Modified antibody
Recombinant antibody-based molecules that trigger both T-cell and B-cell immune responses are disclosed. The recombinant molecules are comprised by at least one targeting unit and at least one antigenic unit connected through a dimerization motif. Also disclosed are nucleic acid molecules encoding the recombinant antibody-based molecule and methods of treating multiple myeloma or lymphoma in a patient using the recombinant antibody-based molecules or the nucleic acid molecules.
1. An isolated nucleic acid encoding a monomer unit of a recombinant antibody-based dimeric molecule, said nucleic acid encoding an antigenic unit, a dimerization motif and a targeting unit operably connected to encode said monomer unit, and wherein said antibody-based dimeric molecule comprises two of said monomer units connected through said dimerization motif, said dimerization motif comprising an Ig hinge region and a Cγ3 domain of each monomer unit, wherein each Ig hinge region contributes to dimerization via disulfide bridging to the other Ig hinge region and each Cγ3 domain contributes to dimerization via hydrophobic interactions to the other Cγ3 domain, and wherein each of said monomer unit comprises a targeting unit for an antigen presenting cell and an antigenic unit, wherein said targeting unit and said antigenic unit in the monomer unit are separated by said dimerization motif and wherein said monomer units each lack a CH2 domain.
2. The nucleic acid of claim 1 , wherein at least one of said targeting units is a single chain fragment variable of Ig (scFv).
3. The nucleic acid of claim 2 , wherein said scFv is anti-HLA, anti-CD14, anti-CD40, or anti-toll-like receptor.
4. The nucleic acid of claim 3 , wherein said anti-HLA is anti-HLA-DP.
5. The nucleic acid of claim 3 , wherein said anti-toll-like receptor is anti-toll-like receptor 2.
6. The isolated nucleic acid of claim 1 , wherein at least one of said targeting unit is a ligand.
7. The isolated nucleic acid of claim 6 , wherein said ligand is soluble CD40 ligand or a chemokine.
8. The isolated nucleic acid of claim 6 , wherein said ligand is a chemokine.
9. The isolated nucleic acid of claim 8 , wherein said chemokine is RANTES or Macrophage Inflammatory Protein 1 alpha.
10. The isolated nucleic acid of claim 8 , wherein said chemokine is MIP-1α.
11. The nucleic acid of claim 1 , wherein at least one of said targeting unit is a bacterial antigen.
12. The nucleic acid of claim 11 , wherein said bacterial antigen is a flagellin.
13. The nucleic acid of claim 1 , wherein said targeting units have the ability to target CD14, CD40, a toll-like receptor, or HLA or HLA-DP.
14. The isolated nucleic acid of claim 1 , wherein said targeting unit have the ability to target a chemokine receptor.
15. The isolated nucleic acid of claim 1 , wherein at least one of said antigenic unit is an antigenic scFv.
16. The isolated nucleic acid of claim 15 , wherein said antigenic scFv has VL and VH chains from a monoclonal Ig produced by myeloma or lymphoma.
17. The nucleic acid of claim 1 , wherein at least one of said antigenic unit is a telomerase or a functional part thereof.
18. The nucleic acid of claim 17 , wherein said telomerase is hTERT.
19. The nucleic acid of claim 1 , wherein at least one of said antigenic units is derived from an infectious agent.
20. The nucleic acid of any one of claim 1 or 19 , wherein at least one of said antigenic unit is derived from a bacterium.
21. The nucleic acid of claim 20 , wherein said bacterium-derived antigenic unit is/are a tuberculosis antigen.
22. The nucleic acid of any one of claim 1 or 19 , wherein at least one of said antigenic unit is derived from a virus.
23. The nucleic acid of claim 22 , wherein said virus-derived antigenic unit is derived from HIV.
24. The nucleic acid of claim 23 , wherein said HIV-derived antigenic unit is derived from gp120.
25. An isolated vector comprising the nucleic acid according to claim 1 .
26. A composition comprising a nucleic acid according to claim 1 or a vector comprising the nucleic acid according to claim 1 , in combination with a physiologically acceptable diluent or carrier.
27. A kit for preparation of a recombinant antibody-based molecule encoded by the nucleic acid according to claim 1 , the kit comprising a nucleic acid according to claim 1 .
28. A method of preparing a recombinant antibody-based molecule comprising:
a. transfecting a vector comprising a nucleic acid encoding a monomer unit of a recombinant antibody-based dimeric molecule into a cell population, wherein the nucleic acid encodes an antigenic unit, a dimerization motif and a targeting unit operably connected to encode said monomer unit, and wherein said antibody-based dimeric molecule comprises two of said monomer units connected through said dimerization motif, said dimerization motif comprising an Iq hinge region and a Cγ3 domain of each monomer unit, wherein each Ig hinge region contributes to dimerization via disulfide bridging to the other Ig hinge region and each Cγ3 domain contributes to dimerization via hydrophobic interactions to the other Cγ3 domain, and wherein each of said monomer unit comprises a targeting unit for an antigen presenting cell and an antigenic unit, wherein said targeting unit and said antigenic unit in the monomer unit are separated by said dimerization motif and wherein said monomer units each lack a CH2 domain;
b. culturing the cell population, wherein the monomers are expressed and homodimerized to generate the recombinant antibody-based molecule;
c. collecting the recombinant antibody-based molecule generated from the cell population; and
d. purifying the expressed antibody-based molecule.