Mono and bispecific antibody binding to hERG1 and hERG1/integrin beta 1
The present invention describes a bispecfic antibody composed of the variable domains (VH and VL) of two antibodies, hERG1 mAb (which binds the extracellular domain S5-P of hERG1) and of β1 integrin mAb TS2/16 or BV7, which bind the extracellular domain of β1 integrin. The present invention relates also to a novel anti-hERG1 molecule bearing a Cys in position 95 of the VH domain. The invention describes also their application for diagnostic and therapeutic purposes in oncology and other fields of medical sciences.
1. A bispecific anti-hERG1 and anti-β 1 integrin antibody (Ab) comprising the Heavy chain Variable (VH) domain and Light chain Variable (VL) domain of an anti-hERG1 Ab which binds the extracellular domain S5-P of hERG 1 and the Heavy chain Variable (VH) domain and Light chain Variable (VL) domain of an anti-β1 integrin Ab which bind the extracellular domain of β1 integrin, wherein the Heavy chain Variable (VH) domain of anti-hERG1 Ab comprises SEQ ID No: 8 wherein the residue at position 95 of SEQ ID No: 8 is Cys; and the Light chain Variable (VL) domain of anti-hERG1 Ab comprises SEQ ID No: 4; and the Heavy chain Variable (VH) domain of anti-β1 integrin Ab comprises SEQ ID No: 26 or SEQ ID No: 46 and a Light chain Variable (VL) domain of anti-β1 integrin Ab comprises SEQ ID No: 24 or SEQ ID No: 48.
2. The bispecific Ab according to claim 1 has a format selected in the group consisting of Tandem scFvs, Diabody format, Single-chain diabodies, Tandem diabodies (TandAbs) and Dual-affinity retargeting molecules (DARTs).
3. The bispecific Ab according to claim 2 comprising a first Heavy chain Variable (VH) domain comprising with SEQ ID NO: 8 wherein the residue at position 95 of SEQ ID NO: 8 is Cys, and a first VL domain comprising SEQ ID NO:4, and a second VH domain comprising SEQ ID NO:26, and a second VL domain comprising SEQ ID NO:24.
4. The bispecific Ab according to claim 3 wherein domains are assembled in the following order: anti-hERG1-Cys VH domain linked by a first linker to anti-β1-integrin VL domain linked by a second linker to anti β1-integrin VH domain linked by a third linker to anti-hERG1-Cys VL domain.
5. The bispecific Ab according to claim 4 wherein the linkers comprise (Gly4Ser)3 motifs.
6. A pharmaceutical composition comprising a bispecific Ab according to claim 1 and a pharmaceutically acceptable ingredient.
7. A polynucleotide encoding the bispecific Ab according to claim 1 .
8. An expression vector or a plasmid comprising the polynucleotide according to claim 7 .
9. A genetically modified microorganism or a cell comprising the expression vector or plasmid according to claim 8 .
10. A kit comprising a bispecific antibody according to claim 1 .
11. The kit of claim 10 , said kit further comprising an anti-hERG1-Cys scFv comprising a VH domain comprising SEQ ID NO: 8 wherein the residue at position 95 of SEQ ID NO: 8 is Cys, and a VL domain comprising SEQ ID NO: 4, said antibody specifically binds hERG1 S5-pore extracellular portion.
12. An anti-hERG1 antibody comprising a Heavy chain Variable (VH) domain comprising SEQ ID NO:8 wherein the residue at position 95 of SEQ ID No: 8 is Cys, and a Light chain Variable (VL) domain comprising SEQ ID NO:4, said antibody specifically binds hERGI S 5 -pore extracellular portion.
13. The anti-hERG1 antibody according to claim 12 which is a fully humanized recombinant Ab, a scFv, Fab, Fv form of simple chain of scFv, diabodies, triabodies, bispecifics, minibodies or phage antibodies.
14. The anti-hERG1 antibody according to claim 13 , wherein said molecule is a scFv and wherein VH and VL are linked by a peptide linker.
15. The anti-hERG1 antibody according to claim 14 , wherein the peptide linker comprises a (Gly4Ser)3 motif.
16. The anti-hERG1 antibody according to claim 12 having SEQ ID No: 10.
17. The anti-hERG1 antibody according to claim 12 which is labelled with a fluorophore or a radionuclide.
18. A pharmaceutical composition comprising the anti-hERG1 antibody according to claim 12 and a pharmaceutically acceptable ingredient.
19. A polynucleotide encoding the anti-hERG1 antibody according to claim 12 .
20. An expression vector or a plasmid comprising the polynucleotide according to claim 19 .
21. A genetically modified microorganism or a cell comprising the expression vector or plasmid according to claim 20 .
22. A kit comprising an anti-hERG1-Cys scFv comprising a VH domain comprising SEQ ID NO: 8 wherein the residue at position 95 of SEQ ID NO: 8 is Cys, and a VL domain comprising SEQ ID NO: 4, said antibody specifically binds hERG1 S5-pore extracellular portion.