first nucleic acid sequence and the second nucleic acid sequence further comprise an immunoglobulin (Ig) signal peptide.
29. The method of claim 28 , wherein the Ig signal peptide comprises an IgE or IgG signal peptide.
30. A method of treating a disease in a subject, the method comprising generating a synthetic antibody in a subject according to the method of claim 1 , wherein the synthetic antibody is specific for an antigen associated with the disease.
31. The method of claim 30 , wherein the synthetic antibody is specific for a self-antigen.
32. The method of claim 31 , wherein the self-antigen is Her2.