Dna antibody constructs and method of using same

Abstract

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 fragment thereof, wherein the recombinant nucleic acid sequence is expressed in the subject to generate the synthetic antibody, wherein the antibody comprises an amino acid sequence at least 80% identical to SEQ ID NO:80. 
     
     
         2 . The method of  claim 1 , wherein the antibody comprises a heavy chain polypeptide, or fragment thereof, and a light chain polypeptide, or fragment thereof. 
     
     
         3 . The method of  claim 2 , wherein the heavy chain polypeptide, or fragment thereof, is encoded by a first nucleic acid sequence and the light chain polypeptide, or 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 the protease of the 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 fragment thereof, the protease cleavage site, and the light chain polypeptide, or fragment thereof, wherein the protease produced by 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 a nucleic acid sequence encoding 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 antibody comprises the amino acid sequence of SEQ ID NO:80. 
     
     
         20 . The method of  claim 1 , wherein the recombinant nucleic acid sequence comprises a nucleic acid sequence at least 90% identical to SEQ ID No: 79. 
     
     
         21 . A method of generating a synthetic antibody in a subject, the method comprising administering to the subject a composition comprising a first recombinant nucleic acid sequence encoding a heavy chain polypeptide, or fragment thereof, and a second recombinant nucleic acid sequence encoding a light chain polypeptide, or fragment thereof, wherein the first recombinant nucleic acid sequence is expressed in the subject to generate a first polypeptide and the second recombinant nucleic acid is expressed in the subject to generate a second polypeptide, wherein the synthetic antibody is generated by the first and second polypeptides, wherein the antibody comprises an amino acid sequence at least 80% identical to SEQ ID NO:80. 
     
     
         22 . The method of  claim 21 , wherein the first recombinant nucleic acid sequence further comprises a first promoter for expressing the first polypeptide in the subject and wherein the second recombinant nucleic acid sequence further comprises a second promoter for expressing the second polypeptide in the subject. 
     
     
         23 . The method of  claim 22 , wherein the first promoter and second promoter are the same. 
     
     
         24 . The method of  claim 23 , wherein the promoter is a cytomegalovirus (CMV) promoter. 
     
     
         25 . The method of  claim 21 , wherein the heavy chain polypeptide comprises a variable heavy region and a constant heavy region 1. 
     
     
         26 . The method of  claim 21 , 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. 
     
     
         27 . The method of  claim 21 , wherein the light chain polypeptide comprises a variable light region and a constant light region. 
     
     
         28 . The method of  claim 21 , wherein the first recombinant nucleic acid sequence and the second recombinant nucleic acid sequence further comprise a Kozak sequence. 
     
     
         29 . The method of  claim 21 , wherein the first recombinant nucleic acid sequence and the second recombinant nucleic acid sequence further comprise a nucleic acid sequence encoding an immunoglobulin (Ig) signal peptide. 
     
     
         30 . The method of  claim 29 , wherein the Ig signal peptide comprises an IgE or IgG signal peptide. 
     
     
         31 . A method of preventing or treating a disease in a subject, the method comprising generating a synthetic antibody in a subject according to the method of  claim 1 . 
     
     
         32 . The method of  claim 31 , wherein the synthetic antibody is specific for a self-antigen. 
     
     
         33 . The method of  claim 32 , wherein the self-antigen is PSMA. 
     
     
         34 . The method of  claim 33 , wherein the recombinant nucleic acid sequence comprises a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:80. 
     
     
         35 . The method of  claim 33 , wherein the recombinant nucleic acid sequence comprises a nucleic acid sequence at least 90% identical to SEQ ID NO:79. 
     
     
         36 . The method of  claim 2 , wherein two leucine to alanine mutations are introduced in the CH2 region of Fc region. 
     
     
         37 . The method of  claim 2 , wherein antibodies generated in the subject are defucosylated. 
     
     
         38 . A synthetic antibody produced by the method of claim 
     
     
         1 . 
     
     
         39 . The product of  claim 38 , wherein the product is single DNA plasmid capable of expressing a functional antibody. 
     
     
         40 . The product of  claim 38 , wherein the product is comprised of two or more distinct DNA plasmids capable of expressing components of a functional antibody that combine in vivo to form a functional antibody. 
     
     
         41 . A method of treating a subject from infection by a pathogen, comprising: administering a nucleotide sequence encoding a synthetic antibody specific for the pathogen, wherein the antibody comprises an amino acid sequence at least 80% identical to SEQ ID NO:80. 
     
     
         42 . The method of  claim 41 , further comprising: administering an antigen of the pathogen to generate an immune response in the subject. 
     
     
         43 . A method of treating a subject from cancer, comprising:
 administering a nucleotide sequence encoding a cancer marker to induce ADCC.   
     
     
         44 . A nucleic acid molecule encoding a synthetic antibody comprising a nucleic acid sequence having at least about 80% identity over an entire length of the nucleic acid sequence set forth in SEQ ID NO:79. 
     
     
         45 . A nucleic acid molecule encoding a synthetic antibody comprising a nucleic acid sequence as set forth in SEQ ID NO:79. 
     
     
         46 . A nucleic acid molecule encoding a synthetic antibody comprising a nucleic acid sequence encoding a protein having at least about 80% identity over an entire length of the amino acid sequence set forth in SEQ ID NO:80. 
     
     
         47 . A nucleic acid molecule encoding a synthetic antibody comprising a nucleic acid sequence encoding a protein comprising an amino acid sequence as set forth in SEQ ID NO:80. 
     
     
         48 . The nucleic acid molecule of  claim 44 , wherein the nucleic acid molecule comprises an expression vector. 
     
     
         49 . A composition comprising the nucleic acid molecule of  claim 44 . 
     
     
         50 . The composition of  claim 49 , further comprising a pharmaceutically acceptable excipient.