Pseudomonas aeruginosa PCRV binding single variable domain antibodies

1. A polypeptide that specifically binds PcrV of Pseudomonas aeruginosa , comprising a first immunoglobulin single variable domain that essentially consists of 4 framework regions and 3 complementarity determining regions (CDR), in which:

CDR1 is the amino acid sequence of SEQ ID NO: 30; and

CDR2 is the amino acid sequence of SEQ ID NO: 49; and

CDR3 is the amino acid sequence of SEQ ID NO: 68.

2. The polypeptide according to claim 1 , that is SEQ ID NO: 12.

3. The polypeptide according to claim 1 , that essentially consists of an immunoglobulin single variable domain selected from a domain antibody (dAb), a V HH , a partially or fully humanized V HH , and a camelized V H .

4. The polypeptide according to claim 1 , comprising two or more immunoglobulin single variable domains that specifically bind PcrV, wherein the first immunoglobulin single variable domain specifically binds a first epitope on PcrV and a second immunoglobulin single variable domain specifically binds a second epitope on PcrV different from the first epitope on PcrV.

5. The polypeptide according to claim 4 , wherein the two or more immunoglobulin single variable domains consist of a domain antibody (dAb), of a V HH , of a partially or fully humanized V HH , or of a camelized V H .

6. The polypeptide according to claim 4 , wherein the second immunoglobulin single variable domain consists of 4 framework regions and 3 complementarity determining regions,

in which:

CDR1 is chosen from the group consisting of the amino acid sequences of SEQ ID NOs: 20-21; and

CDR2 is chosen from the group consisting of the amino acid sequences of SEQ ID NOs: 38-39; and

CDR3 is chosen from the group consisting of the amino acid sequences of SEQ ID NOs: 57-58,

wherein the second immunoglobulin single variable domain specifically binds full length PcrV having SEQ ID NO: 159 and shows 30-90% binding or below 30% binding to chimera 7 having SEQ ID NO: 205 as compared to its binding to full length PcrV; and

wherein the second immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 1 and 2 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 1 and 2.

7. The polypeptide according to claim 4 , wherein the second immunoglobulin single variable domain consists of 4 framework regions and 3 complementarity determining regions,

in which:

CDR1 is chosen from the group consisting of the amino acid sequences of SEQ ID NOs: 22-28;

and

CDR2 is chosen from the group consisting of the amino acid sequences of SEQ ID NOs: 40-47;

and

CDR3 is chosen from the group consisting of the amino acid sequences of SEQ ID NOs: 59-66,

wherein the second immunoglobulin single variable domain specifically binds full length PcrV having SEQ ID NO: 159 and shows 30-90% binding or below 30% binding to chimera 4 having SEQ ID NO: 202 and chimera 6 having SEQ ID NO: 204 as compared to its binding to full length PcrV; and

wherein the second immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 3-10 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 3-10.

8. The polypeptide according to claim 4 , wherein the first immunoglobulin single variable domain does not cross-block the binding to PcrV of the second immunoglobulin single variable domain and/or wherein the first immunoglobulin single variable is not cross-blocked from binding to PcrV by the second immunoglobulin single variable domain and wherein one of (i) to (viii):

(i) the first immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 3-10 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 3-10; and the second immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12;

(ii) the first immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 1-2 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 1-2; and the second immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12;

(iii) the first immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12 and/or is cross-blocked from binding to PcrV by at least one of immunoglobulin single variable domains with SEQ ID NOs: 11-12; and the second immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 3-10 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 3-10;

(iv) the first immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 11-12; and the second immunoglobulin single variable domain cross-blocks the binding to PcrV of at least one of the immunoglobulin single variable domains with SEQ ID NOs: 1-2 and/or is cross-blocked from binding to PcrV by at least one of the immunoglobulin single variable domains with SEQ ID NOs: 1-2;

(v) the first immunoglobulin single variable domain specifically binds full length PcrV having SEQ ID NO: 159 and shows 30-90% binding or below 30% binding to chimera 4 having SEQ ID NO: 202 and chimera 6 having SEQ ID NO: 204 as compared to its binding to full length PcrV having SEQ ID NO: 159; and the second immunoglobulin single variable domain is selected from the polypeptide of claim 1 ;

(vi) the first immunoglobulin single variable domain specifically binds full length PcrV having SEQ ID NO: 159 and shows 30-90% binding or below 30% binding to chimera 7 having SEQ ID NO: 205 as compared to its binding to full length PcrV having SEQ ID NO: 159; and the second immunoglobulin single variable domain is selected from the polypeptide of claim 1 ;

(vii) the first immunoglobulin single variable domain is selected from the polypeptide of claim 1 ; and the second immunoglobulin single variable domain specifically binds full length PcrV having SEQ ID NO: 159 and shows 30-90% binding or below 30% binding to chimera 4 having SEQ ID NO: 202 and chimera 6 having SEQ ID NO: 204 as compared to its binding to full length PcrV having SEQ ID NO: 159;

(viii) the first immunoglobulin single variable domain is selected from the polypeptide of claim 1 ; and the second immunoglobulin single variable domain specifically binds full length PcrV having SEQ ID NO: 159 and shows 30-90% binding or below 30% binding to chimera 7 having SEQ ID NO: 205 as compared to its binding to full length PcrV having SEQ ID NO: 159.

9. The polypeptide according to claim 4 , which is selected from any of SEQ ID NOs: 124, 128, 129, 131, 132, 133, 134, and 138.

10. The polypeptide according to claim 4 , further comprising one or more other groups, residues, moieties or binding units, optionally linked via one or more peptidic linkers.

11. The polypeptide according to claim 10 in which said one or more other groups, residues, moieties or binding units are chosen from the group consisting of domain antibodies (dAbs), V HH S, partially or fully humanized V HHS , and camelized V HS .

12. The polypeptide according to claim 10 , in which said one or more other groups, residues, moieties or binding units provide the polypeptide with increased half-life, compared to the corresponding polypeptide without said one or more other groups, residues, moieties, or binding units.

13. The polypeptide according to claim 12 , in which said one or more other groups, residues, moieties or binding units that provide the polypeptide with increased half-life is chosen from the group consisting of serum proteins, fragments of serum proteins, binding units that can bind to serum proteins, an Fc portion, and small proteins that can bind to serum proteins.

14. The polypeptide according to claim 12 , in which said one or more other binding units that provides the polypeptide with increased half-life are chosen from the group consisting of binding units that can bind to serum albumin, human serum albumin, immunoglobulin, and IgG.

15. The polypeptide according to claim 12 , in which said one or more other binding units that provides the polypeptide with increased half-life are chosen from the group consisting of binding units that can bind to serum albumin, human serum albumin, serum immunoglobulin, and IgG.

16. A composition comprising at least one polypeptide according to claim 1 .

17. The composition according to claim 16 , which is a pharmaceutical composition, that further comprises at least one pharmaceutically acceptable carrier, diluent, excipient and/or adjuvant, and that optionally comprises one or more further pharmaceutically active compounds.

18. The polypeptide according to claim 1 , further comprising one or more other groups, residues, moieties or binding units, optionally linked via one or more peptidic linkers.

19. The polypeptide according to claim 18 in which said one or more other groups, residues, moieties or binding units are chosen from the group consisting of domain antibodies (dAbs), V HH S, partially or fully humanized V HHS , and camelized V HS .

20. The polypeptide according to claim 19 , in which said one or more other groups, residues, moieties or binding units provide the polypeptide with increased half-life, compared to the corresponding polypeptide without said one or more other groups, residues, moieties or binding units.

21. The polypeptide according to claim 20 , in which said one or more other groups, residues, moieties or binding units that provide the polypeptide with increased half-life is chosen from the group consisting of serum proteins, fragments of serum proteins, binding units that can bind to serum proteins, an Fc portion, and small proteins that can bind to serum proteins.

22. A composition comprising at least one polypeptide according to claim 4 .

23. The composition according to claim 22 , which is a pharmaceutical composition, that further comprises at least one pharmaceutically acceptable carrier, diluent, excipient and/or adjuvant, and that optionally comprises one or more further pharmaceutically active compounds.

24. A pharmaceutical device suitable for the pulmonary delivery of at least one polypeptide, comprising at least one polypeptide according to claim 1 , selected from an inhaler for liquids, a nebulizer, metered dose inhaler, aerosol and a dry powder inhaler.

25. A method for preparing a polypeptide, said method comprising linking the polypeptide according to claim 1 that specifically binds a first epitope on PcrV to a second immunoglobulin single variable domain that specifically binds a second epitope on PcrV different from the first epitope on PcrV, and optionally one or more linkers.