TW202204399A - Isolated anti-pcrv antibody, method of producing the same, nucleic acid molecule that encodes the same, host cell and pharmaceutical composition comprising the same and use thereof - Google Patents

Abstract

The present application provides antibodies including antigen-binding fragment thereof that specifically recognizing Pseudomonas PcrV. Also provided are methods of making and using these antibodies.

Description

Isolated anti-PcrV antibody, method for its preparation, nucleic acid molecule encoding it, host cell and pharmaceutical composition comprising the same, and use thereof

[Sequence Listing]

The contents of the Sequence Listing in computer readable form (CRF) as filed herein are incorporated herein by reference in their entirety.

The present invention relates to an antibody that specifically recognizes Pseudomonas aeruginosa ( Pseudomonas aeruginosa , PA) PcrV, its preparation method and use, including the method for treating and preventing Pseudomonas infection with the antibody.

Pseudomonas aeruginosa is an obligate aerobic gram-negative bacillus that exists widely in nature. Although its pathogenicity is generally low, it is a pathogen that causes opportunistic infections that often occur in patients with various underlying medical conditions such as cancer, diabetes, and immunodeficiency, and in patients taking immunosuppressive drugs middle. Pseudomonas aeruginosa infection is prone to occur in patients with mucocutaneous rupture, and patients with chronic structural lung disease (eg, chronic obstructive pulmonary disease or cystic fibrosis) are also at considerable risk for infection. Pseudomonas aeruginosa can cause pneumonia, urinary tract infections, sepsis and other conditions, often with serious consequences. Up to 10% of nosocomial infections are caused by Pseudomonas aeruginosa, and mortality in patients with Pseudomonas aeruginosa bacteremia approaches 40%. Clinically, Pseudomonas aeruginosa infection is considered one of the most difficult infections to treat, not only because of its low sensitivity to existing antibiotics, but also because it easily develops resistance to multiple antibiotics. Therefore, strategies to develop antibiotics have limited advantages in combating P. aeruginosa infections.

Pseudomonas aeruginosa is one of the leading causes of hospital-acquired infections, especially in mechanically ventilated patients, and it is the leading cause of death in cystic fibrosis patients. The Pseudomonas aeruginosa III secretion system (T3SS) is a major virulence factor associated with severe disease that injects bacterial toxins directly into the cytoplasm of host cells. Pseudomonas aeruginosa exerts its high cytotoxicity by injecting toxin into eukaryotic cells through the type III exotoxin secretion system (T3SS). PcrV is a protein that constitutes the type III exotoxin secretion system, consisting of 294 residues (NCBI accession number: AAC45935, SEQ ID NO: 80), and the operon sequence encoding the protein is publicly available (US 6,551,795, Yahr, TL et al., J. Bacteriol. , 1997, vol. 179, p. 7165). The PcrV protein is located at the apex of the T3SS injectable complex, is required for T3SS function, and is a well-validated target in animal models of immunoprophylaxis against Pseudomonas aeruginosa. The T3SS of Pseudomonas aeruginosa is a well-validated target for intervention in infections caused by this opportunistic pathogen. In animal models, both active vaccination with T3SS component proteins and passive immunotherapy targeting PcrV can strongly alleviate disease caused by Pseudomonas aeruginosa. Therefore, control of PcrV may become a therapeutic approach to control Pseudomonas aeruginosa infection (T. Sawa et al., Nature Medicine , 1999, vol. 5, p. 392), and multiple PcrV with neutralizing activity have been reported. Strain antibodies (Shime N et al., J. Immunol. 2001, vol. 167, p. 5880, Imamura Y et al., Eur. Respir. J. , 2007, Vol. 29, p. 965) and monoclonal antibodies (WO2002064161A2, Karine Faure et al., J. Immune. Based. Therapies and Vaccines , 2003, Vol. 1, Dara W. Frank et al., J. Infect. Disease , 2002, Vol. 186, p. 64). However, polyclonal antibodies are difficult to humanize and use as pharmaceutical compositions due to the difficulty in improving their antigenicity. Warrener et al. described an antibody against PcrV, named V2L2-MD (Warrener et al., 2014, Antimicrob. Agents Chemother ., 58, 4384–4391). An anti-PcrV monoclonal antibody PEGylated Fab fragment based on the PcrV-specific murine monoclonal antibody MAB166 was ineffective in preventing respiratory tract infections caused by Pseudomonas aeruginosa in mechanically ventilated patients. Although MAB166 effectively blocks P. aeruginosa T3SS in vitro, it requires considerably higher amounts of antibody to be protective in animal models. The present invention provides a novel anti-PcrV monoclonal antibody, which exhibits effective inhibitory effects both in vitro and in vivo.

The disclosures in all publications, patents, patent applications, and published patent applications mentioned herein are incorporated by reference in their entirety.

The present invention provides an isolated anti-PcrV antibody that can specifically bind to the PcrV epitope of Pseudomonas, and a method for treating Pseudomonas infection.

In one aspect, the invention provides an isolated anti-PcrV antibody, wherein the anti-PcrV antibody comprises a heavy chain variable domain ( _{VH )} comprising: a heavy chain complementarity determining region (HC-CDR) 1 comprising SYWMH (SEQ ID NO: 1), or a variant thereof comprising up to 3 amino acid substitutions; an HC-CDR2 comprising RINEX ₁ EX ₂ SX ₃ SYADSVKG (SEQ ID NO: 50), or a variant comprising up to 3 amino acid substitutions , wherein X ₁ is D, T, N, L, I, S, V, A or H, X ₂ is S, T, N, D, G or R, X ₃ is I or T; and a HC-CDR3 comprising DGX ₁ X ₂ X ₃ X ₄ X ₅ DX ₆ (SEQ ID NO: 51), or a variant thereof comprising up to ₃ amino acid substitutions, wherein X is P, Y or _A and X is Y or W, _X3 is D, T or N, _X4 is S, T or _A , _X5 is L, F or M, X6 is I, V, A, S, L or W; and the light chain can be A variable domain ( _{VL )} comprising: a light chain complementarity determining region (LC-CDR) 1 comprising RASQX ₁ VX ₂ X ₃ NLA (SEQ ID NO: 47), or comprising up to 3 amino acid substitutions Variants of , wherein X ₁ is S, N, D or G, X ₂ is S, K, N, R or T, and X ₃ is N, S, D or G; an LC-CDR 2 containing X ₁ ASSRAT ( SEQ ID NO: 85), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is D, N, H, A or S; and an LC-CDR3 comprising QQYGX ₁ X ₂ PX ₃ T (SEQ ID NO: 85) NO: 86), or a variant thereof comprising at most 3 amino acid substitutions, wherein X ₁ is S, A, T, E, D, H, N, Q, G, L, Y, M, R or V, X ₂ is S, Q, V, E, T, D, M, Y, G, H, L, N, A, F or P, X ₃ is I, L or V.

In some embodiments, there is provided an isolated anti-PcrV antibody comprising a heavy chain variable domain ( _{VH )} comprising: an HC-CDR1 comprising SYWMH (SEQ ID NO. : 1), or a variant comprising at most 3 amino acid substitutions; a HC-CDR2 comprising RINEX1EX2SISYADSVKG (SEQ ID NO: 45), or a variant comprising at most 3 amino acid substitutions, wherein X1 is D, N, I, L or V, X2 is S, T, R, G or N; and one HC-CDR3, including DGPYDX1X2DI (SEQ ID NO: 46), or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is S , A or T, X2 is F or _L ; and _VL comprising: an LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO: 47), or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is N, G, D, or S, X2 is K, R, S, N, or T, and X3 is N, G, S, or D; an LC-CDR2, comprising X1ASSRAT (SEQ ID NO: 48), or at most 3 amino acid substitution variants, wherein X1 is D, N, H or A; and an LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO: 49), or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is S, T, E, H, N, A, D, M or L, X2 is S, Q, E, T, D, G, H, L, N, V or Y, and X3 is I, L or V .

In some embodiments, an isolated anti-PcrV antibody is provided, wherein the anti-PcrV antibody binds to Pseudomonas PcrV with a Kd value of about 0.1 pM to 1 nM. In some embodiments, an isolated anti-PcrV antibody is provided, comprising a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of any one of SEQ ID NOs: 52-64 and _VL comprising LC-CDR1, LC-CDR2 and LC-CDR3 in a _VL having the amino acid sequence of any one of SEQ ID NOs: 65-79.

In some embodiments, any one of the isolated anti-PcrV antibodies as described above, the isolated anti-PcrV antibody comprises: _VH , the _VH comprises: a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1 or its A variant comprising up to 3 amino acid substitutions; a HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 2-9 or a variant comprising up to 3 amino acid substitutions; and a HC-CDR3 comprising SEQ ID NOs : any amino acid sequence in 10-13 or a variant thereof comprising at most 3 amino acid substitutions; and a _VL comprising _: an LC-CDR1 comprising any of the amino acid sequences in SEQ ID NOs: 14-25 or It comprises a variant of at most 3 amino acid substitutions; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 26-29 or a variant thereof comprising at most 3 amino acid substitutions; an LC-CDR3 comprising SEQ ID NOs : any amino acid sequence from 30 to 44 or a variant thereof comprising up to 3 amino acid substitutions.

In some embodiments, any one of the isolated anti-PcrV antibodies described above, the isolated anti-PcrV antibodies comprising: (i _{) a VH comprising} : an HC- comprising the amino acid sequence of SEQ ID NO: 1 CDR1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 30 CDR3, or a _VL variant comprising up to 5 amino acid substitutions in LC-CDRs; (ii _{) a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence HC-CDR2 of SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL , the V _L comprises: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30, or an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30. _VL variants comprising up to 5 amino acid substitutions in the CDRs; (iii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence SEQ ID NO: 3 HC-CDR2, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL comprising _: one comprising amino acids LC-CDR1 of sequence SEQ ID NO: 15, one LC-CDR2 comprising amino acid sequence SEQ ID NO: 27, and one LC-CDR3 comprising amino acid sequence SEQ ID NO: 31, or at most 5 in LC-CDRs Amino acid substituted _VL variants; (iv _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; or at most 5 amino acids in the HC-CDRs A base-acid-substituted _VH variant; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 An LC-CDR3 of amino acid sequence SEQ ID NO: 39, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (v _{) a VH comprising} : one comprising the amino acid sequence of SEQ ID NO: 1 HC-CDR1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH comprising up to 5 amino acid substitutions in the HC-CDRs Variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and one comprising the amino acid sequence of SEQ ID NO: 28 The LC-CDR3 of 32, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (vi _{) a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising at most 5 amino acid substitutions in the HC-CDRs; and a _VL , the _VL comprises: an LC-CDR1 comprising amino acid sequence SEQ ID NO: 17, an LC-CDR2 comprising amino acid sequence SEQ ID NO: 27, and an LC-CDR3 comprising amino acid sequence SEQ ID NO: 33, Or _VL variants comprising up to 5 amino acid substitutions in the LC-CDRs; (vii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence SEQ ID An HC-CDR2 of NO: 2, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _{VL comprising} : an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 18, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 34, or in LC-CDRs _VL variants comprising up to 5 amino acid substitutions; (viii _{) VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; or in HC-CDRs _VH variants of up to 5 amino acid substitutions; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (ix _{) a VH comprising} : an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 1 HC-CDR1, one HC-CDR2 comprising amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising amino acid sequence SEQ ID NO: 11; or HC-CDRs comprising at most 5 amino acid substitutions A _VH variant; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 19, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 The LC-CDR3 of NO: 35, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (x _{) a VH comprising} : an HC- CDR1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 36 CDR3, or _VL variants comprising up to 5 amino acid substitutions in LC-CDRs; (xi _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence HC-CDR2 of SEQ ID NO: 4, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL , the V _L comprises: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a An LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xii) V _H , the _VH comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11 or a _VH variant comprising at most 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 22, and a LC-CDR1 comprising the amino acid sequence of SEQ ID NO. LC-CDR2 of: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xiii) _VH , the _VH Comprising: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11; or in HC-CDRs and a _VL comprising _: one LC-CDR1 comprising the amino _acid sequence SEQ ID NO: 23, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27 , and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xiv _{) a VH comprising} : an LC-CDR3 comprising the amino acid sequence HC-CDR1 of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or at most 5 amino acids in the HC-CDRs A substituted _VH variant; and a _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 21, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence The LC-CDR3 of SEQ ID NO:40, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (xv _{) a VH comprising} : a VL comprising the amino acid sequence of SEQ ID NO:1 HC-CDR1, an HC-CDR comprising the amino acid sequence of SEQ ID NO: 6 2, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising at most 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; LC-CDR1 of ID NO: 24, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 41, or at most 5 amino acid substitutions in the LC-CDRs (xvi _{) VH} comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one comprising the amino acid sequence SEQ ID NO: 9 HC-CDR3 of sequence SEQ ID NO: 11; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: an LC comprising the amino acid sequence of SEQ ID NO:25 - CDR1, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xvii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one comprising the amino acid sequence SEQ ID NO: 11 or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one comprising amino acid an LC-CDR2 of sequence SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xviii) _VH , The _VH comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17 and one comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR2, and one comprising the amino acid sequence of SEQ ID NO: 44 LC-CDR3, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xix _{) a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL , The _VL comprises: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39, or _VL variants comprising up to 5 amino acid substitutions in the LC-CDRs; or (xx _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence SEQ ID HC-CDR2 of NO: 9, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _{VL comprising} : an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or in LC-CDRs _VL variants containing up to 5 amino acid substitutions.

In some embodiments, any one of the isolated anti-PcrV antibodies as described above, the isolated anti-PcrV antibody comprises: a _VH comprising the amino _acid sequence of any one of SEQ ID NOs: 52-64, or comprising a A variant sequence having at least 90% sequence homology to the amino acid sequence of any one of SEQ ID NOs: _52-64 ; and a _VL comprising the amino acid sequence of any one of SEQ ID NOs: 65-79, or comprising A variant sequence having at least 90% sequence homology to any of the amino acid sequences of SEQ ID NOs: 65-79. In some embodiments, the isolated PcrV antibody comprises: (i) a _VH comprising the amino acid sequence of SEQ ID NO:52 and a _VL comprising the amino acid sequence of SEQ ID NO:65; (ii) a VH comprising the amino acid sequence of SEQ ID NO:65; : a _VH of 56 and a _VL comprising the amino acid sequence of SEQ ID NO:65; (iii) a _VH comprising the amino acid sequence of SEQ ID NO:53 and a VL of the amino acid sequence of SEQ ID NO:66; (iv) a _VL comprising the amino acid sequence of SEQ ID NO:66 The _VH of the sequence SEQ ID NO:62 and the _VL comprising the amino acid sequence SEQ ID NO:74; (v) the _VH comprising the amino acid sequence SEQ ID NO:54 and the _VL comprising the amino acid sequence SEQ ID NO:67; ( vi) a _VH comprising the amino acid sequence of SEQ ID NO:55 and a _VL comprising the amino acid sequence of SEQ ID NO:68; (vii) a VH comprising the amino acid sequence of SEQ ID NO:56 and a _VH comprising the amino acid sequence of SEQ ID NO:69 _VL ; (viii) a _VH comprising the amino acid sequence of SEQ ID NO:64 and a _VL comprising the amino acid sequence of SEQ ID NO:78; (ix) a _VH comprising the amino acid sequence of SEQ ID NO:56 and a VL comprising the amino acid sequence of SEQ ID NO:56 The _VL of NO: 70; (x) the _VH comprising the amino acid sequence SEQ ID NO:57 and the _VL comprising the amino acid sequence SEQ ID NO:71; (xi) the _VH comprising the amino acid sequence SEQ ID NO:54 and the VL comprising the amino acid sequence SEQ ID NO:54 VL of amino acid sequence SEQ ID NO: 72; (xii) V _H of amino acid sequence of SEQ ID NO: 56 and _VL of amino _acid sequence of SEQ ID NO: 73; (xiii) V of amino acid sequence of SEQ ID NO: 58 V _H and VL comprising the amino acid sequence of SEQ ID NO: 74; (xiv) V _H comprising the amino acid sequence of SEQ ID NO: 59 and _VL comprising the amino _acid sequence of SEQ ID NO: 75; (xv) comprising the amino acid sequence of SEQ ID The _VH of NO:57 and the _VL comprising the amino acid sequence of SEQ ID NO:76; (xvi) the _VH comprising the amino acid sequence of SEQ ID NO:60 and the _VL comprising the amino acid sequence of SEQ ID NO:77; (xvii) the _VH of amino acid sequence SEQ ID NO: 60 and _VL comprising amino acid sequence SEQ ID NO: 78; (xviii) comprising amino acid sequence SEQ ID NO: 78 The _VH of ID NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:79; (xix) the _VH comprising the amino acid sequence of SEQ ID NO:61 and the _VL comprising the amino acid sequence of SEQ ID NO:74; or (xx) ) comprising the _VH of the amino acid sequence of SEQ ID NO:63 and the _VL of the amino acid sequence of SEQ ID NO:78.

In some embodiments, there is provided, for example, an isolated anti-PcrV antibody that specifically binds PcrV that competitively binds to PcrV with any of the isolated anti-PcrV antibodies described above. In some embodiments, an isolated anti-PcrV antibody is provided that specifically binds to the same epitope as any of the isolated anti-PcrV antibodies described above.

In some embodiments, any of the isolated anti-PcrV antibodies described above, the anti-PcrV antibody comprises an Fc fragment. In some embodiments, the anti-PcrV antibody is a full-length IgG antibody. In some embodiments, the anti-PcrV antibody is a full-length IgGl or IgG4 antibody. In some embodiments, the anti-PcrV antibody is chimeric, fully human or humanized. In some embodiments, the anti-PcrV antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab', F(ab)'2, Fab'-SH, single chain antibody (scFv), Fv fragment, dAb, Fd or diabodies.

In some embodiments, an isolated nucleic acid molecule encoding any of the anti-PcrV antibodies described above is provided. In some embodiments, a vector is provided comprising any of the nucleic acid molecules described above. In some embodiments, a host cell is provided comprising any anti-PcrV antibody, any nucleic acid molecule, or any vector as described above. In some embodiments, there is provided a method of preparing an anti-PcrV antibody, comprising: a) culturing any of the host cells described above under conditions effective to express the anti-PcrV antibody; and b) obtaining an expressed anti-PcrV antibody in the host cell PcrV antibodies.

In some embodiments, there is provided a method of treating a disease or disorder in an individual in need thereof, comprising administering to the individual an effective amount of any of the anti-PcrV antibodies described above, or comprising any of the anti-PcrV antibodies described above pharmaceutical composition. In some embodiments, use of any of the anti-PcrV antibodies described above, or a pharmaceutical composition comprising any of the anti-PcrV antibodies described above, in the manufacture of a medicament for the treatment of a disease or disorder is provided. In some embodiments, the disease or disorder is a pathogenic infection. In some embodiments, the infection is a Gram-negative bacterial infection. In some embodiments, the bacterium is Pseudomonas aeruginosa. In some embodiments, the disease or disorder comprises one or more symptoms caused by Pseudomonas aeruginosa infection. In some embodiments, the symptoms include one or more of fever, chills, fatigue, muscle and joint pain, joint swelling, headache, diarrhea, rash, pus from wounds, bacteremia, acute pneumonia, or intra-abdominal infection.

In some embodiments, any of the methods of treatment described above, the method further comprises administering one or more therapeutic agents. In some embodiments, the at least one therapeutic drug is an antibiotic. In some embodiments, the antibiotic is one or more of imipenem, tobramycin, ciprofloxacin, meropenem, or atreonam.

Also provided are pharmaceutical compositions, kits and articles of manufacture comprising any of the anti-PcrV antibodies, nucleic acids, vectors, isolated host cells described above.

One aspect of the present invention provides anti-PcrV antibodies. Through a combination of scFv phage library screening, affinity maturation, and appropriately designed biochemical and biological experiments, we have identified highly potent antibody molecules capable of binding PcrV that inhibit lysis of RBC, A549 and U937 cells, and also Provides therapeutic and prophylactic protection against Pseudomonas aeruginosa in vivo. The results presented here show that our antibodies bind to different regions or epitopes of PcrV compared to the known anti-PcrV antibody V2L2-MD, and surprisingly, demonstrate in various biological assays that they are better than those already known. Known anti-PcrV antibodies are more effective.

Anti-PcrV antibodies provided by the present invention include, for example, full-length anti-PcrV antibodies, anti-PcrV single chain antibodies (scFvs), anti-PcrV Fc fusion proteins, multispecific (eg bispecific) anti-PcrV antibodies, anti-PcrV immunoconjugates things and the like.

Anti-PcrV antibodies with specific sequences are also provided, as well as antibodies that compete with these antibodies or bind to the same epitope.

Also provided are nucleic acids encoding anti-PcrV antibodies, compositions comprising anti-PcrV antibodies, and methods of making and using anti-PcrV antibodies. [definition]

As used herein, "treatment" or "treating" is a method of obtaining beneficial or desired results, including clinical results. For the purposes of the present invention, the beneficial or desired clinical outcome includes, but is not limited to, one or more of the following: alleviation of one or more symptoms caused by the disease, reduction in the extent of the disease, stabilization of the disease (eg, prevention or delay of disease progression) , to prevent or delay the spread of disease (eg, systemic spread of pathogenic bacteria), to prevent or delay disease recurrence, to delay or slow disease progression, to improve disease state, to alleviate disease (in part or in whole), to reduce the need for one or more other drugs to treat disease dose, delay disease progression, improve or enhance quality of life, gain weight, and/or prolong survival. Also, "treatment" includes reduction of the pathological outcome of infection (eg, host cell lysis or necrosis). The methods of the present invention contemplate any one or more aspects of these treatments.

The term "prevent" and similar words, such as "prevented", "preventing", "prevention", etc., means a prevention, inhibition or reduction of a disease or condition (eg, pathogen infection) method of occurrence or likelihood of recurrence. It also refers to delaying the onset or recurrence of a disease or disorder, or delaying the onset or recurrence of symptoms of a disease or disorder. As used herein, "prevention" and similar terms also include reducing the intensity, impact, symptoms and/or burden of a disease or disorder before it develops or recurs. As used herein, "prevention" and similar terms also include reducing the risk and susceptibility to the occurrence or recurrence of a disease or disorder, such as infection by a pathogen.

The term "antibody" includes full-length antibodies and antigen-binding fragments thereof. Full-length antibodies include two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variable regions in both chains typically include 3 hypervariable loops known as complementarity determining regions (CDRs) (light chain (LC) CDRs include LC-CDR1, LC-CDR2 and LC-CDR3, heavy chain (HC) ) CDRs include HC-CDR1, HC-CDR2 and HC-CDR3). The CDR boundaries of the antibodies or antigen-binding fragments disclosed herein can be defined or recognized by the conventions of Kabat, Chothia or Al-Lazikani (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). The three CDR regions of the heavy or light chain are inserted between flanking segments called framework regions (FRs), which are more conserved than the CDR regions and form the scaffolding that supports the hypervariable loops. The constant regions of the heavy and light chains are not involved in antigen binding, but exhibit a variety of effector functions. Antibodies are classified based on the amino acid sequence of their heavy chain constant region. The five main classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, which are characterized by heavy chains of the alpha, delta, epsilon, gamma, and mu types, respectively. Several major antibody classes are divided into subclasses such as IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), IgA1 (α1 heavy chain n), or IgA2 (α2 heavy chain).

As used herein, the term "antigen-binding fragment" refers to an antibody fragment including, eg, a diabody, Fab, Fab', F(ab')2, Fv fragment, disulfide stabilized Fv fragment (dsFv), (dsFv) ₂ , bispecific dsFv (dsFv-dsFv'), disulfide stabilized diabodies (ds diabodies), single chain antibodies (scFv), scFv dimers (bivalent diabodies) chain antibodies), multispecific antibodies, single domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody that is capable of binding an antigen but does not contain an intact antibody structure, consisting of antibody fragments comprising one or more CDRs Antibody Fragments. Antigen-binding fragments are capable of binding the same antigen as the parent antibody or parent antibody fragment (eg, parent scFv). In some embodiments, an antigen-binding fragment may include one or more CDRs from a particular human antibody grafted into framework regions from one or more different human antibodies.

As used herein, the term "epitope" refers to a specific group of atoms or amino acids on an antigen to which an antibody or antibody portion binds. If two antibodies or antibody portions exhibit competitive binding to an antigen, they are likely to bind to the same epitope on the antigen.

As described herein, when the first antibody inhibits binding of the second antibody to the PcrV target by at least 50% (eg, at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%), the primary antibody "competes" with the secondary antibody for binding to the PcrV target, and vice versa. PCT publication WO 03/48731 describes a high-throughput antibody "epitope classification" method based on cross-competition.

As used herein, the term "specifically binds", "specifically recognizes" or "specifically for" refers to a measurable and reproducible interaction, eg, binding of an antibody to a target can be The target is determined to be present in a heterogeneous population of molecules, including biomolecules. For example, the ability of an antibody to specifically recognize a target (which may be an epitope) means that the antibody binds to that target with higher affinity, avidity, easier and/or longer lasting than other targets. In some embodiments, an antibody that specifically recognizes an antigen reacts with one or more epitopes of the antigen with a binding affinity that is at least 10 times greater than its binding affinity to other targets.

As used herein, an "isolated" anti-PcrV antibody refers to an anti-PcrV antibody that is (1) unrelated to the naturally-occurring protein, (2) free from other proteins of the same origin, (3) made from a different species cells expressed, or (4) not found in nature.

As used herein, the term "isolated nucleic acid" refers to nucleic acid of genomic, cDNA or synthetic origin, or a combination thereof. According to its source, the "isolated nucleic acid" (1) has nothing to do with all or part of the polynucleotides in the "isolated nucleic acid" found in nature, and (2) may be related to the polynucleotides to which it is not linked in the natural state is operably linked, or (3) does not exist in nature as part of a longer sequence.

As used herein, the term "CDR" or "complementarity determining region" refers to the non-contiguous antigen binding sites found within the variable domains of heavy and light chain polypeptides. In the literature Kabat et al. , J. Biol. Chem. 252:6609-6616 (1977); Kabat et al. , US Dept. of Health and Human Services, "Sequences of proteins of immunological interest"(1991); Chothia et al . al. , J. Mol. Biol. 196:901-917 (1987); Al-Lazikani B. et al. , J. Mol. Biol. , 273: 927-948 (1997); MacCallum et al. , J. Mol. Biol. 262:732-745 (1996); Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Lefranc MP et al. , Dev. Comp. Immunol. , 27: 55-77 ( 2003); and Honegger and Plückthun, J. Mol. Biol. , 309:657-670 (2001), where these definitions, when compared with each other, include the overlap or subsection of amino acid residues. set. However, any definition used to refer to the CDRs of an antibody or grafted antibody or variant thereof is included within the scope of the terms as defined and used herein. The positions of amino acid residues included in the CDRs defined by the various references cited above are listed in Table 1 for comparison. Algorithms and binding interfaces for CDR prediction are known in the art and include, for example, Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Ehrenmann F. et al. , Nucleic Acids Res. , 38: D301-D307 (2010); and Adolf-Bryfogle J. et al. , Nucleic Acids Res. , 43: D432-D438 (2015). The contents of the references cited in this paragraph are incorporated herein by reference in their entirety for the purposes of the present invention and for possible inclusion in one or more claims herein. [Table 1] CDR Definition Kabat ¹ Chothia ² MacCallum ³ IMGT ⁴ AHo ⁵ _VH CDR1 31-35 26-32 30-35 27-38 25-40 _VH CDR2 50-65 53-55 47-58 56-65 58-77 _VH CDR3 95-102 96-101 93-101 105-117 109-137 _VL CDR1 24-34 26-32 30-36 27-38 25-40 _VL CDR2 50-56 50-52 46-55 56-65 58-77 _VL CDR3 89-97 91-96 89-96 105-117 109-137 ¹ Amino acid residue numbering refers to the nomenclature in Kabat et al. above ² Amino acid residue numbering refers to the nomenclature in Chothia et al. above ³ Amino acid residue numbering refers to the nomenclature in MacCallum et al. above ⁴ Amino acid residue numbering Refer to the nomenclature in Lefranc et al. above. ⁵ Amino acid residue numbering refers to the nomenclature in Honegger and Plückthun, above.

The term "chimeric antibody" means that a portion of the heavy and/or light chain is identical to or has homology to corresponding sequences in an antibody from a particular species or class or subclass of antibodies, and the chain(s) have Antibodies whose remainder is identical or homologous to corresponding sequences in antibodies from another genus or genus of other antibody classes or subclasses, and fragments of such antibodies, so long as they have the biological activity of the invention (see U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

"Fv" is the smallest antibody fragment that contains complete antigen recognition and binding sites. The fragment is a dimer formed by the tight non-covalent linkage of one heavy chain variable domain and one light chain variable domain. The folding of these two domains derives 6 hypervariable loops (3 loops in each of the light and heavy chains) that provide the antibody with amino acid residues for binding to the antigen and confer the antibody with the antigen specificity of binding. However, even a single variable domain (or half of an Fv fragment, which contains only 3 CDRs specific for an antigen) has the ability to recognize and bind an antigen, albeit with a lower affinity than an intact binding site.

"Single-chain Fv", also abbreviated "sFv" or "scFv", are antibody fragments comprising _VH and _VL antibody domains linked into a single polypeptide chain. In some embodiments, the scFv polypeptide further comprises a linking polypeptide between the _VH and _VL domains that allows the scFv to form a desirable structure for antigen binding. For an overview of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

The term "diabodies" refers to a small antibody fragment prepared by constructing a scFv fragment (see above) using a short linker (eg, 5-10 residues) between the V _H and _VL domains, This allows the variable domains to pair between chains rather than within chains, resulting in a bivalent fragment, ie a fragment with two antigen binding sites. Bispecific diabodies are heterodimers of two "crossover" scFv fragments, where the _VH and _VL domains of the two antibodies are located on different polypeptide chains. Diabodies are fully described in EP 404,097; WO 93/11161; Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993).

"Humanized" forms of non-human (eg, rodent) antibodies are chimeric antibodies that include minimal sequence from the non-human antibody. In most cases, humanized antibodies are human immunoglobulins (receptor antibodies) in which the hypervariable region (HVR) residues of the receptor antibody are derived from non-human species such as mouse, rat, rabbit or Non-human primate hypervariable region residues that have desirable antibody specificity, affinity and properties are substituted (donor antibody). In certain instances, residues in human immunoglobulin framework regions (FRs) are replaced by corresponding non-human residues. Additionally, humanized antibodies may include residues that are not present in either the recipient antibody or the donor antibody. These modifications can further improve the performance of the antibody. Typically, a humanized antibody will contain substantially all, at least one, usually two variable domains, wherein all or substantially all of the hypervariable loops correspond to the hypervariable loops of the non-human immunoglobulin, and all or substantially all of the hypervariable loops All framework regions above are human immunoglobulin sequences. Human antibodies optionally also include at least a portion of an immunoglobulin constant region (Fc), typically a human immunoglobulin constant region. For details see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).

The "percent (%) amino acid sequence identity" or "homology" of the polypeptide and antibody sequences identified herein is defined as: candidate sequences are The percentage of identical amino acid residues in the polypeptide sequences to be compared. Percent amino acid sequence identity can be determined by a variety of alignment means within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. However, for purposes herein, percent amino acid sequence identity values were calculated using the sequence alignment computer program MUSCLE (Edgar, RC, Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, RC, BMC Bioinformatics 5(1) :113, 2004) generated.

The term "Fc receptor" or "FcR" is used to describe a receptor that binds the Fc region of an antibody. In some embodiments, the FcR described herein is an FcR that binds an IgG antibody, a gamma receptor, including receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and variants of these receptors splicing form. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences and differ primarily in the cytoplasmic domain. The cytoplasmic domain of the activating receptor FcγRIIA contains an immunoreceptor tyrosine activation motif (ITAM). The cytoplasmic domain of the inhibitory receptor FcyRIIB contains an immunoreceptor tyrosine inhibitory motif (ITIM) (see M. in Daëron , Annu. Rev. Immunol. 15:203-234 (1997)). The term also includes allotypes, such as FcyRIIIA allotypes: FcyRIIIA-Phe158, FcyRIIIA-Val158, FcyRIIA-R131 and/or FcyRIIA-H131. In Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991) and Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med . 126 FcRs are described in : 330-41 (1995). The term FcR in the present invention encompasses other types of FcRs, including FcRs to be identified in the future. The term FcR also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgGs to the neonate (Guyer et al., J. Immunol . 117:587 (1976) and Kim et al., J. Immunol . 24:249 (1994) )).

The term "FcRn" refers to the neonatal Fc receptor (FcRn). FcRn is structurally similar to the major histocompatibility complex (MHC), consisting of an α chain non-covalently bound to β2 microglobulin. The various functions of the neonatal Fc receptor FcRn are described in Ghetie and Ward (2000) Annu. Rev. Immunol. 18, 739-766. FcRn plays an important role in passive transport of immunoglobulin IgGs from mother to newborn and regulation of serum IgG levels. FcRn acts as a salvage receptor that binds and transports endocytosed IgG in intact form within and between cells and protects them from default degradation pathways.

The "CH1 domain" of a human IgG Fc region typically extends from amino acid 118 to amino acid 215 (EU numbering system).

The "hinge region" is generally defined as extending from Glu 216 to Pro 230 of human IgGl (Burton, Molec. Immunol. 22:161-206 (1985)). The hinge regions of other IgG isotypes can be aligned to the IgGl sequence by placing the first and last cysteine residues that form the inter-heavy chain disulfide bond in the same position as IgGl.

The "CH2 domain" of a human IgG Fc region typically extends from amino acid 231 to amino acid 340. The CH2 domain is unique in that it does not pair tightly with another region, but instead has two N-terminally linked branched sugar chains inserted between the two CH2 domains of an intact native IgG molecule. It has been speculated that carbohydrates may serve as an alternative to domain-to-domain pairing, helping to keep the CH2 domain stable. Burton, Molec Immunol. 22:161-206 (1985).

The "CH3" domain includes an extension within the Fc region from the C-terminal residue to the CH2 domain (from amino acid 341 to the C-terminus of the antibody sequence, usually amino acid residue 446 or 447 of IgG).

"Functional Fc fragments" have "effector functions" possessed by native Fc region sequences. Exemplary "effector functions" include C1q binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors ( Such as B cell receptor; BCR) and so on. Such effector functions typically require binding of an Fc region to a binding domain (eg, antibody variable region) and can be assessed using a variety of experimental methods well known in the art.

Antibodies of IgG Fc variants with "altered" FcR binding affinity or ADCC activity have enhanced or reduced FcR binding activity and/or ADCC activity compared to the parent polypeptide or a polypeptide comprising the native Fc sequence. An Fc variant that exhibits "enhanced binding" to an FcR has a higher binding affinity (eg, a lower apparent Kd or IC50 to at least one FcR) than the parent polypeptide or a polypeptide comprising a native IgG Fc sequence value). In some embodiments, the binding capacity is increased 3-fold, eg, 5, 10, 25, 50, 60, 100, 150, 200, or even up to 500-fold or the binding capacity is increased by 25% to 1000% compared to the parent polypeptide. Fc variants that exhibit "reduced binding" to FcRs, with lower affinity (eg, higher apparent Kd or IC50) for at least one FcR than the parent polypeptide value). Compared to the parent polypeptide, its binding capacity is reduced by 40% or more.

"Antibody-dependent cell-mediated cytotoxicity" or "ADCC" is a form of cytotoxicity that refers to the presence of secreted Ig on certain cytotoxic cells such as natural killer cells (NK), neutrophils, and macrophages. Fc receptors (FcRs) on phagocytes) enable these cytotoxic effector cells to specifically bind antigen-bearing target cells and subsequently kill the target cells using cytotoxins. Antibodies "arm" cytotoxic cells and are required for this killing. Among the major cell types mediating ADCC, NK cells express only FcγRIII, while monocytes express FcγRI, FcγRII and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991) . To assess ADCC activity of target molecules, in vitro ADCC assays can be performed, as described in US Pat. Nos. 5,500,362 or 5,821,337. Suitable effector cells for such experiments include peripheral blood mononuclear cells (PBMCs) and natural killer cells (NKs). Alternatively, or in addition, ADCC activity of target molecules can also be assessed in vivo, eg, as described in animal models as disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).

A polypeptide comprising an Fc region variant that exhibits "enhanced ADCC activity" or is capable of mediating ADCC effects more efficiently in the presence of human effector cells than a wild-type IgG Fc polypeptide or parent polypeptide comprising an Fc region When the polypeptide of the variant contains substantially the same amount of the wild-type IgG Fc polypeptide (or the parent polypeptide) in the experiment, it can mediate ADCC more efficiently in vitro or in vivo. Such variants are typically identified using any in vitro ADCC assay method known in the art, eg, assays or methods for identifying ADCC activity, eg, in animal models and the like. In some embodiments, such variants are 5- to 100-fold, eg, 25- to 50-fold, more efficient at mediating ADCC compared to wild-type Fc (or the parent polypeptide).

"Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (subclasses with appropriate structures) that bind cognate antigens. To assess complement activation, CDC experiments can be performed as described in Gazzano-Santoro et al. , J. Immunol. Methods 202:163 (1996). Polypeptide variants with altered Fc region amino acid sequences and increased or decreased C1q binding capacity are described in US Patent No. 6,194,551 B1 and WO 99/51642. The contents of these patent publications are expressly incorporated herein by reference. See also Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

Unless otherwise specified, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate to each other and that encode the same amino acid sequence. Nucleotide sequences encoding proteins or RNA may also include introns, eg, nucleotide sequences encoding proteins include introns in some forms.

The term "operably linked" refers to a functional linkage between a regulatory sequence and a heterologous nucleotide sequence, thereby allowing expression of the latter. For example, a first nucleotide sequence is operably linked to a second nucleotide sequence when the first nucleotide sequence is in a functional relationship with the second nucleotide sequence. For example, a promoter is operably linked to a coding sequence if it affects the transcription or expression of the coding sequence. Typically, operably linked DNA sequences are contiguous and, where necessary, two protein-coding regions may be joined in the same reading frame.

"Homologous" refers to sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. Two DNA molecules are homologous if the same position of the two compared sequences is the same base group or amino acid monomer subunit, eg, both DNA molecules are adenine at the same position. The percent homology between two sequences is a function of the ratio of the number of matches or homologous positions shared by the two sequences to the total number of positions multiplied by 100. For example, two sequences are 60% homologous if 6 out of 10 positions are matched or homologous. For example, the DNA sequences ATTGCC and TATGGC share 50% homology. Generally, when aligning two sequences, the alignment is made with the aim of obtaining maximum homology.

An "effective amount" of an anti-PcrV antibody or composition disclosed herein refers to an amount sufficient to achieve a particular purpose. An "effective amount" can be determined empirically and by known methods relevant to the purpose.

The term "therapeutically effective amount" refers to an amount of an anti-PcrV antibody or composition described herein that is effective to "treat" a disease or disorder in a subject. In the case of a Pseudomonas aeruginosa infection, a therapeutically effective amount of an anti-PcrV antibody or composition disclosed herein can reduce the number of infected cells, inhibit (ie, to some extent slow and preferably stop) the spread of the infection, and/ Or provide some relief of one or more symptoms associated with the infection. The anti-PcrV antibodies or compositions disclosed herein are capable of inhibiting P. aeruginosa growth and/or killing P. aeruginosa upon infection, and the anti-PcrV antibodies may be cytostatic and/or cytotoxic. In some embodiments, a therapeutically effective amount refers to an amount that inhibits infection in a patient. In some embodiments, a therapeutically effective amount refers to an amount that completely clears the infection in a patient.

As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" refers to a material that has no biological activity or other undesirable properties, eg, that can be incorporated into a pharmaceutical composition administered to a patient without May cause significant adverse biological reactions, or, do not interact in a deleterious manner with any other components contained in the composition. A pharmaceutically acceptable carrier or excipient preferably meets the required criteria for toxicology or manufacturing testing and/or is included in the Inactive Ingredient Guidelines prepared by the US Food and Drug Administration.

Embodiments of the invention described herein should be understood to include embodiments that "consist of" and/or "consist essentially of."

Reference herein to "about" is a value or parameter, including (and describing) variations to that value or parameter itself. For example, a description referring to "about X" includes a description of "X".

As used herein, reference to "not" a value or parameter generally means and describes "other than" a certain value or parameter. For example, the method cannot be used to treat type X infections, meaning that the method is often used to treat types other than type X infections.

As used herein and in the claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. [Anti-PcrV Antibody]

In one aspect, the invention provides anti-PcrV antibodies that specifically bind to PcrV. The anti-PcrV antibodies include, but are not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibody molecules described herein comprising heavy and/or light chain CDRs. In one aspect, the invention provides isolated antibodies that bind to PcrV. Contemplated anti-PcrV antibodies include, for example, full-length anti-PcrV antibodies (eg, full-length IgG1, IgG2, or IgG4), anti-PcrV single-chain antibodies, multispecific (eg, bispecific) anti-PcrV antibodies, anti-PcrV immunoconjugates, and and so on. In some embodiments, the anti-PcrV antibody is a Fab, Fab', F(ab)'2, Fab'-SH, single chain antibody (scFv), Fv fragment, dAb, Fd, nanobody, diabody ) or linear antibodies. In some embodiments, an antibody that specifically binds to PcrV refers to an antibody that binds to PcrV with an affinity that is at least 10 times greater than its non-target binding affinity (including, for example, 10, 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ , 10 ⁶ ). , or 10 ⁷ times). In some embodiments, non-target refers to an antigen that is not PcrV. Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence activated cell sorting (FACS) assay or radioimmunoprecipitation assay (RIA). Kd values can be determined by methods known in the art, such as surface plasmon resonance (SPR) techniques or biolayer interferometry (BLI).

In some embodiments, the anti-PcrV antibody or antigen-binding fragment (a) that specifically binds to P. aeruginosa PcrV promotes, mediates, or enhances opsonophagocytic killing (OPK) of Pseudomonas, and/or (b) Disruption of the activity of the type III toxin secretion system.

While anti-PcrV antibodies comprising human sequences are discussed broadly herein (eg, human heavy and light chain variable domains comprising human CDR sequences), non-human anti-PcrV antibodies are also contemplated. In some embodiments, the non-human anti-PcrV antibody includes human CDR sequences and non-human framework region sequences of the anti-PcrV antibodies described herein, in some embodiments, the non-human framework region sequences include any for use as described herein The one or more human CDR sequences described generate sequences for heavy and/or light chain variable domains, including, for example, mammalian, e.g., mouse, rat, rabbit, porcine, bovine (e.g., bovine, bull, buffalo), deer , sheep, goats, chickens, cats, dogs, ferrets, primates (eg, marmosets, macaques), etc. In some embodiments, non-human anti-PcrV antibodies include anti-PcrV antibodies generated by grafting one or more human CDR sequences described herein into non-human framework regions (eg, murine or chicken framework region sequences).

The complete amino acid sequence of an exemplary PcrV protein includes or consists of the amino acid sequence SEQ ID NO:80. In some embodiments, the anti-PcrV antibodies described herein specifically recognize an epitope in Pseudomonas PcrV. In some embodiments, the anti-PcrV antibody is specific for Pseudomonas PcrV and does not cross-react with other species or types of non-Pseudomonas proteins.

In some embodiments, the anti-PcrV antibody includes an antibody heavy chain constant region and an antibody light chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgGl type heavy chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgG2-type heavy chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgG3-type heavy chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgG4-type heavy chain constant region. In some embodiments, the IgG refers to human IgG. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:82. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:83. In some embodiments, the anti-PcrV antibody comprises a lambda light chain constant region. In some embodiments, the anti-PcrV antibody comprises a kappa light chain constant region. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:81. In some embodiments, the anti-PcrV antibody includes an antibody heavy chain variable domain and an antibody light chain variable domain.

In some embodiments, any one of the isolated anti-PcrV antibodies described above, the isolated anti-PcrV antibody comprises a heavy chain variable domain ( _{VH )} comprising: a heavy chain complementarity determining region (HC- CDR) 1, comprising SYWMH (SEQ ID NO: 1) or a variant thereof comprising up to 3 amino acid substitutions; a HC-CDR2 comprising RINEX ₁ EX ₂ SX ₃ SYADSVKG (SEQ ID NO: 50), or up to 3 amino acid substitution variants, wherein X ₁ is D, T, N, L, I, S, V, A or H, X ₂ is S, T, N, D, G or R, and X ₃ is I or T; and one HC-CDR3 comprising DGX ₁ X ₂ X ₃ X ₄ X ₅ DX ₆ (SEQ ID NO: 51), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is P, Y or A , X ₂ is Y or W, X ₃ is D, T or N, X ₄ is S, T or A, X ₅ is L, F or M, and X ₆ is I, V, A, S, L or W; and a light chain variable domain ( _{VL )} comprising: a light chain complementarity determining region (LC-CDR) 1 comprising RASQX ₁ VX ₂ X ₃ NLA (SEQ ID NO: 47), or comprising at most ₃ amino _acid substitution variants, wherein X1 is S, N, D or G, X2 is S, K, N, R or T, and _X3 is N, S, D or G; an LC-CDR2 comprising X ₁ ASSRAT (SEQ ID NO: 85), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is D, N, H, A or S; and an LC-CDR3 comprising QQYGX ₁ X ₂ PX ₃ T (SEQ ID NO: 86), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is S, A, T, E, D, H, N, Q, G, L, Y, M, R or V, _X2 is S, Q, V, E, T, D, M, Y, G, H, L, N, A, F or P, and _X3 is I, L or V.

In some embodiments, any one of the isolated anti-PcrV antibodies described above, the isolated anti-PcrV antibody comprises a heavy chain variable domain ( _{VH )} comprising: a HC-CDR1 comprising SYWMH (SEQ ID NO: 1), or a variant comprising at most 3 amino acid substitutions; a HC-CDR2 comprising RINEX1EX2SISYADSVKG (SEQ ID NO: 45), or a variant comprising at most 3 amino acid substitutions, wherein X1 is D, N, I, L or V, X2 is S, T, R, G or N; and one HC-CDR3, including DGPYDX1X2DI (SEQ ID NO: 46), or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is S, A or T, X2 is F or _L ; and _VL comprising: an LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO: 47), or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is N, G, D or S, X2 is K, R, S, N or T, and X3 is N, G, S or D; an LC-CDR2 comprising X1ASSRAT (SEQ ID NO: 48), or its A variant comprising up to 3 amino acid substitutions, wherein X1 is D, N, H or A; and an LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO: 49), or a variant comprising up to 3 amino acid substitutions, wherein X1 is S, T, E, H, N, A, D, M or L, X2 is S, Q, E, T, D, G, H, L, N, V or Y, and X3 is I, L or V.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising: a HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1 or comprising up to 3 (eg 1, 2 or 3) amino acids Substituted variants; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 2-9 or variants comprising up to 3 (eg 1, 2 or 3) amino acid substitutions; an HC-CDR3 comprising A variant comprising the amino acid sequence of any one of SEQ ID NOs: 10-13 or comprising up to 3 (eg 1, 2 or 3) amino acid substitutions; and a _VL comprising _: an LC-CDR1 comprising An amino acid sequence of any one of SEQ ID NOs: 14-25 or a variant comprising up to 3 (e.g. 1, 2 or 3) amino acid substitutions; an LC-CDR2 comprising any of the amino acids of SEQ ID NOs: 26-29 Sequences or variants comprising up to 3 (e.g. 1, 2 or 3) amino acid substitutions; an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 30-44 or up to 3 (e.g. 1, 2) or 3) amino acid substitution variants.

In some embodiments, the anti-PcrV antibody, the isolated anti-PcrV antibody comprises: _VH , the _VH comprises: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1; a HC-CDR2 comprising An amino acid sequence of any one of SEQ ID NOs: 2-9; and an HC-CDR3 comprising an amino acid sequence of any of SEQ ID NOs: 10-13; and a _VL comprising: an LC-CDR1 comprising SEQ ID NOs _: 1 ID NOs: any amino acid sequence in 14-25; one LC-CDR2, comprising any amino acid sequence in SEQ ID NOs: 26-29; one LC-CDR3, comprising any amino acid sequence in SEQ ID NOs: 30-44.

_{and _} A _VL comprising LC-CDR1, LC-CDR2 and LC-CDR3 in a _VL having the amino acid sequence of any of SEQ ID NOs: 65-79.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of any one of SEQ ID NOs: 52-64, or comprising at least the amino acid sequence of any one of SEQ ID NOs: 52-64 Variant sequences of 90% (eg at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% ₎ sequence homology, and _VL comprising The amino acid sequence of any of SEQ ID NOs: 65-79, or a variant sequence comprising at least 90% sequence homology to any of the amino acid sequences of SEQ ID NOs: 65-79. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of any one of SEQ ID NOs: _{52-64, and a VL comprising} SEQ ID NOs: 65-79 any amino acid sequence.

In some embodiments, the anti-PcrV antibody is a full-length antibody. In some embodiments, the anti-PcrV antibody comprises an IgGl constant region. In some embodiments, the IgG1 refers to human IgG1. In some embodiments, the anti-PcrV antibody comprises an IgG4 constant region. In some embodiments, the IgG4 refers to human IgG4. In some embodiments, the anti-PcrV antibody heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:82. In some embodiments, the anti-PcrV antibody heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:83. In some embodiments, the anti-PcrV antibody light chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:81.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC comprising the amino acid sequence of SEQ ID NO: 26 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:30. In some embodiments, an anti-PcrV antibody that can specifically compete with an antibody is provided, the antibody comprising: _VH , the _VH comprising: one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one comprising HC-CDR2 of amino acid sequence SEQ ID NO: 2, and a HC-CDR3 comprising amino acid sequence of SEQ ID NO: 10; and _VL comprising _: an LC-CDR1 comprising amino acid sequence of SEQ ID NO: 14 , an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30. In some embodiments, there is provided an anti-PcrV antibody that binds to the same epitope as an antibody, the antibody comprising: _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 14 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:26, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:30.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:52, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:52) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 65, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:65. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:52 and a _VL comprising the amino acid sequence of SEQ ID NO:65. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 52; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 52 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 65.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC comprising the amino acid sequence of SEQ ID NO: 26 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:30. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 14 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:26, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:30. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 14 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:26, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:30.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:56) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 65, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:65. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:65. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 56; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 56 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 65.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 15, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 31, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:31. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 15 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:31. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 15 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:31.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:53, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:53) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 66, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:66. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:53 and a _VL comprising the amino acid sequence of SEQ ID NO:66. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in the _VH having the amino acid sequence of SEQ ID NO: 53; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 53 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 66.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39, or a V at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 23 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 23 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:62, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:62) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 74, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:74. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:62 and a _VL comprising the amino acid sequence of SEQ ID NO:74. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in the _VH having the amino acid sequence of SEQ ID NO: 62; and a _VL comprising amino acids HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 74.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 16, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 32, or an LC-CDR3 comprising at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, an LC comprising the amino acid sequence of SEQ ID NO: 28 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:32. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:32. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 16 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:32.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 54, or comprising at least 90% (e.g. at least 91%, 92% of the amino acid sequence of SEQ ID NO: 54) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 67, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:67. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:54 and a _VL comprising the amino acid sequence of SEQ ID NO:67. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in the _VH having the amino acid sequence of SEQ ID NO: 54; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 54 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 67.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 33, or a V LC-CDRs comprising at most 5 amino acid substitutions _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:33. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:33. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:33.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 55, or comprising at least 90% (eg, at least 91%, 92% amino acid sequence SEQ ID NO: 55) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 68, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO: 68. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:55 and a _VL comprising the amino acid sequence of SEQ ID NO:68. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 55; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 55 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 68.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 18, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 34, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 18, an LC comprising the amino acid sequence of SEQ ID NO: 28 - CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:34. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 18 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:34. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 18 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:34.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:56) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 69, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:69. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:69. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 56; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 56 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 69.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:64, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:64) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 78, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO: 78. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:64 and a _VL comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 64; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 64 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 78.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 19, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 35, or an LC-CDRs comprising at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 19, an LC comprising the amino acid sequence of SEQ ID NO: 28 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:35.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:56) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 70, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:70. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:70. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 56; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 56 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 70.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 20, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 36, or a LC-CDR3 comprising at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 20 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 20 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:36.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:57, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:57) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 71, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO: 71. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:57 and a _VL comprising the amino acid sequence of SEQ ID NO:71. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 57; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 57 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 71.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 21, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37, or a V substituted at most 5 amino acids in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, an LC comprising the amino acid sequence of SEQ ID NO: 28 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 21 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 21 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:37.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 54, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO: 54) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 72, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:72. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:54 and a _VL comprising the amino acid sequence of SEQ ID NO:72. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 54; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 54 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 72.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 22, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38, or a V substituted at most 5 amino acids in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 22, an LC comprising the amino acid sequence of SEQ ID NO: 28 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 22 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 22 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:38.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:56) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 73, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO: 73. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:73. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 56; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 56 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 73.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39, or a V at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 23 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 23 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:58, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:58) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 74, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:74. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:58 and a _VL comprising the amino acid sequence of SEQ ID NO:74. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 58; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 58 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 74.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 21, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 40, or a V substituted at most 5 amino acids in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, an LC comprising the amino acid sequence of SEQ ID NO: 28 - CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:40. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:40. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:40.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 59, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO: 59) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 75, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:75. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:59 and a _VL comprising the amino acid sequence of SEQ ID NO:75. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 59; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 59 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 75.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 24, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 41, or a V substituted at most 5 amino acids in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 24, an LC comprising the amino acid sequence of SEQ ID NO: 28 - CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:41. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 24 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:41. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 24 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:28, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:41.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:57, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:57) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 76, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:76. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:57 and a _VL comprising the amino acid sequence of SEQ ID NO:76. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 57; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 57 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 76.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 25, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42, or an LC-CDR3 comprising at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 25, an LC comprising the amino acid sequence of SEQ ID NO: 29 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:42. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 25 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:29, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:42. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 25 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:29, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:42.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 60, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO: 60) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO _: 77, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO: 77. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:60 and a _VL comprising the amino acid sequence of SEQ ID NO:77. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 60; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 60 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 77.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 60, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO: 60) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 78, or comprising A variant sequence with at least 90% sequence homology thereto. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:60 and a _VL comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 60; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 60 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 78.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 44, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC comprising the amino acid sequence of SEQ ID NO: 27 - CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:44. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:44. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:44.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 56, or comprising at least 90% (eg, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence SEQ ID NO: 79, or comprising the amino acid sequence SEQ ID NO _: 79 79 Variant sequences with at least 90% sequence homology. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:79. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 56; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 56 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO:79.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC comprising the amino acid sequence of SEQ ID NO: 27 -CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 23 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; and _VL comprising _: an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 23 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:39.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 60, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO: 60) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 74, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO:74. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:60 and a _VL comprising the amino acid sequence of SEQ ID NO:74. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 61; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 61 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 74.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12, or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO _: LC-CDR1 of : 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or a V with at most 5 amino acid substitutions in the LC-CDRs _L variant.

In some embodiments, the anti-PcrV antibody comprises _{: a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC comprising the amino acid sequence of SEQ ID NO: 27 - CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43. In some embodiments, there is provided an anti-PcrV antibody that can specifically compete with an antibody comprising: a _VH , the _VH comprising: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43. In some embodiments, there is provided an anti-PcrV antibody that binds the same epitope as an antibody comprising _{: a VH comprising} : an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12; and _VL comprising _: an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 17 CDR1, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and one LC-CDR3 comprising the amino acid sequence of SEQ ID NO:43.

In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:63, or comprising at least 90% (e.g. at least 91%, 92% with the amino acid sequence of SEQ ID NO:63) , 93%, 94%, 95%, 96%, 97%, 98% or 99%) sequence homology variant sequences, and _VL comprising the amino acid sequence of SEQ ID NO _: 78, or comprising A variant sequence having at least 90% sequence homology to the amino acid sequence SEQ ID NO: 78. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino acid sequence of SEQ ID NO:63 and a _VL comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, the anti-PcrV antibody comprises a _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in a _VH having the amino acid sequence of SEQ ID NO: 63; and a _VL comprising amino acids having amino acid sequence SEQ ID NO: 63 HC-CDR1, HC-CDR2 and HC-CDR3 in _VL of sequence SEQ ID NO: 78.

In some embodiments, competition assays can be used to identify monoclonal antibodies that compete with the anti-PcrV antibodies described herein for binding to PcrV. Competition experiments can determine whether two antibodies bind the same epitope by recognizing the same or spatially overlapping epitopes or by competitively inhibiting the binding of one antibody to the antigen by the other. In certain embodiments, such competing antibodies bind the same epitope as the antibodies described herein. Some exemplary competition experiments include, but are not limited to, routine experiments as mentioned in Harlow and Lane (1988) Antibodies: A Laboratory Manual ch. 14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.). Detailed exemplary methods for resolving epitopes bound by antibodies are described in Morris (1996) "Epitope Mapping Protocols," in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, N.J.). In some embodiments, each antibody is said to bind the same epitope if it blocks 50% or more of the binding of the other antibody. In some embodiments, the antibody that competes with an anti-PcrV antibody described herein is a chimeric, humanized or fully human antibody.

Exemplary anti-PcrV antibody sequences are shown in Tables 2 and 3. Those skilled in the art will recognize that there are a variety of known algorithms for predicting the positions of CDRs and for defining antibody light and heavy chain variable regions. Antibodies comprising the CDRs, _VH and/or _VL sequences of anti-PcrV antibodies as described herein, but based on prediction algorithms other than those exemplified in the table below are also within the scope of the invention. Table 2 Exemplary anti-PcrV antibody CDR sequences Antibody name HC-CDR1 HC-CDR2 HC-CDR3 PA49 SYWMH (SEQ ID NO: 1) RINEDETSISYADSVKG (SEQ ID NO: 2) DGPYDSFDI (SEQ ID NO: 10) 6G12 SYWMH (SEQ ID NO: 1) RINEDETSISYADSVKG (SEQ ID NO: 2) DGPYDSLDI (SEQ ID NO: 11) 10B1 SYWMH (SEQ ID NO: 1) RINEVENSISYADSVKG (SEQ ID NO: 3) DGPYDSLDI (SEQ ID NO: 11) 11E9 SYWMH (SEQ ID NO: 1) RINENEGSISYADSVKG (SEQ ID NO: 7) DGPYDTLDI (SEQ ID NO: 13) 1D6 SYWMH (SEQ ID NO: 1) RINEDESSISYADSVKG (SEQ ID NO: 4) DGPYDSLDI (SEQ ID NO: 11) 2A5 SYWMH (SEQ ID NO: 1) RINELESSISYADSVKG (SEQ ID NO: 5) DGPYDSLDI (SEQ ID NO: 11) 4A2 SYWMH (SEQ ID NO: 1) RINEDETSISYADSVKG (SEQ ID NO: 2) DGPYDSLDI (SEQ ID NO: 11) 7H5 SYWMH (SEQ ID NO: 1) RINEDERSISYADSVKG (SEQ ID NO: 9) DGPYDTLDI (SEQ ID NO: 13) 4H9 SYWMH (SEQ ID NO: 1) RINEDETSISYADSVKG (SEQ ID NO: 2) DGPYDSLDI (SEQ ID NO: 11) 5B4 SYWMH (SEQ ID NO: 1) RINENESSISYADSVKG (SEQ ID NO: 6) DGPYDSLDI (SEQ ID NO: 11) 5E6 SYWMH (SEQ ID NO: 1) RINEDESSISYADSVKG (SEQ ID NO: 4) DGPYDSLDI (SEQ ID NO: 11) 6G3 SYWMH (SEQ ID NO: 1) RINEDETSISYADSVKG (SEQ ID NO: 2) DGPYDSLDI (SEQ ID NO: 11) 10D8 SYWMH (SEQ ID NO: 1) RINENEGSISYADSVKG (SEQ ID NO: 7) DGPYDSLDI (SEQ ID NO: 11) 2B4 SYWMH (SEQ ID NO: 1) RINEIETSISYADSVKG (SEQ ID NO: 8) DGPYDSLDI (SEQ ID NO: 11) 5C7 SYWMH (SEQ ID NO: 1) RINENESSISYADSVKG (SEQ ID NO: 6) DGPYDSLDI (SEQ ID NO: 11) 5F8 SYWMH (SEQ ID NO: 1) RINEDERSISYADSVKG (SEQ ID NO: 9) DGPYDSLDI (SEQ ID NO: 11) 6G4 SYWMH (SEQ ID NO: 1) RINEDERSISYADSVKG (SEQ ID NO: 9) DGPYDSLDI (SEQ ID NO: 11) 9F3 SYWMH (SEQ ID NO: 1) RINEDETSISYADSVKG (SEQ ID NO: 2) DGPYDSLDI (SEQ ID NO: 11) 9C7 SYWMH (SEQ ID NO: 1) RINENEGSISYADSVKG (SEQ ID NO: 7) DGPYDALDI (SEQ ID NO: 12) 5F3 SYWMH (SEQ ID NO: 1) RINEDERSISYADSVKG (SEQ ID NO: 9) DGPYDALDI (SEQ ID NO: 12) Antibody name HC-CDR1 HC-CDR2 HC-CDR3 PA49 RASQSVSSNLA (SEQ ID NO: 14) DASSRAT (SEQ ID NO: 26) QQYGSSPLT (SEQ ID NO: 30) 6G12 RASQSVSSNLA (SEQ ID NO: 14) DASSRAT (SEQ ID NO: 26) QQYGSSPLT (SEQ ID NO: 30) 10B1 RASQGVNNNLA (SEQ ID NO: 15) HASSRAT (SEQ ID NO: 27) QQYGTVPIT (SEQ ID NO: 31) 11E9 RASQGVRNNLA (SEQ ID NO: 23) HASSRAT (SEQ ID NO: 27) QQYGLQPIT (SEQ ID NO: 39) 1D6 RASQGVKNNLA (SEQ ID NO: 16) NASSRAT (SEQ ID NO: 28) QQYGEYPLT (SEQ ID NO: 32) 2A5 RASQNVKNNLA (SEQ ID NO: 17) HASSRAT (SEQ ID NO: 27) QQYGAEPIT (SEQ ID NO: 33) 4A2 RASQDVRNNLA (SEQ ID NO: 18) NASSRAT (SEQ ID NO: 28) QQYGTNPLT (SEQ ID NO: 34) 7H5 RASQNVKNNLA (SEQ ID NO: 17) HASSRAT (SEQ ID NO: 27) QQYGNEPIT (SEQ ID NO: 43) 4H9 RASQGVSSNLA (SEQ ID NO: 19) NASSRAT (SEQ ID NO: 28) QQYGHQPIT (SEQ ID NO: 35) 5B4 RASQDVRDNLA (SEQ ID NO: 20) HASSRAT (SEQ ID NO: 27) QQYGETPIT (SEQ ID NO: 36) 5E6 RASQNVKGNLA (SEQ ID NO: 21) NASSRAT (SEQ ID NO: 28) QQYGHDPVT (SEQ ID NO: 37) 6G3 RASQDVTNNLA (SEQ ID NO: 22) NASSRAT (SEQ ID NO: 28) QQYGDLPIT (SEQ ID NO: 38) 10D8 RASQGVRNNLA (SEQ ID NO: 23) HASSRAT (SEQ ID NO: 27) QQYGLQPIT (SEQ ID NO: 39) 2B4 RASQNVKGNLA (SEQ ID NO: 21) NASSRAT (SEQ ID NO: 28) QQYGNQPIT (SEQ ID NO: 40) 5C7 RASQSVNNNLA (SEQ ID NO: 24) NASSRAT (SEQ ID NO: 28) QQYGTTPIT (SEQ ID NO: 41) 5F8 RASQNVSNNLA (SEQ ID NO: 25) AASSRAT (SEQ ID NO: 29) QQYGMGPIT (SEQ ID NO: 42) 6G4 RASQNVKNNLA (SEQ ID NO: 17) HASSRAT (SEQ ID NO: 27) QQYGNEPIT (SEQ ID NO: 43) 9F3 RASQNVKNNLA (SEQ ID NO: 17) HASSRAT (SEQ ID NO: 27) QQYGTHPIT (SEQ ID NO: 44) 9C7 RASQGVRNNLA (SEQ ID NO: 23) HASSRAT (SEQ ID NO: 27) QQYGLQPIT (SEQ ID NO: 39) 5F3 RASQNVKNNLA (SEQ ID NO: 17) HASSRAT (SEQ ID NO: 27) QQYGNEPIT (SEQ ID NO: 43) Table 3 Exemplary _VH and _VL sequences SEQ ID NO describe sequence 52 PA49 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFRSYWMHWVRQAPGKGLVWVSRINEDETSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCARDGPYDSFDIWGQGTMVTVSS 56 6G12, 4A2, 4H9, 6G3, 9F3 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEDETSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 53 10B1 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEVENSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 62 11E9 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINENEGSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDTLDIWGQGTMVTVSS 54 1D6, 5E6 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEDESSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 55 2A5 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINELESSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 64 7H5 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEDERSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDTLDIWGQGTMVTVSS 57 5B4, 5C7 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINENESSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 58 10D8 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINENEGSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 59 2B4 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEIETSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 60 5F8, 6G4 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEDERSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDSLDIWGQGTMVTVSS 61 9C7 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINENEGSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDALDIWGQGTMVTVSS 63 5F3 _VH QVQLVESGGGLVQPGGSLRLSCAASGFSFSSYWMHWVRQAPGKGLVWVSRINEDERSISYADSVKGRFTISRDNAKNTLYLQMNGLRAEDTAVYYCVRDGPYDALDIWGQGTMVTVSS 65 PA49,6G12 V _L EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQHKPGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGGGTKLEIK 66 10B1 V _L EIVMTQSPATLSVSPGERATLSCRASQGVNNNLAWYQHKPGQAPRLLIYHASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGTVPITFGGGTKLEIK 74 11E9, 10D8, 9C7 V _L EIVMTQSPATLSVSPGERATLSCRASQGVRNNLAWYQHKPGQAPRLLIYHASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGLQPITFGGGTKLEIK 67 1D6 _VL EIVMTQSPATLSVSPGERATLSCRASQGVKNNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGEYPLTFGGGTKLEIK 68 2A5 V _L EIVMTQSPATLSVSPGERATLSCRASQNVKNNLAWYQHKPGQAPRLLIYHASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGAEPITFGGGTKLEIK 69 4A2 V _L EIVMTQSPATLSVSPGERATLSCRASQDVRNNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGTNPLTFGGGTKLEIK 78 7H5, 6G4, 5F3 V _L EIVMTQSPATLSVSPGERATLSCRASQNVKNNLAWYQHKPGQAPRLLIYHASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGNEPITFGGGTKLEIK 70 4H9 _VL EIVMTQSPATLSVSPGERATLSCRASQGVSSNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGHQPITFGGGTKLEIK 71 5B4 V _L EIVMTQSPATLSVSPGERATLSCRASQDVRDNLAWYQHKPGQAPRLLIYHASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGETPITFGGGTKLEIK 72 5E6 _VL EIVMTQSPATLSVSPGERATLSCRASQNVKGNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGHDPVTFGGGTKLEIK 73 6G3 _VL EIVMTQSPATLSVSPGERATLSCRASQDVTNNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGDLPITFGGGTKLEIK 75 2B4 V _L EIVMTQSPATLSVSPGERATLSCRASQNVKGNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGNQPITFGGGTKLEIK 76 5C7 V _L EIVMTQSPATLSVSPGERATLSCRASQSVNNNLAWYQHKPGQAPRLLIYNASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGTTPITFGGGTKLEIK 77 5F8 V _L EIVMTQSPATLSVSPGERATLSCRASQNVSNNLAWYQHKPGQAPRLLIYAASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGMGPITFGGGTKLEIK 79 9F3 _VL EIVMTQSPATLSVSPGERATLSCRASQNVKNNLAWYQHKPGQAPRLLIYHASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGTHPITFGGGTKLEIK Table 4 Exemplary sequences SEQ ID NO: describe sequence 80 full-length PcrV MEVRNLNAARELFLDELLAASAAPASAEQEELLALLRSERIVLAHAGQPLSEAQVLKALAWLLAANPSAPPGQGLEVLREVLQARRQPGAQWDLREFLVSAYFSLHGRLDEDVIGVYKDVLQTQDGKRKALLDELKALTAELKVYSVIQSQINAALSAKQGIRIDAGGIDLVDPTLYGYAVGDPRWKDSPEYALLSNLDTFSGKLSIKDFLSGSPKQSGELKGLSDEYPFEKDNNPVGNFATTVSDRSRPLNDKVNEKTTLLNDTSSRYNSAVEALNRFIQKYDSVLRDILSAI 81 light chain constant region RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 82 IgG1 heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 83 IgG4 heavy chain constant region ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK [binding affinity]

Binding affinity can be expressed in terms of Kd, Koff, Kon or Ka. As used herein, the term "Koff" refers to the rate constant of dissociation of an antibody from an antigen/antibody complex, as determined by a kinetic selection device. The term "Kon" refers to the binding rate constant for the binding of an antibody to an antigen to form an antigen/antibody complex. As used herein, the dissociation constant "Kd" refers to the dissociation constant of a specific antibody-antigen interaction, and refers to the antigen concentration required when the antigen occupies half of all antibody binding sites and reaches equilibrium in the antibody molecule solution, which is equal to Koff/ Kon. The determination of Kd assumes that all bound molecules are in solution. In cases where the antibody is attached to the cell wall, such as in yeast expression systems, the corresponding dissociation rate constant is expressed in terms of EC50, which is a good approximation of Kd. The affinity binding constant Ka is the inverse of the dissociation constant Kd.

The equilibrium dissociation constant (Kd) can be used as an indicator of the affinity of the reacting antibody moiety to the antigen. For example, simple analysis by Scatchard method using antibodies labeled with various labels, and Biacore instrument (manufactured by Amersham Biosciences), analysis of interactions between biomolecules by surface plasmon resonance according to the user manual or an accompanying kit . The Kd values obtained using these methods are expressed in units of M. Antibodies that specifically bind to the target may have, for example, ≤ ^10-7 M, ≤ ^10-8 M, ≤ ^10-9 M, ≤ ^10-10 M, ≤ ^10-11 M, ≤ ^10-12 M ^, or ≤ 10- Kd value of ¹³ M.

The binding specificity of an antibody can be determined experimentally by methods known in the art. These methods include, but are not limited to, Western blots, ELISA-, RIA-, ECL-, IRMA-, EIA-, BIAcore tests, and peptide scanning, among others.

In some embodiments, the anti-PcrV antibody specifically binds to a PcrV target with a Kd value of ^10-7 M to ^10-13 M (eg, ^10-7 M to ^10-13 M, ^10-8 M to ^10-13 M, 10 ^-9 M to 10 ^-13 M or 10 ^-10 M to 10 ^-12 M). Thus, in some embodiments, the Kd value for binding between the anti-PcrV antibody and PcrV is 10 ⁻⁷ M to 10 ⁻¹³ M, 1×10 ⁻⁷ M to 5×10 ⁻¹³ M, 10 ⁻⁷ M to 10 ⁻¹² M, 10 ⁻⁷ M to 10 ⁻¹¹ M, 10 ⁻⁷ M to 10 ⁻¹⁰ M, 10 ⁻⁷ M to 10 ⁻⁹ M, 10 ⁻⁸ M to 10 ⁻¹³ M, 1×10 ⁻⁸ M to 5×10 ⁻¹³ M, 10 ⁻⁸ M to 10 ⁻¹² M, 10 ⁻⁸ M to 10 ⁻¹¹ M, 10 ⁻⁸ M to 10 ⁻¹⁰ M, 10 ⁻⁸ M to 10 ⁻⁹ M, 5× 10 ⁻⁹ M to 1×10 ⁻¹³ M, 5×10 ⁻⁹ M to 1×10 ⁻¹² M, 5×10 ⁻⁹ M to 1×10 ⁻¹¹ M, 5×10 ⁻⁹ M-1×10 ⁻¹⁰ M, 10 ⁻⁹ M to 10 ⁻¹³ M, 10 ⁻⁹ M to 10 ⁻¹² M, 10 ⁻⁹ M to 10 ⁻¹¹ M, 10 ⁻⁹ M to 10 ⁻¹⁰ M, 5×10 ⁻¹⁰ M to 1×10 ⁻¹³ M, 5×10 ⁻¹⁰ M to 1×10 ⁻¹² M, 5×10 ⁻¹⁰ M to 1×10 ⁻¹¹ M, 10 ⁻¹⁰ M to 10 ⁻¹³ M, 1×10 ^{− 10} M to 5×10 ⁻¹³ M, 1×10 ⁻¹⁰ M to 1×10 ⁻¹² M, 1×10 ⁻¹⁰ M to 5×10 ⁻¹² M, 1×10 ⁻¹⁰ M to 1×10 ⁻¹¹ M, 10 ⁻¹¹ M to 10 ⁻¹³ M, 1×10 ⁻¹¹ M to 5×10 ⁻¹³ M, 10 ⁻¹¹ M to 10 ⁻¹² M, 10 ⁻¹² M to 10 ⁻¹³ M. In some embodiments, the Kd value for binding between the anti-PcrV antibody and PcrV is ^10-7M to ^10-13M .

In some embodiments, the Kd value for binding between the anti-PcrV antibody and the non-target is higher than the Kd value for the anti-PcrV antibody and the target, and in some embodiments cited herein, the anti-PcrV antibody binds to the target (eg, PcrV) The binding affinity is higher than the binding affinity of the PcrV antibody to the non-target. In some embodiments, non-target refers to a non-PcrV antigen. In some embodiments, the anti-PcrV antibody (against PcrV) has a Kd value for binding to a non-PcrV target that is at least 10 times the Kd value for binding between the anti-PcrV antibody and the target PcrV, eg, 10-100-fold, 100-1000-fold, 10-fold ³ -10 ⁴ times, 10 ⁴ -10 ⁵ times, 10 ⁵ -10 ⁶ times, 10 ⁶ -10 ⁷ times, 10 ⁷ -10 ⁸ times, 10 ⁸ -10 ⁹ times, 10 ⁹ -10 ¹⁰ times, 10 ¹⁰ -10 ¹¹ times, 10 ¹¹ -10 ¹² times. [nucleic acid]

Nucleic acid molecules encoding anti-PcrV antibodies are also contemplated. In some embodiments, a nucleic acid (or set of) encoding a full-length anti-PcrV antibody is provided, including any of the full-length anti-PcrV antibodies described herein. In some embodiments, the nucleic acid (or set of nucleic acids) of the anti-PcrV antibodies described herein may also include nucleic acid sequences encoding polypeptide tags (eg, protein purification tags, His-tags, HA tags).

Also contemplated herein are isolated host cells comprising anti-PcrV antibodies, isolated nucleic acids encoding anti-PcrV antibody polypeptide components, or vectors comprising nucleic acids encoding anti-PcrV antibody polypeptide components described herein.

The invention also includes variants of these nucleic acid sequences. For example, variants include nucleotide sequences that hybridize to a nucleic acid sequence encoding an anti-PcrV antibody of the invention at least under moderately stringent hybridization conditions.

The present invention also provides vectors into which the nucleic acid sequences of the present invention can be inserted.

Briefly, a natural or synthetic nucleic acid encoding an anti-PcrV antibody is inserted into a suitable expression vector such that the nucleic acid is operably linked to 5' and 3' regulatory elements, including, for example, promoters (e.g., lymphocyte-specific). promoter) and 3' untranslated region (UTR), which can express anti-PcrV antibodies (eg full-length anti-PcrV antibodies). The vectors are suitable for replication and integration in eukaryotic host cells. Typical colonization and expression vectors contain transcriptional and translational terminators, initiation sequences, and promoters that regulate the expression of the nucleic acid sequence of interest.

The nucleic acids of the present invention can also be used for nucleic acid immunization and gene therapy using standard gene delivery protocols. Nucleic acid delivery methods are known in the art. See, eg, U.S. Pat. Nos. 5,399,346, 5,580,859, 5,589,466, incorporated herein by reference in their entirety. In some embodiments, the present invention also provides gene therapy vectors.

Nucleic acids can be cloned into many types of vectors. For example, nucleic acids can be cloned into vectors including, but not limited to, plasmids, phagemids, phage derivatives, animal viruses, and cosmids. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

In addition, expression vectors can be provided to cells in the form of viral vectors. Viral vector technology is well known in the art and is described, for example, in Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and other virology or molecular biology manuals. Viruses that can be used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses. In general, suitable vectors include an origin of replication functional in at least one organism, promoter sequences, convenient restriction endonuclease sites, and one or more selectable markers (see, eg, WO 01/96584; WO 01/29058; and US Pat. No. 6,326,193)

Numerous virus-based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. The selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art. The recombinant virus is then isolated and delivered to the subject's cells in vivo or in vitro. Many retroviral systems are known in the art. In some embodiments, adenoviral vectors are used. Many adenoviral vectors are known in the art. In some embodiments, lentiviral vectors are used. Vectors derived from retroviruses, such as lentiviruses, are suitable tools for long-term gene transfer because they allow long-term stable integration of the transgene and propagation in progeny cells. Lentiviral vectors have additional advantages over tumor-derived retroviruses such as mouse leukemia virus because they can transduce non-dividing cells such as hepatocytes. At the same time, it has the additional advantage of low immunogenicity.

Other promoter elements, eg, enhancers, regulate transcription initiation frequency. Typically they are located 30-110 bp upstream of the initiation site, although many promoters have recently been found to also contain functional elements downstream of the initiation site. The spacing between promoter elements is generally flexible so that promoter function is maintained when elements are interchanged or moved relative to each other. In the thymidine kinase (tk) promoter, activity begins to decline only when the spacing between promoter elements increases to 50 bp.

An example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. The promoter sequence is a strong constitutive promoter sequence that can drive high-level expression of any polynucleotide sequence to which it is operably linked. Another example of a suitable promoter is the elongation factor 1α (EF-1α) promoter. However, other constitutive promoters can also be used, including, but not limited to, simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus long terminal repeat (HIV-LTR) promoter promoter, MoMuLV promoter, avian leukemia virus promoter, Epstein-Barr virus immediate early promoter, Rous sarcoma virus promoter, and human gene promoters such as, but not limited to, actin promoter, myosin promoter promoter, hemoglobin promoter and creatine kinase promoter. Furthermore, the present invention should not be limited to the use of only constitutive promoters. Inducible promoters are also contemplated by the present invention. The use of an inducible promoter provides a molecular switch that turns on the expression of a polynucleotide sequence to which it is operably linked when such expression is desired and turns off expression when it is not needed. Inducible promoters include, but are not limited to, metallothionein promoters, glucocorticoid promoters, progesterone promoters, and tetracycline promoters.

In some embodiments, the expression of the anti-PcrV antibody is inducible. In some embodiments, the nucleic acid sequence encoding an anti-PcrV antibody is operably linked to an inducible promoter, including any of the inducible promoters described herein. [inducible promoter]

The use of an inducible promoter provides a molecular switch that turns on expression of a polynucleotide sequence operably linked to it when expression is desired, and turns off expression when expression is not desired. Exemplary inducible promoters suitable for use in eukaryotic cells include, but are not limited to, hormone regulatory elements (see, e.g., Mader, S. and White, JH (1993) Proc. Natl. Acad. Sci. USA 90:5603-5607 ), synthetic ligand regulatory elements (see Spencer, DM et al (1993) Science 262: 1019-1024), and ionizing radiation regulatory elements (see Manome, Y. et al. (1993) Biochemistry 32: 10607-10613; Datta, R. et al. (1992) Proc. Natl. Acad. Sci. USA 89: 1014-10153). See Gingrich et al. (1998) Annual Rev. Neurosci 21:377-405 for other exemplary inducible promoters suitable for use in in vivo or in vitro mammalian systems. In some embodiments, the inducible promoter system used to express the anti-PcrV antibody is the Tet system. In some embodiments, the inducible promoter system for expressing anti-PcrV antibodies is the E. coli lac suppression system.

An exemplary inducible promoter system employed in the present invention is the Tet system. The system is based on the Tet system described by Gossen et al. (1993). In an exemplary embodiment, the polynucleotide of interest is under the control of a promoter comprising one or more Tet operon (TetO) sites. In the inactive state, the Tet repressor (TetR) binds to the TetO site and represses transcription from the promoter. In the activated state, eg, in the presence of an inducer such as tetracycline (Tc), anhydrotetracycline, doxycycline (Dox), or an active analog thereof, the inducer releases TetR from TetO, causing transcription to occur. Doxycycline is a member of the tetracycline antibiotic family, its chemical name is 1-dimethylamino-2,4a,5,7-pentahydroxy-11-methyl-4,6-dioxy-1,4a, 11,11a,12,12a-hexahydrotetraene-3-carboxamide.

In one embodiment, TetR is codon optimized for expression in mammalian cells, such as mouse or human cells. Due to the degeneracy of the genetic code, most amino acids are encoded by more than one codon, allowing a large number of variations in the sequence of a given nucleic acid without any change in the amino acid sequence it encodes. However, many organisms differ in codon usage, also known as "codon bias" (ie, the preference for using a particular codon for a given amino acid). Codon bias is often associated with the presence of dominant tRNA species for a particular codon, which in turn increases the efficiency of mRNA translation. Coding sequences derived from a particular species (eg, prokaryotes) can thus be tailored by codon optimization to improve their expression in different species (eg, eukaryotes).

Other specific variants of the Tet system include the "Tet-Off" and "Tet-On" systems below. In the Tet-off system, transcription is inactivated in the presence of Tc or Dox. In this system, a tetracycline-regulated transcriptional activator (tTA) composed of TetR fused to the strong transcriptional activation domain of herpes simplex virus VP16 regulates the expression of target nucleic acids under the transcriptional control of a tetracycline-responsive promoter element (TRE). The TRE element consists of a TetO sequence fused in tandem to a promoter (usually the minimal promoter sequence derived from the human cytomegalovirus immediate early promoter). In the absence of Tc or Dox, tTA binds TRE and activates transcription of target genes. In the presence of Tc or Dox, tTA cannot bind TRE and target genes cannot be expressed.

In contrast, in the Tet-On system, transcription is activated in the presence of Tc or Dox. The Tet-On system is based on the reverse tetracycline-regulated transcriptional activator rtTA. Like tTA, rtTA is a fusion protein consisting of the TetR repressor and the VP16 transactivation domain. However, a 4-amino acid change in the DNA-binding region of TetR altered the binding properties of rtTA so that it could only recognize the tetO sequence on the target transgene TRE in the presence of Dox. Therefore, in the Tet-On system, rtTA can activate the transcription of TRE-regulated target genes only in the presence of Dox.

Another inducible promoter system is the lac repressor system of E. coli ( see Brown et al., Cell 49:603-612 (1987)). The Lac repressor system functions by regulating the transcription of a polynucleotide of interest operably linked to a promoter comprising the lac operon (lacO). The Lac repressor (lacR) binds to LacO, thereby preventing transcription of the target polynucleotide. Expression of the polynucleotide of interest is induced by a suitable inducer, eg, isopropyl-β-D thiogalactopyranoside (IPTG).

To assess expression of a polypeptide or portion thereof, the expression vector to be introduced into a cell may also contain a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from a population of cells transfected or infected with the viral vector. In other aspects, selectable markers can be carried on separate DNA fragments and used in co-transfection experiments. Either the selectable marker gene or the reporter gene can be flanked by appropriate regulatory sequences to enable expression in the host cell. Useful selectable markers include, for example, antibiotic resistance genes such as neo and the like.

Reporter genes can be used to identify potentially transfected cells and evaluate the function of regulatory sequences. Typically, a reporter gene is a gene not present in or expressed by the recipient organism or tissue that encodes a polypeptide whose expression is manifested by some readily detectable property, such as enzymatic activity. After the DNA is introduced into the recipient cells, the expression of the reporter gene is detected at the appropriate time. Suitable reporter genes may include genes encoding luciferase, beta-galactosidase, chloramphenicol acetyltransferase, secreted alkaline phosphatase or green fluorescent protein ( eg , Ui-Tel et al. , 2000 FEBS Letters 479: 79-82). Suitable expression systems are well known and can be prepared by known techniques or obtained commercially. In general, a construct with the smallest 5' flanking region capable of showing the highest level of expression of the reporter gene is designated as a promoter. Such promoter regions can be linked to reporter genes and used to assess the ability of certain substances in regulating promoter-driven transcription.

In some embodiments, nucleic acids encoding any of the full-length anti-PcrV antibodies described herein are provided. In some embodiments, the nucleic acid includes one or more nucleic acid sequences encoding full-length anti-PcrV antibody heavy and light chains. In some embodiments, each of the one or more nucleic acid sequences is contained in a separate vector. In some embodiments, at least some of the nucleic acid sequences are contained in the same vector. In some embodiments, all nucleic acid sequences are contained in the same vector. The vector can be selected from, eg, mammalian expression vectors and viral vectors (eg, vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses).

Methods for introducing genes into cells and expressing them are known in the art. In the context of expression vectors, the vectors can be readily introduced into host cells, such as mammalian cells, bacteria, yeast or insect cells, by any method known in the art. For example, expression vectors can be introduced into host cells by physical, chemical or biological methods.

Physical methods of introducing polynucleotides into host cells include calcium phosphate precipitation, lipofection, biolistic methods, microinjection, electroporation, and the like. Methods for preparing cells comprising vectors and/or exogenous nucleic acids are well known in the art. See, eg, Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). In some embodiments, the polynucleotide is introduced into the host cell by calcium phosphate transfection.

Biological methods for introducing a polynucleotide of interest into host cells include the use of DNA and RNA vectors. Viral vectors, especially retroviral vectors, have become the most widely used method of inserting genes into mammalian cells, such as human cells. Other viral vectors can be derived from lentiviruses, poxviruses, herpes simplex virus type 1, adenoviruses and adeno-associated viruses, among others. See, eg, U.S. Pat. Nos. 5,350,674 and 5,585,362.

Chemical methods for introducing polynucleotides into host cells include colloidal dispersion systems such as polymer complexes, nanocapsules, microspheres, magnetic beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed gels Agglomerates and liposomes. An exemplary colloidal system used as a delivery vehicle in vivo and in vitro is liposomes (eg, artificial membrane vesicles).

Where non-viral delivery systems are used, exemplary delivery vehicles are liposomes. Consider using lipid formulations to introduce nucleic acids into host cells (in vitro, ex vivo, or in vivo). In another aspect, the nucleic acid can be bound to a lipid. Nucleic acids bound to lipids can be encapsulated into the aqueous interior of liposomes, dispersed in the lipid bilayer of liposomes, linked to liposomes by linking molecules bound to liposomes and oligonucleotides, and embedded in liposomes, Complexes are formed with liposomes, dispersed in lipid-containing solutions, mixed with lipids, associated with lipids, suspended in lipids, contained in or mixed with micelles, or otherwise associated with lipids. The lipid, lipid/DNA or lipid/expression vector-related composition in solution is not limited to any particular structure. For example, they may exist as bilayers, as micelles, or as "collapsed" structures. They can also simply be dispersed in solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances that can be naturally occurring or synthetic lipids. For example, lipids include lipid droplets that are naturally present in the cytoplasm, as well as a class of compounds containing long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, aminoalcohols, and aldehydes.

Regardless of the method used to introduce the exogenous nucleic acid into the host cell or otherwise expose the cell to the inhibitors of the invention, a variety of experiments can be performed in order to confirm the presence of the recombinant DNA sequence in the host cell. Such experiments include, for example, "molecular biology" experiments well known to those skilled in the art. Examples include Southern and Northern blotting, RT-PCR and PCR; "biochemical" experiments, such as the detection of the presence or absence of a particular polypeptide, such as by immunological methods (ELISAs and Western blots) or by the experiments described herein Identification is within the scope of the present invention. [Preparation of anti-PcrV antibody]

In some embodiments, the anti-PcrV antibody is or is derived from a monoclonal antibody. In some embodiments, the anti-PcrV antibody comprises _VH and _VL from a monoclonal antibody, or a variant thereof. In some embodiments, the anti- _PcrV antibody further comprises CH1 and _CL regions from a monoclonal antibody, or a variant thereof. Monoclonal antibodies can be prepared, for example, using methods known in the art, including hybridoma cell methods, phage display methods, or using recombinant DNA methods. Additionally, exemplary phage display methods are described herein and in the Examples below.

In the hybridoma cell method, a hamster, mouse, or other suitable host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that specifically bind to the immunizing agent. Alternatively, lymphocytes can be immunized in vitro. The immunizing agent can include a polypeptide or fusion protein of the protein of interest. Typically, peripheral blood lymphocytes (PBLs) are used if cells of human origin are desired, while splenocytes or lymph node cells are used if cells of non-human mammalian origin are desired. Lymphocytes are fused with an immortal cell line using an appropriate fusion agent, such as polyethylene glycol, to form hybridoma cells. Immortal cell lines are usually transformed mammalian cells, especially myeloma cells of rodent, bovine and human origin. Rat or mouse myeloma cell lines are typically used. Hybridoma cells can be cultured in a suitable medium, preferably containing one or more substances that inhibit the growth or survival of unfused immortal cells. For example, if the parental cells lack the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT), the hybridoma cell culture medium typically includes hypoxanthine, aminopterin, and thymidine (HAT medium), which prevents HGPRT Defective cell growth.

In some embodiments, the immortalized cell line fuses efficiently, ensures stable expression of the antibody at high levels by selected antibody-producing cells, and is sensitive to certain media, such as HAT media. In some embodiments, the immortalized cell line is a mouse myeloma cell line, available from, eg, the Salk Cell Collection, San Diego, California, and the American Type Culture Collection, Manassas, Virginia. Human myeloma and mouse-human hybrid myeloma cell lines are also described for the production of human monoclonal antibodies.

The culture medium in which the hybridoma cells are cultured can then be assayed for the presence or absence of monoclonal antibodies directed against the polypeptide. The binding specificity of monoclonal antibodies produced by hybridoma cells can be determined by immunoprecipitation or in vitro binding experiments such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or analytical methods are known in the art. The binding affinity of a monoclonal antibody can be determined by Scatchard analysis, eg, as described in Munson and Pollard, Anal. Biochem., 107:220 (1980).

After the desired hybridoma cells are identified, the target colony can be sub-selected by limiting dilution and cultured by standard methods. Suitable media for this purpose include, for example, modified Eagle's medium (DMEM) and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in the mammalian body in the form of ascites fluid.

Subcolonized secreted monoclonal antibodies can be isolated or purified from culture medium or ascites by conventional immunoglobulin purification methods, such as protein A-sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis or affinity layers analysis.

In some embodiments, according to any of the anti-PcrV antibodies described herein, the anti-PcrV antibody comprises a sequence selected from a clone of an antibody library (eg, a phage library displaying scFv or Fab fragments). The colonization can be identified by screening a combinatorial library of antibody fragments with the desired activity. For example, various methods are known in the art for generating phage display libraries and screening these libraries for antibodies with desired binding properties. These methods are reviewed, for example, in Hoogenboom et al. , Methods in Molecular Biology 178:1-37 (O'Brien et al. , ed., Human Press, Totowa, NJ, 2001), and in, for example, McCafferty et al. , Nature 348:552-554; Clackson et al. , Nature 352: 624-628 (1991); Marks et al. , J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al. , J. Mol. Biol. 338(2): 299-310 (2004); Lee et al. , J. Mol. Biol . 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al. , J. Immunol . Methods 284(1-2): 119-132 (2004) are further described.

In some phage display methods, the repertoires of the _VH and _VL genes are separately cloned by polymerase chain reaction (PCR) and randomly recombined in a phage library, followed by screening for antigen-binding phages, such as Winter et al . al. , Ann. Rev. Immunol. , 12: 433-455 (1994). Phages typically display antibody fragments as scFv fragments or as Fab fragments. Immunization-derived library phages provide high-affinity antibodies to the immunogen without the need for construction of hybridoma cells. Alternatively, natural repertoires (eg, from humans) can be selected to provide a single source of antibodies against multiple non-self-antigens and self-antigens without any immunization, as in Griffiths et al. , EMBO J , 12: 725-734 (1993 ) described in. Finally, natural libraries can also be prepared by cloning non-rearranged V-gene fragments from stem cells, using PCR primers containing random sequences to encode the CDR3 hypervariable region and rearranging in vitro, as described by Hoogenboom and Winter, J Mol. Biol ., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example, US Pat. /0292936 and 2009/0002360.

The anti-PcrV antibody is prepared by phage display screening the library for the anti-PcrV antibody portion that can specifically bind to the target PcrV. The library may be a human scFv phage display library with at least 1 x 10 ⁹ (eg at least 1 x 10 ⁹ , 2.5 x 10 ⁹ , 5 x 10 ⁹ , 7.5 x 10 ⁹ , 1 x 10 ¹⁰ , 2.5 x 10 ¹⁰ , 5 × 10 ¹⁰ , 7.5 × 10 ¹⁰ or 1 × 10 ¹¹ ) diversity of unique human antibody fragments. In some embodiments, the library is a human native library constructed from DNA extracted from PMBCs and spleen of healthy subjects, comprising all human heavy and light chain subfamilies. In some embodiments, the library is a natural human library constructed from DNA extracted from PMBCs isolated from patients with various diseases, such as patients with autoimmune diseases, cancer patients, and patients with infectious diseases. In some embodiments, the library is a semi-synthetic human library in which the heavy chain CDR3s are completely random, with all amino acids (except cysteine) present at any given position with equal probability. (See, eg, Hoet, RM et al. , Nat. Biotechnol. 23(3):344-348, 2005). In some embodiments, the semi-synthetic human library has heavy chain CDR3s ranging from 5 to 24 in length (eg, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23 or 24) amino acids. In some embodiments, the library is a fully synthetic phage display library. In some embodiments, the library is a non-human phage display library.

Phage colonization with high affinity for the target PcrV can be screened by iterative binding of the phage to the target PcrV bound to a solid support (such as beads for solution panning or mammalian cells for cell panning). cells), followed by removal of unbound phage, and elution of specifically bound phage. Subsequently, the bound phage are eluted for colonization and used to infect suitable host cells, such as E. coli XL1-Blue, for expression and purification. Colonization can be performed by multiple rounds of panning (eg, 2, 3, 4, 5, 6, or more), eg, solution panning, cell panning, or a combination of both, to enrich for phage that specifically bind PcrV. Specific binding of enriched phage colonies to the target PcrV can be detected by any method known in the art, including, for example, ELISA and FACS.

Monoclonal antibodies can also be prepared by recombinant DNA methods, eg, as described in U.S. Patent No. 4,816,567. DNA encoding the monoclonal antibodies of the present invention can be readily isolated and sequenced by conventional methods (eg, by oligonucleotide probes that specifically bind to genes encoding the light and heavy chains of murine antibodies). Hybridoma cells as described above or PcrV-specific phage colonization of the present invention can be used as a source of such DNA. After isolation, the DNA can be placed in an expression vector, which is then transfected into host cells, such as simian COS cells, Chinese hamster ovary cancer (CHO) cells, or non-immunoglobulin-producing myeloma cells, obtained in recombinant hosts. Monoclonal antibodies synthesized in cells. The DNA can also be modified, for example, by substituting homologous non-human sequences with coding sequences for human heavy and light chain constant structural and/or framework regions (US Pat. No. 4,816,567; Morrison et al., supra), or by substituting non-human sequences. All or part of the coding sequence for an immunoglobulin polypeptide is covalently linked to the immunoglobulin coding sequence. Such non-immunoglobulin polypeptides can replace the constant regions of the antibodies of the invention, or can replace an antigen binding site in the variable domains of the antibodies of the invention, to form chimeric bivalent antibodies.

The antibody may be a monovalent antibody. Methods of making monovalent antibodies are known in the art. For example, a recombinant expression method involving immunoglobulin light chains and modified heavy chains. Heavy chains are typically truncated anywhere in the Fc region to prevent cross-linking of the heavy chains to each other. Alternatively, relevant cysteine residues are substituted with other amino acid residues or deleted to prevent cross-linking.

In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce antibody fragments, particularly Fab fragments, can be accomplished using any method known in the art.

Antibody variable domains with the desired binding specificities (antibody-antigen binding sites) can be fused to immunoglobulin constant regions. Preferably, the fusion is to an immunoglobulin heavy chain constant region, which includes at least part of the hinge, CH2 and CH3 regions. In some embodiments, the first heavy chain constant region (CH1) comprising the site necessary for light chain binding is present in at least one fusion. The DNA encoding the immunoglobulin heavy chain fusion, and if desired, the DNA encoding the immunoglobulin light chain, is inserted into a separate expression vector and co-transfected into a suitable host organism. [Fully and Humanized Antibodies]

The anti-PcrV antibody (eg, full-length anti-PcrV antibody) can be a humanized antibody or a fully human antibody. Humanized forms of non-human (eg, mouse) antibody portions are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (eg, Fv, Fab, Fab', F(ab') ₂ , scFv, or other antigen-binding subsequences), which typically include minimal sequence derived from non-human immunoglobulins. Humanized antibodies include human immunoglobulins, immunoglobulin chains, or fragments thereof (acceptor antibodies) in which residues of the acceptor CDRs are replaced by non-human (donor antibody) CDRs with the desired specificity, affinity, and properties Residue substitutions such as mouse, rat or rabbit CDRs. In some embodiments, human immunoglobulin Fv framework region residues are replaced with corresponding non-human residues. Humanized antibodies may also contain amino acid residues that are neither in the acceptor antibody nor in the sequences of the introduced CDRs or framework regions. Typically, a humanized antibody comprises at least one, usually two variable domains, wherein all or substantially all of the CDR regions correspond to the CDR regions of a non-human immunoglobulin and all or substantially all of the framework regions are human immunoglobulin consensus sequences .

Typically, humanized antibodies contain one or more amino acid residues introduced from non-human sources. Those non-human amino acid residues are often referred to as "transferred in" residues, usually from a "transferred in" variable domain. According to some embodiments, humanization can be performed substantially as follows by Winter and colleagues (Jones et al. , Nature, 321: 522-525 (1986); Riechmann et al. , Nature, 332: 323-327 ( 1988); Verhoeyen et al. , Science, 239: 1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of human antibodies. Thus, such "humanized" antibody portions (US Patent No. 4,816,567), which are substantially less than fully human antibodies, have variable domains that have been replaced by corresponding sequences from non-human sources. In practice, humanized antibody portions are typically human antibody portions in which some CDR residues and possibly some framework region residues are replaced by residues from analogous sites in rodent antibodies.

Fully human antibodies are an alternative to humanization. For example, it is currently possible to generate transgenic animals (eg, mice) that, upon immunization, produce complete libraries of fully human antibodies without producing endogenous immunoglobulins. For example, homozygous deletion of the antibody heavy chain joining region (JH) gene in chimeric and germline mutant mice has been reported to completely inhibit endogenous antibody production. Transfer of human germline immunoglobulin gene arrays into such germline mutant mice produces fully human antibodies upon antigenic stimulation, see, e.g., akobovits et al. , PNAS USA, 90:2551 (1993); Jakobovits et al ., Nature, 362:255-258 (1993); Bruggemann et al. , Year in Immunol., 7:33 (1993); US Patent Nos. 5,545,806, 5,569,825, 5,591,669; 5,545,807; Alternatively, fully human antibodies can be prepared by introducing human immunoglobulin loci into transgenic animals (eg, mice in which the endogenous immunoglobulin genes have been partially or fully silenced). Following antigenic stimulation, fully human antibody production can be found to be very similar to its production in humans in all respects, including gene rearrangement, assembly, and antibody libraries. and 5,661,016, and Marks et al., Bio/Technology, 10: 779-783 (1992); Lonberg et al., Nature, 368: 856-859 (1994); Morrison, Nature, 368: 812-813 (1994); Fishwild et al., Nature Biotechnology, 14: 845-851 (1996); Neuberger, Nature Biotechnology, 14: 826 (1996); Lonberg and Huszar, Intern. Rev. Immunol., 13: 65-93 (1995).

Fully human antibodies can also be produced by in vitro activation of B cells (see US Patents 5,567,610 and 5,229,275) or by using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Techniques of Cole et al. and Boerner et al. It can also be used to prepare fully human monoclonal antibodies. See Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985) and Boerner et al., J. Immunol., 147(1): 86-95 (1991). [Anti-PcrV antibody variant]

In some embodiments, the amino acid sequences of the anti-PcrV antibody variants provided herein (eg, full-length anti-PcrV antibodies) are also contemplated. For example, it may be desirable to improve the binding affinity and/or other biological activities of the antibody. The amino acid sequences of antibody variants can be prepared by introducing appropriate modifications in the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and/or insertions and/or substitutions of residues in the amino acid sequence of the antibody. The final construction can be accomplished by any combination of deletions, insertions and substitutions of amino acid residues to have the desired characteristics. For example, antigen binding.

In some embodiments, anti-PcrV antibody variants with one or more amino acid substitutions are provided. Target sites for substitution mutations include hypervariable regions (HVRs) and framework regions (FRs). Amino acid substitutions can be introduced into the antibody of interest and the product screened for the desired activity, e.g., improved biological activity, maintained/improved antigen binding capacity, reduced immunogenicity, or improved opsonophagocytic killing (OPK) of pathogenic bacteria, e.g., Pseudomonas aeruginosa.

Conservative substitutions are shown in Table 5 below Table 5 Conservative substitutions original residue Exemplary substitution better replacement Ala (A) Val; Leu; Ile Val Arg (R) Lys; Gln; Asn Lys Asn (N) Gln; His; Asp, Lys; Arg Gln Asp (D) Glu; Asn Glu Cys (C) Ser; Ala Ser Gln (Q) Asn; Glu Asn Glu (E) Asp; Gln Asp Gly (G) Ala Ala His (H) Asn; Gln; Lys; Arg Arg Ile (I) Leu; Val; Met; Ala; Phe; Norleucine Leu Leu (L) Norleucine; Ile; Val; Met; Ala; Phe Ile Lys (K) Arg; Gln; Asn Arg Met (M) Leu; Phe; Ile Leu Phe (F) Trp; Leu; Val; Ile; Ala; Tyr Tyr Pro (P) Ala Ala Ser (S) Thr Thr Thr (T) Val; Ser Ser Trp (W) Tyr; Phe Tyr Tyr (Y) Trp; Phe; Thr; Ser Phe Val (V) Ile; Leu; Met; Phe; Ala; Norleucine Leu

Amino acids are divided into different categories based on their side chain properties: Hydrophobic amino acids: Norleucine Norleucine, Methionine Met, Ala Ala, Valine Val, Leu Leu, Ile Isoleucine; Neutral hydrophilic amino acids: cysteine Cys, serine Ser, threonine Thr, asparagine Asn, glutamine Gln; Acidic amino acids: aspartic acid Asp, glutamic acid Glu; Basic amino acids: Histidine His, Lysine Lys, Arginine Arg; Amino acids that affect chain orientation: Glycine Gly, Proline Pro; Aromatic amino acids: tryptophan Trp, tyrosine Tyr, phenylalanine Phe.

Substitutions of non-conservative amino acids include substitution of one of the above classes for another class.

An exemplary substitutional variant is an affinity matured antibody, which can be conveniently generated using, for example, phage display-based affinity maturation techniques. Briefly, one or more CDR residues are mutated, variant antibody moieties are displayed on phage, and variants are screened for specific biological activities (eg, based on RBC cell lysis inhibition assays or binding affinity). Changes (eg, substitutions) in the HVRs regions can be made to obtain improved RBC cleavage-based inhibition assays or antibody affinity. Changes can be made in "hot spots" of the HVR, ie residues encoded by codons that are hypermutated during somatic maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), And/or at specific critical residues (SDRs), the resulting variants _VH and _VL were tested for binding affinity. Methods for the construction and reselection of affinity maturation from secondary libraries have been described in, for example, Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press , Totowa, NJ, (2001)).

In some affinity maturation embodiments, diversity is introduced into the variable genes selected for affinity maturation by any of a variety of methods (eg, error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis). Then create a secondary library. The library is screened to identify antibody variants with the desired affinity. Another approach to introducing diversity includes HVR-mediated approaches, in which several HVR residues (eg, 4-6 residues at a time) are randomized. HVR residues involved in antigen binding are specifically identified, eg, using alanine scanning mutagenesis or modeling. Usually the CDR-H3 and CDR-L3 regions are particularly important targets.

In some embodiments, substitutions, insertions or deletions may occur within one or more of the HVRs, so long as such changes do not substantially reduce the ability of the antibody to bind antigen. For example, conservative changes (eg, conservative substitutions provided herein) can be made in HVRs that do not substantially reduce binding affinity. These changes may occur outside of HVR "hot spots" or SDRs. In some embodiments of the variant VH and VL sequences provided above, each HVR is either unchanged or contains no more than 1, 2 or 3 amino acid substitutions.

A useful method for identifying amino acid residues or regions of an antibody that can be targeted for mutation is called "alanine scanning mutagenesis", as described in Cunningham and Wells (1989) Science, 244:1081-1085. In this method, one or a group of target residues (eg, charged residues such as arginine, aspartic acid, histidine, lysine, and glutamic acid) are replaced by neutral or negatively charged amino acids (eg, , alanine or glutamic acid) to determine whether antibody-antigen interactions are affected. Further substitutions can be introduced at the amino acid position to demonstrate that the position is functionally sensitive to the initial substitution. Alternatively or additionally, the contact sites between the antibody and the antigen are identified by the crystal structure of the antigen-antibody complex. These contact site residues and adjacent residues can be targeted or eliminated as substitution candidates. Variants are screened to determine if they have the desired properties.

Insertions of amino acid sequences, including fusions at the amino and/or carboxy termini, ranging in length from 1 residue to polypeptides containing 100 or more residues, including intrasequence insertions of 1 or more amino acid residues base. Examples of terminal insertions include antibodies with a methionine residue at the N-terminus. Other insertional variants of the antibody molecule include the fusion of an enzyme (eg, ADEPT) to the N-terminus or C-terminus of the antibody molecule or a polypeptide that increases the serum half-life of the antibody. [Fc region variant]

In some embodiments, one or more amino acid modifications are introduced into the Fc region of an antibody described herein (eg, a full-length anti-PcrV antibody or anti-PcrV antibody fusion protein), resulting in an Fc region variant. In some embodiments, the Fc region variant has enhanced ADCC potency, typically associated with Fc binding receptors (FcRs). In some embodiments, the Fc region variant has reduced ADCC potency. There are many examples of changes or mutations in Fc sequences affecting their potency, for example, WO 00/42072 and Shields et al. J Biol. Chem. 9(2): 6591-6604 (2001) describe enhanced binding to FcRs or Attenuated antibody variants. The contents of these publications are incorporated herein by reference.

Antibody-dependent cell-mediated cytotoxicity (ADCC) is the mechanism of action of therapeutic antibodies against tumor cells. ADCC is a cell-mediated immune defense in which effector cells of the immune system actively lyse target cells (eg, infected cells) when antigens on the surface of the target cell membrane are bound by specific antibodies (eg, anti-PcrV antibodies). Usually ADCC effects involve NK cells activated by antibodies. NK cells express the Fc receptor CD16. This receptor recognizes and binds to the Fc portion of an antibody molecule bound to the surface of the target cell. The most common Fc receptors on the surface of NK cells are CD16 or FcγRIII. Binding of Fc receptors to the Fc region of an antibody results in activation of NK cells, release of cytolytic particles, and subsequent apoptosis of target cells.

In some embodiments, the invention also provides anti-PcrV antibody variants (eg, full-length anti-PcrV antibody variants) comprising an Fc region with some, but not all, effector functions, such that it has an extended half-life in vivo, however specific Effector functions (eg, CDC or ADCC) are nonessential or deleterious, and such anti-PcrV antibodies make ideal candidates for the present invention. The reduction/elimination of CDC and/or ADCC activity is confirmed by performing cytotoxicity assays in vitro and/or in vivo. For example, an Fc receptor (FcR) binding assay confirms that the antibody lacks FcγR binding (and thus may lack ADCC activity) but retains FcRn binding. Among the major cells mediating ADCC, NK cells express only FcγRIII, while monocytes express FcγRI, FcγRII and FcγRIII. The expression of FcRs on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet Annu. Rev. Immunol. 9:457-492 (1991). A non-limiting example of in vitro assessment of ADCC activity of target molecules is described in US Pat. No. 5,500,362 (see, e.g., Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83:7059-7063 ( 1986)) and Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985); US Pat. No. 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, nonradioactive assays can be used (see, eg, ACTI™ Flow Cytometry Nonradioactive Cytotoxicity Assay (CellTechnology, Inc. Mountain View, Calif.) and CYTOTOX 96™ Nonradioactive Cytotoxicity Assay (Promega, Madison, Wis. )). Effector cells used in such assays include peripheral blood mononuclear cells (PBMCs) and natural killer cells (NKs). Alternatively, additionally, ADCC activity of the target molecule is detected in vivo, eg, in animal models, as described in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). A C1q binding assay can also be performed to confirm that the antibody cannot bind to C1q and thus lack CDC activity. See, eg, WO2006/029879 and WO 2005/100402 for C1q and C3c binding ELISAs. To assess complement activation, CDC assays can be performed (see, eg, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance/half-life are determined using methods known in the art (see, eg, Petkova, S. B. et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

Antibodies with reduced effector function include substitution of one or more residues at residues 238, 265, 269, 270, 297, 327 and 329 of the Fc region (U.S. Pat. No. 6,737,056). These Fc variants include Fc variants with substitutions of two or more residues at positions 265, 269, 270, 297 and 327, including the Fc variant termed "DANA", which has residues 265 and 297 substituted with alanine (US Pat. No. 7,332,581).

Such antibody variants with increased or decreased binding to FcRs have been described (see eg US Pat. No. 6,737,056; WO 2004/056312 and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)).

In some embodiments, alterations in the Fc region result in altered (ie, enhanced or reduced) opsonization, as described in Moore et al., MAbs . 2(2): 181-189 (2010).

In some embodiments, there is provided an anti-PcrV antibody (eg, full-length anti-PcrV antibody) variant comprising an Fc region variant with one or more amino acid substitutions capable of extending half-life and/or enhancing interaction with Fc receptors (FcRn ) combination. Antibodies with extended half-life and improved FcRn binding are described in US2005/0014934A1 (Hinton et al.). The Fc region of these antibodies contains one or more amino acid substitutions that enhance the binding of the Fc region to FcRn. These Fc variants comprise 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or One or more substitutions at residue 434, eg, substitution of residue 434 in the Fc region (US Pat. No. 7,371,826).

See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821 and WO 94/29351 for examples of other Fc region variants.

The present invention contemplates anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) comprising any of the Fc variants described herein, or a combination thereof. [glycosylated variants]

In some embodiments, the anti-PcrV antibodies provided herein (eg, full-length anti-PcrV antibodies) are altered to increase or decrease the degree of glycosylation of the anti-PcrV antibodies. The addition or deletion of glycosylation sites on the anti-PcrV antibody can be conveniently accomplished by altering the amino acid sequence of the anti-PcrV antibody or polypeptide portion thereof to thereby add or remove one or more glycosylation sites.

Where the anti-PcrV antibody contains an Fc region, the sugar attached to it can be changed. Natural antibodies produced by mammalian cells typically contain branched biantennary oligosaccharides, usually N-linked to the Fc region CH2 domain Asn297, see eg Wright et al., TIBTECH 15:26-32 (1997) . The oligosaccharides may comprise a variety of carbohydrates such as mannose, N-acetylglucosaminide (GlcNAc), galactose and sialic acid, and trehalose linked to GlcNAc in the "stem" portion of the biantennary oligosaccharide structure. In some embodiments, the anti-PcrV antibodies of the invention can be oligosaccharide-modified to generate anti-PcrV antibody variants with certain improved properties.

The N-glycans linked to the CH2 domain of the Fc region are heterogeneous. Antibodies or Fc fusion proteins produced in CHO cells are fucosylated by fucosyltransferase activity, see Shoji-Hosaka et al., J. Biochem. 2006, 140:777-83. Typically, a small fraction of naturally occurring afucosylated IgGs can be detected in human serum. N-glycosylation of the Fc region is important for its binding to Fc R; non-fucosylated N-glycans enhance the binding capacity of Fc to Fc RIIIa. Enhanced binding to FcRIIIa results in enhanced ADCC effect, which is beneficial in certain antibody therapeutic applications requiring cytotoxicity.

In some embodiments, enhanced effector functions may be detrimental when Fc-mediated cytotoxicity is not desired. In some embodiments, the Fc fragment or CH2 domain is aglycosylated. In some embodiments, glycosylation is prevented by mutating the N-glycosylation site in the CH2 domain.

In some embodiments, variants of anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) are provided that comprise an Fc region in which the carbohydrate structures attached to the Fc region have reduced or lack of fucose, which may enhance ADCC function. Specifically, provided herein are anti-PcrV antibodies that have reduced fucose relative to the same anti-PcrV antibodies produced by wild-type CHO cells. That is, they are characterized by lower amounts of fucose than antibodies produced by native CHO cells (eg, CHO cells that produce native glycosylated forms, CHO cells that contain the native FUT8 gene). In some embodiments, the N-linked glycans of the anti-PcrV antibody have less than 50%, 40%, 30%, 20%, 10% or 5% fucose. For example, the anti-PcrV antibody may have a fucose content of 1%-80%, 1%-65%, 5%-65%, or 20%-40%. In some embodiments, the N-linked glycans of the anti-PcrV antibody do not contain fucose, ie, wherein the anti-PcrV antibody is completely free of fucose, or free of fucose or defucosylated. The fucose content was calculated by calculating the average content of fucose within the sugar chain attached to Asn297 relative to the sum of all sugar structures (such as complex, hybrid or mannose structures) attached to Asn297 measured by MALDI-TOF mass spectrometry. The amount is determined as described in WO 2008/077546. Asn297 refers to the asparagine residue at position 297 in the Fc region (EU Fc region residue numbering system). However, due to minor sequence changes in the antibody, Asn297 can also be located ±3 amino acids upstream or downstream of position 297, i.e. between positions 294 and 300. These fucosylated variants may have enhanced ADCC function. See, eg, US Patent Publication Nos. US 2003/0157108 (Presta, L.), US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "defucosylated" or "fucose-deficient" antibody variants include US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002 US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; WO 2003/085119; 053742; WO2002/031140; Okazaki et al. J. Mol. Biol. 336: 1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004). Cell lines capable of producing defucosylated antibodies include Lec13 CHO cells lacking protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US Pat Appl No US 2003/0157108 A1, Presta, L; and WO 2004/056312 A1, Adams et al., especially Example 11), and gene knockout cell lines, such as the alpha-1,6-fucosyltransferase gene, FUT8 knockout CHO cells (see Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4): 680-688 (2006) ; and WO2003/085107).

Anti-PcrV antibody (eg, full-length anti-PcrV antibody) variants further provide bisected oligosaccharides, eg, wherein a biantennary oligosaccharide attached to the Fc region of an anti-PcrV antibody is bisected by GlcNAc. Such anti-PcrV antibody (eg, full-length anti-PcrV antibody) variants may have reduced fucosylation and/or enhanced ADCC function. Examples of such antibody variants are in WO 2003/011878 (Jean-Mairet et al.); US Pat. No. 6,602,684 (Umana et al.); US 2005/0123546 (Umana et al.), and Ferrara et al. , Biotechnology and Bioengineering, 93(5): 851-861 (2006). Also provided are variants of anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) having at least one galactose residue in the oligosaccharide linked to the Fc region. Such anti-PcrV antibody variants may have enhanced CDC function. Such variants are described, for example, in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO 1999/22764 (Raju, S.).

In some embodiments, the anti-PcrV antibody (eg, full-length anti-PcrV antibody) variant comprises an Fc region capable of binding to FcγRIII. In some embodiments, the anti-PcrV antibody (eg, full-length anti-PcrV antibody) variant comprising an Fc region has ADCC activity in the presence of human effector cells (eg, T cells), or with other antibodies having a human wild-type IgGl Fc region Enhanced ADCC activity in the presence of human effector cells compared to the same anti-PcrV antibody (eg, full-length anti-PcrV antibody). [Cysteine Engineering Variant]

In some embodiments, it is desirable to prepare cysteine-engineered anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) in which one or more amino acid residues are substituted with cysteine residues. In some embodiments, the substituted residues occur at sites accessible to the anti-PcrV antibody. By substituting cysteine for those residues, the reactive sulfhydryl group is located at an accessible site of the anti-PcrV antibody and can be used to couple the anti-PcrV antibody to other moieties, such as drug moieties or linker-drug moieties, to prepare anti-PcrV immunoconjugates as further described herein. Cysteine-engineered anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) can be prepared as described, eg, in U.S. Pat. No. 7,521,541. [derivative]

In some embodiments, the anti-PcrV antibodies provided herein (eg, full-length anti-PcrV antibodies) can be further modified to include other non-proteinaceous moieties known in the art and readily available. Suitable moieties for derivatizing anti-PcrV antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymers, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1, 3-dioxolane, poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyamino acid (homopolymer or random copolymer), dextran or poly(n-ethylene pyrrolidone) polyethylene glycol, propylene glycol homopolymers, propylene oxide/ethylene oxide copolymers, polyoxyethylated polyols (eg, glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde has advantages in manufacturing due to its stability in water. The polymers can be of any molecular weight and can be branched or unbranched. The number of polymers attached to the anti-PcrV antibody can vary, and if more than one polymer is attached, they can be the same or different molecules. In general, the amount and/or type of polymer used for derivatization can be determined based on considerations including, but not limited to, the need to improve the properties or function of the anti-PcrV antibody, whether the anti-PcrV antibody derivative is used under specific conditions treatment, etc.

In some embodiments, conjugates of anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) to non-protein moieties are provided, possibly selectively heated by exposure to radiation. In some embodiments, the non-protein moiety is a carbon nanotube (Kam et al. , Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation can be of any wavelength, including, but not limited to, wavelengths that do not harm normal cells, but at which wavelengths can heat the non-protein moiety to a temperature that kills cells close to the anti-PcrV antibody non-protein moiety conjugate. [pharmaceutical composition]

Also provided herein are compositions (e.g., pharmaceutical compositions, such as pharmaceutical compositions, such as pharmaceutical compositions, Also referred to here as preparations). In some embodiments, there is provided a pharmaceutical composition comprising any of the anti-PcrV antibodies described herein and a pharmaceutically acceptable carrier.

Suitable anti-PcrV antibodies can be obtained by mixing an anti-PcrV antibody of the desired purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Antibody preparations, prepared in the form of lyophilized preparations or liquid preparations. Acceptable carriers, excipients or stabilizers are non-toxic to recipients at the dosages and concentrations used, and include buffers such as: phosphate, citric acid, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (eg, ten octaalkyldimethylbenzylammonium chloride; hexamethylammonium chloride; benzalkonium chloride; benzethonium chloride; phenol; butanol or benzyl alcohol; alkyl parabens such as parabens methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than 10 residues) polypeptides; proteins such as serum albumin Proteins, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, aspartamine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose, or dextrin; chelating agents such as EDTA; carbohydrates such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes such as zinc-protein complexes and/or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG); exemplary formulations are described in WO98/56418 and expressly incorporated herein by reference. Lyophilized formulations suitable for subcutaneous administration are described in WO97/04801. Such lyophilized formulations can be reconstituted into high protein concentration formulations with suitable diluents, and the reconstituted formulations can be administered to the individual to be treated herein by subcutaneous administration. Cationic liposomes or liposomes can be used to deliver the anti-PcrV antibodies of the present invention to cells.

In addition to the anti-PcrV antibody (eg, full-length anti-PcrV antibody), the formulations described herein may also include one or more other active substances necessary for the treatment of specific conditions, preferably substances with complementary activities and no adverse reactions to each other. For example, in addition to anti-PcrV antibodies, it may be desirable to further include antineoplastic, growth inhibitory, cytotoxic or chemotherapeutic drugs. These molecules are present in combination in amounts effective for the intended purpose. The effective amount of these other substances depends on the amount of anti-PcrV antibody in the formulation, the type of disease or disorder or treatment, and other factors as discussed above. These drugs are generally used at the same dose and route of administration as described herein, or at 1% to 99% of the currently used dose.

The anti-PcrV antibody (eg, full-length anti-PcrV antibody) can also be entrapped in microcapsules prepared, for example, by coacervation techniques and interfacial polymerization, such as in colloidal drug delivery systems (eg, liposomes, albumin microspheres, microcapsules, respectively). Hydroxymethylcellulose or gelatin-microcapsules and poly(methyl methacrylate) microcapsules in emulsions, nanoparticles and nanocapsules) or in macroemulsions. Sustained release formulations can be prepared.

Sustained-release formulations of anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) can be prepared. Suitable examples of sustained release formulations include semipermeable matrices of solid hydrophobic polymers containing the antibody (or fragments thereof), which matrices are in the form of shaped articles, eg, films or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (eg, poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactic acid (US Pat. No. 3,773,919), L-glutamine Acid and L-glutamic acid ethyl ester copolymer, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymer such as LUPRON DEPOT ^TM (composed of lactic acid-glycolic acid copolymer and leuprolide acetate) injectable microspheres) and poly-D(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid can release molecules for more than 100 days, certain hydrogels can release proteins for shorter periods of time. When encapsulated antibodies remain in the body for long periods of time, they can denature or aggregate upon exposure to humidity at 37 °C, potentially resulting in loss of biological activity or changes in immunogenicity. According to the corresponding mechanism, rational strategies can be designed to stabilize anti-PcrV antibodies. For example, if the aggregation mechanism is found to be intermolecular SS bond formation through thiodisulfide exchange, then specific polymers can be developed by modifying sulfhydryl residues, lyophilizing in acidic solutions, controlling water content, using appropriate additives, and Matrix composition to achieve stabilization.

In some embodiments, the anti-PcrV antibody (eg, full-length anti-PcrV antibody) is formulated with citrate, sodium chloride, acetate, succinate, glycine, polysorbate 80 (Tween 80), or any of the foregoing in the combined buffer. In some embodiments, the anti-PcrV antibody is formulated in a buffer comprising from about 100 mM to about 150 mM glycine. In some embodiments, the anti-PcrV antibody is formulated in a buffer comprising from about 50 mM to about 100 mM NaCl. In some embodiments, the anti-PcrV antibody is formulated in a buffer comprising from about 10 mM to about 50 mM acetate. In some embodiments, the anti-PcrV antibody is formulated in a buffer comprising from about 10 mM to about 50 mM succinic acid. In some embodiments, the anti-PcrV antibody is formulated in a buffer comprising about 0.005% to about 0.02% polysorbate 80. In some embodiments, the anti-PcrV antibody is formulated in a buffer having a pH between 5.1 and 5.6. In some embodiments, the anti-PcrV antibody is formulated in a buffer comprising 10 mM citric acid, 100 mM NaCl, 100 mM glycine, and 0.01% polysorbate 80, wherein the pH of the formulation is 5.5.

Formulations for in vivo administration must be sterile. This can be easily achieved, for example, by applying sterile filtration membrane filtration. [Therapeutic or preventive method using anti-PcrV antibody]

In certain embodiments, a method of treating a Pseudomonas infection in an individual is provided, the method comprising administering to the individual an effective amount of a composition comprising any of the anti-PcrV antibodies described herein. In some embodiments, the method of treating a Pseudomonas infection further provides a therapeutic or prophylactic effect for a disease and/or disorder associated with a Pseudomonas infection. In certain aspects, there is provided a method of preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising any of the anti-PcrV antibodies described herein. In some embodiments, an anti-PcrV antibody (any of the anti-PcrV antibodies described above) or a pharmaceutical composition (any of the above-described pharmaceutical compositions comprising an anti-PcrV antibody) is used in the manufacture of a medicament for the treatment of a disease or disorder use in.

Diseases and/or conditions associated with Pseudomonas infection include, but are not limited to, fever, chills, fatigue, muscle and joint pain, joint swelling, headache, diarrhea, rash, wound discharge, bacteremia, acute pneumonia, intra-abdominal Infect. Further exemplary diseases include, but are not limited to, respiratory tract infections, bacteremia, septic shock, septic arthritis, enteritis, skin and soft tissue infections (eg, burn wound infections), urinary tract infections, intestinal infections, ulcerative keratitis, Chronic suppurative otitis media, mastoiditis, sinusitis and endocarditis. In some embodiments, the method of treating or preventing Pseudomonas infection reduces mortality from Pseudomonas infection.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody. In some embodiments, the anti-PcrV antibody specifically binds a linear epitope in Pseudomonas PcrV. In some embodiments, the anti-PcrV antibodies described herein specifically bind to a non-linear epitope in Pseudomonas PcrV.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising any of the amino acid sequences of SEQ ID NOs: 2-9, and an HC-CDR2 comprising any of the amino acids of SEQ ID NOs: 10-13 HC-CDR3 of the sequence; and _VL , the _VL comprising: an LC-CDR1 comprising any amino acid sequence in SEQ ID NOs: 14-25, a LC-CDR1 comprising any amino acid sequence in SEQ ID NOs: 26-29 LC-CDR2, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 30-44. In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: a HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 2-9, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NOs: HC-CDR3 of any amino acid sequence in 10-13; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 14-25, one comprising SEQ ID NOs: 26- An LC-CDR2 comprising the amino acid sequence of any of 29, and an LC-CDR3 comprising the amino acid sequence of any of SEQ ID NOs: 30-44. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 2-9, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 1 HC-CDR3 of any one of the amino acid sequences of SEQ ID NOs: 10-13; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 14-25, one comprising SEQ ID An LC-CDR2 comprising the amino acid sequence of any of NOs: 26-29, and an LC-CDR3 comprising the amino acid sequence of any of SEQ ID NOs: 30-44.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises the amino acid sequence of any one of SEQ ID NOs: 52-64; and _VL , the _VL comprises the amino acid sequence of any one of SEQ ID NOs: 65-79. In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are _VH comprising the amino _acid sequence of any one of SEQ ID NOs: _52-64 ; and _VL comprising the amino acid sequence of any one of SEQ ID NOs: 65-79. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody At position, the antibody comprises a _VH comprising the amino _acid sequence of any one of SEQ ID NOs: _52-64 ; and a _VL comprising the amino acid sequence of any one of SEQ ID NOs: 65-79.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 10 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 26, and one comprising the amino acid sequence SEQ ID NO: 26 30 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR3 of: 10; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26 LC-CDR3 of SEQ ID NO: 30.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:52 and the _VL comprising the amino acid sequence of SEQ ID NO:65. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:52 and a _VL comprising the amino acid sequence of SEQ ID NO:65.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 10, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 26, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 30, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 52, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 52; and V _L , the _VL comprises the amino acid sequence of SEQ ID NO: 65, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 65.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 26, and one comprising the amino acid sequence SEQ ID NO: 26 30 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26 LC-CDR3 of SEQ ID NO: 30. In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:65. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:65.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 26, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 30, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 56, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 56; and V _L , the _VL comprises the amino acid sequence of SEQ ID NO: 65, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 65.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 31 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 15, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 31.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody comprising The _VH of the amino acid sequence SEQ ID NO:53 and the _VL comprising the amino acid sequence SEQ ID NO:66. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:53 and a _VL comprising the amino acid sequence of SEQ ID NO:66.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 15, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 31, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 53, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 53; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 66, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 66.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 13 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and one comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of 39. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7 HC-CDR3 of: 13; and _VL comprising _: one LC-CDR1 comprising amino acid sequence SEQ ID NO: 23, one LC-CDR2 comprising amino acid sequence SEQ ID NO: 27, and one comprising amino acid sequence LC-CDR3 of SEQ ID NO: 39.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:62 and the _VL comprising the amino acid sequence of SEQ ID NO:74. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:62 and a _VL comprising the amino acid sequence of SEQ ID NO:74.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 13, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 23, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 39, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 62, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 62; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 74, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 74. In some embodiments, this method can prevent Pseudomonas infection in an individual.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 16, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence SEQ ID NO: 28 32 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 32.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:54 and the _VL comprising the amino acid sequence of SEQ ID NO:67. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:54 and a _VL comprising the amino acid sequence of SEQ ID NO:67.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 16, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 32, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:54, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:54; and V _L , the _VL comprises the amino acid sequence of SEQ ID NO: 67, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 67.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 5, and one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence SEQ ID NO: 27 LC-CDR3 of 33. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 33.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:55 and the _VL comprising the amino acid sequence of SEQ ID NO:68. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:55 and a _VL comprising the amino acid sequence of SEQ ID NO:68.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 5, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 33, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:55, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:55; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 68, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 68.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 18, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and one comprising the amino acid sequence of SEQ ID NO: 28 34 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 18, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 34.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:69. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:69.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 18, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 34, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:56; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 69, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 69.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 13 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and one comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of 43. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9 HC-CDR3 of: 13; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 43.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:64 and the _VL comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody The antibody antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:64 and a _VL comprising the amino acid sequence of SEQ ID NO:78.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 13, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 43, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:64, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:64; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 78, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 78.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 19, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence SEQ ID NO: 28 35 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 19, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 35.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:70. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:70.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 19, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 35, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:56; and V _L , the _VL comprises the amino acid sequence of SEQ ID NO: 70, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 70.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and one comprising the amino acid sequence of SEQ ID NO: 27 36 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6 and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 27, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 36.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:57 and the _VL comprising the amino acid sequence of SEQ ID NO:71. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:57 and a _VL comprising the amino acid sequence of SEQ ID NO:71.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 6, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 20, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 36, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:57, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:57; and V _L , the _VL comprises the amino acid sequence of SEQ ID NO: 71, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 71.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and one comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of 37. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 37.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:54 and the _VL comprising the amino acid sequence of SEQ ID NO:72. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:54 and a _VL comprising the amino acid sequence of SEQ ID NO:72.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 21, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 37, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:54, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:54; and V _L , the _VL comprises the amino acid sequence of SEQ ID NO: 72, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 72.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 22, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and one comprising the amino acid sequence of SEQ ID NO: 28 38 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 22, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 38.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:73. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:73.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 22, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 38, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:56; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 73, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 73.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of 39. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 39.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:58 and the _VL comprising the amino acid sequence of SEQ ID NO:74. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:58 and a _VL comprising the amino acid sequence of SEQ ID NO:74.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid-substituted variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 23, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 39, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 58, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 58; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 74, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 74.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and one comprising the amino acid sequence of SEQ ID NO: 28 40 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 40.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:59 and the _VL comprising the amino acid sequence of SEQ ID NO:75. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:59 and a _VL comprising the amino acid sequence of SEQ ID NO:75.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 8, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 21, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 40, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:59, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:59; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 75, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 75.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 24, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and one comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of 41. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 24, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR3 of SEQ ID NO: 41.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:57 and the _VL comprising the amino acid sequence of SEQ ID NO:76. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:57 and a _VL comprising the amino acid sequence of SEQ ID NO:76.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 6, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 24, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 28, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 41, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:57, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:57; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 76, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 76.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 25, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and one comprising the amino acid sequence of SEQ ID NO: 29 42 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 25, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 29 LC-CDR3 of SEQ ID NO: 42.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody comprising A _VH comprising the amino acid sequence of SEQ ID NO: 60 and a _VL comprising the amino acid sequence of SEQ ID NO: 77 are included. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:60 and a _VL comprising the amino acid sequence of SEQ ID NO:77.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 25, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 29, and one comprising the amino acid sequence SEQ ID NO: 29 LC-CDR3 of SEQ ID NO: 42, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:60, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:60; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 77, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 77.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and one comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of 43. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 43.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:60 and the _VL comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:60 and a _VL comprising the amino acid sequence of SEQ ID NO:78.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 43, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:60, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:60; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 78, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 78.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 11 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and one comprising the amino acid sequence of SEQ ID NO: 27 44 LC-CDR3. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR3 of: 11; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 44.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:79. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:56 and a _VL comprising the amino acid sequence of SEQ ID NO:79.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 44, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO:56, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO:56; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 79, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 79.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 12 _-CDR3 ; and _VL comprising: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of 39. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7 HC-CDR3 of: 12; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 39.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:61 and the _VL comprising the amino acid sequence of SEQ ID NO:74. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:61 and a _VL comprising the amino acid sequence of SEQ ID NO:74.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 12, or at most 5 amino acid-substituted variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 23, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 39, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 61, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 61; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 74, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 74.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Include a _VH comprising: one HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 9, and one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 12 _-CDR3 ; and _VL comprising: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and one comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of 43. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody comprises a _VH comprising: an HC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 1, a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 7 HC-CDR3 of: 12; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 LC-CDR3 of SEQ ID NO: 43.

In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody competes with an antibody that Included are the _VH comprising the amino acid sequence of SEQ ID NO:63 and the _VL comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, a method of treating or preventing a Pseudomonas infection in an individual is provided, comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody binds to the same table as the following antibody position, the antibody includes a _VH comprising the amino acid sequence of SEQ ID NO:63 and a _VL comprising the amino acid sequence of SEQ ID NO:78.

In some embodiments, there is provided a method of treating or preventing a Pseudomonas infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-PcrV antibody, wherein the anti-PcrV antibody comprises _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 12, or at most 5 amino acid substitution variants; and _VL comprising _: one LC-CDR1 comprising the amino acid sequence SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence LC-CDR3 of SEQ ID NO: 43, or a variant comprising up to 5 amino acid substitutions. In some embodiments, the anti-PcrV antibody comprises a _VH comprising the amino _acid sequence of SEQ ID NO: 63, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 63; and V _L , the _VL comprising the amino acid sequence of SEQ ID NO: 78, or a variant sequence having at least 90% sequence homology with the amino acid sequence of SEQ ID NO: 78.

In some embodiments, any of the methods of treatment or prevention described herein, the anti-PcrV antibody comprises an antibody heavy chain constant region and an antibody light chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgGl heavy chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgG2 heavy chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgG3 heavy chain constant region. In some embodiments, the anti-PcrV antibody comprises an IgG4 heavy chain constant region. In some embodiments, the IgG is human IgG. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:82. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:83. In some embodiments, the anti-PcrV antibody comprises a lambda light chain constant region. In some embodiments, the anti-PcrV antibody comprises a kappa light chain constant region. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence of SEQ ID NO:81. In some embodiments, the anti-PcrV antibody comprises one antibody heavy chain variable region and one antibody light chain variable region.

In some embodiments, any of the methods of treatment or prevention described herein further provides a therapeutic or prophylactic effect of a disease and/or disorder associated with a Pseudomonas infection. In some embodiments, the methods prevent Pseudomonas infection in the individual.

In some embodiments, the individual is a mammal (eg, human, non-human primate, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, etc.). In some embodiments, the individual is a human. In some embodiments, the individual is a clinical patient, a clinical trial volunteer, an experimental animal, and the like. In some embodiments, the individual is less than 60 years old (including, for example, less than 50, 40, 30, 25, 20, 15 or 10 years old). In some embodiments, the individual is older than 60 years (including, for example, older than 70, 80, 90, or 100 years old).

In some embodiments, the individual has one or more risk factors associated with P. aeruginosa infection. For example, in some embodiments, the individual has exposed or disrupted the mucous layer of the skin. In some embodiments, the individual has one or more burns. In some embodiments, the individual has one or more surgical wounds. In some embodiments, the individual has a skin disorder. In some embodiments, the individual has a foreign body implanted, such as, but not limited to, a ventilator or catheter. In some embodiments, the individual is diagnosed with or genetically predisposed to an immunodeficiency disorder, including but not limited to HIV infection, AIDS, and/or neutropenia. In some embodiments, the individual has received one or more forms of chemotherapy. In some embodiments, the individual has received one or more forms of adrenal glucocorticoid therapy. In some embodiments, the individual has received one or more forms of chemotherapy. In some embodiments, the individual is diagnosed with or genetically predisposed to developing cancer, diabetes, and/or chronic structural lung disease (eg, cystic fibrosis or COPD). In some embodiments, the individual is diagnosed with or genetically predisposed to a dysbiosis of the digestive system and/or other organs. In some embodiments, the individual has one or more risk factors associated with one or more of the diseases or conditions described above.

In some embodiments, the invention provides a method of delivering an anti-PcrV antibody, such as any of the anti-PcrV antibodies described herein, eg, an isolated anti-PcrV antibody, into cells infected with a pathogen in an individual, the The method includes administering to the individual a composition comprising an anti-PcrV antibody.

In some embodiments, according to any of the methods described herein, the method further comprises administering one or more additional therapeutic drugs. In some embodiments, the at least one therapeutic drug is an antibiotic. In some embodiments, the antibiotic is a penicillin, a cephalosporin, a carbapenem, a fluoroquinolone, an aminoglycoside, a monoamine, a polymyxin, a β-lactamase containing Antibiotic compositions of inhibitors, or any combination thereof. In some embodiments, the antibiotic is cefepime, ceftazidime, cefpirome, imipenem, meropenem, ticarcillin, guaracillin, azlocillin, carbenicillin, mezlocillin, atrox Nan, tobramycin, gentamicin, amikacin, ciprofloxacin, levofloxacin, cefotaxone-sulbactam, guaracillin-tazobactam, fosfomycin, or any combination thereof. In some embodiments, the antibiotic is one or more of imipenem, tobramycin, ciprofloxacin, meropenem, or atreonam. In some embodiments, the antibiotic is one or more of gentamicin, ampicillin, or kanamycin.

Diagnostic methods for infectious pathogens expressing PcrV and clinical descriptions of these diseases are known in the art. These methods include, but are not limited to, eg, immunohistochemistry, PCR, and fluorescence in situ hybridization (FISH).

In some embodiments, the anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) and/or compositions are used in combination with a second, third, or fourth agent (including, eg, antibiotics) to treat or prevent association with PcrV-expressing pathogens disease or condition. [Dosage and method of administration of anti-PcrV antibody]

The dosage of an anti-PcrV antibody (eg, isolated anti-PcrV antibody) composition administered to an individual (eg, a human) may vary depending on the particular composition, mode of administration, and type of disease being treated. In some embodiments, the amount of the composition (eg, a composition comprising an isolated anti-PcrV antibody) is effective to produce an objective response (eg, a partial response or a complete response) in treating or preventing a Pseudomonas infection. In some embodiments, the amount of the anti-PcrV antibody composition is sufficient to produce a complete response in the individual. In some embodiments, the amount of the anti-PcrV antibody composition is sufficient to produce a partial response in the individual. In some embodiments, the administered dose of the anti-PcrV antibody composition (eg, when administered alone) is sufficient to produce greater than 20%, 25%, 30%, 35%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85% or 90% overall response rate. An individual's response to the therapeutic or prophylactic methods described herein can be determined, for example, by detection of Pseudomonas using Gram stain or other phenotypic tests.

In some embodiments, the amount of a composition (eg, a composition comprising an isolated anti-PcrV antibody) is sufficient to prolong progression-free survival in an individual. In some embodiments, the amount of the composition is sufficient to prolong overall survival of the individual. In some embodiments, the amount of the composition (eg, when administered alone) is sufficient to produce a clinical benefit of greater than 50%, 60%, 70%, or 77% in a population of individuals treated with an anti-PcrV antibody composition.

In some embodiments, the amount of a composition (eg, a composition comprising an isolated anti-PcrV antibody), when used alone or in combination with a second, third, and/or fourth agent, is the same as before treatment in the same subject Compared to the corresponding organ load of other subjects unreachabiliated, it is sufficient to reduce the intensity of the pseudogenic organic load at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 100%. The magnitude of this therapeutic effect can be measured using standard methods, such as in vitro assays of purified enzymes, cell-based assays, animal models, or human trials.

In some embodiments, the amount of anti-PcrV antibody (eg, full-length anti-PcrV antibody) in the composition is less than the amount that causes a toxic effect (ie, an effect above a clinically acceptable level of toxicity) when the composition is administered to an individual level, or at a level where potential side effects can be managed or tolerated.

In some embodiments, following the same dosing regimen, the amount of the composition approaches the maximum tolerated dose (MTD) of the composition. In some embodiments, the amount of the composition is above 80%, 90%, 95% or 98% of the MTD.

In some embodiments, the amount of anti-PcrV antibody (eg, full-length anti-PcrV antibody) in the composition is in the range of 0.001 μg to 1000 μg.

In some embodiments, the composition or method further comprises one or more antibiotics. In some embodiments, the antibiotic in the composition (eg, imipenem, tobramycin, ciprofloxacin, meropenem, atreonam, ticarcillin, guaracillin, azlocillin, carbenicillin , mezlocillin, gentamicin or amikacin) in the range of 0.001 µg to 1000 µg.

In any of the above embodiments, the effective amount of the PcrV antibody (eg, full-length anti-PcrV antibody) in the composition is in the range of 0.1 μg/kg to 100 mg/kg, calculated on body weight.

In any of the above embodiments, the antibiotic (eg, imipenem, tobramycin, ciprofloxacin, meropenem, atreonam, ticarcillin, guaracillin, azlocillin) in the composition , carbenicillin, mezlocillin, gentamicin, or amikacin), based on body weight, in the range of 0.1 µg/kg to 100 mg/kg.

Anti-PcrV antibody compositions can be administered to an individual (eg, a human) by a variety of routes including, for example, intravenous, intraarterial, intraperitoneal, intrapulmonary, oral, inhalation, intravascular, intramuscular Injection, intratracheal, subcutaneous, intraocular, intrathecal, mucosal or transdermal. In some embodiments, an extended release formulation of the composition is used. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered via the portal vein. In some embodiments, the composition is administered arterially. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrahepatically. In some embodiments, the composition is administered by hepatic arterial infusion. In some embodiments, the composition is administered to a site remote from the first lesion. [Products and kits]

In some embodiments of the invention, there is provided an article of manufacture comprising a substance that can be used to treat or prevent a Pseudomonas infection in an individual, or to deliver an anti-PcrV antibody (eg, a full-length anti-PcrV antibody) to cells attached to PcrV-expressing pathogens. The article of manufacture may include a container and a label or package insert on or accompanying the container. Suitable containers include, for example, bottles, vials, syringes, and the like. Containers can be made from a variety of materials, such as glass or plastic. Typically, the container contains a composition effective to treat the disease or condition described herein and has a sterile port (eg, the container may be an IV bag or a vial with a hypodermic needle pierceable cap). At least one active substance in the composition is the anti-PcrV antibody of the present invention. The label or package insert identifies the specific condition for which the composition can be used to treat. The label or package insert further includes instructions for administering the anti-PcrV antibody composition to the patient. Articles of manufacture and kits including combination therapy are contemplated herein.

Package insert refers to the instructions, usually contained in commercial packages of therapeutic products, that contain information regarding the indications, usage, dosage, administration, contraindications and/or warnings associated with the use of these therapeutic products. In some embodiments, the package insert states that the composition can be used to treat bacterial infections. In some embodiments, the package insert states that the composition can be used to treat a Pseudomonas infection.

Additionally, the article of manufacture can further comprise a second container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Green's solution, or dextrose solution. Other materials required from a commercial and user standpoint may also be included, including other buffers, diluents, filters, needles and syringes.

Kits are also provided that can be used for various purposes, such as for treating or preventing Pseudomonas infection in an individual, or for delivering anti-PcrV antibodies (eg, full-length anti-PcrV antibodies) to cells to which PcrV-expressing pathogens are attached, Optionally combined with the product. Kits of the invention include one or more containers comprising an anti-PcrV antibody composition (or single-dose form and/or article of manufacture) and, in some embodiments, further comprising another agent (eg, an agent described herein) and/or instructions for use consistent with any of the methods described herein. The kit may further include instructions for selecting an individual suitable for treatment. Instructions for use that accompany the kits of the present invention are typically written instructions on a label or package insert (eg, paper sheets included in the kit), machine-readable instructions (eg, instructions on a magnetic or optical storage disc) is also acceptable.

For example, in some embodiments, the kit includes a composition comprising an anti-PcrV antibody (eg, a full-length anti-PcrV antibody). In some embodiments, the kit comprises: a) a composition comprising any one of the anti-PcrV antibodies described herein, and b) an effective amount of at least one other agent capable of enhancing the effect of the anti-PcrV antibody (eg, therapeutic effect, detection effect). In some embodiments, the kit includes: a) a composition comprising any of the anti-PcrV antibodies described herein, and b) instructions for administering the anti-PcrV antibody composition to an individual for the treatment of a Pseudomonas infection in the individual . In some embodiments, the kit comprises: a) a composition comprising any one of the anti-PcrV antibodies described herein, and b) an effective amount of at least one other agent capable of enhancing the effect of the anti-PcrV antibody (e.g., therapeutic effect, testing effect) and c) instructions for administering to the individual anti-PcrV antibody compositions and other substances for the treatment of Pseudomonas infection in the individual. The anti-PcrV antibody and other substances can be present in separate containers or in the same container. For example, the kit can include a particular composition or two or more compositions, one of which includes an anti-PcrV antibody and the other of which includes another agent.

In some embodiments, the kit comprises a nucleic acid (or set of) encoding an anti-PcrV antibody (eg, a full-length anti-PcrV antibody). In some embodiments, the kit comprises: a) a (or panel) encoding anti-PcrV antibody, and b) a host cell expressing the nucleic acid (or panel). In some embodiments, the kit comprises: a) a nucleic acid (or set of) encoding an anti-PcrV antibody, and b) instructions for use for: i) expressing an anti-PcrV antibody in a host cell, ii) preparing an anti-PcrV antibody A composition of a PcrV antibody, and iii) administering to an individual a composition comprising an anti-PcrV antibody to treat or prevent a Pseudomonas infection in the individual. In some embodiments, the kit comprises: a) a nucleic acid (or set of nucleic acids) encoding an anti-PcrV antibody, b) a host cell expressing the nucleic acid (or set of nucleic acids), and c) instructions for use, suitable for: i ) expressing the anti-PcrV antibody in a host cell, ii) preparing a composition comprising the anti-PcrV antibody, and iii) administering the composition comprising the anti-PcrV antibody to an individual to treat or prevent Pseudomonas infection in the individual.

The kits of the present invention are packaged in a suitable form. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (eg, sealed Mylar or plastic bags), and the like. The kit may optionally provide additional components such as buffers and instructional information. Accordingly, the present invention also provides articles of manufacture including vials (eg, sealed vials), bottles, jars, flexible packaging, and the like.

Instructions for use of anti-PcrV antibody compositions generally include information such as dosage, administration period and route of administration. Containers can be unit dose, bulk (eg, multi-dose) or subunit dose. For example, a kit is provided comprising a sufficient dose of an anti-PcrV antibody (eg, a full-length anti-PcrV antibody) as described herein to provide a long-term effective treatment of an individual, eg, one week, 8 days, 9 days, 10 days, 11 days , 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months or more. The kit may also contain multiple unit doses of the anti-PcrV antibody, the pharmaceutical composition, and instructions for use, and packaged in an amount sufficient for storage and use in pharmacies, eg, hospital pharmacies and compounding pharmacies.

Those skilled in the art will recognize several possible embodiments within the scope and spirit of the invention. The present invention will now be described in more detail with reference to the following non-limiting examples. The following examples further illustrate the invention, but should not be construed to limit its scope in any way. [detailed description] Example 1: Preparation of recombinant Pseudomonas PcrV and screening of single-chain antibodies (scFv) against PcrV [Preparation of recombinant Pseudomonas PcrV]

The full-length sequence of PcrV (PAO1) gene was synthesized (Shanghai Jierui), and it was sub-selected into the expression vector pET using the restriction enzyme recognition sites NdeI and BamHI. Label PcrV with a His-tag or other common tags. Construction and production of expression vectors pET-6His-PcrV and pET-6His-Avi-PcrV. In these constructions, "His" represents the His tag, and "Avi" represents the Avi tag. Recombinant PcrV, including pET-6His-PcrV, pET-6His-Avi-PcrV, were expressed and purified according to the manufacturer's instructions. Briefly, E. coli cells were transformed with the expression vector, induced with IPTG, and grown overnight at 25°C, 220rpm. Subsequently, E. coli cells were sonicated, cell debris was pelleted and removed, and proteins were isolated by centrifugation.

His-tagged proteins were purified using a nickel column (Ni Sepharose purification) according to the manufacturer's instructions. In particular, immobilized metal affinity chromatography (IMAC) analysis was performed using Qiagen Ni-NTA Superflow cartridges. The column was first equilibrated with buffer A1 (50 mM Na ₃ PO ₄ , 0.15 M NaCl, pH 7.2) at a flow rate of 150 cm/h. The supernatant of the medium (adjusted to pH 7.2) was passed through the column at a rate of 150 cm/h at room temperature. Next, buffer A1 (6 column volumes) was used to equilibrate the column at a flow rate of 150 cm/h. The column was washed with 10 column volumes of 50 mM PB buffer (0.15M NaCl and 0.2M imidazole, pH 7.2) and the eluate was collected. [Preparation of Biotinylated PcrV Antigen]

6His-Avi-PcrV was biotinylated using biotin ligase B0101A (GENECOPOEIA ^™ ) according to the manufacturer's instructions. Briefly, buffer A/B and ligase were added to 6His-Avi-PcrV, followed by incubation at 30°C for 1 hour. Biotinylated PcrV is hereinafter designated Bhavi-PcrV. Biotinylation efficiency was detected by ELISA. Briefly, the concentration of 6His-Avi-PcrV in the supernatant was quantified by ELISA by incubation with magnetic beads (Dynabeads™ MyOne™ Streptavidin T1) to remove Bhavi-PcrV, using a known concentration of 6His-Avi-PcrV as a standard . The biotinylation labeling efficiency was determined to be 90%. [Screening anti-PcrV single chain antibody (scFv)]

Construction of yeast scFv antibody display library: RNAs were collected from 2000 human blood samples, cDNA was obtained by reverse transcription, and _VH and _VK fragments were amplified with _VH- and _VK -specific primers. After gel extraction and purification, scFvs were generated by linking _VH and _VK via linkers. These scFvs were cloned into the yeast display plasmid PYD1 and then electroporated into yeast to construct a library producing yeast scFv antibody display. [Screening of anti-PcrV single chain antibodies (scFv) by PcrV binding]

Isolation of scFvs recognizing PcrV from a yeast display library. Briefly, cells expressing anti-PcrV scFv antibodies were enriched using MACS magnetic bead sorting. Bhavi-PcrV was mixed with magnetic beads (Dynabeads™ MyOne™ Streptavidin T1) overnight according to the manufacturer's instructions to allow biotinylated PcrV to coat the magnetic beads. The scFv antibody yeast pool is then mixed with magnetic bead-coated PcrV to enrich for yeast displaying PcrV antibodies, and the unbound is washed away in an elution step. Then, to screen for yeast-displayed antibodies with high affinity to PcrV, enriched yeast cells were labeled with PcrV protein and sorted by FACS. FACS-mediated screening was repeated for 2-3 cycles. The screened yeast bank cells were placed on agar and single colonies were picked and subjected to further FACS analysis. The scFv gene was extracted from a yeast colony positive for PcrV binding, fused with a 6-His tag, and subcolonized into a prokaryotic expression vector. Subsequently, the His-tagged scFvs were purified using the nickel column described above. A panel of positive scFv antibodies was obtained after screening and tested for their ability to inhibit red blood cell (RBC) lysis by Pseudomonas aeruginosa. [Analysis of candidate anti-PcrV scFv antibodies by inhibition of RBC cleavage]

The cloned monoclonal scFv antibodies were screened and their biological activities were assessed by RBC lysis assay. Briefly, red blood cells (RBCs) were prepared by centrifuging fresh human whole blood or rabbit whole blood, adding EDTA and washing several times with phosphate buffered saline (PBS). The washed RBCs were diluted in DMEM medium containing 10% fetal bovine serum (FBS, Gibco) to a final concentration of 2.5% (v/v) and added to 96-well circles together with purified anti-PcrV antibody diluted in PBS. in the bottom plate. Pseudomonas aeruginosa strain 57/66 (O6) was grown to logarithmic growth phase in 2× YT medium (Oxford), collected by centrifugation and resuspended in DMEM medium containing FBS until the absorbance at 600nm (OD600) was 0.15. 10 μl of the bacterial suspension was added to the RBC-antibody mixture, mixed by shaking, and incubated for 3 hours at 37°C, 5% CO ₂ . The culture plate was briefly centrifuged (1000 rpm, 1 min) to precipitate intact RBCs, the supernatant was transferred to a 96-well flat bottom plate, and the OD405 value was measured to detect lysis, and the relative amount of lysis inhibition was calculated and plotted. The IC50 values of the antibodies were also determined. Example 2: Preparation and characterization of full-length human anti-PcrV antibody [Preparation of full-length human anti-PcrV antibody]

The most promising scFv antibodies were reconstituted into human IgGl antibody molecules with human IgGl heavy chain constant regions and human kappa light chain constant regions. _VL and _VH were amplified from prokaryotic expression vectors and then constructed into eukaryotic expression vectors pTT5-L (containing kappa constant region) and pTT5-H1 (containing IgG1 heavy chain constant region), respectively. The plasmids expressing light chain or heavy chain were extracted, co-transfected into 293F cells, cultured at 37°C, 8% CO ₂ , and 120 rpm for 5 days, and the culture medium was purified by Protein A affinity chromatography.

Briefly, a Protein A column was first equilibrated with 6 column volumes of 50 mM PBS buffer (pH 7.2) containing 0.15 M NaCl at a flow rate of 150 cm/h. The culture supernatant (adjusted to pH 7.2) was flowed through the column at a flow rate of 150 cm/h. After further equilibration, 50 mM citric acid-sodium citrate buffer (pH 3.5) was used to collect the eluate containing anti-PcrV antibodies. The full-length antibody was functionally characterized for its ability to inhibit RBC lysis (see Example 1) and A549/U937 cell lysis, and to improve survival in a mouse model of acute pneumonia, following the methods in Example 1 and as described below. [Analysis of candidate anti-PcrV antibodies by A549 or U937 cleavage inhibition]

To test the ability to inhibit Pseudomonas aeruginosa-induced cytotoxicity and cytolysis, candidate anti-PcrV antibodies were added to the human bronchial epithelial cell line A549 or the human lymphoma cell line U937, which were treated with 10% FBS. DMEM medium was seeded in white 96-well plates (Nunc Nunclon Delta). Pseudomonas aeruginosa strain PA103 (O11) in logarithmic growth phase was added at a multiplicity of infection of 10 (MOI=10), incubated at 37°C, 5% CO for ₂ hours, and then equilibrated at room temperature for 20 minutes. Membrane integrity was examined by quantifying lactate dehydrogenase released from lysed cells using the CytoTox-ONE kit (Promega). The relative amount of lysis inhibition was plotted accordingly. The IC50 values of the antibodies were also determined. [Analysis of candidate anti-PcrV antibodies by mouse acute pneumonia model]

To examine the ability of candidate anti-PcrV antibodies to improve survival in a mouse model of acute pneumonia. In the prophylactic model, 7-8 week old BALB/c mice (Vitronilever) were injected intraperitoneally (ip) with candidate antibodies or PBS at a dose of 1, 5, 10 or 25 mg/kg. To establish an acute pneumonia model, BALB/c mice were intranasally inoculated with lethal doses or twice lethal as described previously (DiGiandomenico et al., 2007, Proc. Natl. Acad. Sci. USA , 104:4624–4629). amount (8×10 ⁵ -1.6×10 ⁶ CFU; 1* or 2* LD90) of Pseudomonas aeruginosa (PA103 strain) suspended in 40 μl of inoculum. Mice survival was recorded 7 days after infection. [PA49 breeding antibody is further optimized as a lead antibody]

Among the prepared full-length antibodies, the cloned monoclonal antibody PA49 was selected as the lead parental antibody based on the ability to inhibit RBC lysis, A549 cell or U937 cell lysis, and the ability to improve the survival rate of the acute pneumonia mouse model. Example 3: Preparation and characterization of optimized full-length anti-PcrV antibodies

Among the prepared full-length antibodies, the cloned monoclonal antibody PA49 was selected as the lead parent antibody for further optimization. In particular, PA49 was optimized to improve its affinity and biological activity. Combinatorial libraries were designed using the "Augmented Binary Substitution" technique (see Sue Townsend, et al, 2015, PNAS vol. 112, 15354-15359). In LC-CDR1, LC-CDR2, HC-CDR1 and HC-CDR2, binary replacement CDR fragment cassettes are inserted in frame to form a library where each position encodes only the parental or human germline residue of interest . HC-CDR3 was also expanded with 1 ± 1 random substitution per colony, that is, 2 sites were randomly selected out of 9 sites (full length of HC-CDR3) for NNK mutation (encoding all 20 amino acids). This technique reduces the amount of non-germline sequences in the CDRs as the parent antibody matures with affinity, thereby reducing the risk of immunogenicity of the final molecule.

Starting from the scFv of PA49, a phage scFv display library containing mutations in the CDR regions was constructed as described above. Mutants with high affinity and low off-rate binding to PcrV were identified by BLI and tested for their ability to inhibit RBC lysis, A549 cell lysis, or U937 cell lysis. scFv antibodies with comparable or improved biological activity compared to the parental scFv resulting from PA49 optimization were used to prepare full-length antibodies. An optimized set of full-length antibodies was obtained after screening.

The binding kinetics and affinity of the optimized anti-PcrV monoclonal antibody and the control antibody V2L2-MD to PcrV were detected by BLI, and the results are shown in Table 6. Table 6: Binding affinity of optimized anti-PcrV mAb colonization to PcrV Optimized Anti-PcrV mAb Colonization Kon(1/Ms) Kdis(1/s) Kd (M) Optimized Anti-PcrV mAb Colonization Kon(1/Ms) Kdis(1/s) Kd (M) 4A2 1.11E+05 <1.0E-07 <1.0E-12 5F8 7.37E+04 <1.0E-07 <1.0E-12 2A5 1.59E+05 <1.0E-07 <1.0E-12 6G12 7.75E+04 <1.0E-07 <1.0E-12 10B1 7.32E+04 <1.0E-07 <1.0E-12 6G3 1.17E+05 <1.0E-07 <1.0E-12 2B4 1.66E+05 <1.0E-07 <1.0E-12 4H9 1.53E+05 <1.0E-07 <1.0E-12 5B4 1.00E+05 <1.0E-07 <1.0E-12 6G4 2.11E+05 <1.0E-07 <1.0E-12 5C7 8.12E+04 <1.0E-07 <1.0E-12 5F3 2.19E+05 <1.0E-07 <1.0E-12 1D6 1.01E+05 1.30E-05 1.29E-10 10D8 2.34E+05 3.83E-05 1.64E-10 9F3 1.68E+05 <1.0E-07 <1.0E-12 9C7 1.80E+05 1.28E-04 7.12E-10 V2L2-MD 7.27E+05 <1.0E-07 <1.0E-12

The optimized full-length antibodies were then functionally characterized for their ability to inhibit RBC lysis, A549 cell lysis, or U937 cell lysis according to the methods described in Example 1 and Example 2. Optimized anti-PcrV antibody inhibits RBC, A549 or U937 cell lysis by Pseudomonas aeruginosa Optimized anti-PcrV antibody inhibits RBC cleavage

The ability of the optimized anti-PcrV mAb to clone to inhibit RBC lysis was compared with the parental PA49 mAb and the control antibody V2L2-MD. The RBC lysis inhibition assay was as described in Example 1.

The results are shown in Figures 1A, 1B and Table 7. Compared with the parental PA49 mAb and the control antibody V2L2-MD, all antibodies optimized by PA49 showed better or equivalent effect in inhibiting RBC cleavage. Table 7: Efficacy of optimized anti-PcrV mAbs to inhibit RBC cleavage Optimized Anti-PcrV mAb Colonization RBC lysis detection IC50 (μg/ml) Optimized Anti-PcrV mAb Colonization RBC lysis detection IC50 (μg/ml) PA49 1.4503 2B4 0.34 6G12 0.72 5C7 0.4824 10B1 0.2903 5F8 0.5113 1D6 0.3845 6G4 0.1331 2A5 0.1891 9F3 0.4572 4A2 0.1774 9C7 0.4942 4H9 0.4077 11E9 0.3023 5B4 0.1939 5F3 0.3796 5E6 0.2270 7H5 0.2510 6G3 0.3306 V2L2-MD 0.5476 10D8 0.2250 [Optimized anti-PcrV antibody inhibits A549 cell lysis or U937 cell lysis]

The ability of the optimized anti-PcrV antibody to inhibit the lysis of A549 or U937 cells was also evaluated. A549 and U937 cell lysis inhibition experiments are shown in Example 2.

The results are shown in Figures 2A and 2B, all antibodies optimized by PA49 showed high efficacy in inhibiting A549 (Figure 2A) or U937 (Figure 2B) cell lysis at doses of 100 μg/ml and lower, And the inhibitory effect was dose-dependent. Example 4: Characterization of Nonspecific Binding and Affinity of Optimized Anti-PcrV Antibodies

Further Characterization of Optimized Antibodies 6G4, 5F3, 9C7, 11E9, 7H5, 6G12, 2A5, 4A2, 5B4, 10D8, 2B4, Control Antibody V2L2-MD, Negative Control Antibody Tildrakizumab, or Positive Control Antibody in BV ELISA or 293 Cell Cross-Reactivity Assays Nonspecific binding of lenzilumab. [Non-specific binding of anti-PcrV antibodies]

The optimized anti-PcrV antibodies were characterized for non-specific binding by measuring cross-reactivity with BV particles and PcrV-negative 293 cells.

Cross-reactivity with BV particles: Optimized antibodies 6G4, 5F3, 9C7, 11E9, 7H5, 6G4, 5F3, 9C7, 11E9, 7H5, Cross-reactivity of 6G12, 2A5, 4A2, 5B4, 10D8, 2B4, control antibody V2L2-MD, negative control antibody Tildrakizumab or positive control antibody Lenzilumab with BV particles. The results are shown in Figure 3, the positive control Lenzilumab had a high level of non-specific binding to BV particles. In contrast, the optimized antibody exhibited low levels of nonspecific binding, similar to V2L2-MD.

Cross-reactivity with 293T cells: Cross-reactivity of optimized antibodies 5F3, 9C7 or V2L2-MD with PcrV-negative 293 cells was detected by FACS. The results are shown in Figures 4A-4C, the positive control Lenzilumab showed high levels of 293 cell binding, the optimized antibodies 5F3 or 9C7 showed low levels of 293 binding similar to the negative control (without any antibody) or V2L2-MD, while Lenzilumab, a positive control antibody specific for GM-CSF on 293 cells, showed higher levels of binding to 293 cells.

Taken together, these results indicate that the optimized antibodies 5F3 and 9C7 have low nonspecific binding similar to the control antibody V2L2-MD. Example 5: Optimized full-length anti-PcrV antibodies broadly neutralize Pseudomonas aeruginosa strains

To examine whether the optimized anti-PcrV antibody has broad-spectrum neutralizing activity against P. aeruginosa strains, the optimized anti-PcrV antibody against several clinically relevant P. aeruginosa strains (O1, O11, O6) was investigated. The ability to induce RBC lysis by these strains represents the majority of clinical cases of Pseudomonas aeruginosa infection. [Inhibition of RBC cleavage by an optimized anti-PcrV antibody]

The optimized anti-PcrV monoclonal antibodies 11E9, 9C7, 5F3 and 7H5 were analyzed for their ability to inhibit RBC lysis by Pseudomonas aeruginosa strain O1-52/66; 5B4, 4A2, 2A5 and 11E9 were analyzed for Pseudomonas aeruginosa strains Inhibitory effect of bacterial strain O11-PA103; the inhibitory effect of 9C7, 5F3, 7H5 or control antibody V2L2-MD on Pseudomonas aeruginosa strain O6-57/66 was analyzed. The RBC lysis inhibition assay was as described in Example 1.

The results are shown in Figures 5A-5C and Tables 8A-8C, all the optimized monoclonal antibodies can effectively inhibit the RBC lysis caused by strain O1-52/66 (Figure 5A) or strain O11-PA103 (Figure 5B), 9C7, 5F3 or 7H5 inhibited RBC lysis by strain O6-57/66 (Fig. 5C) better than the control antibody V2L2-MD. Table 8A: Inhibitory potency of optimized anti-PcrV antibodies against RBC lysis by different P. aeruginosa strains Pseudomonas aeruginosa strains Monoclonal antibody selection O1-52/66 IC50 (µg/mL) 11E9 0.2056 9C7 0.3422 5F3 0.3247 7H5 0.444 Table 8B: Inhibitory potency of optimized anti-PcrV antibodies on RBC lysis by different P. aeruginosa strains Pseudomonas aeruginosa strains Monoclonal antibody selection O11-PA103 IC50 (µg/mL) 5B4 0.0925 4A2 0.0799 2A5 0.1444 11E9 0.1973 Table 8C: Inhibitory potency of optimized anti-PcrV antibodies on RBC lysis by different P. aeruginosa strains Pseudomonas aeruginosa strains Monoclonal antibody selection O6-57/66 IC50 (µg/mL) 9C7 0.3746 5F3 0.3796 7H5 0.2880 V2L2-MD 0.5476

To further demonstrate whether the optimized antibodies could broadly neutralize P. aeruginosa strains, the binding ability of these antibodies to different forms of PcrV mutants was tested. Briefly, 100 PcrV sequences were randomly selected from the Genebank database. Compared to the wild type sequence (SEQ ID No: 80), 13% of the selected sequences contained R, G or K at position 225 (S in the wild type sequence SEQ ID No: 80).

The optimized 9C7, 5F3, 4A2, 2A5, 5B4, 11E9, or 7H5 antibodies had affinity and specificity for and strong binding to these PcrV mutants (data not shown). These results further demonstrate that the optimized anti-PcrV antibody has broad-spectrum neutralizing activity against Pseudomonas aeruginosa strains that may express various forms of PcrV mutants. Example 6: Optimized anti-PcrV antibodies for the prevention of Pseudomonas aeruginosa infection

Improved survival in a mouse model of pneumonia and an intraperitoneal infection demonstrated that the optimized anti-PcrV antibody protected prophylactically from P. aeruginosa infection. [Anti-PcrV antibody improves survival in mouse pneumonia model]

The parental anti-PcrV antibody PA49, the optimized antibody 6G12, 4A2, 10D8, 7H5, 11E9, 6G4, 2A5, 9C7, 5F3 or the control antibody V2L2-MD were evaluated for their ability to improve survival in a mouse model of pneumonia, compared with HIV-10E8 (negative control) for comparison. HIV-10E8 is a previously described antibody that broadly neutralizes HIV (see Huang J, et al, 2012, Nature 491:406).

In the prophylactic model, 7-8 week old BALB/c mice (Vitronilever) were injected intraperitoneally (ip) 24 hours before infection with the antibody at a dose of 1 or 10 mg/kg based on mouse body weight. To induce acute pneumonia, BALB/c mice were intranasally inoculated with a lethal dose (1*LD90) or twice the lethal dose (2*LD90) (8×10 ⁵ ~ 1.6×10 ⁶ CFU) of aeruginosa suspended in 40 μl of inoculum Pseudomonas (strain PA103). Mice survival was recorded 7 days after infection, and the results were represented by Kaplan-Meier survival curves.

The results are shown in the Kaplan-Meier survival curves in Figures 6A-6C, compared with the negative control group, the lethal dose (1*LD90 or 2*LD90; 8×10 ⁵ ~ 1.6×10 ⁶ CFU) of Pseudomonas aeruginosa (PA103 strain), all anti-PcrV antibodies significantly improved the survival rate of acute pneumonia model mice. Differences in survival were calculated using the logarithmic test. [Organ Burden in Pneumonia Model Mice Administered with Anti-PcrV Antibody]

The optimized anti-PcrV antibodies 4A2, 9C7, 11E9, 5F3, 7H5 or 2A5 were further evaluated for their ability to reduce organ burden in an acute pneumonia model. HIV-10E8 served as a negative control.

In the prophylactic model, 7-8 week old BALB/c mice (Vitronilever) were injected intraperitoneally (ip) with the antibody at a dose of 10 mg/kg based on mouse body weight 24 hours before infection. To induce acute pneumonia, BALB/c mice were intranasally inoculated with a half-lethal dose (0.5*LD90; ³ x 105 CFU) of Pseudomonas aeruginosa (strain PA103) suspended in 40 μl of inoculum. Twenty-four hours after infection, mice were sacrificed, and lungs, spleens, and kidneys were extracted, homogenized, diluted and placed on agar to measure viable units (CFU), which represents the P. aeruginosa load in each organ sample.

The results are shown in Figure 7. Compared with the negative control HIV-10E8, all the optimized anti-PcrV antibodies showed higher efficacy in reducing the organ load of the lung, spleen and kidney of the pneumonia model mice. Differences in survival were calculated using the logarithmic test. [Using anti-PcrV antibody to improve the survival rate of intraperitoneal infection model mice]

Using HIV-10E8 as a negative control, the ability of the optimized anti-PcrV antibodies 6G4 or 10D8 to improve survival in intraperitoneal infection model mice was evaluated. A mouse intraperitoneal infection model was constructed as described previously (see Warrener et al., 2014, Antimicrob. Agents Chemother ., 58, 4384–4391).

In the prophylactic model, 7-8 week old BALB/c mice (Vitronilever) were injected intraperitoneally (ip) with the antibody at a dose of 10 mg/kg based on mouse body weight 24 hours before infection. To induce intraperitoneal infection, BALB/c mice were intraperitoneally inoculated with 4-fold lethal dose (4*LD90 = 1×10 ⁶ CFU) of Pseudomonas aeruginosa (strain O6-57/66) suspended in 300 μl of inoculum. Mice survival was recorded 7 days after infection.

The results are shown in Figure 8. When inoculated with 4 times the lethal dose (4* LD ₉₀ ) of Pseudomonas aeruginosa (strain 57/66), compared with the negative control group, all anti-PcrV antibodies were at a dose of 10 mg/kg antibody It can significantly improve the survival rate of intraperitoneal infection model mice. Differences in survival were calculated using the logarithmic test.

Taken together, these results demonstrate that the anti-PcrV antibodies disclosed herein can prevent Pseudomonas aeruginosa infection in a mouse model of pneumonia and a mouse model of intraperitoneal infection. Example 7: Optimized anti-PcrV antibodies for the treatment of Pseudomonas aeruginosa infection

The improved survival rate of pneumonia model mice proved that the optimized anti-PcrV antibody could treat Pseudomonas aeruginosa infection. [Optimized anti-PcrV antibody improves the survival rate of pneumonia model mice]

The optimized anti-PcrV antibodies 4A2, 2A5, 9C7, 6G12 and PA49 were evaluated for their ability to improve survival in an acute pneumonia model. HIV-10E8 served as a negative control.

In a therapeutic model, to induce acute pneumonia, 7-8 week old BALB/c mice (Vitronilever) were intranasally inoculated with a lethal dose (1*LD90 = ^8x105 CFU) of sham aeruginosa suspended in 40 μl of inoculum Monomonas (PA103 strain). One hour after infection, antibodies or PBS were injected intravenously at a dose of 2 mg/kg based on mouse body weight. Mice survival was recorded 7 days after infection.

The results are shown in Figure 9. When inoculated with a lethal dose (1*LD90) of Pseudomonas aeruginosa (PA103 strain), compared with the negative control group, all anti-PcrV antibodies significantly improved the survival of acute pneumonia model mice Rate. Compared with the parental antibody PA49, the optimized antibody 4A2, 2A5, 9C7 or 6G12 also significantly improved the survival rate of acute pneumonia model mice. Differences in survival were calculated using the logarithmic test. The results indicate that the anti-PcrV antibodies described herein can be used to treat Pseudomonas aeruginosa infection. Example 8: Optimized Anti-PcrV Antibody and Antibiotic Combination Treatment of Pseudomonas aeruginosa Infection Anti-PcrV antibody combined with antibiotics improves survival in a mouse model of intraperitoneal infection

The ability of the optimized anti-PcrV antibody 9C7 or 4A2 to improve survival in a mouse model of intraperitoneal infection was evaluated, compared with the antibiotic meropenem, and combined treatment of 9C7 and meropenem or 4A2 and meropenem. HIV-10E8 served as a negative control.

Anti-PcrV antibodies were tested in a prophylactic model, 7-8 week old BALB/c mice (Vitronilever) were injected intraperitoneally with antibodies 24 hours before infection or with antibiotics 2 hours after infection. Specifically, mice were administered with optimized anti-PcrV antibodies to clone 9C7 or 4A2 (5 mg/kg), meropenem (6 mg/kg) or a combination of 9C7 (5 mg/kg) and meropenem (6 mg/kg) or 4A2 ( 5 mg/kg) and meropenem (6 mg/kg). To induce intraperitoneal infection, BALB/c mice were intraperitoneally inoculated with a 3-fold lethal dose (3*LD90= ^7x105 CFU) of Pseudomonas aeruginosa (strain 57/66) suspended in 300 μl of inoculum. Mice survival was recorded 5 days after infection.

The results are shown in Figure 10. When inoculated with a 3-fold lethal dose (3*LD90) of Pseudomonas aeruginosa (strain 57/66), the optimized antibodies 9C7 (5mg/kg) or 4A2 (5mg/kg) were used alone ) or the antibiotic meropenem (6 mg/kg) did not provide 100% protection against intraperitoneal infection in model mice. 9C7 (5mg/kg) combined with antibiotic meropenem (6mg/kg) or 4A2 (5mg/kg) combined with antibiotic meropenem (6mg/kg) when antibodies were combined with antibiotics compared to antibodies alone or antibiotics alone /kg) combined use significantly improved the survival rate of intraperitoneal infection model mice (p < 0.05). Differences in survival were calculated using the logarithmic test. These results demonstrate the clinical potential of the anti-PcrV antibodies described herein in combination with antibiotics in neutralizing P. aeruginosa. Example 9: Pharmacokinetics of optimized anti-PcrV antibodies

To study the in vivo pharmacokinetics of the optimized anti-PcrV antibodies, the levels of 5F3, 9C7, 7H5 or the control antibody V2L2-MD in rat plasma were measured over time.

Pharmacokinetics in rats: 32 healthy adult rats (about 0.2 kg body weight) were equally divided into two groups, and the average body weight of each group was similar. One group received 5F3, 9C7, 7H5 or V2L2-MD intravenously at 30 mg/kg, and the other group received 5F3, 9C7, 7H5 or V2L2-MD intravenously at 3 mg/kg. Blood samples were collected at 0 hours, 0.5 hours, 2 hours, 8 hours, 1 day, 3 days, 7 days, 11 days, 17 days, 23 days, 31 days, 41 days, and 52 days after injection, respectively. After centrifugation, the concentration of antibodies in plasma was determined by ELISA. In ELISA experiments, 96-well plates were coated with synthetic PcrV. The next day, after washing with PBST, blocking with 200 µL of PBS-milk for 1 hour, and washing with PBST, plasma was added to the plate and incubated at 37°C for 1 hour. The 96-well plate was washed 6 times with 0.1% TBST, then 100 µL of goat anti-human Fc antibody AP (diluted 1:3000 in PBS) was added to each well and incubated for 1 hour. After washing 6 times with 0.1% TBST, 50µL of pNPP was added to each well, and the color was developed at 37˚C for 10-20min. The signal was read at a wavelength of 410 nm with a microplate reader.

The results are shown in Figures 11A and 11B, at both high and low doses (3 mg/kg or 30 mg/kg) intravenously, the half-lives of 5F3, 9C7 and 7H5 were all longer than that of the control antibody V2L2-MD, indicating that the Compared with the antibody V2L2-MD, the optimized antibody showed more stable pharmacokinetics at different doses.

without.

The results shown in Figures 1A-1B show the ability of the lead-optimized antibody to inhibit RBC cleavage, compared to the parental PA49 monoclonal antibody and the control antibody V2L2-MD.

The results shown in FIG. 2A are the ability of the lead-optimized antibody to inhibit the lysis of A549 cells, and the results shown in FIG. 2B are the ability of the lead-optimized antibody to inhibit the lysis of U937 cells.

The results shown in Figure 3 are the cross-reactivity of the lead-optimized antibody to BV particles detected by ELISA, compared to the control antibody V2L2-MD.

Figures 4A-4C show the cross-reactivity of the optimized antibodies 5F3, 9C7 or the control antibody V2L2-MD with PcrV-negative HEK293 cells detected by FACS.

The results shown in Figure 5A show the inhibitory ability of optimized antibodies 11E9, 9C7, 5F3 or 7H5 on O1-52/66-induced RBC cleavage; Figure 5B shows the results of optimized antibodies 5B4, 4A2, 2A5 or 11E9 on O11-PA103 Inhibitory ability of RBC lysis; the results shown in Figure 5C are the inhibitory ability of optimized antibody 9C7, 5F3, 7H5 or the control antibody V2L2-MD to O6-57/66-induced RBC lysis.

Results shown in Figures 6A-6C show the ability of the parental PA49 monoclonal antibody and the optimized antibodies 6G12, 4A2, 10D8, 7H5, 11E9, 6G4, 2A5, 9C7 or 5F3 to prophylactically increase survival in a mouse model of acute pneumonia. Figure 6A shows the ability of PA49 or 6G12 to prophylactically improve survival in a mouse acute pneumonia model when the inoculum of Pseudomonas aeruginosa (PA103 strain) was 2*LD90 (~1.6×10 ⁶ CFU) The results shown in Figure 6B show that when the inoculum of Pseudomonas aeruginosa (PA103 strain) was 1*LD90 (~8×10 ⁵ CFU), 4A2, 10D8, 7H5 or 11E9 prophylactically prevented acute pneumonia in mice The ability to improve the survival rate; the results shown in Figure 6C are when the inoculum of Pseudomonas aeruginosa (PA103 strain) is 2*LD90 (~1.6×10 ⁶ CFU), 6G4, 2A5, 9C7 or 5F3 in mice with acute pneumonia The ability to prophylactically increase survival in the model.

The results shown in Figure 7 are optimized for the ability of antibodies 4A2, 9C7, 11E9, 5F3, 7H5 or 2A5 to reduce organ burden prophylactically in a mouse model of acute pneumonia.

The results shown in Figure 8 are the ability of the optimized anti-PcrV antibody 6G4 or 10D8 to prophylactically increase the survival rate in a mouse intraperitoneal infection model when the inoculum of Pseudomonas aeruginosa (57/66) is 4*LD90.

The results shown in Figure 9 show that the optimized anti-PcrV antibodies 4A2, 2A5, 9C7, 6G12 or PA49 therapeutically improved the survival rate in a mouse acute pneumonia model when the inoculation amount of Pseudomonas aeruginosa (PA103) was 1*LD90 Ability.

The results shown in Figure 10 are when the inoculum of Pseudomonas aeruginosa (57/66) was 3*LD90, the optimized anti-PcrV antibody 9C7 or 4A2 was administered alone or in combination with meropenem (MEM) in the abdominal cavity of mice The ability to prophylactically increase survival in an intra-infection model.

The results shown in Figures 11A-11B are the pharmacokinetics of the optimized antibody 5F3, 9C7, 7H5 or the control antibody V2L2-MD in rats at an intravenous dose of 3 mg/kg or 30 mg/kg, respectively.

The results shown in Figures 12A-12B are sequence alignments of anti-PcrV antibody variable domain sequences. Complementarity determining regions (Kabat definition) are indicated. The results shown in Figure 12A are sequence alignments of heavy chain variable domain sequences. The results shown in Figure 12B are sequence alignments of light chain variable domain sequences.

Claims (28)

An isolated anti-PcrV antibody comprising: a heavy chain variable domain ( _{VH )} comprising: a heavy chain complementarity determining region (HC-CDR) 1 comprising SYWMH (SEQ ID NO: 1) or a variant thereof comprising at most 3 amino acid substitutions; a HC-CDR2 comprising RINEX ₁ EX ₂ SX ₃ SYADSVKG (SEQ ID NO: 50), or a variant thereof comprising at most 3 amino acid substitutions, wherein X ₁ is D, T, N, L, I, S, V, _A , or H, X2 is S, T, N, D, G, or R, and _X3 is I or T; and one HC-CDR3, including DGX ₁ X ₂ X ₃ X ₄ X ₅ DX ₆ (SEQ ID NO: 51), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is P, Y or A, X ₂ is Y or W, and X ₃ is D, T or N, _X4 is S, T or _A , _X5 is L, F or M, X6 is I, V, A, S, L or W; and a light chain variable domain ( _VL ), The _VL comprises: a light chain complementarity determining region (LC-CDR) 1 comprising RASQX ₁ VX ₂ X ₃ NLA (SEQ ID NO: 47), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is S, N, D or G, X2 is S, K, N, R or T, _X3 is N, S, D or G; an LC _- CDR2 comprising X1 _ASSRAT (SEQ ID NO: 85), or a variant thereof comprising up to 3 amino acid substitutions, wherein X ₁ is D, N, H, A or S; and an LC-CDR3 comprising QQYGX ₁ X ₂ PX ₃ T (SEQ ID NO: 86), or Variants comprising up to ₃ amino acid substitutions wherein X is S, _A , T, E, D, H, N, Q, G, L, Y, M, R or V and X is S, Q, V , E, T, D, M, Y, G, H, L, N, A, F or P, X ₃ is I, L or V. An isolated anti-PcrV antibody comprising: a _VH comprising: a HC-CDR1 comprising _SYWMH (SEQ ID NO: 1) or a variant thereof comprising at most 3 amino acid substitutions; a HC-CDR2, comprising RINEX1EX2SISYADSVKG (SEQ ID NO: 45) or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is D, N, I, L or V, and X2 is S, T, R, G or N; and a HC-CDR3 comprising DGPYDX1X2DI (SEQ ID NO: 46) or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is S, A or T, X2 is F or L; and _VL , the _VL Include: an LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO: 47) or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is N, G, D or S, and X2 is K, R, S, N or T , X3 is N, G, S or D; an LC-CDR2 comprising X1ASSRAT (SEQ ID NO: 48) or a variant thereof comprising up to 3 amino acid substitutions, wherein X1 is D, N, H or A; and a LC-CDR3, comprising QQYGX1X2PX3T (SEQ ID NO: 49) or a variant comprising up to 3 amino acid substitutions, wherein X1 is S, T, E, H, N, A, D, M or L, and X2 is S, Q, E, T, D, G, H, L, N, V or Y, and X3 is I, L or V. The isolated anti-PcrV antibody of claim 1 or 2, wherein the anti-PcrV antibody binds to Pseudomonas PcrV with a Kd value of about 0.1 pM to 1 nM. An isolated anti-PcrV antibody, comprising _{: VH comprising} : a HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, or a variant thereof comprising at most 3 amino acid substitutions; a HC-CDR2 comprising An amino acid sequence of any one of SEQ ID NOs: 2-9, or a variant thereof comprising up to 3 amino acid substitutions; and a HC-CDR3 comprising an amino acid sequence of any one of SEQ ID NOs: 10-13, or a variant thereof comprising up to 3 amino acid substitution variants; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 14-25, or a variant comprising up to 3 amino acid substitutions; an LC - a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 26-29, or a variant thereof comprising at most 3 amino acid substitutions; an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 30-44, or its Variants containing up to 3 amino acid substitutions. An isolated anti-PcrV antibody, comprising: _VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 in the _VH having any of the amino acid sequences of SEQ ID NOs: 52-64; and _VL comprising LC-CDR1, LC-CDR2 and LC-CDR3 in _VL having the amino acid sequence of any of SEQ ID NOs: 65-79. The isolated anti-PcrV antibody of any one of claims 1 to 5, comprising: (i _{) a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid HC-CDR2 of sequence SEQ ID NO: 2, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 10; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL , the _VL comprises: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30, or an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 30. - _VL variants comprising up to 5 amino acid substitutions in the CDRs; (ii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence SEQ ID NO: 2 HC-CDR2, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising at most 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: a comprising LC-CDR1 of amino acid sequence SEQ ID NO: 14, one LC-CDR2 comprising amino acid sequence SEQ ID NO: 26, and one LC-CDR3 comprising amino acid sequence SEQ ID NO: 30, or at most 5 in LC-CDRs amino acid-substituted _VL variants; (iii _{) a VH comprising} : one HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: The LC-CDR1 of 15, the LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and the LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 31, or a _VL comprising at most 5 amino acid substitutions in the LC-CDRs Variants; (iv _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one comprising the amino acid sequence SEQ ID NO: 7 HC-CDR3 of NO: 13; or at most 5 amino acids in HC-CDRs and _VL comprising _: one LC-CDR1 comprising the amino _acid sequence SEQ ID NO: 23, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence The LC-CDR3 of SEQ ID NO:39, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (v _{) a VH comprising} : a VL comprising the amino acid sequence of SEQ ID NO:1 HC-CDR1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 32 LC-CDR3, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (vi _{) a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising HC-CDR2 of amino acid sequence SEQ ID NO: 5, and one HC-CDR3 comprising amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in HC-CDRs; and _VL , all Said _VL comprises: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 33, or _VL variants comprising up to 5 amino acid substitutions in the LC-CDRs; (vii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising the amino acid sequence SEQ ID NO: 1 2 HC-CDR2, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising at most 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: a an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 18, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 34, or at most LC-CDRs 5 amino acid substituted _VL variants; (vii i _{) a VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 13 HC-CDR3; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, and one comprising the amino acid sequence LC-CDR2 of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (ix) _VH , where Said _VH comprises: an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 19, and a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 28 LC-CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 35, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (x _{) a VH comprising} : a HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or at most HC-CDRs 5 amino acid substituted _VH variants; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 36, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xi _{) a VH comprising} : a VL comprising the amino acid sequence of SEQ ID NO: : 1 HC-CDR1, one HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 4, and one HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or V containing at most 5 amino acid substitutions in the HC-CDRs _H variant; and _VL comprising _: a package An LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 37, or at most LC-CDRs 5 amino acid substituted _VL variants; (xii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and a HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: a HC-CDR3 comprising the amino acid sequence of SEQ ID NO LC-CDR1 of : 22, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38, or a V substituted at most 5 amino acids in the LC-CDRs _L variant; (xiii _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one comprising the amino acid sequence SEQ ID NO: 7 HC-CDR3 of ID NO: 11; or a _VH variant comprising up to 5 amino acid substitutions in the HC-CDRs; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO:23 , an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 39, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xiv _{) VH} comprising: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 8, and one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 11 -CDR3; or a _VH variant comprising up to 5 amino acid substitutions in HC-CDRs; and a _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21, and a LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 21 LC-CDR2 of ID NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 40, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xv) _VH comprising: one HC-CDR1 comprising the amino _acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 6, and one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 11 CDR3; or a _VH variant comprising up to 5 amino acid substitutions in HC-CDRs; and a _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 24, one comprising the amino acid sequence of SEQ ID The LC-CDR2 of NO: 28, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 41, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xvi) _VH , the V _H comprises: one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 11; _VH variants comprising up to 5 amino acid substitutions in the CDRs; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 25, an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 29 CDR2, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 42, or a _VL variant comprising at most 5 amino acid substitutions in the LC-CDRs; (xvii) _VH comprising: an amino _acid comprising HC-CDR1 of sequence SEQ ID NO: 1, one HC-CDR2 comprising amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising amino acid sequence SEQ ID NO: 11; or at most 5 in HC-CDRs An amino acid substituted _VH variant; and a _VL comprising _: one LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, one LC-CDR2 comprising the amino acid sequence SEQ ID NO: 27, and one comprising the amino acid sequence The LC-CDR3 of sequence SEQ ID NO:43, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (xviii _{) a VH comprising} : one comprising the amino acid sequence of SEQ ID NO:1 the HC-CDR1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; or at most 5 amino acid substitutions in the HC-CDRs and _VL comprising _: an LC-CDR1 comprising the amino _acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 The LC-CDR3 of ID NO: 44, or a _VL variant comprising up to 5 amino acid substitutions in the LC-CDRs; (xix _{) a V H comprising} : an HC comprising the amino acid sequence of SEQ ID NO: 1 - CDR1, one HC-CDR2 comprising the amino acid sequence SEQ ID NO: 7, and one HC-CDR3 comprising the amino acid sequence SEQ ID NO: 12; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL comprising _: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC comprising the amino acid sequence of SEQ ID NO: 39 - CDR3, or a _VL variant comprising up to 5 amino acid substitutions in LC-CDRs; or (xx _{) VH comprising} : one HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, one comprising HC-CDR2 of amino acid sequence SEQ ID NO: 9, and one HC-CDR3 comprising amino acid sequence of SEQ ID NO: 12; or _VH variants comprising up to 5 amino acid substitutions in the HC-CDRs; and _VL , the Said _VL comprises: an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 43, or _VL variants containing up to 5 amino acid substitutions in the LC-CDRs. The isolated anti-PcrV antibody of any one of claims 1 to 6, comprising: _VH , the _VH comprising any amino acid sequence of SEQ ID NOs: 52-64, or comprising the same amino acid sequence as SEQ ID NOs: 52 - a variant sequence having at least 90% sequence homology to any of the amino acid sequences in 64; and a _VL comprising the amino acid sequence of any one of SEQ ID NOs _: 65-79, or comprising an amino acid sequence with SEQ ID NOs: 65 - Variant sequences having at least 90% sequence homology in any of the amino acid sequences of 79. The isolated anti-PcrV antibody of claim 7, comprising: (i) a V _H comprising the amino acid sequence of SEQ ID NO: 52 and a _VL comprising the amino acid sequence of SEQ ID NO: 65; (ii) comprising the amino acid sequence of SEQ ID NO: 65 The _VH of NO:56 and the _VL comprising the amino acid sequence of SEQ ID NO:65; (iii) the _VH comprising the amino acid sequence of SEQ ID NO:53 and the _VL comprising the amino acid sequence of SEQ ID NO:66; (iv) the VH comprising the amino acid sequence of SEQ ID NO:66; The _VH of the amino acid sequence of SEQ ID NO:62 and the _VL of the amino acid sequence of SEQ ID NO:74; (v) the _VH of the amino acid sequence of SEQ ID NO:54 and the _VL of the amino acid sequence of SEQ ID NO:67; (vi) a V _H comprising the amino acid sequence of SEQ ID NO: 55 and a VL comprising the amino acid sequence of SEQ ID NO: 68; (vii) a V _H comprising the amino acid sequence of SEQ ID NO: 56 and a V _H comprising the amino acid sequence of SEQ ID NO: 69 (viii) the _VH comprising the amino acid sequence of SEQ ID NO: 64 and the _VL comprising the amino acid sequence SEQ ID NO: 78; (ix ₎ the _VH comprising the amino acid sequence SEQ ID NO: 56 and the amino acid sequence SEQ ID NO: 56 The _VL of ID NO:70; (x) the _VH comprising the amino acid sequence of SEQ ID NO:57 and the _VL comprising the amino acid sequence of SEQ ID NO:71; (xi) the _VH comprising the amino acid sequence of SEQ ID NO:54 and the A VL comprising the amino acid sequence of SEQ ID NO: 72; (xii) a V _H comprising the amino acid sequence of SEQ ID NO: 56 and a VL comprising the amino _acid sequence of SEQ ID NO: 73; (xiii) a _VL comprising the amino acid sequence of SEQ ID NO: 58 The V _H and the VL comprising the amino acid sequence SEQ ID NO: 74; (xiv) the V _H comprising the amino acid sequence SEQ ID NO: 59 and the _VL comprising the amino _acid sequence SEQ ID NO: 75; (xv) the amino acid sequence SEQ ID NO: 75 _VH of ID NO: 57 and containing amino acids _VL of sequence SEQ ID NO: 76; (xvi) V _H comprising amino acid sequence SEQ ID NO: 60 and _VL comprising amino acid sequence SEQ ID NO: 77; (xvii) V comprising amino acid sequence SEQ ID NO: 60 _H and the VL comprising the amino acid sequence of SEQ ID NO: 78; (xviii) the V _H comprising the amino acid sequence of SEQ ID NO: 56 and the _VL comprising the amino _acid sequence of SEQ ID NO: 79; (xix) the amino acid sequence of SEQ ID NO: : a _VH of 61 and a _VL comprising the amino acid sequence of SEQ ID NO:74; or (xx) a _VH comprising the amino acid sequence of SEQ ID NO:63 and a _VL comprising the amino acid sequence of SEQ ID NO:78. An isolated anti-PcrV antibody that specifically binds to PcrV, which competitively binds to PcrV with the isolated anti-PcrV antibody described in any one of claims 1 to 8, or described in any one of claims 1 to 8 The isolated anti-PcrV antibodies specifically bind to the same epitope. The isolated anti-PcrV antibody of any one of claims 1 to 9, wherein the anti-PcrV antibody comprises an Fc fragment. The isolated anti-PcrV antibody of claim 10, wherein the anti-PcrV antibody is a full-length IgG antibody. The isolated anti-PcrV antibody of claim 10, wherein the anti-PcrV antibody is a full-length IgGl or IgG4 antibody. The isolated anti-PcrV antibody of any one of claims 1 to 12, wherein the anti-PcrV antibody is chimeric, fully human or humanized. The isolated anti-PcrV antibody of any one of claims 1 to 9, wherein the anti-PcrV antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab', F(ab)'2, Fab'-SH , single chain antibodies (scFv), Fv fragments, dAbs, Fd or diabodies in the group consisting of. A nucleic acid molecule encoding the anti-PcrV antibody of any one of claims 1 to 14. A vector comprising the nucleic acid molecule of claim 15. An isolated host cell comprising the isolated anti-PcrV antibody of any one of claims 1 to 14, the nucleic acid molecule of claim 15 or the vector of claim 16. A method for preparing an isolated anti-PcrV antibody, comprising the following steps: a) culturing the host cell of claim 17 under conditions effective to express anti-PcrV antibodies; and b) Obtaining the expressed anti-PcrV antibody in the host cell. A pharmaceutical composition comprising the isolated anti-PcrV antibody described in any one of claims 1 to 14, the nucleic acid molecule described in claim 15, the vector described in claim 16, or the isolated antibody described in claim 17 a host cell; and a pharmaceutically acceptable carrier. A use of the pharmaceutical composition of claim 19 for the preparation of a medicament for treating a disease or disorder in an individual in need. The use as claimed in claim 20, wherein the disease or disorder is a pathogenic infection. The use of claim 21, wherein the infection is a Gram-negative bacterial infection. The use according to claim 22, wherein the Gram-negative bacteria is Pseudomonas aeruginosa. The use of any one of claims 20 to 23, wherein the disease or disorder comprises one or more symptoms caused by Pseudomonas aeruginosa infection. The use of claim 24, wherein the symptoms include fever, chills, fatigue, muscle and joint pain, joint swelling, headache, diarrhea, rash, pus from wounds, bacteremia, acute pneumonia, intra-abdominal infection, respiratory tract One of infection, septic shock, septic arthritis, enteritis, skin and soft tissue infection, urinary tract infection, intestinal infection, ulcerative keratitis, chronic suppurative otitis media, mastoiditis, sinusitis and endocarditis or more. The use of any one of claims 20 to 25, wherein the pharmaceutical composition is further administered in combination with one or more therapeutic agents. The use of claim 26, wherein at least one of the therapeutic agents is an antibiotic. The use of claim 27, wherein the antibiotic is one or more of imipenem, tobramycin, ciprofloxacin, meropenem and atreonam.

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