Fusion proteins comprising an anti-CD40 antibody and HIV antigenic peptides
The present invention includes compositions and methods for the expression, secretion and use of novel compositions for use as, e.g., vaccines and antigen delivery vectors, to delivery antigens to antigen presenting cells. In one embodiment, the vector is an anti-CD40 antibody, or fragments thereof, and one or more antigenic peptides linked to the anti-CD40 antibody or fragments thereof, including humanized antibodies.
1. An antibody-antigen fusion protein comprising an anti-CD40 antibody comprising:
a heavy chain variable domain (VH) which comprises in sequence hypervariable regions CDR1H, CDR2H and CDR3H, the CDR1H having the amino acid sequence GFTFSDYYMY (SEQ ID NO.:45), the CDR2H having the amino acid sequence YINSGGGSTYYPDTVKG (SEQ ID NO.:46), and the CDR3H having the amino acid sequence RGLPFHAMDY (SEQ ID NO.:47); and
a light chain variable domain (VL) which comprises in sequence hypervariable regions CDR1L, CDR2L and CDR3L, the CDR1L having the amino acid sequence SASQGISNYLN (SEQ ID NO.:41), the CDR2L having the amino acid sequence YTSILHS (SEQ ID NO.:42), and the CDR3L having the amino acid sequence QQFNKLPPT (SEQ ID NO.:43); and
one or more HIV antigenic peptides, wherein at least one of the viral antigenic peptide(s) comprises an antigenic peptide of the pol HIV antigen.
2. The fusion protein of claim 1 , wherein the one or more HIV antigenic peptides comprise one or more antigenic peptides of gag, nef, and env HIV antigens.
3. The fusion protein of claim 2 , wherein the one or more antigenic peptides comprise a HIV polypeptide with an amino acid sequence corresponding to one or more of SEQ ID NOS:1-5.
4. The fusion protein of claim 1 , wherein the anti-CD40 antibody is a humanized antibody.
5. A vaccine comprising the fusion protein of claim 1 .
6. A nucleic acid expression vector encoding the fusion protein of claim 1 .
7. A method of generating antigen-loaded, activated dendritic cells comprising: obtaining previously isolated patient dendritic cells and exposing the dendritic cells to activating amounts of the fusion protein of claim 1 .
8. A method of producing a vaccine of claim 5 , the method comprising introducing a flexible linker between an anti-CD40 antibody and the one or more HIV antigenic peptides wherein the anti-CD40 antibody comprises:
a heavy chain variable domain (VH) which comprises in sequence hypervariable regions CDR1H, CDR2H and CDR3H, the CDR1H having the amino acid sequence GFTFSDYYMY (SEQ ID NO.:45), the CDR2H having the amino acid sequence YINSGGGSTYYPDTVKG (SEQ ID NO.:46), and the CDR3H having the amino acid sequence RGLPFHAMDY (SEQ ID NO.:47); and
a light chain variable domain (VL) which comprises in sequence hypervariable regions CDR1L, CDR2L and CDR3L, the CDR1L having the amino acid sequence SASQGISNYLN (SEQ ID NO.:41), the CDR2L having the amino acid sequence YTSILHS (SEQ ID NO.:42), and the CDR3L having the amino acid sequence QQFNKLPPT (SEQ ID NO.:43).
9. A method of making an anti-CD40 antibody-antigen fusion protein, the method comprising expressing the fusion protein of claim 1 in a host cell and isolating the protein from the host cell.
10. The fusion protein of claim 1 , wherein the fusion protein further comprises one or more flexible linkers between the anti-CD40 antibody and the one or more antigenic peptides.
11. The fusion protein of claim 10 , wherein the one or more flexible linkers comprises a polypeptide with an amino acid sequence corresponding to one or more of: SEQ ID NO:11-14 and 145.