Biological products
A multivalent antibody fusion protein which comprises an immunoglobulin moiety, for example a Fab or Fab′ fragment, with a first specificity for an antigen of interest, and further comprises two single domain antibodies (dAb) with specificity for a second antigen of interest, wherein the two single domain antibodies are linked by a disulfide bond. There is also provided particular dual specificity antibody fusion proteins comprising a Fab or Fab′ fragment and one or more single domain antibodies which may be stabilized by a disulfide bond therebetween.
1. A divalent antibody fusion protein, comprising:
(i) an immunoglobulin moiety with a specificity for an antigen of interest,
wherein the immunoglobulin moiety is a Fab or Fab′ fragment having a heavy chain and a light chain; and
(ii) two single domain antibodies (dAb), a VH dAb and a VL dAb, that together bind to the same human serum albumin,
wherein the VH dAB comprises complementary determining region (CDR)—H1 having the amino acid sequence set forth in SEQ ID NO:56, CDR-H2 having the amino acid sequence set forth in SEQ ID NO:57, and CDR-H3 having the amino acid sequence set forth in SEQ ID NO:58;
wherein the VL dAB comprises CDR-L1 having the amino acid sequence set forth in SEQ ID NO:59, CDR-L2 having the amino acid sequence set forth in SEQ ID NO:60, and CDR-L3 having the amino acid sequence set forth in SEQ ID NO:61;
wherein (i) the VH dAb is connected directly or via a linker to the C-terminus of the Fab or Fab′ heavy chain and the VL dAb is connected directly or via a linker to the C-terminus of the Fab or Fab′ light chain, or (ii) the VH dAb is connected directly or via a linker to the C-terminus of the Fab or Fab′ light chain and the VL dAb is connected directly or via a linker to the Fab or Fab′ heavy chain; and
wherein the VH dAb and the VL dAb are linked by a disulfide bond between two engineered cysteine residues at positions VH44 and VL100.
2. The divalent antibody fusion protein of claim 1 , wherein the VH dAb and the VL dAb are humanised.
3. The divalent antibody fusion protein of claim 1 , wherein the Fab or Fab′ is fully human or humanised.
4. The divalent antibody fusion protein of claim 1 , wherein the VH dAb is connected to the C-terminus of the Fab or Fab′ heavy chain directly or via a linker and the VL dAb is connected to the C-terminus of the Fab or Fab′ light chain directly or via a linker.
5. The divalent antibody fusion protein of claim 1 , wherein the VH dAb is connected to the C-terminus of the Fab or Fab′ heavy chain and the VL dAb is connected to the C-terminus of the Fab or Fab′ light chain via a linker having the amino acid sequence set forth in any one of SEQ ID NOs:13 or 45.
6. The divalent antibody fusion protein of claim 1 , wherein the VH dAb is fused to the C-terminus of the heavy chain constant region (CH1) of the Fab or Fab′ and the VL dAb is fused to the C-terminus of the light chain constant region of the Fab or Fab′ or wherein the VL dAb antibody is fused to the C-terminus of the heavy chain constant region (CH1) of the Fab or Fab′ and the VH dAb antibody is fused to the C-terminus of the light chain constant region of the Fab or Fab′.
7. A pharmaceutical composition comprising the divalent antibody fusion protein of claim 1 .
8. The divalent antibody fusion protein of claim 1 , wherein the divalent antibody fusion protein contains one CL and one CH1 domain, wherein the CL and the CH1 domains are in the Fab or Fab′ fragment.