Combinations of antibodies and bispecific antibodies comprising antigen binding that specifically recognizes Pseudomonas PCRV and PSL

Abstract

The present invention provides a bispecific antibody that specifically recognizes PcrV and / or Psl from Pseudomonas aeruginosa, a pharmaceutical composition comprising an antibody that specifically recognizes non-overlapping epitopes of PcrV and / or Psl from Pseudomonas genus, and methods for preparing and using the same.

Description

Technical Field

[0001] Submission of Sequence Listing in ASCII Text File The content in the ASCII text file submitted below is hereby incorporated by reference in its entirety into this specification. Sequence Listing in computer-readable form (CRF) (File name: 202011036860_Sequence Listing.TXT, Recording date: November 03, 2020; Size: 276 KB)

[0002] The present invention relates to a pharmaceutical composition comprising a bispecific antibody that specifically recognizes PcrV and / or Psl derived from Pseudomonas aeruginosa (PA), an antigen-binding molecule that specifically recognizes a non-overlapping epitope of Pseudomonas PcrV, and / or an antigen-binding molecule that specifically recognizes Pseudomonas Psl, a method for preparing the same, and uses thereof, which uses include a method for treating and preventing Pseudomonas infections.

Background Art

[0003] Pseudomonas aeruginosa is an obligate aerobic Gram-negative bacillus that widely exists in nature. Pseudomonas aeruginosa usually has low pathogenicity but is an opportunistic pathogen, so Pseudomonas aeruginosa infections often occur in patients with various existing diseases such as cancer, diabetes, and immunodeficiency, or in patients receiving drugs with immunosuppressive effects. Patients with broken skin and mucous membranes are easily infected with Pseudomonas aeruginosa, and there is also a significant risk for patients with chronic structural lung diseases (such as chronic obstructive pulmonary disease and cystic fibrosis). Pseudomonas aeruginosa often causes pneumonia, urinary tract infections, sepsis, etc., and often brings serious consequences. In up to 10% of hospitals, infections are caused by Pseudomonas aeruginosa, and the mortality rate of patients with Pseudomonas aeruginosa bacteremia is approaching 40%. In the clinical field, Pseudomonas aeruginosa infection is regarded as one of the most difficult infections to treat. This is because Pseudomonas aeruginosa itself not only has low sensitivity to existing antibiotics but also tends to have high resistance to various antibiotics. Therefore, the strategy of developing antibiotics has limited merits in dealing with Pseudomonas aeruginosa infections.

[0004] Pseudomonas aeruginosa is one of the main causes of nosocomial infections, especially in mechanically ventilated patients, and is the main cause of death in patients with cystic fibrosis. The Pseudomonas aeruginosa type III exotoxin secretion system (T3SS) is a major virulence factor associated with the severity of the disease and can directly inject bacterial toxins into the cytoplasm of host cells. Pseudomonas aeruginosa injects toxins into eukaryotic cells via the type III exotoxin secretion system (T3SS) to exert its high cytotoxicity. PcrV is a protein that constitutes the type III exotoxin secretion system, consists of 294 residues (NCBI accession number: AAC45935, SEQ ID NO: 71), and the operon sequence encoding the protein has been disclosed (US 6,551,795, Yahr, T. L. et al., J. Bacteriol., 1997, vol. 179, p. 7165). The PcrV protein located at the tip of the T3SS injectisome complex is essential for the function of T3SS and is a well-validated target in animal models of immunoprevention strategies targeting Pseudomonas aeruginosa. The T3SS of Pseudomonas aeruginosa is a well-validated target for intervening in infections caused by this opportunistic pathogen. In animal models, both active vaccination with T3SS component proteins and passive immunotherapy targeting PcrV can potently reduce diseases caused by Pseudomonas aeruginosa. Control of PcrV may lead to therapeutic means (T. Sawa et al., Nature Medicine, 1999, vol. 5, p. 392) for controlling Pseudomonas aeruginosa infection, so polyclonal antibodies with neutralizing activity against PcrV (Shime N et al., J. Immunol. 2001, vol. 16 (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) have been reported. However, since it is difficult to improve the antigenicity of polyclonal antibodies, it is difficult to humanize them and use them as pharmaceutical compositions. Antibodies against PcrV called V2L2-MD are described in Warrener et al., 2014, Antimicrob. Agents Chemother., 58, 4384-4391. The pegylated Fab fragment of an anti-PcrV monoclonal antibody based on the PcrV-specific mouse monoclonal antibody MAB166 is inactive against the prevention of Pseudomonas aeruginosa respiratory infections in mechanically ventilated patients. MAB166 can effectively block the Pseudomonas aeruginosa T3SS in vitro, but requires a relatively high antibody dose for protection in animal models. The present invention provides a novel anti-PcrV monoclonal antibody that shows effective inhibition of PcrV in vitro and in vivo.

[0005] One important component of the Pseudomonas aeruginosa biofilm matrix is the Psl polysaccharide, which is produced by proteins encoded by the polysaccharide synthesis locus. Psl can be free extracellularly or bound to the cell surface. The structure of extracellularly free Psl is composed of pentasaccharide repeats containing D-mannose, L-rhamnose, and D-glucose. Psl has both structural and protective functions during biofilm formation and is also known to be able to protect biofilms from antibiotics (by chemical binding) and the immune system (by an unknown mechanism), making it a potential ideal target for new therapeutic options (Ray VA et al. Anti-Psl Targeting of Pseudomonas aeruginosa Biofilms for Neutrophil-Mediated Disruption. Sci Rep. 2017). (DiGiandomenico, A et al. Identification of broadly protective human antibodies to Pseudomonas aeruginosa exopolysaccharide Psl by phenotypic screening. J Exp Med 209, 1273-1287; Valerie A. Ray, et al, Anti-Psl targeting of Pseudomonas aeruginosa biofilms for neutrophil mediated disruption, Scientific Reports 7, Article number: 16065 (2017)) describes human monoclonal antibodies (mAbs) that target Psl, such as Wapr-001, Wapr-016, Cam-003 or its derivative Psl0096. MedI3902 (also called MEDI3902) is a bivalent bispecific human immunoglobulin G1 (IgG1) κ monoclonal antibody (mAb) that can selectively bind to the PcrV protein and Psl extracellular polysaccharide on the surface of Pseudomonas aeruginosa. MedI3902 has a high protective effect in a mouse model of Pseudomonas aeruginosa infection.See, for example, PCT Publication Nos. WO2013 / 070615, WO2014 / 074528, PCT Application No. PCT / US2015 / 029063, and PCT Application No. PCT / US2015 / 036576.

[0006] The contents of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety. SUMMARY OF THE INVENTION

[0007] The present invention provides a bispecific molecule that specifically recognizes Pseudomonas PcrV and specifically recognizes Pseudomonas P sl, a bispecific molecule that specifically recognizes a non-overlapping epitope of Pseudomonas PcrV, and a pharmaceutical composition comprising an antigen-binding protein that specifically recognizes a non-overlapping epitope of Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Psl. The present invention also provides methods of using them for the prevention and treatment of Pseudomonas infections.

[0008] In one aspect of the present invention, a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein the first antigen-binding domain comprises (a) a heavy chain variable domain (V H ) and a light chain variable domain (V L ), and the V H comprises a heavy chain complementarity determining region (HC-CDR) 1 of DX1X2MS (SEQ ID NO:20), where X1 is N or Y and X2 is Y, H or P, an HC-CDR2 of X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G, and an HC-CDR3 of GRFSTX1SX2HFX3RAVYGMDV (SEQ ID NO:21), where X1 is L, S, N or D, X2 is S or A, and X3 is F or Y, and the V Lcomprises a light chain complementarity determining region (LC-CDR) 1 containing RASQGISSYLA (SEQ ID NO:10), an LC-CDR2 containing AASTLQS (SEQ ID NO:11), and an LC-CDR3 containing QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T, or (b) V H and V L and said V H comprises an HC-CDR1 containing SYWMH (SEQ ID NO:22), an HC-CDR2 containing RINEX1EX2SISYADSVKG (SEQ ID NO:44), where X1 is D or N and X2 is T, G or R, and an HC-CDR3 containing DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T and X2 is F or L, and said V L comprises an LC-CDR1 containing RASQX1VX2X3NLA (SEQ ID NO:46), where X1 is S, G or N, X2 is S, R or K, and X3 is S or N, an LC-CDR2 containing X1ASSRAT (SEQ ID NO:42), where X1 is D or H, and an LC-CDR3 containing QQYGX1X2PX3T (SEQ ID NO:47), where X1 is S, L or N, X2 is S, Q or E, and X3 is L or I, or (c) V H and V L and said V H comprises an HC-CDR1 containing X1X2X3MS (SEQ ID NO:17), where X1 is D or S, X2 is Y or N, and X3 is P, H, Y or S, an HC-CDR2 containing X1ISESGGSTX2X3ADSVKG (SEQ ID NO:18), where X1 is G or V, X2 is N or Y, and X3 is D or Y, and GRFX1X2X3X4X5X6FX7RAVYGMDV (SEQ comprises HC-CDR3 containing [[ID=]], where X1 is S or C, X2 is T, G, D, Y, Q or A, X3 is S, D, N, E, L, A or Y, X4 is S, T, Y or A, X5 is S, H, Q, A, R, K, G, E, Y or D, X6 is H or C, and X7 is F or Y, and said V L comprises LC-CDR1 containing RASQGIX1SYLA (SEQ ID NO:209), where X1 is S or R, LC-CDR2 containing AASTLQS (SEQ ID NO:11), and LC-CDR3 containing QQLX1SYPLX2 (SEQ ID NO:210), where X1 is S, N or K, and X2 is S or T, or (d) V H and V L comprises, and said V H comprises HC-CDR1 containing SYWMH (SEQ ID NO:22), HC-CDR2 containing RINEX1EX2SISYADSVKG (SEQ ID NO:211), where X1 is D, N, I, L or V, and X2 is S, T, R, G or N, and HC-CDR3 containing DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T, and X2 is F or L, and said V L comprises RASQX1VX2X3NLA (SEQ ID NO:212), where X1 is N, G, D or S, X2 is K, R, S, N or T, and X3 is N, G, S or D, LC-CDR1, LC-CDR2 containing X1ASSRAT (SEQ ID NO:213), where X1 is D, N, H or A, and LC-CDR3 containing QQYGX1X2PX3T (SEQ ID NO:214), where 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, to provide a bispecific molecule. In some embodiments, said second antigen-binding domain is (a) V H and V L comprises, and said V Hcomprises an HC-CDR1 comprising SSGDYWG (SEQ ID NO:48), an HC-CDR2 comprising SIHNX1GSTYYNPSLKG (SEQ ID NO:81), where X1 is S or Q, and an HC-CDR3 comprising QFGSETYYX1GIX2P (SEQ ID NO:82), where X1 is T, N or P and X2 is D or Q, said V L comprises an LC-CDR1 comprising RSSQSLLHSX1GYNYLD (SEQ ID NO:83), where X1 is N or R, an LC-CDR2 comprising LGSNRAS (SEQ ID NO:70), and an LC-CDR3 comprising MQALQTPYT (SEQ ID NO:74), or (b) V H and V L comprises, said V H comprises an HC-CDR1 comprising IHSVH (SEQ ID NO:50), an HC-CDR2 comprising TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 comprising DGX1S (SEQ ID NO:84), where X1 is D or T, said V L comprises an LC-CDR1 comprising RASQGISSWLA (SEQ ID NO:68), an LC-CDR2 comprising HASTLES (SEQ ID NO:72), and an LC-CDR3 comprising LQAX1SLPHT (SEQ ID NO:85), where X1 is N, F, S or K, or (c) V H and V L comprises, said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75, or (d) V H and V L comprises, said VH comprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:51, an HC-CDR2 containing the amino acid sequence SEQ ID NO:56, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:64, wherein said V L comprises an amino acid sequence SEQ ID NO:69, an LC-CDR2 containing the amino acid sequence SEQ ID NO:73, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:80.

[0009] In one aspect of the present invention, a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein said second antigen-binding domain comprises (a) V H and V L wherein said V H comprises an HC-CDR1 containing SSGDYWG (SEQ ID NO:48), an HC-CDR2 containing SIHNX1GSTYYNPSLKG (SEQ ID NO:81), where X1 is S or Q, and an HC-CDR3 containing QFGSETYYX1GIX2P (SEQ ID NO:82), where X1 is T, N or P and X2 is D or Q, and said V L comprises RSSQSLLHSX1GYNYLD (SEQ ID NO:83), where X1 is N or R, an LC-CDR2 containing LGSNRAS (SEQ ID NO:70), and an LC-CDR3 containing MQALQTPYT (SEQ ID NO:74), or (b) V H and V L wherein said V H comprises an HC-CDR1 containing IHSVH (SEQ ID NO:50), an HC-CDR2 containing TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 containing DGX1S (SEQ ID NO:84), where X1 is D or T, and said V Lcomprises LC-CDR1 comprising RASQGISSWLA (SEQ ID NO:68) and HASTLES (SEQ ID NO:72) comprising LC-CDR2, and LC-CDR3 comprising LQAX1SLPHT (SEQ ID NO:85), where X1 is N, F, S or K, or (c) V H and V L comprising, said V H comprises HC-CDR1 comprising amino acid sequence SEQ ID NO:49, HC-CDR2 comprising amino acid sequence SEQ ID NO:54, and HC-CDR3 comprising amino acid sequence SEQ ID NO:61, said V L comprises LC-CDR1 comprising amino acid sequence SEQ ID NO:67, LC-CDR2 comprising amino acid sequence SEQ ID NO:71, and LC-CDR3 comprising amino acid sequence SEQ ID NO:75, or (d) V H and V L comprising, said V H comprises HC-CDR1 comprising amino acid sequence SEQ ID NO:51, HC-CDR2 comprising amino acid sequence SEQ ID NO:56, and HC-CDR3 comprising amino acid sequence SEQ ID NO:64, said V L comprises LC-CDR1 comprising amino acid sequence SEQ ID NO:69, LC-CDR2 comprising amino acid sequence SEQ ID NO:73, and LC-CDR3 comprising amino acid sequence SEQ ID NO:80, to provide a bispecific molecule.

[0010] In some embodiments, said first antigen-binding domain comprises V H and V L comprising, said V H comprises HC-CDR1 comprising amino acid sequence SEQ ID NO:1, HC-CDR2 comprising amino acid sequence SEQ ID NO:4, and HC-CDR3 comprising amino acid sequence SEQ ID NO:6, said V LIt includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:10, an LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:12. In some embodiments, the first antigen-binding domain is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:2, an HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:7. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:10, an LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:13. In some embodiments, the first antigen-binding domain is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:22, an HC-CDR2 containing the amino acid sequence SEQ ID NO:23, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:26. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:30, an LC-CDR2 containing the amino acid sequence SEQ ID NO:33, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:35. In some embodiments, the first antigen-binding domain is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:22, an HC-CDR2 containing the amino acid sequence SEQ ID NO:24, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:27. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:31, an LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:36.

[0011] In some embodiments, the second antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:48, an HC-CDR2 having the amino acid sequence SEQ ID NO:52, and an HC-CDR3 having the amino acid sequence SEQ ID NO:57, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:65, an LC-CDR2 having the amino acid sequence SEQ ID NO:70, and an LC-CDR3 having the amino acid sequence SEQ ID NO:74. In some embodiments, the second antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:50, an HC-CDR2 having the amino acid sequence SEQ ID NO:55, and an HC-CDR3 having the amino acid sequence SEQ ID NO:62, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:68, an LC-CDR2 having the amino acid sequence SEQ ID NO:72, and an LC-CDR3 having the amino acid sequence SEQ ID NO:76.

[0012] In some embodiments, according to any of the bispecific molecules described herein, the first antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:1, an HC-CDR2 having the amino acid sequence SEQ ID NO:4, and an HC-CDR3 having the amino acid sequence SEQ ID NO:6, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:10, an LC-CDR2 having the amino acid sequence SEQ ID NO:11, and an LC-CDR3 having the amino acid sequence SEQ ID NO:12, and the second antigen-binding domain comprises V H and V L wherein the VH comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74. In some embodiments, said first antigen-binding domain comprises V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:12, and said second antigen-binding domain comprises V H and V L and said V H comprises the amino acid sequence SEQ ID NO:49 for HC-CDR1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75. In some embodiments, said first antigen-binding domain comprises V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and said V Lcomprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:10, an LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:12, and the second antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:50, an HC-CDR2 containing the amino acid sequence SEQ ID NO:55, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:62, and the V L comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:68, an LC-CDR2 containing the amino acid sequence SEQ ID NO:72, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:76.

[0013] In some embodiments, the first antigen-binding domain comprises V H and V L wherein the V H comprises the amino acid sequence SEQ ID NO:91 or 161, the V L comprises the amino acid sequence SEQ ID NO:112 or 182, the second antigen-binding domain comprises V H and V L wherein the V H comprises the amino acid sequence SEQ ID NO:103 or 173, and the V L comprises the amino acid sequence SEQ ID NO:117 or 187. In some embodiments, the first antigen-binding domain comprises V H and V L wherein the V H comprises the amino acid sequence SEQ ID NO:91 or 161, the V L comprises the amino acid sequence SEQ ID NO:112 or 182, the second antigen-binding domain comprises V H and V L wherein the V H comprises the amino acid sequence SEQ ID NO:109 or 179, and the V LIt includes the amino acid sequence SEQ ID NO: 120 or 190. In some embodiments, the first antigen-binding domain is V H and V L and the V H includes the amino acid sequence SEQ ID NO: 91 or 161, the V L includes the amino acid sequence SEQ ID NO: 112 or 182, and the second antigen-binding domain is V H and V L and the V H includes the amino acid sequence SEQ ID NO: 107 or 177, and the V L includes the amino acid sequence SEQ ID NO: 119 or 189.

[0014] In some embodiments, the first antigen-binding domain is a Fab arm that specifically recognizes PcrV, the second antigen-binding domain is a single-chain variable region fragment (scFv) that specifically recognizes Psl, the bispecific molecule further includes an Fc region containing CH2 and CH3 domains, the single-chain variable region fragment (scFv) is interconnected with the Fab arm via a first polypeptide linker (L1) and with the Fc region via a second polypeptide linker (L2), and the bispecific molecule is bivalent for binding to each of PcrV and Psl. In some embodiments, the first antigen-binding domain is a single-chain variable region fragment (scFv) that specifically recognizes PcrV, the second antigen-binding domain is a Fab arm that specifically recognizes Psl, the bispecific molecule further includes an Fc region containing CH2 and CH3 domains, the single-chain variable region fragment (scFv) is interconnected with the Fab arm via a first polypeptide linker (L1) and with the Fc region via a second polypeptide linker (L2), and the bispecific molecule is bivalent for binding to each of PcrV and Psl. In some embodiments, the bispecific molecule has the amino acid sequence SEQ ID NO: 135, 136 or 159, and / or SEQ It contains any one amino acid sequence of ID NOs: 195 to 208.

[0015] In some embodiments, the bispecific molecule comprises an Fc region, and the Fc region is selected from the group consisting of the Fc regions of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, and IgD. In some embodiments, the Fc region comprises a variable Fc region. In some embodiments, the Fc region is glycosylated. In some embodiments, the Fc region is deglycosylated. In some embodiments, the Fc region has reduced fucosylation or is afucosylated. In some embodiments, the variable Fc region comprises a substitution at position 297. In some embodiments, the substitution at position 297 is 297Q. In some embodiments, the variable Fc region comprises substitutions at one or more of the sites of positions 239, 282, 289, 297, 312, 324, 330, 335, 337, 339, 356, 359, 361, 383, 384, 398, 400, 440, 422, and 442 according to the EU index number.

[0016] In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising any one amino acid sequence of SEQ ID NOs: 137 to 152 and 160, and / or a light chain comprising the amino acid sequence SEQ ID NO: 135, 136, or 159.

[0017] In one aspect of the present invention, a pharmaceutical composition comprising (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV, and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises (a) a heavy chain variable domain (V H ) and a light chain variable domain (V L ), and the V Hcomprises a heavy chain complementarity determining region (HC-CDR) 1 of DX1X2MS (SEQ ID NO:20), where X1 is N or Y and X2 is Y, H or P; a HC-CDR2 of X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G; and a HC-CDR3 of GRFSTX1SX2HFX3RAVYGMDV (SEQ ID NO:21), where X1 is L, S, N or D and X2 is S or A; and HC-CDR3 is F or Y; L comprises a light chain complementarity determining region (LC-CDR) 1 comprising RASQGISSYLA (SEQ ID NO:10), a LC-CDR2 comprising AASTLQS (SEQ ID NO:11), and a LC-CDR3 comprising QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T; or (b) V H and V L The V H comprises an HC-CDR1 comprising SYWMH (SEQ ID NO:22), an HC-CDR2 comprising RINEX1EX2SISYADSVKG (SEQ ID NO:44), where X1 is D or N and X2 is T, G or R, and an HC-CDR3 comprising DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T and X2 is F or L; L comprises an LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO:46), where X1 is S, G or N, X2 is S, R or K, and X3 is S or N; an LC-CDR2 comprising X1ASSRAT (SEQ ID NO:42), where X1 is D or H; and an LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO:47), where X1 is S, L or N, X2 is S, Q or E, and X3 is L or I; or (c) V H and V L The V Hcontains X1X2X3MS (SEQ ID NO:17), where X1 is D or S, X2 is Y or N, X3 is P, H, Y or S, the HC-CDR1, and X1ISESGGSTX2X3ADSVKG (SEQ ID NO:18), where X1 is G or V, X2 is N or Y, X3 is D or Y, the HC-CDR2, and GRFX1X2X3X4X5X6FX7RAVYGMDV (SEQ ID NO:38), where X1 is S or C, X2 is T, G, D, Y, Q or A, X3 is S, D, N, E, L, A or Y, X4 is S, T, Y or A, X5 is S, H, Q, A, R, K, G, E, Y or D, X6 is H or C, X7 is F or Y, the HC-CDR3, and the V L contains RASQGIX1SYLA (SEQ ID NO:209), where X1 is S or R, the LC-CDR1, AASTLQS (SEQ ID NO:11), the LC-CDR2, and QQLX1SYPLX2 (SEQ ID NO:210), where X1 is S, N or K, X2 is S or T, the LC-CDR3, or (d)V H and V L contains, the V H contains SYWMH (SEQ ID NO:22), the HC-CDR1, RINEX1EX2SISYADSVKG (SEQ ID NO:211), where X1 is D, N, I, L or V, X2 is S, T, R, G or N, the HC-CDR2, and DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T, X2 is F or L, the HC-CDR3, and the V LIt provides a pharmaceutical composition comprising LC-CDR1 containing RASQX1VX2X3NLA (SEQ ID NO:212), where X1 is N, G, D or S, X2 is K, R, S, N or T, X3 is N, G, S or D; LC-CDR2 containing X1ASSRAT (SEQ ID NO:213), where X1 is D, N, H or A; and LC-CDR3 containing QQYGX1X2PX3T (SEQ ID NO:214), where 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, X3 is I, L or V.

[0018] In one aspect of the present invention, there is provided a method for treating and / or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject in an effective amount (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises (a) a heavy-chain variable domain (V H ) and a light-chain variable domain (V L ), and the V H comprises a heavy-chain complementarity-determining region (HC-CDR) 1 containing DX1X2MS (SEQ ID NO:20), where X1 is N or Y, and X2 is Y, H or P; an HC-CDR2 containing X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G; and an HC-CDR3 containing GRFSTX1SX2HFX3RAVYGMDV (SEQ ID NO:21), where X1 is L, S, N or D, X2 is S or A, and X3 is F or Y, and the V Lcomprises a light chain complementarity determining region (LC-CDR) 1 comprising RASQGISSYLA (SEQ ID NO:10), an LC-CDR2 comprising AASTLQS (SEQ ID NO:11), and an LC-CDR3 comprising QQLSSYPLX1 (SEQ ID NO:19), wherein X1 is S or T, or (b) V H and V L comprises, wherein said V H comprises an HC-CDR1 comprising SYWMH (SEQ ID NO:22), an HC-CDR2 comprising RINEX1EX2SISYADSVKG (SEQ ID NO:44), wherein X1 is D or N and X2 is T, G or R, and an HC-CDR3 comprising DGPYDX1X2DI (SEQ ID NO:45), wherein X1 is S, A or T and X2 is F or L, and said V L comprises an LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO:46), wherein X1 is S, G or N, X2 is S, R or K, and X3 is S or N, an LC-CDR2 comprising X1ASSRAT (SEQ ID NO:42), wherein X1 is D or H, and an LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO:47), wherein X1 is S, L or N, X2 is S, Q or E, and X3 is L or I, or (c) V H and V L comprises, wherein said V Hcomprises HC-CDR1 containing X1X2X3MS (SEQ ID NO:17), where X1 is D or S, X2 is Y or N, and X3 is P, H, Y or S, HC-CDR2 containing X1ISESGGSTX2X3ADSVKG (SEQ ID NO:18), where X1 is G or V, X2 is N or Y, and X3 is D or Y, and HC-CDR3 containing GRFX1X2X3X4X5X6FX7RAVYGMDV (SEQ ID NO:38), where X1 is S or C, X2 is T, G, D, Y, Q or A, X3 is S, D, N, E, L, A or Y, X4 is S, T, Y or A, X5 is S, H, Q, A, R, K, G, E, Y or D, X6 is H or C, and X7 is F or Y, said V L comprises LC-CDR1 containing RASQGIX1SYLA (SEQ ID NO:209), where X1 is S or R, LC-CDR2 containing AASTLQS (SEQ ID NO:11), and LC-CDR3 containing QQLX1SYPLX2 (SEQ ID NO:210), where X1 is S, N or K, and X2 is S or T, or (d)V H and V L comprises, said V H comprises HC-CDR1 containing SYWMH (SEQ ID NO:22), HC-CDR2 containing RINEX1EX2SISYADSVKG (SEQ ID NO:211), where X1 is D, N, I, L or V, and X2 is S, T, R, G or N, and HC-CDR3 containing DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T, and X2 is F or L, said V LIt provides a method for treating and / or preventing a disease or medical condition in a subject in need thereof, comprising an LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO:212), wherein X1 is N, G, D or S, X2 is K, R, S, N or T, X3 is N, G, S or D; an LC-CDR2 comprising X1ASSRAT (SEQ ID NO:213), wherein X1 is D, N, H or A; and an LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO:214), 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, X3 is I, L or V.

[0019] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is (a) V H and V L comprising, wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:2, an HC-CDR2 comprising the amino acid sequence SEQ I D NO:5, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:7, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:13, or (b) V H and V L comprising, wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:12, or (c) V H and V L comprising, wherein said V Hcomprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:22, an HC-CDR2 containing the amino acid sequence SEQ ID NO:23, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:26, wherein said V L comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:30, an LC-CDR2 containing the amino acid sequence SEQ ID NO:33, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:35, or (d) V H and V L comprising, wherein said V H comprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:22, an HC-CDR2 containing the amino acid sequence SEQ ID NO:24, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:27, wherein said V L comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:31, an LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:36.

[0020] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is (a) V H and V L comprising, wherein said V H comprises an HC-CDR1 containing SSGDYWG (SEQ ID NO:48), an HC-CDR2 containing SIHNX1GSTYYNPSLKG (SEQ ID NO:81), where X1 is S or Q, and an HC-CDR3 containing QFGSETYYX1GIX2P (SEQ ID NO:82), where X1 is T, N or P and X2 is D or Q, wherein said V L comprises an LC-CDR1 containing RSSQSLLHSX1GYNYLD (SEQ ID NO:83), where X1 is N or R, an LC-CDR2 containing LGSNRAS (SEQ ID NO:70), and an LC-CDR3 containing MQALQTPYT (SEQ ID NO:74), or (b) V H and V L comprising, wherein said V Hcomprises an HC-CDR1 comprising IHSVH (SEQ ID NO:50), an HC-CDR2 comprising TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 comprising DGX1S (SEQ ID NO:84), wherein X1 is D or T, and said V L comprises an LC-CDR1 comprising RASQGISSWLA (SEQ ID NO:68), an LC-CDR2 comprising HASTLES (SEQ ID NO:72), and an LC-CDR3 comprising LQAX1SLPHT (SEQ ID NO:85), wherein X1 is N, F, S or K, or (c) V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75, or (d) V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:51, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:56, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:64, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:69, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:73, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:80.

[0021] In one aspect of the present invention, a pharmaceutical composition comprising (i) an antigen-binding protein that specifically recognizes Pseudomonas Psl and (ii) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV, wherein the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises (a) V H and V L wherein said V H comprises an HC-CDR1 comprising SSGDYWG (SEQ ID NO:48), an HC-CDR2 comprising SIHNX1GSTYYNPSLKG (SEQ ID NO:81), where X1 is S or Q, and an HC-CDR3 comprising QFGSETYYX1GIX2P (SEQ ID NO:82), where X1 is T, N or P and X2 is D or Q, and said V L comprises an LC-CDR1 comprising RSSQSLLHSX1GYNYLD (SEQ ID NO:83), where X1 is N or R, an LC-CDR2 comprising LGSNRAS (SEQ ID NO:70), and an LC-CDR3 comprising MQALQTPYT (SEQ ID NO:74), or (b) V H and V L wherein said V H comprises an HC-CDR1 comprising IHSVH (SEQ ID NO:50), an HC-CDR2 comprising TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 comprising DGX1S (SEQ ID NO:84), where X1 is D or T, and said V L comprises an LC-CDR1 comprising RASQGISSWLA (SEQ ID NO:68), an LC-CDR2 comprising HASTLES (SEQ ID NO:72), and an LC-CDR3 comprising LQAX1SLPHT (SEQ ID NO:85), where X1 is N, F, S or K, or (c) V H and V L wherein said V Hcomprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, wherein said V L comprises an amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75, or (d) V H and V L wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:51, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:56, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:64, wherein said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:69, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:73, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:80, and provides a pharmaceutical composition.

[0022] In one aspect of the present invention, there is provided a method for treating and / or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject in need thereof an effective amount of (i) an antigen-binding protein that specifically recognizes Pseudomonas Psl and (ii) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV, wherein the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises (a) V H and V L wherein said V H comprises an HC-CDR1 comprising SSGDYWG (SEQ ID NO:48), an HC-CDR2 comprising SIHNX1GSTYYNPSLKG (SEQ ID NO:81), where X1 is S or Q, and an HC-CDR3 comprising QFGSETYYX1GIX2P (SEQ ID NO:82), where X1 is T, N or P and X2 is D or Q, wherein said VL comprises LC-CDR1 containing RSSQSLLHSX1GYNYLD (SEQ ID NO:83), where X1 is N or R, LC-CDR2 containing LGSNRAS (SEQ ID NO:70), and LC-CDR3 containing MQALQTPYT (SEQ ID NO:74), or (b) V H and V L comprises, wherein said V H comprises HC-CDR1 containing IHSVH (SEQ ID NO:50), HC-CDR2 containing TIISSGTTTTYAQSFQD (SEQ ID NO:55), and HC-CDR3 containing DGX1S (SEQ ID NO:84), where X1 is D or T, and said V L comprises RASQGISSWLA (SEQ ID NO:68) as LC-CDR1, HASTLES (SEQ ID NO:72) as LC-CDR2, and LQAX1SLPHT (SEQ ID NO:85) as LC-CDR3, where X1 is N, F, S or K, or (c) V H and V L comprises, wherein said V H comprises HC-CDR1 with amino acid sequence SEQ ID NO:49, HC-CDR2 with amino acid sequence SEQ ID NO:54, and HC-CDR3 with amino acid sequence SEQ ID NO:61, and said V L comprises LC-CDR1 with amino acid sequence SEQ ID NO:67, LC-CDR2 with amino acid sequence SEQ ID NO:71, and LC-CDR3 with amino acid sequence SEQ ID NO:75, or (d) V H and V L comprises, wherein said V H comprises HC-CDR1 with amino acid sequence SEQ ID NO:51, HC-CDR2 with amino acid sequence SEQ ID NO:56, and HC-CDR3 with amino acid sequence SEQ ID NO:64, and said V LProvided is a method for treating and / or preventing a disease or disorder in a subject in need thereof, comprising an LC-CDR1 comprising the amino acid sequence SEQ ID NO:69, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:73, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:80.

[0023] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:2, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:5, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:7, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:13.

[0024] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:12. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V Hcomprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:23, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:26, wherein said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, an amino acid sequence SEQ ID NO:33 for LC-CDR2, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:35. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:27, said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:36.

[0025] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and said V Hcomprises HC-CDR1 containing the amino acid sequence SEQ ID NO:50, HC-CDR2 containing the amino acid sequence SEQ ID NO:55, and HC-CDR3 containing the amino acid sequence SEQ ID NO:62, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:68, LC-CDR2 containing the amino acid sequence SEQ ID NO:72, and LC-CDR3 containing the amino acid sequence SEQ ID NO:76.

[0026] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:1, HC-CDR2 containing the amino acid sequence SEQ ID NO:4, and HC-CDR3 containing the amino acid sequence SEQ ID NO:6, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:12, and the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:48, HC-CDR2 containing the amino acid sequence SEQ ID NO:52, and HC-CDR3 containing the amino acid sequence SEQ ID NO:57, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:65, LC-CDR2 containing the amino acid sequence SEQ ID NO:70, and LC-CDR3 containing the amino acid sequence SEQ ID NO:74.

[0027] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises V H and V L wherein said VH comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:1, HC-CDR2 containing the amino acid sequence SEQ ID NO:4, and HC-CDR3 containing the amino acid sequence SEQ ID NO:6, wherein said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:12, and wherein said antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:49, HC-CDR2 containing the amino acid sequence SEQ ID NO:54, and HC-CDR3 containing the amino acid sequence SEQ ID NO:61, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:67, LC-CDR2 containing the amino acid sequence SEQ ID NO:71, and LC-CDR3 containing the amino acid sequence SEQ ID NO:75.

[0028] In some embodiments, said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V H and V L comprises, and said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:2, HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and HC-CDR3 containing the amino acid sequence SEQ ID NO:7, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:13, and wherein said antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and said V Hcomprises HC-CDR1 containing the amino acid sequence SEQ ID NO:48, HC-CDR2 containing the amino acid sequence SEQ ID NO:52, and HC-CDR3 containing the amino acid sequence SEQ ID NO:57, wherein said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:65, LC-CDR2 containing the amino acid sequence SEQ ID NO:70, and a LC-CDR3 containing the amino acid sequence SEQ ID NO:74.

[0029] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:1, HC-CDR2 containing the amino acid sequence SEQ ID NO:4, and HC-CDR3 containing the amino acid sequence SEQ ID NO:6, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:12. The antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:22, HC-CDR2 containing the amino acid sequence SEQ ID NO:24, and the amino acid sequence SEQ ID NO:27 for HC-CDR3, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:31, LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and LC-CDR3 containing the amino acid sequence SEQ ID NO:36. And the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L wherein said V Hcomprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, wherein said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:74.

[0030] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said V H comprises the amino acid sequence SEQ ID NO:91, said V L comprises the amino acid sequence SEQ ID NO:112, and the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises V H and V L and said V H comprises the amino acid sequence SEQ ID NO:103, said V L comprises the amino acid sequence SEQ ID NO:117. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said V H comprises the amino acid sequence SEQ ID NO:92, said V L comprises the amino acid sequence SEQ ID NO:113, and the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises V H and V L and said V H comprises the amino acid sequence SEQ ID NO:103, said V L comprises the amino acid sequence SEQ ID NO:117. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said VH contains the amino acid sequence SEQ ID NO:91, and said V L contains the amino acid sequence SEQ ID NO:112, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L contains the amino acid sequence SEQ ID NO:107, and said V H contains the amino acid sequence SEQ ID NO:119. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V L and V H contains the amino acid sequence SEQ ID NO:91, and said V L contains the amino acid sequence SEQ ID NO:112, and the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H contains the amino acid sequence SEQ ID NO:96, and said V L contains the amino acid sequence SEQ ID NO:115, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L contains the amino acid sequence SEQ ID NO:103, and said V H contains the amino acid sequence SEQ ID NO:117, and said V L contains the amino acid sequence SEQ ID NO:115, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L contains the amino acid sequence SEQ ID NO:103, and said V H contains the amino acid sequence SEQ ID NO:117, and said V L contains the amino acid sequence SEQ ID NO:117.

[0031] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV and / or the antigen-binding protein that specifically recognizes Pseudomonas Psl are administered simultaneously. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV and / or the antigen-binding protein that specifically recognizes Pseudomonas Psl are administered sequentially.

[0032] In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is about 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, or 1:5. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is about 2:1 or 1:1. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, or 1:5. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 2:1 or 1:1. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1:1 or 1:1:2.

[0033] In some embodiments, provided is a method of treating and / or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject in need an effective amount of any of the pharmaceutical compositions and / or bispecific molecules described herein.

[0034] In some embodiments, according to any of the methods described herein, the disease or disorder is a pathogenic infection. In some embodiments, the infection is a Gram-negative bacterial infection. In some embodiments, the pathogen is Pseudomonas aeruginosa. In some embodiments, the disease or disorder includes one or more of the symptoms caused by Pseudomonas aeruginosa infection. In some embodiments, the symptoms include fever, chills, fatigue, muscle and joint pain, joint swelling, headache, diarrhea, skin rash, wound pus, bacteremia, acute pneumonia, intra-abdominal infection, respiratory infection, septic shock, suppurative arthritis, enteritis, skin and soft tissue infection, urinary tract infection, intestinal infection, ulcerative keratitis, chronic suppurative otitis media, mastoiditis, sinusitis, and endocarditis.

[0035] In some embodiments, there is provided an isolated nucleic acid molecule encoding any one of the above-described antigen-binding proteins that recognize PcrV, the antigen-binding proteins that recognize Psl, and the bispecific molecule. In some embodiments, there is provided a vector comprising any one of the nucleic acid molecules. In some embodiments, there is provided a host cell comprising any one of the above-described antigen-binding proteins or bispecific molecules, any one of the nucleic acid molecules, or any one of the vectors. In some embodiments, a) culturing any one of the above-described host cells under conditions effective to express an anti-PcrV antibody, an anti-Psl antibody, or a bispecific molecule that recognizes PcrV and / or Psl, and b) obtaining from the host cell the expressed anti-PcrV antibody, anti-Psl antibody, or bispecific molecule that recognizes PcrV and / or Psl. A method for preparing an anti-PcrV antibody, an anti-Psl antibody, or a bispecific molecule that recognizes PcrV and / or Psl is provided. Also provided are pharmaceutical compositions, kits, and products comprising any one of the above-described bispecific molecules, anti-PcrV antibodies and / or anti-Psl antibodies, nucleic acids, vectors, and isolated host cells.

[0036] BRIEF DESCRIPTION OF THE DRAWINGS ​

[0037]

Figure 1A

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[0038] In one aspect of the invention, a bispecific molecule that specifically recognizes PcrV and / or Psl is provided. In one aspect of the invention, a combination of an antigen-binding protein that specifically recognizes PcrV and / or an antigen-binding protein that specifically recognizes Psl is provided. By using a combination of screening of a scFv phage library, affinity maturation, and appropriately designed biochemical and biological assays, highly effective antigen-binding proteins that bind to PcrV and antibody molecules that bind to Psl were identified. The results herein show that, compared to the administration of a single antibody molecule alone, a combination of these antigen-binding sites, either (i) in the form of a pharmaceutical composition or (ii) in the form of a bispecific antibody, can cumulatively or synergistically inhibit the lysis of red blood cells and A549 cells by Pseudomonas aeruginosa and provide stronger treatment and prophylaxis against Pseudomonas aeruginosa in vivo.

[0039] Also provided are nucleic acids encoding PcrV and Psl binding proteins and domains, bispecific molecules, compositions comprising PcrV and Psl binding proteins or bispecific molecules, and methods for manufacturing and using PcrV and Psl binding proteins and bispecific molecules.

[0040] Definitions As described herein, "treatment" or "treating" is a method of obtaining a result that includes a beneficial or desired clinical outcome. In view of the objectives of the present invention, said beneficial or desired clinical outcome includes, but is not limited to, one or more of alleviating one or more symptoms caused by a disease, reducing the degree of the disease, stabilizing the disease (e.g., preventing or delaying the worsening of the disease), preventing or delaying the spread of the disease (e.g., systemic spread of a pathogen), preventing or delaying the recurrence of the disease, delaying or retarding the progression of the disease, improving the condition of the disease, alleviating the disease (partially or completely), reducing the amount of one or more other drugs required for the treatment of the disease, delaying the progression of the disease, improving or enhancing the quality of life, increasing body weight, and / or extending the survival period. At the same time, "treatment" also includes a reduction in the pathological consequences of an infectious disease (e.g., lysis or necrosis of host cells). The methods of the present invention contemplate any one or more of these treatments.

[0041] The term "prevent", and similar words such as "prevented", "preventing", "prevention" or "prophylactic", denote a method of preventing, inhibiting or reducing the likelihood of the occurrence or recurrence of a disease or medical condition (e.g., infection by a pathogen). It further refers to delaying the occurrence or recurrence of a particular disease or medical condition, or delaying the occurrence or recurrence of the symptoms of a particular disease or medical condition. As used herein, "prevention" and similar words further include reducing the intensity, impact, symptoms and / or burden of a disease or medical condition prior to its occurrence or recurrence. As used herein, "prevention" and similar words further include reducing the risk and susceptibility to the occurrence or recurrence of a disease or medical condition, such as infection by a pathogen.

[0042] As described herein, an "antigen-binding protein" broadly refers to a protein that includes a portion that specifically binds to an antigen or target. Examples of antigen-binding proteins are antibodies and antibody fragments.

[0043] As described herein, the term "antibody" is used broadly and includes, but is not limited to, various antibody structures such as monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, and antigen-binding fragments thereof, as long as they exhibit the desired antigen-binding activity. A full-length antibody includes two heavy chains and two light chains. The heavy-chain variable region and the light-chain variable region are responsible for binding to the antigen. The variable regions in the two chains generally include three highly variable loops. Those loops are referred to as complementarity-determining regions (CDRs) (light-chain (LC) CDRs include LC-CDR1, LC-CDR2, and LC-CDR3, and heavy-chain (HC) CDRs include HC-CDR1, HC-CDR2, and HC-CDR3). The CDR boundaries of the antibodies or antigen-binding fragments described in the present invention are defined or recognized according to 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 chain or light chain are the framework regions Inserted between the flanking regions, referred to as framework regions (FRs), the framework regions (FRs) have a higher degree of conservation than the CDR regions and form struts that support highly variable loops. The constant regions of the heavy chains and the constant regions of the light chains do not participate in antigen binding but exhibit a variety of effector functions. Antibodies are classified based on the amino acid sequences of their heavy chain constant regions. The five main classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, which are characterized by having α, δ, ε, γ, and μ heavy chain types, respectively. Some of the main classes are further classified into subclasses, such as IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), IgA1 (α1 heavy chain), or IgA2 (α2 heavy chain).

[0044] As described herein, the term "antigen-binding fragment" refers to an antibody fragment that includes, for example, a diabody, Fab, Fab’, F(ab’)2, Fv fragment, disulfide-stabilized Fv fragment (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv’), disulfide-stabilized diabody (ds diabody), single-chain antibody (scFv), scFv dimer (bivalent diabody), multispecific antibody consisting of antibody fragments containing one or more CDRs, single-domain antibody, nanobody, domain antibody, bivalent domain antibody, or any other antibody fragment that can bind to an antigen but does not contain a complete antibody structure. The antigen-binding fragment further contains a fusion protein including the antibody fragment. The antigen-binding fragment can bind to the same antigen as the parent antibody or parent antibody fragment (e.g., parent scFv). In some embodiments, the antigen-binding fragment can include one or more CDRs from a specific human antibody transplanted into a framework region from one or more different human antibodies.

[0045] As described herein, the term "bispecific antibody" refers to an antibody having binding specificity for two different antigens or epitopes in one molecule. The manufacturing process of bispecific antibodies includes the design of the whole molecule, the synthesis and cloning of nucleotide sequences in each domain, the expression in mammalian cells, and the purification of the final product.

[0046] As described herein, the term "antigen-binding domain" refers to the portion of an antigen-binding molecule that specifically binds to an antigen. More specifically, the term "antigen-binding domain" refers to a portion of an antibody that specifically binds to a part or all of an antigen and includes complementary regions. In the case of a large antigen, the antigen-binding molecule may bind only to a specific part of the antigen. The specific part is called an antigen epitope. For example, the antigen-binding domain may be provided by one or more variable domains (also called variable regions). Preferably, the antigen-binding domain includes a variable domain of the antibody light chain (V L ) and a variable domain of the antibody heavy chain (V H ). In one aspect, the antigen-binding domain can bind to the antigen and block or partially block the function of the antigen. The antigen-binding domain that specifically binds to PcrV or Psl includes antibodies and fragments thereof as further defined herein.

[0047] As described 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. When two types of antibodies or antibody portions show competitive binding to an antigen, they may bind to the same epitope on the antigen.

[0048] As described herein, a first antibody "competes" with a second antibody for binding to a PcrV target if it inhibits the binding of the second antibody to the target PcrV by at least 50% (e.g., at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%) in the presence of an equimolar concentration of the first antibody, or vice versa. The PCT publication WO03 / 48731 describes a method for "epitope classification" of high-throughput antibodies based on antibody cross-competition.

[0049] As described herein, the terms "specifically binds", "specifically recognizes" or "is specific for" refer to a measurable and reproducible interaction, e.g., a binding between a target and an antibody that determines the presence of the target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody can specifically recognize a certain target (which may be an epitope) means that the binding between the antibody and the target has a higher affinity, a higher binding force, is easier and has a longer duration compared to the binding between the antibody and other targets. In some embodiments, an antibody that specifically recognizes an antigen reacts with one or more antigenic determinants of the antigen with a binding affinity that is at least 10-fold greater than its binding affinity for other targets.

[0050] As described herein, an "isolated" antibody refers to an antibody that is (1) not related to naturally occurring proteins, (2) does not contain other proteins from the same source, (3) is expressed by heterologous cells, or (4) does not exist in nature.

[0051] As described herein, the term "isolated nucleic acid" refers to a nucleic acid derived from genomic, cDNA or synthetic sources, or a combination thereof. Depending on its origin, the "isolated nucleic acid" is (1) not related to all or part of the polynucleotide in a "isolated nucleic acid" found in nature, (2) operably linked to a polynucleotide that is not linked to it in its natural state, or (3) does not exist naturally as part of a larger sequence.

[0052] As described herein, the term "CDR" or "complementary determining region" refers to the non - contiguous antigen - binding sites found in the variable regions of heavy - chain polypeptides and light - chain polypeptides. Kabat et al., J. Biol. Chem. 252:6609 - 6616(1977); Kabat et al., U.S. Dept. of Health and Human Services, “Sequences of proteins “of immunological interest” (1991); Chothia et 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 M. P. et al., Dev. Comp. Immunol., 27:55-77 (2003); and Honegger and Plückthun, J. Mol. Biol., 309:657-670 (2001) have already described these specific regions, and when compared to each other, these definitions include overlapping or subsets of amino acid residues. However, any definition scheme for the CDRs of an antibody or a grafted antibody or variants thereof is intended to be within the scope of the terms defined and used herein. The amino acid residues including the CDRs defined in each of the above references are listed in Table 1 for comparison. CDR prediction algorithms and interfaces are known in the art and are described, for example, in the references including 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 content of the references cited in this paragraph is incorporated herein by reference in its entirety as used in this application and as may be included in one or more claims of the present invention.

[0053]

Table 1

[0054] The term "chimeric antibody" refers to an antibody in which part of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species, or an antibody belonging to a particular antibody class or subclass, and the remainder of these chains is identical or homologous to the corresponding sequence in an antibody derived from another species, or an antibody belonging to another antibody class or subclass. Such antibody fragments need only have the biological activity of the present invention (see U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

[0055] "Fv" is the smallest antibody fragment that contains the intact antigen recognition and binding site. The fragment is a dimer in which one heavy chain variable domain and one light chain variable domain are linked by a tight non-covalent bond. From the folding of these two domains, six highly variable loops (three loops each in the light and heavy chains) are generated, and the highly variable loops provide the amino acid residues for antigen binding and confer antigen binding specificity on the antibody. However, even a single variable domain (or a half Fv fragment containing only the three antigen-specific CDRs) has the ability to recognize and bind an antigen, although its affinity is lower than that of the intact binding site.

[0056] "Single-chain Fv", also referred to as "sFv" or "scFv", refers to an antibody fragment comprising a V H antibody domain and a V L antibody domain linked in a single polypeptide chain. In some embodiments, the scFv polypeptide comprises a V H and a V LIt further includes a linking polypeptide between them. For an overview of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenberg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0057] The term "diabodies" refers to small antibody fragments prepared by constructing an scFv fragment (see the content of the previous paragraph) between V H and V L using a short linker (e.g., 5-10 residues). By this, inter-chain pairing rather than intra-chain pairing of the variable domains is achieved, generating a bivalent fragment, i.e., a fragment having two antigen-binding sites. Bispecific diabodies are heterodimers of two "crossover" scFv fragments, in which the V domains and V H domains of the two antibodies are located on different polypeptide chains. Diabodies have already been fully described in EP404,097; WO93 / 11161; Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993). L

[0058] ​The "humanized" form of a non-human (e.g., rodent) antibody is a chimeric antibody that contains a minimal sequence derived from the non-human antibody. Most of the humanized antibody is human immunoglobulin (the recipient antibody), and the residues derived from the hypervariable regions (HVRs) of the recipient antibody are replaced with hypervariable regions derived from a non-human species (the donor antibody) of mouse, rat, rabbit, or non-human primate that have the desired antibody specificity, affinity, and performance. In certain instances, residues in the framework regions (FRs) of the human immunoglobulin are replaced with the corresponding non-human residues. Additionally, the humanized antibody may contain 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 contains at least one, and generally two, variable domains, in which 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 framework regions are human immunoglobulin sequences. A human antibody optionally also contains at least a portion of the immunoglobulin constant region (Fc), typically the constant region of a human immunoglobulin. 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).

[0059] The "percent amino acid sequence identity (%)" or "homology" of the polypeptide and antibody sequences identified in this specification is defined as the percentage of identical amino acid residues in the candidate sequence and the polypeptide sequence to be compared, considering conservative substitutions as part of the sequence identity when comparing the sequences. The percent amino acid sequence identity can be determined by various comparison methods within the skill in the art, such as available computer software like BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. One of ordinary skill in the art can determine appropriate parameters for measurement and comparison, including any algorithm necessary to achieve maximum comparison over the entire length of the sequences being compared. However, for the purposes of the present invention, the value of the percent amino acid sequence identity is generated using the sequence comparison computer program MUSCLE (Edgar, R.C., Nucleic Acids Research 32(5):1792 - 1797, 2004; Edgar, R.C., BMC Bioinformatics 5(1):113, 2004).

[0060] The term "Fc receptor" or "FcR" is used to describe a receptor that binds to the Fc region of an antibody. In some embodiments, the FcR described in the present invention is an FcR that binds to an IgG antibody (a type of γ receptor), including receptors of the FcγRI, FcγRII, and FcγRIII subclasses, as well as allelic variants and alternative splicing forms of these receptors. The FcγRII receptor includes FcγRIIA (an activating receptor) and FcγRIIB (an inhibitory receptor). FcγRIIA (activating receptor) and FcγRIIB (inhibitory receptor) have similar amino acid sequences and differ mainly in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine - based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine - based inhibitory motif (ITIM) (M.in Da▲e▼ (see Ron, Annu.Rev.Immunol.15:203-234(1997)). The term further includes allotypes, such as the FcγRIIIA allotypes: FcγRIIIA-Phe158, FcγRIIIA-Val158, FcγRIIA-R131 and / or FcγRIIA-H131. Ravetch and Kinet, Annu.Rev.Immunol 9:457-92(1991) and Capel et al., Immunomethods 4:25-34(1994); and de In Haas et al., J.Lab.Clin.Med.126:330-41(1995), it was described for FcRs. The term FcR in the present invention contains other types of FcRs including FcRs to be specified in the future. The term FcR also includes the neonatal receptor FcRn 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)).

[0061] The term "FcRn" refers to the neonatal Fc receptor (FcRn). FcRn is structurally similar to the major histocompatibility complex (MHC) and is composed of an α chain non-covalently bound to β2-microglobulin. Multiple 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 the passive transport of immunoglobulin IgGs from mother to neonate and the regulation of serum IgG levels. As a rescue receptor, FcRn binds and transports IgG endocytosed in its complete form intracellularly and intercellularly, and rescues them from the default degradation pathway.

[0062] The "CH1 domain" of the human IgG heavy chain constant region usually extends from amino acid position 118 to amino acid position 215 (EU numbering system).

[0063] The "hinge region" is usually defined as the range extending from Glu at position 216 to Pro at position 230 of human IgG1 (Burton, Molec. Immunol. 22: 161 - 206 (1985)). By placing the first and last cysteine residues that form the disulfide bond between the heavy chains at the same positions as in IgG1, the hinge regions of other IgG isotypes can be compared with the IgG1 sequence.

[0064] The "CH2 domain" of the human IgG Fc region usually extends from the amino acid at position 231 to the amino acid at position 340. The CH2 domain is unique in that it does not pair closely with another domain. Instead, branched sugar chains linked at two N - termini are inserted between the two CH2 domains of the fully - formed native IgG molecule. The sugar can serve as a substitute for domain - to - domain pairing and is presumed to contribute to the stability of the CH2 domain. Burton, Molec Immunol. 22: 161 - 206 (1985).

[0065] The "CH3" domain includes the portion within the Fc region that extends from the C - terminal residue to the CH2 domain (from the amino acid at position 341 to the C - terminal of the antibody sequence, usually the amino acid residue at position 446 or 447 of IgG).

[0066] A "functional Fc fragment" has the "effector functions" possessed by the sequence of the native Fc region. Exemplary "effector functions" include C1q binding, complement - dependent cytotoxicity (CDC), Fc receptor binding, antibody - dependent cell - mediated cytotoxicity (ADCC), phagocytosis, down - regulation of cell - surface receptors (e.g., B - cell receptor; BCR), etc. Such effector functions usually require a combination of the Fc region and a binding domain (e.g., the antibody variable domain) and can be evaluated using various experimental methods known in the art.

[0067] Antibodies with an IgG Fc variant having an "altered" FcR binding affinity or ADCC activity have an increased or decreased FcR binding activity and / or ADCC activity as compared to the parent polypeptide or a polypeptide containing the native Fc sequence. An Fc variant showing "enhanced binding" to FcR has a higher affinity (e.g., a lower apparent Kd or IC50 value) for at least one FcR than the parent polypeptide or a polypeptide containing the native IgG Fc sequence. In some embodiments, the binding ability is increased by about 3-fold compared to the parent polypeptide, e.g., 5, 10, 25, 50, 60, 100, 150, 200-fold, up to 500-fold, or the binding ability is improved by any of 25% to 1000%. An Fc variant showing "reduced binding" to FcR has a lower affinity (e.g., a higher apparent Kd or IC50 value) for at least one FcR than the parent polypeptide. Its binding ability is reduced by 40% or more compared to the parent polypeptide.

[0068] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" is a form of cytotoxicity in which secreted Ig binds to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer cells (NK), neutrophils, and macrophages), and these cytotoxic effector cells are specifically bound to target cells carrying the antigen, and then the target cells are killed using cytotoxins. Antibodies "arm" cytotoxic cells, which is necessary for their toxicity. In the main cell types involved in ADCC, NK cells express only FcγRIII, and monocytes express FcγRI, FcγRII, and FcγRIII. Ravetch And Kinet, Annu. Rev. Immunol 9:457-92 (1991), summarizes FcR expression in hematopoietic cells in Table 3 on page 464. To evaluate the ADCC activity of the molecule of interest, in vitro ADCC experiments can be performed, as described in U.S. Patent No. 5,500,362 or 5,821,337. Effector cells suitable for such experiments include peripheral blood mononuclear cells (PBMC) and natural killer cells (NK). Optionally, the ADCC activity of the molecule of interest may be evaluated in vivo, for example, as described in the animal model disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).

[0069] A polypeptide containing a mutant Fc region, as compared to a polypeptide containing a wild-type IgG Fc or a parental polypeptide, exhibits "enhanced ADCC activity" or can more effectively mediate the ADCC effect in the presence of human effector cells. The polypeptide containing the mutant Fc region can more effectively mediate ADCC in either in vitro or in vivo situations when the amount is substantially the same as that of a polypeptide containing a wild-type IgG Fc (or a parental polypeptide) during the experiment. Usually, such mutants are identified by any in vitro ADCC experimental method known in the art and are used in experiments and methods for identifying the activity of ADCC, such as those using animal models. In some embodiments, the effect of mediating ADCC by such mutants is improved by 5 to 100 times, for example, 25 to 50 times, as compared to wild-type Fc (or a parental polypeptide).

[0070] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of a typical complement pathway is initiated by the binding of the first component of the complement system (C1q) to an antibody (a subclass with an appropriate structure) that binds to a homologous antigen. To evaluate complement activation, for example, a CDC experiment as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996) may be performed. U.S. Patent No. 6,194,551 B1 and WO99 / 51642 describe polypeptide variants having an altered Fc region amino acid sequence and an increased or decreased C1q binding ability. The contents of these patent publications are hereby expressly incorporated by reference. Also refer to Idusogie et al. J. Imm unol. 164:4178-4184 (2000).

[0071] Unless otherwise indicated, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate to each other and encode the same amino acid sequence. A nucleotide sequence encoding a protein or RNA may further include introns. For example, a nucleotide sequence encoding a protein may include introns in some forms.

[0072] The term "operably linked" refers to a functional linkage between a regulatory sequence and a heterologous nucleotide sequence, whereby the latter is expressed. For example, when a first nucleotide sequence and a second nucleotide sequence are in a functional relationship, the first nucleotide sequence and the second nucleotide sequence are operably linked. For example, when a promoter affects the transcription or expression of a coding sequence, the promoter and the coding sequence are operably linked. Usually, operably linked DNA sequences are continuous, and if necessary, two protein-coding regions can be linked in the same open reading frame.

[0073] "Homologous" refers to sequence similarity or identity between two polypeptides or two nucleic acid molecules. When the positions of both of the two sequences being compared are the same base or amino acid monomer subunit, for example, when the same position of two DNA molecules is adenine in both cases, the two DNA molecules are homologous at that position. The percentage homology between two sequences is a function that multiplies by 100 the ratio of the number of shared matches or homologous positions to the total number of positions in the two sequences. For example, in two sequences, if 6 out of 10 positions are matched or homologous, the homology of the two sequences is 60%. For example, the homology of the DNA sequences ATTGCC and TATTGC is 50%. Usually, when comparing two sequences, the comparison is made to obtain the maximum homology.

[0074] The "effective amount" of an antibody or composition described herein refers to an amount sufficient to achieve a particular purpose. The "effective amount" can be determined by experience and by known methods related to said purpose.

[0075] The term "therapeutically effective amount" refers to the amount of an antibody or composition described herein that can effectively "treat" a disease or disorder of an individual. In the case of Pseudomonas aeruginosa infection, the therapeutically effective amount of an antibody or composition described herein is an amount that can reduce the number of infected cells, inhibit the spread of the infection (i.e., alleviate or preferably stop to some extent), and / or alleviate one or more symptoms associated with the infection. When infected, an antibody or composition described herein can inhibit the growth of Pseudomonas aeruginosa and / or kill Pseudomonas aeruginosa, and the antibody can be cell growth inhibitory or cytotoxic. In some embodiments, the therapeutically effective amount refers to an amount that inhibits the infection in a patient. In some embodiments, the therapeutically effective amount refers to an amount that completely eradicates the infection in a patient.

[0076] As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" refers to a material that has no biological activity or other undesirable properties. For example, such a material can be added to a pharmaceutical composition to be administered to a patient so as not to cause adverse biological effects or interact with any other optional components included in the composition in a harmful manner. Pharmaceutically acceptable vectors or excipients preferably meet the criteria required for toxicology or manufacturing tests and / or are included in the Inactive Ingredients Guide compiled by the U.S. Food and Drug Administration.

[0077] The examples of the present application described herein should be understood to include examples "consisting of" and / or "consisting essentially of".

[0078] As used herein, "about" refers to a numerical value or parameter and includes (and describes) variants of the numerical value or parameter itself. For example, the description of "about X" includes the description of "X".

[0079] As used herein, the description of a numerical value or parameter as "not" usually means "other than" the numerical value or parameter. For example, the statement that the method cannot be used for the treatment of X-type infections means that the method is usually used for the treatment of types of infections other than X-type infections.

[0080] Unless otherwise clearly stated, the singular forms "a", "one" and "the" used in this specification and the claims include plural objects.

[0081] A bispecific antibody that recognizes Pseudomonas PcrV and Pseudomonas Psl In one aspect of the present invention, there is provided a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl.

[0082] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas aeruginosa PcrV comprises (a) a heavy-chain variable domain (V H ) and a light-chain variable domain (V L ), wherein the V H comprises a heavy-chain complementarity-determining region (HC-CDR) 1 of DX1X2MS (SEQ ID NO:20), where X1 is N or Y and X2 is Y, H, or P, an HC-CDR2 of X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G, and an HC-CDR3 of GRFSTX1SX2HFX3RAVYGMDV (SEQ ID NO:21), where X1 is L, S, N, or D, X2 is S or A, and X3 is F or Y, and the V L comprises a light-chain complementarity-determining region (LC-CDR) 1 of RASQGISSYLA (SEQ ID NO:10), an LC-CDR2 of AASTLQS (SEQ ID NO:11), and an LC-CDR3 of QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T, or (b) V H and V L , wherein the V H comprises an HC-CDR1 of X1X2X3MS (SEQ ID NO:17), where X1 is D or S, X2 is Y or N, and X3 is P, H, Y, or S, an HC-CDR2 of X1ISESGGSTX2X3ADSVKG (SEQ ID NO:18), where X1 is G or V, X2 is N or Y, and X3 is D or Y, and an HC-CDR3 of GRFX1X2X3X4X5X6FX7RAVYGMDV (SEQ ID NO:38), where X1 is S or C, X2 is T, G, D, Y, Q, or A, X3 is S, D, N, E, L, A, or Y, X4 is S, T, Y, or A, X5 is S, H, Q, A, R, K, G, E, Y, or D, X6 is H or C, and X7 is F or Y, and the V LIt comprises an LC-CDR1 containing RASQGIX1SYLA (SEQ ID NO:209), where X1 is S or R, an LC-CDR2 containing AASTLQS (SEQ ID NO:11), and an LC-CDR3 containing QQLX1SYPLX2 (SEQ ID NO:210), where X1 is S, N or K and X2 is S or T.

[0083] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises (a) V H and V L wherein said V H comprises an HC-CDR1 containing DNX1MS (SEQ ID NO:14), where X1 is Y or H, an HC-CDR2 containing X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G, and an HC-CDR3 containing GRFSTX1SSHFX2RAVYGMDV (SEQ ID NO:16), where X1 is L or S and X2 is F or Y, and said V L comprises an LC-CDR1 containing RASQGISSYLA (SEQ ID NO:10), an LC-CDR2 containing AASTLQS (SEQ ID NO:11), and an LC-CDR3 containing QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T.

[0084] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises (a) V H and V L wherein said V H comprises an HC-CDR1 containing SYWMH (SEQ ID NO:22), an HC-CDR2 containing RINEX1EX2SISYADSVKG (SEQ ID NO:44), where X1 is D or N and X2 is T, G or R, and an HC-CDR3 containing DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T and X2 is F or L, and said VL comprises LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO:46), wherein X1 is S, G or N, X2 is S, R or K, X3 is S or N, LC-CDR2 comprising X1ASSRAT (SEQ ID NO:42), wherein X1 is D or H, and LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO:47), wherein X1 is S, L or N, X2 is S, Q or E, X3 is L or I, or (b) V H and V L comprises, said V H comprises HC-CDR1 comprising SYWMH (SEQ ID NO:22), HC-CDR2 comprising RINEX1EX2SISYADSVKG (SEQ ID NO:211), wherein X1 is D, N, I, L or V, X2 is S, T, R, G or N, and HC-CDR3 comprising DGPYDX1X2DI (SEQ ID NO:45), wherein X1 is S, A or T, X2 is F or L, said V L comprises LC-CDR1 comprising RASQX1VX2X3NLA (SEQ ID NO:212), wherein X1 is N, G, D or S, X2 is K, R, S, N or T, X3 is N, G, S or D, LC-CDR2 comprising X1ASSRAT (SEQ ID NO:213), wherein X1 is D, N, H or A, and LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO:214), 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, X3 is I, L or V.

[0085] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is (a) V H and V L comprises, said V Hcomprises an HC-CDR1 containing SYWMH (SEQ ID NO:22), an HC-CDR2 containing RINEX1EX2SISYADSVKG (SEQ ID NO:39), wherein X1 is D or N and X2 is T or G, and an HC-CDR3 containing DGPYDX1LDI (SEQ ID NO:40), wherein X1 is S or A, and said V L comprises an LC-CDR1 containing RASQX1VX2X3NLA (SEQ ID NO:41), wherein X1 is S or G, X2 is S or R, and X3 is S or N, an LC-CDR2 containing X1ASSRAT (SEQ ID NO:42), wherein X1 is D or H, and an LC-CDR3 containing QQYGX1X2PX3T (SEQ ID NO:43), wherein X1 is S or L, X2 is S or Q, and X3 is L or I.

[0086] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and said V H comprises an HC-CDR1 containing SSGDYWG (SEQ ID NO:48), an HC-CDR2 containing SIHNX1GSTYYNPSLKG (SEQ ID NO:81), wherein X1 is S or Q, and an HC-CDR3 containing QFGSETYYX1GIX2P (SEQ ID NO:82), wherein X1 is T, N or P, and X2 is D or Q, and said V L comprises an LC-CDR1 containing RSSQSLLHSX1GYNYLD (SEQ ID NO:83), wherein X1 is N or R, an LC-CDR2 containing LGSNRAS (SEQ ID NO:70), and an LC-CDR3 containing MQALQTPYT (SEQ ID NO:74). In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and said V Hcomprises an HC-CDR1 comprising SSGDYWG (SEQ ID NO:48), an HC-CDR2 comprising SIHNX1GSTYYNPSLKG (SEQ ID NO:125), where X1 is S, K or Q, and an HC-CDR3 comprising QFGSETYYX1GIX2P (SEQ ID NO:126), where X1 is N, S, V, T or P and X2 is D, Y, C, H, S, R, A, E, G, K, W, V or Q, said V L comprises an LC-CDR1 comprising RSSQSLLHSX1GYNYLD (SEQ ID NO:127), where X1 is N, A, V, F, R, G, H, Q, W or P, an LC-CDR2 comprising LGSNRAS (SEQ ID NO:70), and an LC-CDR3 comprising MQALQTPX1T (SEQ ID NO:128), where X1 is R or Y.

[0087] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas aeruginosa Psl is V H and V L comprising, said V H comprises an HC-CDR1 comprising IHSVH (SEQ ID NO:50), an HC-CDR2 comprising TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 comprising DGX1S (SEQ ID NO:84), where X1 is D or T, said V L comprises an LC-CDR1 comprising RASQGISSWLA (SEQ ID NO:68), an LC-CDR2 comprising HASTLES (SEQ ID NO:72), and an LC-CDR3 comprising LQAX1SLPHT (SEQ ID NO:85), where X1 is N, F, S or K. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas aeruginosa Psl is V H and V L comprising, said V Hcomprises an HC-CDR1 containing IHSVH (SEQ ID NO:50), an HC-CDR2 containing TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 containing X1X2X3X4 (SEQ ID NO:129), where X1 is D, Y or N, X2 is G or A, X3 is D or T, and X4 is S, A or T, and said V L comprises an LC-CDR1 containing RASQGISSWLA (SEQ ID NO:68), an LC-CDR2 containing HASTLES (SEQ ID NO:72), and an LC-CDR3 containing LQAX1SLPHT (SEQ ID NO:130), where X1 is N, D, Y, F, P, G, K, H, A, C, E, Q, R, S, T, V, W or L.

[0088] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H comprises an HC-CDR1 containing any one amino acid sequence of SEQ ID NOs: 1-3, an HC-CDR2 containing amino acid sequence 4 or 5, and an HC-CDR3 containing any one amino acid sequence of SEQ ID NOs: 6-9, and said V L comprises an LC-CDR1 containing amino acid sequence SEQ ID NO:10, an LC-CDR2 containing amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing amino acid sequence SEQ ID NO:12 or 13.

[0089] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H comprises an HC-CDR1 containing amino acid sequence SEQ ID NO:1, an HC-CDR2 containing amino acid sequence SEQ ID NO:4, and an HC-CDR3 containing amino acid sequence SEQ ID NO:6, and said V LIt comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:10, an LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:12. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and the V H comprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:2, an HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:7. The V L comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:10, an LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:13. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and the V H comprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:2, an HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:8. The V L comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:10, an LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:12. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and the V H comprises an HC-CDR1 containing the amino acid sequence SEQ ID NO:3, an HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:9. The V Lcomprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:13.

[0090] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising any one of the amino acid sequences of SEQ ID NOs:23-25, and an HC-CDR3 comprising any one of the amino acid sequences of SEQ ID NOs:26-29, and said V L comprises an LC-CDR1 comprising any one of the amino acid sequences of SEQ ID NOs:30-32, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:33 or 34, and an LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NOs:35-37.

[0091] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:23, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:26, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:33, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:35. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V Hcomprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:27, wherein said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:36. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:25, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:27, wherein said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:37. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:25, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:28, wherein said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:37. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V Hcomprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:28, wherein said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, and the amino acid sequence SEQ ID NO:34-containing LC-CDR2 and LC-CDR3 comprising the amino acid sequence SEQ ID NO:36. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, HC-CDR2 comprising the amino acid sequence SEQ ID NO:25, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:26, and said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:37. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:26, and said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:36. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein said V Hcomprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, HC-CDR2 comprising the amino acid sequence SEQ ID NO:23, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:29, wherein said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, LC-CDR2 comprising the amino acid sequence SEQ ID NO:33, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:35.

[0092] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and comprises, wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, HC-CDR2 comprising the amino acid sequence SEQ ID NO:52 or 53, and HC-CDR3 comprising any one amino acid sequence of SEQ ID NOs:57-60, and said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:65 or 66, LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:74.

[0093] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and comprises, wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:50, HC-CDR2 comprising the amino acid sequence SEQ ID NO:55, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:62 or 63, and said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:68, LC-CDR2 comprising the amino acid sequence SEQ ID NO:72, and LC-CDR3 comprising any one amino acid sequence of SEQ ID NOs:76-79.

[0094] In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and includes the V H comprising an HC-CDR1 having the amino acid sequence SEQ ID NO:48, an HC- CDR2 having the amino acid sequence SEQ ID NO:52, and an HC-CDR3 having the amino acid sequence SEQ ID NO:57, and the V L comprising an LC-CDR1 having the amino acid sequence SEQ ID NO:65, an LC-CDR2 having the amino acid sequence SEQ ID NO:70, and an LC-CDR3 having the amino acid sequence SEQ ID NO:74. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and includes the V H comprising an HC-CDR1 having the amino acid sequence SEQ ID NO:48, an HC-CDR2 having the amino acid sequence SEQ ID NO:53, and an HC-CDR3 having the amino acid sequence SEQ ID NO:58, and the V L comprising an LC-CDR1 having the amino acid sequence SEQ ID NO:66, an LC-CDR2 having the amino acid sequence SEQ ID NO:70, and an LC-CDR3 having the amino acid sequence SEQ ID NO:74. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and includes the V H comprising an HC-CDR1 having the amino acid sequence SEQ ID NO:48, an HC-CDR2 having the amino acid sequence SEQ ID NO:52, and an HC-CDR3 having the amino acid sequence SEQ ID NO:59, and the V L comprising an LC-CDR1 having the amino acid sequence SEQ ID NO:65, an LC-CDR2 having the amino acid sequence SEQ ID NO:70, and an LC-CDR3 having the amino acid sequence SEQ ID NO:74. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and VL comprises the V H comprises HC-CDR1 having the amino acid sequence SEQ ID NO:48, HC-CDR2 having the amino acid sequence SEQ ID NO:53, and HC-CDR3 having the amino acid sequence SEQ ID NO:60, and the V L comprises LC-CDR1 having the amino acid sequence SEQ ID NO:65, LC-CDR2 having the amino acid sequence SEQ ID NO:70, and LC-CDR3 having the amino acid sequence SEQ ID NO:74. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L comprises the V H comprises HC-CDR1 having the amino acid sequence SEQ ID NO:49, HC-CDR2 having the amino acid sequence SEQ ID NO:54, and HC-CDR3 having the amino acid sequence SEQ ID NO:61, and the V L comprises LC-CDR1 having the amino acid sequence SEQ ID NO:67, LC-CDR2 having the amino acid sequence SEQ ID NO:71, and LC-CDR3 having the amino acid sequence SEQ ID NO:75. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L comprises the V H comprises HC-CDR1 having the amino acid sequence SEQ ID NO:50, HC-CDR2 having the amino acid sequence SEQ ID NO:55, and HC-CDR3 having the amino acid sequence SEQ ID NO:62, and the V L comprises LC-CDR1 having the amino acid sequence SEQ ID NO:68, LC-CDR2 having the amino acid sequence SEQ ID NO:72, and LC-CDR3 having the amino acid sequence SEQ ID NO:76. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L comprises the V Hcomprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:50, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:55, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:63, wherein said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:68, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:72, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:77. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L , wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:50, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:55, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:63, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:68, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:72, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:78. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L , wherein said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:50, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:55, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:63, and said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:68, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:72, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:79. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L , wherein said V Hcomprises HC-CDR1 containing the amino acid sequence SEQ ID NO:51, HC-CDR2 containing the amino acid sequence SEQ ID NO:56, and HC-CDR3 containing the amino acid sequence SEQ ID NO:64, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:69, LC-CDR2 containing the amino acid sequence SEQ ID NO:73, and LC-CDR3 containing the amino acid sequence SEQ ID NO:80.

[0095] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H contains the amino acid sequence SEQ ID NO:91 or 161, and said V L contains the amino acid sequence SEQ ID NO:112 or 182. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H contains HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:91 or 161, and said V L contains LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:112 or 182.

[0096] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L and said V H contains the amino acid sequence SEQ ID NO:92 or 162, and said V L contains the amino acid sequence SEQ ID NO:113 or 183. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V Lcomprising said V H is a V having the amino acid sequence SEQ ID NO:92 or 162 H and comprises HC-CDR1, HC-CDR2 and HC-CDR3 in said V L is a V having the amino acid sequence SEQ ID NO:113 or 183 L and comprises LC-CDR1, LC-CDR2 and LC-CDR3 in said V

[0097] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L comprising said V H comprises the amino acid sequence SEQ ID NO:93 or 163, and said V L comprises the amino acid sequence SEQ ID NO:112 or 182. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L comprising said V H is a V having the amino acid sequence SEQ ID NO:93 or 163 H and comprises HC-CDR1, HC-CDR2 and HC-CDR3 in said V L is a V having the amino acid sequence SEQ ID NO:112 or 182 L and comprises LC-CDR1, LC-CDR2 and LC-CDR3 in said V

[0098] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L comprising said V H comprises the amino acid sequence SEQ ID NO:94 or 164, and said V L comprises the amino acid sequence SEQ ID NO:113 or 183. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L comprising said V Hhaving the amino acid sequence SEQ ID NO:94 or 164 H and wherein the V L is the amino acid sequence SEQ ID NO. : V with 113 or 183 L The LC-CDR1, LC-CDR2 and LC-CDR3 in

[0099] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is H and V L The V H comprises the amino acid sequence SEQ ID NO: 95 or 165, L comprises the amino acid sequence SEQ ID NO: 114 or 184. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises the amino acid sequence SEQ ID NO: 114 or 184. H and V L The V H having the amino acid sequence SEQ ID NO:95 or 165 H and wherein the V L having the amino acid sequence SEQ ID NO:114 or 184 L The LC-CDR1, LC-CDR2 and LC-CDR3 in

[0100] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is H and V L The V H comprises the amino acid sequence SEQ ID NO: 96 or 166, L comprises the amino acid sequence SEQ ID NO: 115 or 185. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises the amino acid sequence SEQ ID NO: 115 or 185. H and V L The V Hcomprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO:96 or 166, and said V H comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO:115 or 185. L In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V L and V

[0101] wherein said V H comprises the amino acid sequence SEQ ID NO:97 or 167, and said V L comprises the amino acid sequence SEQ ID NO:116 or 186. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO:97 or 167, and said V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO:116 or 186. H In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V L comprises the amino acid sequence SEQ ID NO:98 or 168, and said V

[0102] comprises the amino acid sequence SEQ ID NO:116 or 186. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V H comprises the amino acid sequence SEQ ID NO:98 or 168, and said V L comprises the amino acid sequence SEQ ID NO:116 or 186. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V His V having the amino acid sequence SEQ ID NO: 98 or 168 H and includes HC-CDR1, HC-CDR2 and HC-CDR3 in the said V L is V having the amino acid sequence SEQ ID NO: 116 or 186 L and includes LC-CDR1, LC-CDR2 and LC-CDR3 in the said V

[0103] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein the said V H includes the amino acid sequence SEQ ID NO: 99 or 169, and the said V L includes the amino acid sequence SEQ ID NO: 115 or 185. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein the said V H includes HC-CDR1, HC-CDR2 and HC-CDR3 in the V having the amino acid sequence SEQ ID NO: 99 or 169 H and the said V L is the V having the amino acid sequence SEQ ID NO : 115 or 185 L and includes LC-CDR1, LC-CDR2 and LC-CDR3 in the said V

[0104] In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein the said V H includes the amino acid sequence SEQ ID NO: 100 or 170, and the said V L includes the amino acid sequence SEQ ID NO: 116 or 186. In some embodiments, the first antigen-binding domain that specifically recognizes Pseudomonas PcrV is V H and V L wherein the said V Hcomprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 100 or 170, and said V H comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 116 or 186. L In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V L and V

[0105] H wherein said V L comprises an amino acid sequence SEQ ID NO: 101 or 171, and said V H comprises an amino acid sequence SEQ ID NO: 115 or 185. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V L H and V L wherein said V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 101 or 171, and said V H comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 115 or 185. L L H L H L

[0106] In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V H comprises an amino acid sequence SEQ ID NO: 102 or 172, and said V L comprises an amino acid sequence SEQ ID NO: 114 or 184. In some embodiments, said first antigen-binding domain that specifically recognizes Pseudomonas PcrV comprises V H and V L wherein said V His V having the amino acid sequence SEQ ID NO: 102 or 172 H and includes HC-CDR1, HC-CDR2 and HC-CDR3 in said V L is V having the amino acid sequence SEQ ID NO: 114 or 184 L and includes LC-CDR1, LC-CDR2 and LC-CDR3 in said V

[0107] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and said V H includes the amino acid sequence SEQ ID NO: 103 or 173, and said V L includes the amino acid sequence SEQ ID NO: 117 or 187. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and said V H includes HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 103 or 173 H and said V L includes LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 117 or 187 L and includes LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 117 or 187

[0108] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and said V H includes the amino acid sequence SEQ ID NO: 104 or 174, and said V L includes the amino acid sequence SEQ ID NO: 118 or 188. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and said V Hhas HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 104 or 174, and said V H has LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO L : 118 or 188. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V L and V

[0109] H wherein said V L comprises an amino acid sequence SEQ ID NO: 105 or 175, and said V H comprises an amino acid sequence SEQ ID NO: 117 or 187. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V L H and V L wherein said V H has HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 105 or 175, and said V H has LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 117 or 187. L L H L H L

[0110] In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V H comprises an amino acid sequence SEQ ID NO: 106 or 176, and said V L comprises an amino acid sequence SEQ ID NO: 117 or 187. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V Hcomprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 106 or 176, and said V H comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 117 or 187. L In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V L and V

[0111] wherein said V H comprises the amino acid sequence SEQ ID NO: 107 or 177, and said V L comprises the amino acid sequence SEQ ID NO: 119 or 189. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 107 or 177, and said V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 119 or 189. H In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V L comprises the amino acid sequence SEQ ID NO: 108 or 178, and said V

[0112] In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V H comprises the amino acid sequence SEQ ID NO: 108 or 178, and said V L comprises the amino acid sequence SEQ ID NO: 119 or 189. In some embodiments, said second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V Hhaving the amino acid sequence SEQ ID NO: 108 or 178 H and wherein the V L having the amino acid sequence SEQ ID NO: 119 or 189 L The LC-CDR1, LC-CDR2 and LC-CDR3 in

[0113] In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas sp. Psl is H and V L The V H comprises the amino acid sequence SEQ ID NO: 109 or 179, L comprises the amino acid sequence SEQ ID NO: 120 or 190. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises the amino acid sequence SEQ ID NO: 120 or 190. H and V L The V H having the amino acid sequence SEQ ID NO: 109 or 179 H and wherein the V L is the amino acid sequence SEQ ID NO. : 120 or 190 V L The LC-CDR1, LC-CDR2 and LC-CDR3 in

[0114] In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas sp. Psl is H and V L The V H comprises the amino acid sequence SEQ ID NO: 110 or 180, L comprises the amino acid sequence SEQ ID NO: 121 or 191. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises the amino acid sequence SEQ ID NO: 122 or 192. H and V L The V His a V having the amino acid sequence SEQ ID NO: 110 or 180 H and includes HC-CDR1, HC-CDR2 and HC-CDR3 in the V L is a V having the amino acid sequence SEQ ID NO: 121 or 191 L and includes LC-CDR1, LC-CDR2 and LC-CDR3 in the V

[0115] In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes the amino acid sequence SEQ ID NO: 110 or 180, and the V L includes the amino acid sequence SEQ ID NO: 122 or 192. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and the V H is a V having the amino acid sequence SEQ ID NO: 110 or 180 and includes HC-CDR1, HC-CDR2 and HC-CDR3 in the V H and the V L is a V having the amino acid sequence SEQ ID NO: 122 or 192 and includes LC-CDR1, LC-CDR2 and LC-CDR3 in the V L In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V

[0116] In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes the amino acid sequence SEQ ID NO: 110 or 180, and the V L includes the amino acid sequence SEQ ID NO: 123 or 193. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl is V H and V L and the V Hcomprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 110 or 180, and said V H comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 123 or 193. L In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V L and V

[0117] wherein said V H comprises an amino acid sequence SEQ ID NO: 111 or 181, and said V L comprises an amino acid sequence SEQ ID NO: 124 or 194. In some embodiments, the second antigen-binding domain that specifically recognizes Pseudomonas Psl comprises V H and V L wherein said V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 111 or 181, and said V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V having the amino acid sequence SEQ ID NO: 124 or 194. H In some embodiments, the present invention provides a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein the first antigen-binding domain comprises (a) V H and V L wherein said V L comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V having the amino acid sequence SEQ ID NO: 111 or 181, and said V

[0118] In some embodiments, the present invention provides a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein the first antigen-binding domain comprises (a) V H and V L wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3 comprising the amino acid sequence SEQ ID NO: 6, and said V Lcomprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, and LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:12, or (b) V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:2, HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and HC-CDR3 containing the amino acid sequence SEQ ID NO:7, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:13, or (c) V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:22, HC-CDR2 containing the amino acid sequence SEQ ID NO:23, and HC-CDR3 containing the amino acid sequence SEQ ID NO:26, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:30, LC-CDR2 containing the amino acid sequence SEQ ID NO:33, and LC-CDR3 containing the amino acid sequence SEQ ID NO:35, or (d) V H and V L wherein said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:22, HC-CDR2 containing the amino acid sequence SEQ ID NO:24, and HC-CDR3 containing the amino acid sequence SEQ ID NO:27, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:31, LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and LC-CDR3 containing the amino acid sequence SEQ ID NO:36, to provide a bispecific molecule.

[0119] In some embodiments, the present invention provides a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein the second antigen-binding domain comprises V H and V L , wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:48, an HC-CDR2 having the amino acid sequence SEQ ID NO:52, and an HC-CDR3 having the amino acid sequence SEQ ID NO:57, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:65, an LC-CDR2 having the amino acid sequence SEQ ID NO:70, and an LC-CDR3 having the amino acid sequence SEQ ID NO:74, or (b) V H and V L , wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:49, an HC-CDR2 having the amino acid sequence SEQ ID NO:54, and an HC-CDR3 having the amino acid sequence SEQ ID NO:61, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:67, an LC-CDR2 having the amino acid sequence SEQ ID NO:71, and an LC-CDR3 having the amino acid sequence SEQ ID NO:75, or (c) V H and V L , wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:51, an HC-CDR2 having the amino acid sequence SEQ ID NO:56, and an HC-CDR3 having the amino acid sequence SEQ ID NO:64, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:69, an LC-CDR2 having the amino acid sequence SEQ ID NO:73, and an LC-CDR3 having the amino acid sequence SEQ ID NO:80, and provides a bispecific molecule.

[0120] In some embodiments, the present invention provides a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein the first antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:1, an HC-CDR2 having the amino acid sequence SEQ ID NO:4, and an HC-CDR3 having the amino acid sequence SEQ ID NO:6, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:10, an LC-CDR2 having the amino acid sequence SEQ ID NO:11, and an LC-CDR3 having the amino acid sequence SEQ ID NO:12, and wherein the second antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:48, an HC-CDR2 having the amino acid sequence SEQ ID NO: 52, and an HC-CDR3 having the amino acid sequence SEQ ID NO:57, and the V L comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:65, an LC-CDR2 having the amino acid sequence SEQ ID NO:70, and an LC-CDR3 having the amino acid sequence SEQ ID NO:74, thus providing a bispecific molecule.

[0121] In some embodiments, the present invention provides a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, wherein the first antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:1, an HC-CDR2 having the amino acid sequence SEQ ID NO:4, and an HC-CDR3 having the amino acid sequence SEQ ID NO:6, and the V Lcomprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:12, and said second antigen-binding domain comprises V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75, and provides a bispecific molecule.

[0122] In some embodiments, the present invention is a bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, said first antigen-binding domain comprising V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:2, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:5, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:7, said V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:13, and said second antigen-binding domain comprises V H and V L and said V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, said V LProvided is a bispecific molecule comprising an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74.

[0123] In some embodiments, the invention is a bispecific molecule comprising a first antigen-binding domain that specifically recognizes PcrV of the genus Pseudomonas and a second antigen-binding domain that specifically recognizes Psl of the genus Pseudomonas, wherein the first antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:2, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:5, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:7, the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:13, and the second antigen-binding domain comprises V H and V L wherein the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75. Provided is a bispecific molecule.

[0124] In some embodiments, according to any of the bispecific molecules described herein, the first antigen-binding domain is a single-chain variable region fragment (scFv) that specifically recognizes Pseudomonas PcrV, the second antigen-binding domain is a Fab arm that specifically recognizes Pseudomonas Psl, the bispecific molecule further comprises an Fc region comprising CH2 and CH3 domains, the single-chain variable region fragment (scFv) is interconnected with the Fab arm via a first polypeptide linker (L1), and is interconnected with the Fc region via a second polypeptide linker (L2). In some embodiments, the bispecific molecule is bivalent for binding to each of PcrV and Psl.

[0125] In some embodiments, according to any of the bispecific molecules described herein, the first antigen-binding domain is a Fab arm that specifically recognizes Pseudomonas PcrV, the second antigen-binding domain is a single-chain variable region fragment (scFv) that specifically recognizes Pseudomonas Psl, the bispecific molecule further comprises an Fc region comprising CH2 and CH3 domains, the single-chain variable region fragment (scFv) is interconnected with the Fab arm via a first polypeptide linker (L1), and is interconnected with the Fc region via a second polypeptide linker (L2). In some embodiments, the molecule is a bispecific molecule that is bivalent for binding to each of PcrV and Psl.

[0126] In some embodiments, according to any of the bispecific molecules described herein, the antigen-binding domain of the single-chain variable region fragment (scFv) that specifically recognizes PcrV or Psl comprises engineered cysteine variants, and by introducing two cysteine variants at the interface of V H and V L a bispecific antibody with stabilized disulfide bonds is obtained.

[0127] The linker can be used to bind the domains and / or regions of the chimeric heavy chain of the bispecific molecule into one continuous molecule. In some embodiments, the bispecific molecule comprises at least two polypeptide linkers, L1 and L2. In some embodiments, the bispecific molecule can comprise additional linkers, such as a flexible linker that interconnects the variable heavy and light chains of, for example, an scFv. In some embodiments, the bispecific molecule can comprise additional linkers, such as a flexible linker that interconnects the variable heavy and light chains of, for example, an scFv, and another linker that connects other binding units to the core structure of the bispecific molecule. An example of other binding units connected to the core structure of the bispecific molecule is shown in FIG. 6.

[0128] Exemplary non-limiting examples of linkers are polypeptide chains comprising at least 4 residues. Such linker moieties are flexible and hydrophilic and form little or no secondary structure of their own (linker moiety or flexible linker moiety). After the assembly of the molecule is complete, the linker of at least 4 amino acids can be used to bind domains and / or regions that are in proximity to each other. Also, long linkers can be used. In some embodiments, the linker can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 100, 125, 150, 175 or 200 residues. When multiple linkers are used to interconnect parts of the molecule, the linkers can be the same or different (e.g., of the same or different length and / or amino acid sequence).

[0129] In some aspects, the polypeptide linker comprises or consists of a glycine-serine (Gly-Ser) linker As described herein, the term "glycine-serine (Gly-Ser) linker" refers to a peptide consisting of glycine and serine residues. An exemplary glycine-serine (Gly-Ser) linker comprises an amino acid sequence represented by the general formula (Gly4Ser) n where n is a positive integer (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10). Preferred glycine-serine (Gly-Ser) linkers are (Gly4Ser)2 and (Gly4Ser)4. Another exemplary glycine-serine (Gly-Ser) linker is (Gly4Ser)3. In other embodiments, two or more glycine-serine (Gly-Ser) linkers are incorporated in series into a single polypeptide linker. In some embodiments, the polypeptide linker comprises at least a portion of a hinge region (e.g., derived from an IgG1, IgG2, IgG3 or IgG4 molecule) and a series of glycine-serine (Gly-Ser) residues (e.g., a glycine-serine (Gly-Ser) linker such as (Gly4Ser) n ).

[0130] In some embodiments, L1 and / or L2 include a hinge portion and a linker portion such as, for example, a linker portion including a glycine-serine (Gly-Ser) linker. In other aspects, L1 and / or L2 include only the hinge portion or only a linker portion such as a linker portion including a glycine-serine (Gly-Ser) linker. In some aspects, L1 and L2 include a glycine-serine (Gly-Ser) linker. In some aspects, the glycine-serine (Gly-Ser) linker portions of L1 and L2 are of the same length, and in other aspects, the glycine-serine (Gly-Ser) linker portions of L1 and L2 are of different lengths. When the bispecific molecule includes an scFv, the heavy and light chains of the scFv can be connected via a flexible linker. In some embodiments, this flexible linker typically does not include a hinge portion and is a glycine-serine (Gly-Ser) linker or other flexible linker. The length and amino acid sequence of the flexible linker interconnecting the domains of the scFv can be selected and optimized.

[0131] In some embodiments, the bispecific molecule comprises a binding unit 1 (BU1) and a binding unit 2 (BU2). In some embodiments, BU1 comprises a Fab domain. In some embodiments, BU2 comprises a scFv. In some embodiments, the polypeptide linker (e.g., L1 and / or L2) comprises a glycine-serine (Gly-Ser) or all-glycine (Gly) linker and a part or modified part of the hinge region. In some aspects, the polypeptide linker (L1) connecting BU1 (e.g., the Fab domain) to another binding domain BU2 (e.g., the scFv) in the bispecific molecule comprises the amino acid sequence EPKSDKTGGGGSGGGGS (SEQ ID NO:153) or EPKSCGKTGGGGSGGGGS (SEQ ID NO:154) or EPKSCGGGGSGGGGS (SEQ ID NO:155). In some aspects, the polypeptide linker (L2) connecting BU2 to the Fc domain in the bispecific molecule comprises the amino acid sequence GGGGSGGGGSEPKSDKTHTCPPCP (SEQ ID NO:156) or GGGGSGGGGSCPPCP (SEQ ID NO:157) or GGGGSGGGGSDKTHTCPPCP (SEQ ID NO:158).

[0132] Regardless of the polypeptide linker used to interconnect binding unit 1 and binding unit 2 and to interconnect binding unit 2 and Fc (e.g., L1 and L2), the bispecific molecule can optionally comprise additional polypeptide linkers. The lengths and sequences of these additional polypeptide linkers are independently selected. For example, the bispecific molecule can further comprise a flexible polypeptide linker (L3) interconnecting the variable heavy and light chains (V HSCFV and V LSCFV ) of the scFv. This flexible polypeptide linker can comprise a glycine-serine (Gly-Ser) linker Generally, this linker does not include a hinge region. In some embodiments, this flexible polypeptide linker (L3) that interconnects the variable heavy and light chains of the scFv comprises the sequence GGGGSGGGGSGGGGSGGGG (SEQ ID NO:87).

[0133] In some embodiments, the bispecific molecule of the invention comprises two heavy-light chain pairs. In some embodiments, the polypeptide sequence of the chimeric heavy chain of said bispecific molecule can comprise a polypeptide sequence comprising an antibody heavy chain variable domain 1 (VH1), a polypeptide sequence comprising an antibody heavy chain constant region 1 (CH1), a polypeptide sequence comprising a first polypeptide linker (L1), a polypeptide sequence comprising a binding unit 2 (BU2) that binds to a second epitope, a polypeptide sequence comprising a second polypeptide linker (L2), and a polypeptide sequence comprising an Fc domain. In some aspects, the Fc domain comprises a CH2 domain and a CH3 domain. In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. The polypeptide sequence of the light chain of the bispecific molecule can comprise a light chain variable domain 1 (VL1) and a light chain constant region (CL). In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, VH1, VL1, CH1, and CL are used to denote a binding unit 1 (BU1) that is part of the portion that binds to the first epitope. In some embodiments, one or more additional binding units (e.g., scFvs) are present at the N-terminus and / or C-terminus of the core of the bispecific molecule. In some embodiments, one or more additional binding units (e.g., scFvs) are present in the hinge region. Thus, the heavy chain of the bispecific molecule can comprise an extended core and, from the N-terminus to the C-terminus, VH1-CH1-L1-(BU) nIt can include a polypeptide sequence such as -L2-CH2-CH3 (n>l). In some embodiments, the bispecific molecule includes a CH1 containing the amino acid sequence SEQ ID No:134. In some embodiments, the bispecific molecule includes a CH2-CH3 containing the amino acid sequence SEQ ID NO:131, 132, or 133. In some embodiments, the bispecific molecule includes a CH2-CH3 containing one or more substitutions at positions 22, 24, 26 according to the amino acid sequence SEQ ID NO:131. In some embodiments, the bispecific molecule includes a CH2-CH3 containing one or more substitutions of M22Y, S24T, T26E among them according to the amino acid sequence SEQ ID NO:131. In some embodiments, the bispecific molecule includes a CH2-CH3 containing one or more substitutions at positions 198, 204 according to the amino acid sequence SEQ ID NO:131. In some embodiments, the bispecific molecule includes a CH2-CH3 containing one or more substitutions of M198L, N204S among them according to the amino acid sequence SEQ ID NO:131.

[0134] In some embodiments, the binding unit 2 (BU2) is a scFv, and the chimeric heavy chain of the bispecific molecule can include a polypeptide sequence containing an antibody heavy chain variable domain (VH1), a polypeptide sequence containing an antibody heavy chain constant region 1 (CH1), a polypeptide sequence containing a first polypeptide linker (L1), a polypeptide sequence containing an antibody light chain variable domain (VL2), a polypeptide sequence containing a flexible linker (L3), a polypeptide sequence containing an antibody heavy chain variable domain (VH2), a polypeptide sequence containing a second polypeptide linker (L2), and a polypeptide sequence containing an Fc domain. In some embodiments, the chimeric heavy chain of the bispecific molecule can include a polypeptide sequence such as VH1-CH1-L1-VL2-L3-VH2-L2-Fc from the N-terminus to the C-terminus. The chimeric heavy chain is a polypeptide chain consisting of an amino acid sequence (e.g., the amino acid sequence of each polypeptide domain). Alternatively, in some embodiments, the chimeric heavy chain of the bispecific molecule is VH1-CH1-L1-VH2-L3-VL2-L2 from the N-terminus to the C-terminus -Fc-like polypeptide sequences can be included. The chimeric heavy chain is a polypeptide chain consisting of an amino acid sequence (e.g., the amino acid sequence of each polypeptide domain). Note that VH1, VL1, CH1, and CL are used to indicate a part of binding unit 1, and VH1 and VL1 are used to indicate the part that binds to the first epitope. VH2 and VL2 are used to indicate the part of binding unit 2 that binds to the second epitope. In some embodiments, one or more additional binding units (e.g., scFvs) are present at the N-terminus and / or C-terminus of the core of the bispecific molecule. In other aspects, one or more additional binding units (e.g., scFvs) are present in the hinge region. The antibody heavy chain variable regions contained within the tandem scFv are represented by VH3, VH4, VH5, and the corresponding antibody light chain variable regions within the tandem scFv are represented by VL3, VL4, VL5. In some embodiments, the bispecific molecule comprises a CH1 containing the amino acid sequence SEQ ID No: 134. In some embodiments, the bispecific molecule comprises a CH2-CH3 containing the amino acid sequence SEQ ID NO: 131, 132, or 133. In some embodiments, the bispecific molecule comprises a CH2-CH3 containing one or more substitutions at positions 22, 24, 26 according to the amino acid sequence SEQ ID NO: 131. In some embodiments, the bispecific molecule comprises a CH2-CH3 containing one or more substitutions of M22Y, S24T, T26E according to the amino acid sequence SEQ ID NO: 131. In some embodiments, the bispecific molecule comprises a CH2-CH3 containing one or more substitutions at positions 198, 204 according to the amino acid sequence SEQ ID NO: 131. In some embodiments, the bispecific molecule comprises a CH2-CH3 containing one or more substitutions of M198L, N204S according to the amino acid sequence SEQ ID NO: 131.

[0135] In some embodiments, the chimeric heavy chain of the bispecific molecule can include a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can include a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule includes a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:137 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule includes a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:139 and a light chain comprising the amino acid sequence SEQ ID NO:135.

[0136] In some embodiments, the chimeric heavy chain of the bispecific molecule can include a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can include a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule includes a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:138 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule includes a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:140 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule includes the amino acid sequence SEQ ID NO:141 and a light chain comprising the amino acid sequence SEQ ID NO:135.

[0137] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:140, and a light chain comprising the amino acid sequence SEQ ID NO:136. a chimeric heavy chain, and a light chain comprising the amino acid sequence SEQ ID NO:136. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:141, and a light chain comprising the amino acid sequence SEQ ID NO:136. In some embodiments, the bispecific molecule comprises an amino acid sequence SEQ ID NO:142, and a light chain comprising the amino acid sequence SEQ ID NO:136.

[0138] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:142, and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:143, and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises an amino acid sequence SEQ ID NO:144, and a light chain comprising the amino acid sequence SEQ ID NO:135.

[0139] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:148 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:146 and a light chain comprising the amino acid sequence SEQ ID NO:135.

[0140] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:145 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:147 and a light chain comprising the amino acid sequence SEQ ID NO:135.

[0141] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:149 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:150 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:151 and a light chain comprising the amino acid sequence SEQ ID NO:135. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence S EQ ID NO:152 and a light chain comprising the amino acid sequence SEQ ID NO:135.

[0142] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus. In some embodiments, the bispecific molecule comprises a chimeric heavy chain comprising the amino acid sequence SEQ ID NO:159 and a light chain comprising the amino acid sequence SEQ ID NO:160.

[0143] In some embodiments, the chimeric heavy chain of the bispecific molecule can comprise a polypeptide sequence such as VH1-CH1-L1-BU2-L2-CH2-CH3 from the N-terminus to the C-terminus. In some embodiments, the light chain of the bispecific molecule can comprise a polypeptide sequence such as VL1-CL from the N-terminus to the C-terminus.

[0144] Exemplary antibody sequences are shown in Tables 2-6, and the CDR numbers follow the Kabat EU index. Those skilled in the art know that there are many known algorithms for predicting the positions of CDRs and defining the light chain variable region and heavy chain variable region of an antibody. The CDRs of anti-PcrV antibodies, anti-Psl antibodies or bispecific antibodies that specifically recognize Pseudomonas PcrV and / or Pseudomonas Psl described herein, V H and / or V L sequences are included, but antibodies not exemplified in the following tables based on prediction algorithms are also within the scope of the present invention. The anti-PcrV antibody sequences or anti-Psl antibody sequences of our International Patent Application Numbers PCT / CN2020 / 100592, PCT / CN2020 / 093702 and PCT / CN2020 / 107666 are incorporated into the present invention.

[0145]

Table 2-1

[0146]

Table 2-2

[0147]

Table 2-3

[0148]

Table 3-1

[0149]

Table 3-2

[0150]

Table 3-3

[0151]

Table 3-4

[0152]

Table 3-5

[0153]

Table 3-6

[0154]

Table 4

[0155]

Table 5

[0156]

Table 6-1

[0157]

Table 6-2

[0158]

Table 6-3

[0159]

Table 6-4

[0160]

Table 6-5

[0161]

Table 6-6

[0162]

Table 6-7

[0163]

Table 6-8

[0164]

Table 6-9

[0165]

Table 6-10

[0166] A combination of antibodies that recognize Pseudomonas PcrV and Pseudomonas Psl In one aspect of the present invention, there is provided a pharmaceutical composition comprising (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas Psl.

[0167] In some embodiments, there is provided a pharmaceutical composition comprising (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises (a) a heavy-chain variable domain (V H ) and a light-chain variable domain (V L ), and the V Hcomprises a heavy chain complementarity determining region (HC-CDR) 1 that is DX1X2MS (SEQ ID NO:20), where X1 is N or Y, and X2 is Y, H, or P; a HC-CDR2 that is X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G; and a HC-CDR3 that is GRFSTX1SX2HFX3RAVYGMDV (SEQ ID NO:16), where X1 is N or Y, and X2 is Y, H, or P. and HC-CDR3, wherein X1 is L, S, N or D, X2 is S or A, and X3 is F or Y; L comprises a light chain complementarity determining region (LC-CDR) 1 comprising RASQGISSYLA (SEQ ID NO:10), a LC-CDR2 comprising AASTLQS (SEQ ID NO:11), and a LC-CDR3 comprising QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T; or (b) V H and V L The V H comprises an HC-CDR1 that is X1X2X3MS (SEQ ID NO:17), where X1 is D or S, X2 is Y or N, and X3 is P, H, Y, or S; an HC-CDR2 that is X1ISESGGSTX2X3ADSVKG (SEQ ID NO:18), where X1 is G or V, X2 is N or Y, and X3 is D or Y; and an HC-CDR2 that is GRFX1X2X3X4X5X6FX7RAVYGMDV (SEQ ID NO:19), where X1 is G or V, X2 is N or Y, and X3 is D or Y. X1 is S or C, X2 is T, G, D, Y, Q or A, X3 is S, D, N, E, L, A or Y, X4 is S, T, Y or A, X5 is S, H, Q, A, R, K, G, E, Y or D, X6 is H or C, and X7 is F or Y; and L RASQGIX1SYLA(SEQ Provided is a pharmaceutical composition comprising an LC-CDR1 containing ID NO:209), wherein X1 is S or R, an LC-CDR2 containing AASTLQS (SEQ ID NO:11), and an LC-CDR3 containing QQLX1SYPLX2 (SEQ ID NO:210), wherein X1 is S, N or K and X2 is S or T.

[0168] In some embodiments, a pharmaceutical composition comprising (i) an antigen-binding protein that specifically recognizes Pseudomonas Psl and (ii) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV, wherein the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises (a) V H and V L wherein the V H comprises an HC-CDR1 containing SSGDYWG (SEQ ID NO:48), an HC-CDR2 containing SIHNX1GSTYYNPSLKG (SEQ ID NO:81), wherein X1 is S or Q, and an HC-CDR3 containing QFGSETYYX1GIX2P (SEQ ID NO:82), wherein X1 is T, N or P and X2 is D or Q, and the V L comprises an LC-CDR1 containing RSSQSLLHSX1GYNYLD (SEQ ID NO:83), wherein X1 is N or R, an LC-CDR2 containing LGSNRAS (SEQ ID NO:70), and an LC-CDR3 containing MQALQTPYT (SEQ ID NO:74), or (b) V H and V L wherein the V H comprises an HC-CDR1 containing IHSVH (SEQ ID NO:50), an HC-CDR2 containing TIISSGTTTTYAQSFQD (SEQ ID NO:55), and an HC-CDR3 containing DGX1S (SEQ ID NO:84), wherein X1 is D or T, and the V L comprises RASQGISSWLA (SEQ ID LC-CDR1 containing NO:68), LC-CDR2 containing HASTLES(SEQ ID NO:72), and LC-CDR3 containing LQAX1SLPHT(SEQ ID NO:85), where X1 is N, F, S or K, or (c)V H and V L containing, said V H contains HC-CDR1 with amino acid sequence SEQ ID NO:49, HC-CDR2 with amino acid sequence SEQ ID NO:54, and HC-CDR3 with amino acid sequence SEQ ID NO:61, said V L contains LC-CDR1 with amino acid sequence SEQ ID NO:67, LC-CDR2 with amino acid sequence SEQ ID NO:71, and LC-CDR3 with amino acid sequence SEQ ID NO:75, or (d)V H and V L containing, said V H contains HC-CDR1 with amino acid sequence SEQ ID NO:51, HC-CDR2 with amino acid sequence SEQ ID NO:56, and HC-CDR3 with amino acid sequence SEQ ID NO:64, said V L contains LC-CDR1 with amino acid sequence SEQ ID NO:69, LC-CDR2 with amino acid sequence SEQ ID NO:73, and LC-CDR3 with amino acid sequence SEQ ID NO:80, to provide a pharmaceutical composition.

[0169] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises a heavy-chain variable domain (V H ) and a light-chain variable domain (V L ), said V Hcomprises a heavy chain complementarity determining region (HC-CDR) 1 containing DX1X2MS (SEQ ID NO:20), where X1 is N or Y and X2 is Y, H or P, an HC-CDR2 containing X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G, and an HC-CDR3 containing GRFSTX1SX2HFX3RAVYGMDV (SEQ ID NO:21), where X1 is L, S, N or D, X2 is S or A, and X3 is F or Y, said V L comprises a light chain complementarity determining region (LC-CDR) 1 containing RASQGISSYLA (SEQ ID NO:10), an LC-CDR2 containing AASTLQS (SEQ ID NO:11), and an LC-CDR3 containing QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T. In some embodiments, said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V H and V L comprises, said V H comprises an HC-CDR1 containing DNX1MS (SEQ ID NO:14), where X1 is Y or H, an HC-CDR2 containing X1ISESGGSTNYADSVKG (SEQ ID NO:15), where X1 is V or G, and an HC-CDR3 containing GRFSTX1SSHFX2RAVYGMDV (SEQ NO:16), where X1 is L or S and X2 is F or Y, said V L comprises an LC-CDR1 containing RASQGISSYLA (SEQ ID NO:10), an LC-CDR2 containing AASTLQS (SEQ ID NO:11), and an LC-CDR3 containing QQLSSYPLX1 (SEQ ID NO:19), where X1 is S or T.

[0170] In some embodiments, said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V H and V L comprises, said V Hcomprises HC-CDR1 containing the amino acid sequence SEQ ID NO:1, HC-CDR2 containing the amino acid sequence SEQ ID NO:4, and HC-CDR3 containing the amino acid sequence SEQ ID NO:6, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:12. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:2, HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and HC-CDR3 containing the amino acid sequence SEQ ID NO:7, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:13. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said V H comprises HC-CDR1 containing the amino acid sequence SEQ ID NO:2, HC-CDR2 containing the amino acid sequence SEQ ID NO:5, and HC-CDR3 containing the amino acid sequence SEQ ID NO:8, and said V L comprises LC-CDR1 containing the amino acid sequence SEQ ID NO:10, LC-CDR2 containing the amino acid sequence SEQ ID NO:11, and LC-CDR3 containing the amino acid sequence SEQ ID NO:12. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and said V Hcomprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:3, HC-CDR2 comprising the amino acid sequence SEQ ID NO:5, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:9, wherein said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and the amino acid sequence SEQ ID NO:13.

[0171] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is (a) V H and V L wherein said V H comprises HC-CDR1 comprising SYWMH (SEQ ID NO:22), HC-CDR2 comprising RINEX1EX2SISYADSVKG (SEQ ID NO:44), where X1 is D or N, and X2 is T, G or R, and HC-CDR3 comprising DGPYDX1X2DI (SEQ ID NO:45), where X1 is S, A or T, and X2 is F or L, and said V L comprises RASQX1VX2X3NLA (SEQ ID NO:46), where X1 is S, G or N, X2 is S, R or K, and X3 is S or N, LC-CDR1, LC-CDR2 comprising X1ASSRAT (SEQ ID NO:42), where X1 is D or H, and LC-CDR3 comprising QQYGX1X2PX3T (SEQ ID NO:47), where X1 is S, L or N, X2 is S, Q or E, and X3 is L or I, or (b) V H and V L wherein said V H comprises HC-CDR1 comprising SYWMH (SEQ ID NO:22), RINEX1EX2SISYADSVKG (SEQ comprising HC-CDR2 containing ID NO: 211), wherein X1 is D, N, I, L or V and X2 is S, T, R, G or N, and HC-CDR3 containing DGPYDX1X2DI (SEQ ID NO: 45), wherein X1 is S, A or T and X2 is F or L, said V L comprises RASQX1VX2X3NLA (SEQ ID NO: 212), wherein X1 is N, G, D or S, X2 is K, R, S, N or T, X3 is N, G, S or D for LC-CDR1, X1ASSRAT (SEQ ID NO: 213), wherein X1 is D, N, H or A for LC-CDR2, and QQYGX1X2PX3T (SEQ ID NO: 214), 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, X3 is I, L or V for LC-CDR3. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L comprising, said V H comprises HC-CDR1 containing SYWMH (SEQ ID NO: 22), HC-CDR2 containing RINEX1EX2SISYADSVKG (SEQ ID NO: 39), wherein X1 is D or N and X2 is T or G, and HC-CDR3 containing DGPYDX1LDI (SEQ ID NO: 40), wherein X1 is S or A, said V L comprises RASQX1VX2X3NLA (SEQ ID NO: 41), wherein X1 is S or G, X2 is S or R, X3 is S or N for LC-CDR1, X1ASSRAT (SEQ ID NO: 42), wherein X1 is D or H for LC-CDR2, and QQYGX1X2PX3T (SEQ ID NO: 4 3), wherein X1 is S or L, X2 is S or Q, X3 is L or I for LC-CDR3.

[0172] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising any one of the amino acid sequences of SEQ ID NOs:23-25, and an HC-CDR3 comprising any one of the amino acid sequences of SEQ ID NOs:26-29, and the V L comprises an LC-CDR1 comprising any one of the amino acid sequences of SEQ ID NOs:30-32, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:33 or 34, and an LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NOs:35-37.

[0173] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:23, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:26, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:30, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:33, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:35. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:27, and the V LIt includes LC-CDR1 containing the amino acid sequence SEQ ID NO:31, LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and LC-CDR3 containing the amino acid sequence SEQ ID NO:36. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H includes HC-CDR1 containing the amino acid sequence SEQ ID NO:22, HC-CDR2 containing the amino acid sequence SEQ ID NO:25, and HC-CDR3 containing the amino acid sequence SEQ ID NO:27, and the V L includes LC-CDR1 containing the amino acid sequence SEQ ID NO:32, LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and LC-CDR3 containing the amino acid sequence SEQ ID NO:37. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H includes HC-CDR1 containing the amino acid sequence SEQ ID NO:22, HC-CDR2 containing the amino acid sequence SEQ ID NO:25, and HC-CDR3 containing the amino acid sequence SEQ ID NO:28, and the V L includes LC-CDR1 containing the amino acid sequence SEQ ID NO:32, LC-CDR2 containing the amino acid sequence SEQ ID NO:34, and LC-CDR3 containing the amino acid sequence SEQ ID NO:37. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H includes HC-CDR1 containing the amino acid sequence SEQ ID NO:22, HC-CDR2 containing the amino acid sequence SEQ ID NO:24, and HC-CDR3 containing the amino acid sequence SEQ ID NO:28, and the V Lcomprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:36. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L . The V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:25, and an amino acid sequence SEQ ID NO:26. The V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:32, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:37. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L . The V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:26. The V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:36. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L . The V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:23, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:29. The V LIt comprises an LC-CDR1 containing the amino acid sequence SEQ ID NO:30, an LC-CDR2 containing the amino acid sequence SEQ ID NO:33, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:35.

[0174] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H comprises an HC-CDR1 containing SSGDYWG (SEQ ID NO:48), an HC-CDR2 containing SIHNX1GSTYYNPSLKG (SEQ ID NO:81), where X1 is S or Q, and an HC-CDR3 containing QFGSETYYX1GIX2P (SEQ ID NO:82), where X1 is T, N or P and X2 is D or Q. The V L comprises an LC-CDR1 containing RSSQSLLHSX1GYNYLD (SEQ ID NO:83), where X1 is N or R, an LC-CDR2 containing LGSNRAS (SEQ ID NO:70), and an LC-CDR3 containing MQALQTPYT (SEQ ID NO:74). In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H comprises an HC-CDR1 containing SSGDYWG (SEQ ID NO:48), an HC-CDR2 containing SIHNX1GSTYYNPSLKG (SEQ ID NO:125), where X1 is S, K or Q, and an HC-CDR3 containing QFGSETYYX1GIX2P (SEQ ID NO:126), where X1 is N, S, V, T or P and X2 is D, Y, C, H, S, R, A, E, G, K, W, V or Q. The V L comprises an LC-CDR1 containing RSSQSLLHSX1GYNYLD (SEQ ID NO:127), where X1 is N, A, V, F, R, G, H, Q, W or P, an LC-CDR2 containing LGSNRAS (SEQ It includes LC-CDR2 containing ID NO:70), and LC-CDR3 containing MQALQTPX1T (SEQ ID NO:128), where X1 is R or Y.

[0175] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl is V H and V L and includes, where the V H includes HC-CDR1 containing IHSVH (SEQ ID NO:50), HC-CDR2 containing TIISSGTTTTYAQSFQD (SEQ ID NO:55), and HC-CDR3 containing DGX1S (SEQ ID NO:84), where X1 is D or T, and the V L includes LC-CDR1 containing RASQGISSWLA (SEQ ID NO:68), LC-CDR2 containing HASTLES (SEQ ID NO:72), and LC-CDR3 containing LQAX1SLPHT (SEQ ID NO:85), where X1 is N, F, S or K. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl is V H and V L and includes, where the V H includes HC-CDR1 containing IHSVH (SEQ ID NO:50), HC-CDR2 containing TIISSGTTTTYAQSFQD (SEQ ID NO:55), and HC-CDR3 containing X1X2X3X4 (SEQ ID NO:129), where X1 is D, Y or N, X2 is G or A, X3 is D or T, X4 is S, A or T, and the V L includes LC-CDR1 containing RASQGISSWLA (SEQ ID NO:68), LC-CDR2 containing HASTLES (SEQ ID NO:72), and LC-CDR3 containing LQAX1SLPHT (SEQ ID NO:130), where X1 is N, D, Y, F, P, G, K, H, A, C, E, Q, R, S, T, V, W or L.

[0176] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52 or 53, and an HC-CDR3 comprising any one of the amino acid sequences of SEQ ID NOs:57-60, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65 or 66, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74.

[0177] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:50, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:55, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:62 or 63, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:68, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:72, and an LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NOs:76-79.

[0178] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, and the V LIt includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:65, an LC-CDR2 containing the amino acid sequence SEQ ID NO:70, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:74. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:48, an HC-CDR2 containing the amino acid sequence SEQ ID NO:53, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:58. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:66, an LC-CDR2 containing the amino acid sequence SEQ ID NO:70, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:74. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:48, an HC-CDR2 containing the amino acid sequence SEQ ID NO:52, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:59. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:65, an LC-CDR2 containing the amino acid sequence SEQ ID NO:70, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:74. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:48, an HC-CDR2 containing the amino acid sequence SEQ ID NO:53, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:60. The V LIt includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:65, an LC-CDR2 containing the amino acid sequence SEQ ID NO:70, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:74. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:49, an HC-CDR2 containing the amino acid sequence SEQ ID NO:54, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:61. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:67, an LC-CDR2 containing the amino acid sequence SEQ ID NO:71, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:75. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:50, an HC-CDR2 containing the amino acid sequence SEQ ID NO:55, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:62. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:68, an LC-CDR2 containing the amino acid sequence SEQ ID NO:72, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:76. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:50, an HC-CDR2 containing the amino acid sequence SEQ ID NO:55, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:63. The V LIt includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:68, an LC-CDR2 containing the amino acid sequence SEQ ID NO:72, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:77. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:50, an HC-CDR2 containing the amino acid sequence SEQ ID NO:55, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:63. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:68, an LC-CDR2 containing the amino acid sequence SEQ ID NO:72, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:78. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:50, an HC-CDR2 containing the amino acid sequence SEQ ID NO:55, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:63. The V L includes an LC-CDR1 containing the amino acid sequence SEQ ID NO:68, an LC-CDR2 containing the amino acid sequence SEQ ID NO:72, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:79. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes an HC-CDR1 containing the amino acid sequence SEQ ID NO:51, an HC-CDR2 containing the amino acid sequence SEQ ID NO:56, and an HC-CDR3 containing the amino acid sequence SEQ ID NO:64. The V L has the amino acid sequence SEQ ID It comprises an LC-CDR1 containing NO:69, an LC-CDR2 containing the amino acid sequence SEQ ID NO:73, and an LC-CDR3 containing the amino acid sequence SEQ ID NO:80.

[0179] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and the V H contains the amino acid sequence SEQ ID NO:91, and the V L contains the amino acid sequence SEQ ID NO:112. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:91, and the V L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:112.

[0180] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and the V H contains the amino acid sequence SEQ ID NO:92, and the V L contains the amino acid sequence SEQ ID NO:113. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:92, and the V L contains the amino acid sequence SEQ ID NO:113 in VL comprises LC-CDR1, LC-CDR2 and LC-CDR3 in

[0181] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V H comprises the amino acid sequence SEQ ID NO:93, and said V L comprises the amino acid sequence SEQ ID NO:112. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V H is V having the amino acid sequence SEQ ID NO:93 H comprising HC-CDR1, HC-CDR2 and HC-CDR3 in L and said V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in

[0182] In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V H comprises the amino acid sequence SEQ ID NO:94, and said V L comprises the amino acid sequence SEQ ID NO:113. In some embodiments, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprising, wherein said V H is V having the amino acid sequence SEQ ID NO:94 H comprising HC-CDR1, HC-CDR2 and HC-CDR3 in L and said V LIt includes LC-CDR1, LC-CDR2 and LC-CDR3 therein.

[0183] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H includes the amino acid sequence SEQ ID NO:95, and the V L includes the amino acid sequence SEQ ID NO:114. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H is the V having the amino acid sequence SEQ ID NO:95 and includes HC-CDR1, HC-CDR2 and HC-CDR3 in V H , and the V L is the V having the amino acid sequence SEQ ID NO:114 and includes LC-CDR1, LC-CDR2 and LC-CDR3 in V L .

[0184] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H includes the amino acid sequence SEQ ID NO:96, and the V L includes the amino acid sequence SEQ ID NO:115. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H is the V having the amino acid sequence SEQ ID NO:96 and includes HC-CDR1, HC-CDR2 and HC-CDR3 in V H , and the V L is the V having the amino acid sequence SEQ ID NO:115 and includes LC-CDR1, LC-CDR2 and LC-CDR3 in V L .

[0185] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises the amino acid sequence SEQ ID NO:97, and the V L comprises the amino acid sequence SEQ ID NO:116. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:97, and the V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:116.

[0186] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises the amino acid sequence SEQ ID NO:98, and the V L comprises the amino acid sequence SEQ ID NO:116. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:98, and the V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:116.

[0187] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises the amino acid sequence SEQ ID NO:99, and the V L comprises the amino acid sequence SEQ ID NO:115. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:99, and the V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:115.

[0188] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises the amino acid sequence SEQ ID NO:100, and the V L comprises the amino acid sequence SEQ ID NO:116. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:100, and the V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:116.

[0189] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises the amino acid sequence SEQ ID NO: 101, and the V L comprises the amino acid sequence SEQ ID NO: 115. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises HC-CDR1, HC-CDR2, and HC-CDR3 in V H having the amino acid sequence SEQ ID NO: 101, and the V L comprises LC-CDR1, LC-CDR2, and LC-CDR3 in V L having the amino acid sequence SEQ ID NO: 115.

[0190] In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises the amino acid sequence SEQ ID NO: 102, and the V L comprises the amino acid sequence SEQ ID NO: 114. In some embodiments, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L and the V H comprises HC-CDR1, HC-CDR2, and HC-CDR3 in V H having the amino acid sequence SEQ ID NO: 102, and the V L comprises LC-CDR1, LC-CDR2, and LC-CDR3 in V L having the amino acid sequence SEQ ID NO: 114.

[0191] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes the amino acid sequence SEQ ID NO: 103, and the V L includes the amino acid sequence SEQ ID NO: 117. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes HC-CDR1, HC-CDR2, and HC-CDR3 in V H having the amino acid sequence SEQ ID NO: 103, and the V L includes LC-CDR1, LC-CDR2, and LC-CDR3 in V L having the amino acid sequence SEQ ID NO: 117.

[0192] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes the amino acid sequence SEQ ID NO: 104, and the V L includes the amino acid sequence SEQ ID NO: 118. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H includes HC-CDR1, HC-CDR2, and HC-CDR3 in V H having the amino acid sequence SEQ ID NO: 104, and the V L includes LC-CDR1, LC-CDR2, and LC-CDR3 in V L having the amino acid sequence SEQ ID NO: 118.

[0193] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V Lcomprising said V H comprises the amino acid sequence SEQ ID NO: 105, said V L comprises the amino acid sequence SEQ ID NO: 117. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprising said V H has the amino acid sequence SEQ ID NO: 105 and comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V H , said V L has the amino acid sequence SEQ ID NO: 117 and comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V L .

[0194] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprising said V H comprises the amino acid sequence SEQ ID NO: 106, said V L comprises the amino acid sequence SEQ ID NO: 117. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprising said V H has the amino acid sequence SEQ ID NO: 106 and comprises HC-CDR1, HC-CDR2 and HC-CDR3 in V H , said V L has the amino acid sequence SEQ ID NO: 117 and comprises LC-CDR1, LC-CDR2 and LC-CDR3 in V L .

[0195] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprising said V H comprises the amino acid sequence SEQ ID NO: 107, said V LIt contains the amino acid sequence SEQ ID NO:119. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl protein contains V H and V L and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:107, and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:107, and the V L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:119. L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:119.

[0196] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl contains V H and V L and the V H contains the amino acid sequence SEQ ID NO:108, and the V L contains the amino acid sequence SEQ ID NO:119. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl contains V H and V L and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:108, and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:108, and the V L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:119. L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:119.

[0197] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl contains V H and V L and the V H contains the amino acid sequence SEQ ID NO:109, and the V LIt contains the amino acid sequence SEQ ID NO:120. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:109, and the V L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:120.

[0198] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H contains the amino acid sequence SEQ ID NO:110, and the V L contains the amino acid sequence SEQ ID NO:121. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H contains the HC-CDR1, HC-CDR2 and HC-CDR3 in V H having the amino acid sequence SEQ ID NO:110, and the V L contains the LC-CDR1, LC-CDR2 and LC-CDR3 in V L having the amino acid sequence SEQ ID NO:121.

[0199] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and the V H contains the amino acid sequence SEQ ID NO:110, and the V L contains the amino acid sequence SEQ ID NO:122. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is VH and V L comprises, and said V H has the amino acid sequence SEQ ID NO:110, and the V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in, and said V L has the amino acid sequence SEQ ID NO:122, and the V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in.

[0200] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and said V H comprises the amino acid sequence SEQ ID NO:110, and said V L comprises the amino acid sequence SEQ ID NO:123. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and said V H has the amino acid sequence SEQ ID NO:110, and the V H comprises HC-CDR1, HC-CDR2 and HC-CDR3 in, and said V L has the amino acid sequence SEQ ID NO:123, and the V L comprises LC-CDR1, LC-CDR2 and LC-CDR3 in.

[0201] In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and said V H comprises the amino acid sequence SEQ ID NO:111, and said V L comprises the amino acid sequence SEQ ID NO:124. In some embodiments, the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and said V H has the amino acid sequence SEQ ID NO:111, and the VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 therein, wherein said V L has an amino acid sequence SEQ ID NO:124 for V L comprising LC-CDR1, LC-CDR2 and LC-CDR3 therein.

[0202] In some embodiments, the pharmaceutical composition comprises (i) an antigen-binding protein that specifically recognizes Pseudomonas Psl, and (ii) an antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV, and the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is about 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 or 1:20. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is about 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:5. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is about 2:1 or about 1:1.

[0203] In some embodiments, the pharmaceutical composition comprises (i) an antigen-binding protein that specifically recognizes Pseudomonas Psl, and (ii) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV, and the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 or 1:20. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:5. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 2:1 or about 1:1.

[0204] In some embodiments, the pharmaceutical composition comprises (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV, and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas Psl, and the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 or 1:20. In some embodiments, Pseudomonas The molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, or 1:5. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is about 2:1 or about 1:1.

[0205] In some embodiments, the pharmaceutical composition comprises (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas Psl, and the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, or 1:20. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, or 1:5. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1 or about 1:2.

[0206] In some embodiments, a pharmaceutical composition comprising an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L wherein the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:12, and the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises V H and V L wherein the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74, provides a pharmaceutical composition. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1 or about 1:2.

[0207] In some embodiments, a pharmaceutical composition comprising an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is VH and V L comprises, wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and said V L comprises LC -CDR1 comprising the amino acid sequence SEQ ID NO:10, LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:12, and the antigen-binding protein that specifically recognizes Pseudomonas Psl comprises V H and V L comprises, wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, and said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:75, and provides a pharmaceutical composition. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1 or about 1:2.

[0208] In some embodiments, a pharmaceutical composition comprising an antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV and an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV comprises V H and V L comprises, wherein said V Hcomprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:2, HC-CDR2 comprising the amino acid sequence SEQ ID NO:5, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:7, wherein said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:13, and wherein said antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L and wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, and wherein said V L comprises LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:74, and provides a pharmaceutical composition. In some embodiments, the molar ratio of said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV to said antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1 or about 1:2.

[0209] A pharmaceutical composition comprising an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas PcrV and an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V H and V L and wherein said V H comprises HC-CDR1 comprising the amino acid sequence SEQ ID NO:2, HC-CDR2 comprising the amino acid sequence SEQ ID NO:5, and HC-CDR3 comprising the amino acid sequence SEQ ID NO:7, and wherein said V Lcomprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:13, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:49, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:54, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:61, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75, and provides a pharmaceutical composition. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1 or about 1:2.

[0210] In some embodiments, a pharmaceutical composition comprising an antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV, an antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV, and an antigen-binding protein that specifically recognizes Pseudomonas Psl, wherein the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V H and V L comprises, and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:1, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:4, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:6, and the V Lcomprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:10, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:11, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:12, and the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V H and V L comprises, and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:22, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:24, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:27, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:31, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:34, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:36, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V H and V L comprises, and the V H comprises an HC-CDR1 comprising the amino acid sequence SEQ ID NO:48, an HC-CDR2 comprising the amino acid sequence SEQ ID NO:52, and an HC-CDR3 comprising the amino acid sequence SEQ ID NO:57, and the V L comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74, and provides a pharmaceutical composition. In some embodiments, the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is about 1:1:1 or 1:1:2.

[0211] Anti-PcrV antigen-binding domain and antigen-binding protein In some embodiments, the antigen-binding domain and the antigen-binding protein that specifically recognize Pseudomonas PcrV include, but are not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibodies comprising the heavy-chain CDRs and / or light-chain CDRs described herein. In one aspect of the invention, the antigen-binding domain or the antigen-binding protein is an isolated antibody that binds to PcrV. Predicted antigen-binding domains or antigen-binding proteins that specifically recognize Pseudomonas PcrV include, for example, whole or fragments of full-length anti-PcrV antibodies (e.g., full-length IgG1, IgG2 or IgG4), anti-PcrV single-chain antibodies, multispecific (e.g., bispecific) anti-PcrV antibodies, anti-PcrV immune complexes, and the like. 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 or diabody. In some embodiments, an antigen-binding domain or antigen-binding protein that specifically binds to PcrV means that the binding affinity of the antigen-binding domain or antigen-binding protein to PcrV is at least 10-fold or more (e.g., 10, 10 2 10 3 10 4 10 5 10 6 10 7 times) greater than the binding affinity of the antigen-binding domain or antigen-binding protein to a non-target. In some embodiments, a non-target refers to an antigen other than PcrV.

[0212] Binding affinity can be measured by methods known in the art, such as ELISA, fluorescence-activated cell sorting (FACS) analysis, or radioimmuno-precipitation analysis (RIA). The Kd value can be measured by methods known in the art, such as surface plasmon resonance (SPR) technology or biolayer interferometry (BLI) technology.

[0213] ​In one aspect, the antigen-binding domain or the antigen-binding protein that specifically binds to Pseudomonas PcrV promotes, mediates, or enhances Pseudomonas aeruginosa opsonophagocytic killing (OPK) and / or (b) disrupts the activity of the type III toxin secretion system.

[0214] As used herein, antigen-binding domains or antigen-binding proteins that include human sequences (e.g., human heavy and light chain variable domains that include human CDR sequences) are described in detail, but non-human antigen-binding domains or antigen-binding proteins are also contemplated. In some embodiments, the non-human antigen-binding domain or antigen-binding protein includes the human CDR sequences and non-human framework region sequences of the antigen-binding domains or antigen-binding proteins described herein. In some embodiments, the non-human framework region sequences are any sequences that can be used to form heavy and / or light chain variable domains using one or more of the human CDR sequences described herein, including mammalian animals such as mice, rats, rabbits, pigs, bovines (e.g., cows, oxen, swine), deer, sheep, goats, chickens, cats, dogs, ferrets, primates (e.g., marmosets, macaques). In some embodiments, the non-human antigen-binding domain or antigen-binding protein includes an antigen-binding domain or antigen-binding protein produced by grafting one or more of the human CDR sequences described herein onto a non-human framework region (e.g., a mouse or chicken framework region sequence).

[0215] The complete amino acid sequence of the exemplary PcrV protein comprises or consists of the amino acid sequence SEQ ID NO:86. In some embodiments, an antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically recognizes one epitope on Pseudomonas PcrV. In some embodiments, the antigen-binding domain or the antigen-binding protein is specific for Pseudomonas PcrV and does not have cross-species reactivity or does not cross-react with other types of non-Pseudomonas proteins.

[0216] In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to a linear epitope on Pseudomonas PcrV. In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to a non-linear epitope on Pseudomonas PcrV. In some embodiments, the antigen-binding domain or antigen-binding protein described herein specifically binds to an epitope on Pseudomonas PcrV, wherein the epitope comprises at least 1, 2, 3, 4, 5, or 6 amino acid residues selected from the group consisting of Gln160, Asp165, Asp170, Asp173, Thr175, and Ser202 in Pseudomonas PcrV according to amino acid sequence SEQ ID NO:86. In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to an epitope on Pseudomonas PcrV, wherein the epitope comprises at least 2 amino acid residues selected from the group consisting of Gln160, Asp165, Asp170, Asp173, Thr175, and Ser202 in Pseudomonas PcrV according to amino acid sequence SEQ ID NO:86. In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to an epitope on Pseudomonas PcrV, wherein the epitope comprises Gln160, Asp165, Asp170, Asp173, Thr175, and Ser202 in Pseudomonas PcrV according to amino acid sequence SEQ ID NO:86 and... It comprises at least 3 amino acid residues selected from the group consisting of r175 and Ser202. In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to an epitope on Pseudomonas PcrV, wherein the epitope comprises at least 4 amino acid residues selected from the group consisting of Gln160, Asp165, Asp170, Asp173, Thr175, and Ser202 in Pseudomonas PcrV according to the amino acid sequence SEQ ID NO:86. In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to an epitope on Pseudomonas PcrV, wherein the epitope comprises at least 5 amino acid residues selected from the group consisting of Gln160, Asp165, Asp170, Asp173, Thr175, and Ser202 in Pseudomonas PcrV according to the amino acid sequence SEQ ID NO:86. In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas PcrV described herein specifically binds to an epitope on Pseudomonas PcrV, wherein the epitope comprises Gln160, Asp165, Asp170, Asp173, Thr175, and Ser202 in Pseudomonas PcrV according to the amino acid sequence SEQ ID NO:86.

[0217] In some embodiments, the bispecific molecule comprises an antibody heavy chain constant region and an antibody light chain constant region. In some embodiments, the bispecific molecule comprises an IgG1 heavy chain constant region. In some embodiments, the bispecific molecule comprises an IgG2 heavy chain constant region. In some embodiments, the bispecific molecule comprises an IgG3 heavy chain constant region. In some embodiments, the bispecific molecule comprises an IgG4 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 SEQ ID NO:89. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:90. In some embodiments, the bispecific molecule comprises a λ light chain constant region. In some embodiments, the anti-PcrV antibody comprises a κ light chain constant region. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:88. ID NO:88. In some embodiments, the bispecific molecule comprises an antibody heavy chain variable domain and an antibody light chain variable domain.

[0218] Anti-Psl antigen-binding domain and antigen-binding protein In some embodiments, the antigen-binding domain and antigen-binding protein that specifically recognize Pseudomonas Psl include, but are not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibodies comprising the heavy-chain CDRs and / or light-chain CDRs described herein. In one aspect of the invention, the antigen-binding domain or the antigen-binding protein is an isolated antibody that binds to Psl. Predicted antigen-binding domains or antigen-binding proteins that specifically recognize Pseudomonas Psl include, for example, whole or fragments of full-length anti-Psl antibodies (e.g., full-length IgG1, IgG2 or IgG4), anti-Psl single-chain antibodies, multispecific (e.g., bispecific) anti-Psl antibodies, anti-Psl immune complexes, and the like. In some embodiments, the anti-Psl antibody is a Fab, Fab’, F(ab)’2, Fab’-SH, single-chain antibody (scFv), Fv fragment, dAb, Fd, nanobody or diabody. In some embodiments, an antigen-binding domain or antigen-binding protein that specifically binds to Psl means that the binding affinity of the antigen-binding domain or antigen-binding protein to Psl is at least 10-fold or more (e.g., 10, 10 2 10 3 10 4 10 5 10 6 10 7 times) greater than the binding affinity of the antigen-binding domain or antigen-binding protein to a non-target. In some embodiments, a non-target refers to an antigen other than Psl.

[0219] Binding affinity can be measured by methods known in the art, such as ELISA, fluorescence-activated cell sorting (FACS) analysis, or radioimmunoprecipitation analysis (RIA). The Kd value can be measured by methods known in the art, such as surface plasmon resonance (SPR) technology or biolayer interferometry (BLI) technology.

[0220] In one aspect, the antigen-binding domain or the antigen-binding protein that specifically binds to Pseudomonas aeruginosa Psl promotes, mediates, or enhances opsonophagocytic killing (OPK) of Pseudomonas aeruginosa and / or inhibits the binding of Pseudomonas aeruginosa to epithelial cells.

[0221] Although this specification describes in detail antigen-binding domains or antigen-binding proteins that include human sequences (e.g., human heavy-chain variable domains and light-chain variable domains that include human CDR sequences), non-human antigen-binding domains or antigen-binding proteins are also contemplated. In some embodiments, the non-human antigen-binding domain or antigen-binding protein includes the human CDR sequences and non-human framework region sequences of the antigen-binding domains or antigen-binding proteins described herein. In some embodiments, the non-human framework region sequences are any sequences that can be used to form a heavy-chain variable domain and / or a light-chain variable domain using one or more of the human CDR sequences described herein, including mammalian animals such as mice, rats, rabbits, pigs, bovines (e.g., cows, oxen, water buffalo), deer, sheep, goats, chickens, cats, dogs, ferrets, primates (e.g., marmosets, macaques), etc. In some embodiments, the non-human antigen-binding domain or antigen-binding protein includes an antigen-binding domain or antigen-binding protein produced by grafting one or more of the human CDR sequences described herein onto a non-human framework region (e.g., a mouse or chicken framework region sequence).

[0222] In some embodiments, the antigen-binding domain or antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl described herein specifically recognizes one epitope on Pseudomonas aeruginosa Psl. In some embodiments, the antigen-binding domain or the antigen-binding protein is specific for Pseudomonas aeruginosa Psl and has no species cross-reactivity or does not cross-react with other types of non-Pseudomonas proteins.

[0223] In some embodiments, the bispecific molecule comprises an antibody heavy chain constant region and an antibody light chain constant region. In some embodiments, the bispecific molecule comprises an IgG1 heavy chain constant region. In some embodiments, the bispecific molecule comprises an IgG2 heavy chain constant region. In some embodiments, the bispecific molecule comprises an IgG3 heavy chain constant region. In some embodiments, the bispecific molecule comprises an IgG4 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 SEQ ID NO:89. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:90. In some embodiments, the bispecific molecule comprises a λ light chain constant region. In some embodiments, the anti-PcrV antibody comprises a κ light chain constant region. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:88. ID NO:88. In some embodiments, the bispecific molecule comprises an antibody heavy chain variable domain and an antibody light chain variable domain.

[0224] Binding affinity Binding affinity can be represented by Kd, Koff, Kon, or Ka. As used herein, the term "Koff" is intended to refer to the dissociation rate constant of the antigen-binding domain from the antigen-binding domain / antigen complex. "Koff" is measured by a kinetic selection device. The term "Kon" is the association rate constant when an antibody binds to an antigen to form an antigen-binding domain / antigen complex. The dissociation constant "Kd" as used herein refers to the dissociation constant of a particular antibody-antigen interaction and refers to the antigen concentration required when, in a solution of antibody molecules, the antigen occupies half of all the antigen-binding domains and equilibrium is reached, and is equal to Koff / Kon. The measurement of Kd is performed on the premise that all binding molecules are in solution. When the antigen-binding domain is linked to the cell wall, for example, in a yeast expression system, the corresponding dissociation rate constant is represented by EC50, which is a suitable approximation of Kd. The affinity constant Ka is the reciprocal of the dissociation constant Kd. In some embodiments, the antigen-binding domain is contained within an antigen-binding protein.

[0225] The equilibrium dissociation constant (Kd) is used as an indicator of the affinity between the antigen-binding domain moiety and the antigen. For example, it can be easily analyzed by performing the Scatchard method using an antibody labeled with various labeling reagents or the biomolecular interaction analysis by surface plasmon resonance using Biacore (manufactured by Amersham Biosciences) according to the instruction manual and the attached kit. The Kd values obtained by these methods are expressed in units of M. The Kd value of an antibody that specifically binds to a target can be, for example, ≤10 -7 M, ≤10 -8 M, ≤10 -9 M, ≤10 -10 M, ≤10 -11 M, ≤10 -12 M or ≤10 -13 M and may be.

[0226] The binding specificity of the antibody can be measured by known methods in the art. These methods include, but are not limited to, Western blots, ELISA-, RIA-, ECL-, IRMA-, EIA-, BIAcore tests, and peptide scans.

[0227] In some embodiments, the antigen-binding domain that specifically recognizes PcrV specifically binds to the PcrV target, and its Kd value is about 10 -7 M to about 10 -13 M (e.g., about 10 -7 M to about 10 -13 M, about 10 -8 M to about 10 -13 M, about 10 -9 M to about 10 -13 M or about 10 -10 M to about 10 -12 M). Therefore, in some embodiments, the Kd value of the binding between the antigen-binding domain that specifically recognizes PcrV and PcrV is about 10 -7 M to about 10 -13 M, about 1×10 -7 M to about 5×10 -13 M, about 10 -7 M to about 10 -12 M, about 10 -7 M to about 10 -11 M, about 10 -7 M to about 10 -10 M, about 10 -7 M to about 10 -9 M, about 10 -8 M to about 10 -13 M, about 1×10 -8 M to about 5×10 -13 M, about 10 -8 M to about 10 -12 M, about 10 -8 M to about 10 -11 M, about 10 -8 M to about 10 -10 M, about 10 -8 M to about 10 -9 M, about 5×10 -9 M to about 1×10 -13 M, about 5×10 -9 M to about 1×10 -12 M, about 5×10-9 M ~ about 1×10 -11 M, about 5×10 -9 M ~ about 1×10 -10 M, about 10 -9 M ~ about 10 -13 M, about 10 -9 M ~ about 10 -12 M, about 10 -9 M ~ about 10 -11 M, about 10 -9 M ~ about 10 -10 M, about 5×10 -10 M ~ about 1×10 -13 M, about 5×10 -10 M ~ about 1×10 -12 M, about 5×10 -10 M ~ about 1×10 -11 M, about 10 -10 M ~ about 10 -13 M, about 1×10 -10 M ~ about 5×10 -13 M, about 1×10 -10 M ~ about 1×10 -12 M, about 1×10 -10 M ~ about 5×10 -12 M, about 1×10 -10 M ~ about 1×10 -11 M, about 10 -11 M ~ about 10 -13 M, about 1×10 -11 M ~ about 5×10 -13 M, about 10 -11 M ~ about 10 -12 M, about 10 -12 M ~ about 10 -13 M. In some embodiments, the Kd value of the binding between the antigen-binding domain that specifically recognizes PcrV and PcrV is about 10 -7 M ~ about 10 -13 M.

[0228] In some embodiments, the antigen-binding domain that specifically recognizes PcrV and non-target The Kd value of the binding between [antigen binding domain that specifically recognizes PcrV] and the target is higher than the Kd value between the antigen binding domain that specifically recognizes PcrV and the target, and in some embodiments cited herein, the binding affinity between the antigen binding domain that specifically recognizes PcrV and the target (e.g., PcrV) is higher than the binding affinity between the antigen binding domain that specifically recognizes PcrV and a non-target. In some embodiments, the non-target refers to a non-PcrV antigen. In some embodiments, the Kd value of the binding between the antigen binding domain that specifically recognizes PcrV and a non-PcrV target is at least about 10 times, e.g., about 10 - 100 times, about 100 - 1000 times, about 10 3 ~10 4 times, about 10 4 ~10 5 times, about 10 5 ~10 6 times, about 10 6 ~10 7 times, about 10 7 ~10 8 times, about 10 8 ~10 9 times, about 10 9 ~10 10 times, about 10 10 ~10 11 times, about 10 11 ~10 12 times.

[0229] In some embodiments, the antigen binding domain that specifically recognizes Psl specifically binds to the Psl target, and its Kd value is about 10 -7 M to about 10 -13 M (e.g., about 10 -7 M to about 10 -13 M, about 10 -8 M to about 10 -13 M, about 10 -9 M to about 10 -13 M, or about 10 -10 M to about 10 -12 M). Therefore, in some embodiments, the Kd value of the binding between the antigen binding domain that specifically recognizes Psl and Psl is about 10 -7 M to about 10 -13M, about 1×10 -7 M to about 5×10 -13 M, about 10 -7 M to about 10 -12 M, about 10 -7 M to about 10 -11 M, about 10 -7 M to about 10 -10 M, about 10 -7 M to about 10 -9 M, about 10 -8 M to about 10 -13 M, about 1×10 -8 M to about 5×10 -13 M, about 10 -8 M to about 10 -12 M, about 10 -8 M to about 10 -11 M, about 10 -8 M to about 10 -10 M, about 10 -8 M to about 10 -9 M, about 5×10 -9 M to about 1×10 -13 M, about 5×10 -9 M to about 1×10 -12 M, about 5×10 -9 M to about 1×10 -11 M, about 5×10 -9 M to about 1×10 -10 M, about 10 -9 M to about 10 -13 M, about 10 -9 M to about 10 -12 M, about 10 -9 M to about 10 -11 M, about 10 -9 M to about 10 -10 M, about 5×10 -10 M to about 1×10 -13 M, about 5×10 -10 M to about 1×10 -12 M, about 5×10 -10 M to about 1×10 -11 M, about 10 -10 M to about 10 -13 M, about 1×10 -10 M to about 5×10 -13 M, about 1×10 -10 M to about 1×10 -12 M, about 1×10 -10 M to about 5×10 -12 M, about 1×10-10 M ~ about 1 × 10 -11 M, about 10 -11 M ~ about 10 -13 M, about 1 × 10 -11 M ~ about 5 × 10 -13 M, about 10 -11 M ~ about 10 -12 M, about 10 -12 M ~ about 10 -13 M. In some embodiments, the Kd value of the binding between an antigen - binding domain that specifically recognizes Psl and Psl is about 10 -7 M ~ about 10 -13 M.

[0230] In some embodiments, the Kd value of the binding between an antigen - binding domain that specifically recognizes Psl and a non - target is higher than the Kd value of the binding between the antigen - binding domain that specifically recognizes Psl and the target, and in some embodiments cited herein, the binding affinity between an antigen - binding domain that specifically recognizes Psl and the target (e.g., Psl) is higher than the binding affinity between the antigen - binding domain that specifically recognizes Psl and the non - target. In some embodiments, the non - target refers to a non - Psl antigen. In some embodiments, the Kd value of the binding between an antigen - binding domain that specifically recognizes Psl and a non - Psl target is at least about 10 times, e.g., about 10 - 100 times, about 100 - 1000 times, about 10 3 ~ 10 4 times, about 10 4 ~ 10 5 times, about 10 5 ~ 10 6 times, about 10 6 ~ 10 7 times, about 10 7 ~ 10 8 times, about 10 8 ~ 10 9 times, about 10 9 ~ 10 10 times, about 10 10 ~ 10 11 times, about 10 11 ~ 10 12 times.

[0231] Nucleic acid Also included are nucleic acid molecules encoding a PcrV binding domain or protein, a Psl binding domain or protein, and a bispecific molecule. In some embodiments, the present There is provided a nucleic acid (or set of nucleic acids) encoding a full-length anti-PcrV antibody or anti-Psl antibody, including any one of the full-length anti-PcrV antibodies or anti-Psl antibodies described in the present specification. In some embodiments, the nucleic acid (or set of nucleic acids) of the antigen-binding domain, antigen-binding protein or bispecific molecule described herein may further include a nucleic acid sequence encoding a peptide tag (e.g., a protein purification tag, His-tag, HA-tag).

[0232] Also included herein are isolated host cells comprising an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl, an isolated nucleic acid encoding the polypeptide component of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Pseudomonas Psl, or a vector comprising a nucleic acid encoding the polypeptide component of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Pseudomonas Psl as described herein.

[0233] The present invention further includes variants of these nucleic acid sequences. For example, the variant includes a nucleotide sequence that hybridizes with the nucleic acid sequence encoding the antigen-binding domain, antigen-binding protein or bispecific molecule of the present invention under at least moderately stringent hybridization conditions.

[0234] The present invention also provides a vector into which the nucleic acid of the present invention is inserted.

[0235] Briefly, for example, an antigen-binding domain, antigen-binding protein or bispecific molecule can be expressed by inserting a natural or synthetic nucleic acid encoding the antigen-binding domain, antigen-binding protein or bispecific molecule into a suitable expression vector such that it is operably linked to 5' and 3' terminal regulatory elements including a promoter (e.g., a lymphocyte-specific promoter) and a 3' untranslated region (UTR). The vector can be suitable for replication and integration in eukaryotic host cells. Typical cloning and expression vectors include transcription and translation terminators, initiation sequences, and promoters useful for regulating the expression of the nucleic acid sequence of interest.

[0236] The nucleic acids described in the present invention can also be used for nucleic acid immunization and gene therapy by using standard gene delivery protocols. Methods of gene delivery are known in the art. See, for example, U.S. Pat. Nos. 5,399,346, 5,580,859, 5,589,466, the entire contents of which are incorporated herein by reference. In some embodiments, the present invention further provides gene therapy vectors.

[0237] The nucleic acids can be cloned into various types of vectors. For example, the nucleic acids can be cloned into vectors including, but not limited to, plasmids, phagemids, phage derivatives, animal viruses and cosmids. Particular vectors of interest include expression vectors, replication vectors, probe generation vectors and sequencing vectors.

[0238] Also, the expression vector can be provided to cells in the form of a viral vector. Viral vector technology is well known in the art, for example, Green and Sambrook (2013, Molecular Cloning: A Laboratory described in manuals, such as the Cold Spring Harbor Laboratory Manual, New York, and other virology or molecular biology handbooks. Viruses used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses. Usually, a suitable vector contains an origin of replication that functions in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers (see, for example, WO 01 / 96584; WO 01 / 29058; and U.S. Pat. No. 6,326,193).

[0239] Many virus-based systems for gene delivery into mammalian cells have already been developed. For example, retroviruses provide a convenient platform for gene delivery systems. The selected gene is inserted into a vector using techniques known in the art and packaged into retroviral particles. The recombinant virus is then isolated and delivered to the cells of a subject in vitro or in vivo. 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 means for achieving long-term gene delivery because the introduced gene allows for long-term stable integration and propagation in sub-cells. Lentiviral vectors have further advantages over retroviruses derived from tumors such as murine leukemia virus because they can transduce non-dividing cells, such as hepatocytes. At the same time, lentiviral vectors also have the further advantage of low immunogenicity.

[0240] Other promoter elements, such as enhancers, regulate the initial frequency of transcription. Usually, the promoter is located in the region 30 - 110 bp upstream of the start site, but recently, many promoters have been found to include functional elements where the start site is located downstream. Since the spacing between promoter elements is usually flexible, the function of the promoter is maintained when the positions of the elements are exchanged or moved relative to each other. In the thymidine kinase (tk) promoter, the spacing between promoter element elements can be increased up to 50 bp before the decrease in activity begins.

[0241] An example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence that can drive high-level expression of any polynucleotide sequence operably linked thereto. Other examples of suitable promoters are the elongation growth factor 1α (EF-1α) promoter. However, other constitutive promoters may also be used, including, but not limited to, the simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, avian leukosis virus promoter, Epstein - Barr virus immediate early promoter, Rous sarcoma virus promoter, and human gene promoters, including, but not limited to, the actin promoter, myosin promoter, hemoglobin promoter, and creatine kinase promoter. Also, the present invention should not be limited to the use of constitutive promoters. Inducible promoters are also contemplated as part of the present invention. By using an inducible promoter, a molecular switch is provided that can turn on the expression of an operably linked polynucleotide sequence when expression is desired and turn off the expression when expression is not desired. Inducible promoters include, but are not limited to, the metallothionein promoter, glucocorticoid promoter, progesterone promoter, and tetracycline promoter.

[0242] In some embodiments, the expression of the antigen-binding domain, antigen-binding protein or bispecific molecule is inducible. In some embodiments, the nucleic acid sequence encoding the antigen-binding domain, antigen-binding protein or bispecific molecule is operably linked to an inducible promoter comprising any of the inducible promoters described herein.

[0243] Inducible promoter By using an inducible promoter, a molecular switch is provided that can turn on the expression of an operably linked polynucleotide sequence when expression is desired and turn off the expression when expression is not desired. Exemplary inducible promoters applicable to eukaryotic cells include, but are not limited to, hormone regulatory elements (see, e.g., Mader, S. and White, J. H. (1993) Proc. Natl. Acad. Sci. USA 90:5603-5607), synthetic ligand regulatory elements (see Spencer, D. M. 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). Other exemplary inducible promoters applicable to in vivo or in vitro mammalian systems can be found in Gingrich et al. (1998) Annual Rev. Neurosci 21:377-405. In some embodiments, the inducible promoter system for expressing the antigen-binding domain, antigen-binding protein or bispecific molecule is the Tet system. In some embodiments, the inducible promoter system for expressing the antigen-binding domain, antigen-binding protein or bispecific molecule is the lac repressor system derived from E. coli.

[0244] An exemplary inducible promoter system for use in the present invention is the Tet system. This system was described by Gossen et al. (1993). In an exemplary instance, a polynucleotide of interest is controlled by a promoter containing one or more Tet operator (TetO) sites. In the non-activated state, the Tet repressor (TetR) binds to the TetO site and inhibits transcription of the promoter. In the activated state, in the presence of an inducer such as, for example, tetracycline (Tc), anhydrotetracycline, doxycycline (Dox) or an active analog thereof, the inducer causes the release of TetR from TetO, thereby enabling transcription. Doxycycline is one of the members of the tetracycline antibiotic family, and its chemical name is 1-dimethylamino-2,4a,5,7,12-pentahydroxy-11-methyl-4,6-dioxy-1,4a,11,11a,12,12a-hexahydrotetracene-3-carboxamide.

[0245] In one embodiment, TetR is codon-optimized for expression in mammalian cells such as, for example, mouse or human cells. Most amino acids are encoded by multiple codons due to the degeneracy of the genetic code, such that a given nucleic acid sequence has many variants, but the amino acid sequence encoded by the nucleic acid is not altered. However, many organisms differ in their codon usage, which is also referred to as "codon bias" (i.e., bias in the use of specific codons for a particular amino acid). Codon bias often correlates with the presence of the dominant species of tRNA for a particular codon, thereby improving the efficiency of mRNA translation. Thus, coding sequences derived from a particular organism (e.g., a prokaryote) can be adjusted through codon optimization for improved expression in a different organism (e.g., a eukaryote).

[0246] Other specific variations of the Tet system include the following "Tet-Off" and "Tet-On" systems. In the Tet-Off system, transcription is inactivated in the presence of Tc or Dox. In this system, the tetracycline-controlled transactivator protein (tTA), which is composed of TetR fused to the strong transactivation domain of VP16 from herpes simplex virus, regulates the expression of the target nucleic acid under the transcriptional control of the tetracycline-responsive promoter element (TRE). TRE is composed of a concatemer of TetO sequences fused to a promoter (usually the minimal promoter sequence derived from the immediate-early promoter of human cytomegalovirus). In the absence of Tc or Dox, tTA binds to TRE and activates the transcription of the target gene. When Tc or Do x is present, tTA cannot bind to TRE and the target gene cannot be expressed.

[0247] Conversely, in the Tet-On system, transcription becomes activated in the presence of Tc or Dox. The Tet-On system is based on the reverse tetracycline-regulated transactivator rtTA. Similar to tTA, rtTA is a fusion protein composed of the TetR repressor and the VP16 transactivation domain. However, four amino acid changes in the DNA-binding region of TetR alter the binding properties of rtTA such that it can recognize only the tetO sequence within the target-introduced gene TRE when Dox is present. Thus, in the Tet-On system, the transcription of the target gene regulated by TRE can be activated only by rtTA in the presence of Dox.

[0248] Another inducible promoter system is the lac repressor system from Escherichia 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 containing the lac operator (lacO). The Lac repressor (lacR) binds to LacO, thereby preventing transcription of the polynucleotide of interest. Expression of the polynucleotide of interest is induced by an appropriate inducer, such as isopropyl-β-D-thiogalactopyranoside (IPTG).

[0249] To assess the expression of a polypeptide or a part thereof, the expression vector to be introduced into cells may further contain a selectable marker gene and / or a reporter gene to facilitate the identification and selection of expressing cells from a cell population transfected or infected with a viral vector. On the other hand, a selectable marker may be carried on a separate DNA fragment and used in co-transfection experiments. Either the selectable marker gene or the reporter gene can be adjacent to appropriate regulatory sequences so as to be expressed in the host cell. Useful selectable markers include, for example, antibiotic resistance genes such as neo and similar genes.

[0250] Reporter genes are used for the identification of potentially transfected cells and the functional evaluation of regulatory sequences. Usually, a reporter gene is a gene that does not exist in or is not expressed by the organism or tissue of the receptor, and the reporter gene encodes a polypeptide that expresses an easily detectable property such as enzyme activity. The expression of the reporter gene is detected at an appropriate time after the DNA has been introduced into the recipient cells. Suitable reporter genes can include genes encoding luciferase, β-galactosidase, chloramphenicol acetyltransferase, secreted alkaline phosphatase, or green fluorescent protein (for example, Ui-Tel et al., 2000 FEBS Letters 479: 79-82). Suitable expression systems are known and can be prepared by known techniques or obtained commercially. Usually, a construct with the smallest 5' flanking region showing the highest expression level of the reporter gene is identified as a promoter. Such a promoter region can be linked to the reporter gene and used to evaluate the ability of several substances to regulate promoter-driven transcription.

[0251] In some embodiments, provided is a nucleic acid encoding any one of the antigen-binding domains, antigen-binding proteins, or bispecific molecules described herein. In some embodiments, the nucleic acid comprises one or more nucleic acid sequences encoding the heavy and light chains of the antigen-binding domain, antigen-binding protein, or bispecific molecule. In some embodiments, each nucleic acid sequence 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 of the nucleic acid sequences are contained in the same vector. The vector can be selected, for example, from mammalian expression vectors and viral vectors (for example, vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses).

[0252] ​Methods for introducing genes into cells and expressing them are known in the art. In the context of expression vectors, vectors can be readily introduced into host cells such as mammalian cells, bacteria, yeast, or insect cells by any method in the art. For example, expression vectors can be introduced into host cells by physical, chemical, or biological methods.

[0253] Physical methods for introducing polynucleotides into host cells include calcium phosphate precipitation, lipofection, gene bombardment, microinjection, electroporation, and the like. Methods for preparing cells containing vectors and / or exogenous nucleic acids are well known in the art. See, for example, Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). In some embodiments, introduction of the polynucleotide into the host cell is performed by calcium phosphate transfection.

[0254] Biological methods for introducing the polynucleotide of interest into host cells include methods using DNA or RNA vectors. Viral vectors, particularly retroviral vectors, are among the most widely used methods for 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. See, for example, U.S. Pat. Nos. 5,350,674 and 5,585,362.

[0255] 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 water-in-oil emulsions, micelles, mixed gel clusters, and liposomes. An exemplary colloidal system used as a delivery vector in vivo and in vitro is liposomes (e.g., artificial membrane vesicles).

[0256] When using a non-viral delivery system, an exemplary delivery vector is a liposome. It is contemplated to introduce nucleic acids into host cells (in vitro, ex vivo, or in vivo) using lipid formulations. On the other hand, the nucleic acids can bind to lipids. Nucleic acids bound to lipids can be encapsulated within the aqueous interior of liposomes, dispersed within the lipid bilayer of liposomes, connected to liposomes via linker molecules that bind to liposomes and oligonucleotides, embedded in liposomes, form complexes with liposomes, be dispersed in a lipid-containing solution, mixed with lipids, bound to lipids, suspended in lipids, be included in or mixed with micelles, or otherwise bind to lipids. Compositions related to lipids, lipid / DNA, or lipid / expression vectors are not limited to specific structures in solution. For example, they can exist in bilayer structures, micelles, or "disrupted" structures. They can also simply be dispersed in solution and may form aggregates of non-uniform size or shape. Lipids are fatty substances and can be naturally occurring or synthetic. For example, lipids contain fat droplets that naturally exist in the cytoplasm and a class of compounds including long-chain aliphatic hydrocarbons and their derivatives such as fatty acids, alcohols, amines, amino alcohols, aldehydes, etc.

[0257] Whether it is a method for introducing exogenous nucleic acids into host cells or a method for exposing cells to the inhibitors of the present application in other ways, various experiments can be conducted to confirm the presence of recombinant DNA sequences in host cells. Such experiments include, for example, "molecular biology" experiments well-known to those skilled in the art. For example, Southern and Northern blotting, RT-PCR and PCR; "biochemistry" experiments, such as detection by the presence or absence of a specific polypeptide, immunological methods (ELISA and Western blots) or identification by the experiments described herein are all included within the scope of the present invention.

[0258] Preparation of antigen-binding proteins and bispecific molecules In some embodiments, the antigen-binding protein (e.g., an antigen-binding protein that specifically recognizes PcrV or Psl) is a monoclonal antibody. In some embodiments, the antigen-binding protein or the bispecific molecule is derived from a monoclonal antibody. In some embodiments, the antigen-binding protein or the bispecific molecule comprises V H and V L , or variants thereof. In some embodiments, the antigen-binding protein or the bispecific molecule further comprises CH1 and CL domains derived from a monoclonal antibody, or variants thereof. Monoclonal antibodies can be prepared using methods known in the art, such as, for example, the hybridoma method, yeast display, phage display methods, or methods using recombinant DNA methods. Also, exemplary yeast display and phage display methods are described herein and in the examples below.

[0259] In the hybridoma method, usually, a hamster, mouse, or other suitable host animal is immunized with an immunizing agent to produce or induce lymphocytes that can produce antibodies that specifically bind to the immunizing agent. Also, lymphocytes can be immunized in vitro. The immunizing agent can include the polypeptide or fusion protein of the protein of interest. Usually, when human-derived cells are required, peripheral blood lymphocytes (PBL) are applied, and when cells derived from non-human mammals are required, spleen cells or lymph node cells are applied. The lymphocytes are fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form hybridoma cells. The immortalized cell line is usually a transformed mammalian cell, particularly a murine, bovine, and human myeloma cell. Usually, a rat or mouse myeloma cell line is applied. The hybridoma cells can be cultured in a suitable medium, and it is preferable that the medium contains one or more substances that inhibit the growth or survival of unfused immortalized cells. For example, when the parental cells lack hypoxanthine-guanine phosphoribosyltransferase (HGPRT "or" HPRT), the hybridoma cell culture medium usually contains hypoxanthine, aminopterin, and thymidine (HAT medium), and this culture medium can prevent the growth of HGPRT-deficient cells.

[0260] In some embodiments, the immortalized cell line fuses efficiently, supports stable and high-level expression of the antibody by the selected antibody-producing cells, and this cell line is sensitive to several media, such as HAT medium. In some embodiments, the immortalized cell line is a mouse myeloma cell line. For example, the mouse myeloma cell line can be obtained from the Salk Cell Collection in San Diego, California, and the American Type Culture Collection in Manassas, Virginia. Also, the use of human myelomas and mouse-human hybrid myeloma cell lines for the preparation of human monoclonal antibodies is also described.

[0261] Moreover, it is possible to measure whether monoclonal antibodies against the polypeptide are present in the medium for culturing hybridoma cells. The binding specificity of the monoclonal antibodies produced by the hybridoma cells can be determined by immunoprecipitation methods or in vitro binding experiments such as, for example, radioimmuno-precipitation assay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or analytical methods are known in the art. The binding affinity of the monoclonal antibodies can be determined, for example, by the Scatchard analysis described in Munson and Pollard, Anal. Biochem., 107:220(1980).

[0262] After the desired hybridoma cells are identified, the target clones can be subcloned by the limiting dilution method and cultured by standard methods. Suitable media for this purpose include, for example, Dulbecco's Modified Eagle Medium (DMEM) and RPMI-1640 medium. Alternatively, the hybridoma cells can grow in the form of ascites in the body of a mammal.

[0263] The monoclonal antibodies secreted by the subclones can be separated or purified from the medium or ascites by conventional immunoglobulin purification methods such as, for example, protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis or affinity chromatography.

[0264] ​In some embodiments, for any one of the antigen-binding proteins or bispecific molecules described herein, the antigen-binding protein or bispecific molecule comprises a cloned sequence selected from an antibody library (e.g., a phage library presenting scFv or Fab fragments). The clone can be identified by screening for antibody fragments having the desired activity in a combinatorial library. For example, in the art, various methods are known for generating phage display libraries and screening these libraries to obtain antibodies having the desired binding properties. Such methods are described, for example, in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O’Brien et al., ed., Human Press, Totowa, N.J., 2001). And further, for example, in 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, N.J., 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).

[0265] In certain phage display methods, V H and V LAll components of the gene are cloned by polymerase chain reaction (PCR) respectively, randomly recombined in a phage library, and then phages capable of binding to the antigen are screened. This is described, for example, in Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994). Phages usually display antibody fragments as scFv fragments or Fab fragments. Library phages derived from immunity provide high-affinity antibodies against immunogens without the need to construct hybridoma cells. Alternatively, a naive repertoire (e.g., of human origin) can be cloned to provide a single source of antibodies against multiple non-self and self antigens without any immunization. This is described, for example, in Griffiths et al., EMBO J, 12: 725-734 (1993). Finally, naive libraries can also clone unrearranged V-gene fragments from stem cells, encode the CDR3 hypervariable region using PCR primers containing random sequences, and can be prepared by a method of in vitro rearrangement. This is described, for example, in Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example, U.S. Pat. No. 5,750,373, and US Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936 and 2009 / 0002360.

[0266] The antigen-binding protein or bispecific molecule can be prepared by applying phage display to screen for antigen-binding moieties that can specifically bind to a target antigen (e.g., PcrV or Psl) in the library. The library can be a human scFv phage display library and can have at least 1×10 9(e.g., at least 1×10 9 , 2.5×10 9 , 5×10 9 , 7.5×10 9 , 1×10 10 , 2.5×10 10 , 5×10 10 , 7.5×10 10 or 1×10 11 ) and is a specific human antibody fragment having a variety of diversities. In some embodiments, the library is a human natural library constructed with DNA extracted from human PMBCs and spleens of healthy subjects and includes all human heavy and light chain subfamilies. In some embodiments, the library is a human natural library constructed with DNA extracted from PMBCs isolated from patients with various diseases, such as patients with autoimmune diseases, cancer patients, and infectious disease patients. In some embodiments, the library is a semi-synthetic human library where the heavy chain CDR3 is completely random and all amino acids (except cysteine) are present at any given position with the same probability. (See, for example, Hoet, R.M. et al., Nat. Biotechnol. 23(3):344 - 348, 2005). In some embodiments, the length of the heavy chain CDR3 of the semi-synthetic human library is from 5 to 24 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24) amino acids in length. In some embodiments, the library is a fully synthetic phage display library. In some embodiments, the library is a non-human phage display library.

[0267] Phage clones having high affinity for a target antigen (e.g., PcrV or Psl) can be screened by repeated binding of the phage to the target antigen, where the target antigen is bound to a solid support (e.g., beads used for solution panning or mammalian cells used for cell panning), and unbound phage are removed and specifically bound phage are eluted. Subsequently, the bound phage clones are eluted and used for infection of a suitable host cell, e.g., E. coli XL1-Blue, for expression and purification. Multiple rounds of panning (e.g., 2, 3, 4, 5, 6 or any number of rounds) such as solution panning, cell panning, or a combination of both can be performed to enrich phage clones that specifically bind to the target antigen. Specific binding of the enriched phage clones to the target antigen can be detected by any method known in the art, including, for example, ELISA and FACS.

[0268] Monoclonal antibodies and bispecific molecules can be prepared, for example, by recombinant DNA methods described in U.S. Patent No. 4,816,567. DNA encoding the monoclonal antibodies described in the present invention can be easily isolated and sequenced by conventional methods (e.g., oligonucleotide probes that specifically bind to genes encoding the light and heavy chains of a mouse antibody). Hybridoma cells or antigen-specific phage clones of the present invention as described above can be used as a source of this DNA. After isolation, the DNA is inserted into an expression vector, and the vector is transfected into, for example, Simian COS cells, Chinese hamster ovary cancer (CHO) cells, or immunoglobulin Transfect host cells such as myeloma cells that do not produce phosphorus to obtain monoclonal antibodies synthesized within the recombinant host cells. The DNA may be modified, for example, by replacing the human heavy and light chain constant regions with the coding sequences, and / or replacing homologous non-human sequences with the framework regions (U.S. Patent No. 4,816,567; Morrison et al., supra), or covalently linking all or part of the coding sequence of the non-immunoglobulin polypeptide to the coding sequence of the immunoglobulin. This non-immunoglobulin polypeptide can replace the constant domain of the antibody of the present invention, or replace the antigen-binding site within the variable domain of the antibody of the present invention to form a chimeric bivalent antibody. In some embodiments, variable domains targeting different epitopes or antigens can be added to generate chimeric bispecific antibodies.

[0269] The antigen-binding protein can be a monovalent antibody. Methods for preparing monovalent antibodies are known in the art. For example, it is an expression method related to the recombination of immunoglobulin light chains and modified heavy chains. Usually, in order to prevent the heavy chains from cross-linking with each other, the heavy chains are cut short at any position in the Fc region. Alternatively, in order to prevent cross-linking, the relevant cysteine residues are replaced or deleted with other amino acid residues.

[0270] In vitro methods are also applicable to the preparation of monovalent antibodies. Digesting the antibody to generate antibody fragments, particularly Fab fragments, can be achieved by any method known in the art.

[0271] Antibody variable domains having the desired binding specificity (antibody-antigen binding site) can be fused to immunoglobulin constant domains. Preferably, they are fused to an immunoglobulin heavy chain constant region comprising at least a portion of the hinge, CH2, and CH3 regions. In some embodiments, a first heavy chain constant region (CH1) containing the sites necessary for light chain binding is present in at least one of the fusions. DNA encoding the immunoglobulin heavy chain fusions and optionally the immunoglobulin light chain is inserted into separate expression vectors and co-transfected into a suitable host organism. In some embodiments, antibody variable domains targeting different epitopes or different antigens can be fused to the immunoglobulin constant domain sequence to generate chimeric bispecific molecules.

[0272] Fully human and humanized antibodies The antigen-binding protein or bispecific molecule may comprise a humanized antibody portion or a fully human antibody portion. Humanized forms of non-human (such as mouse) antibody portions are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (e.g., Fv, Fab, Fab’, F(ab’)2, scFv, or other antigen-binding subsequences of an antibody), and typically contain a minimal sequence derived from a non-human immunoglobulin. A humanized antibody comprises a human immunoglobulin, immunoglobulin chain, or fragment thereof (the recipient antibody), where residues from the recipient CDRs are replaced by CDR residues of a non-human (donor antibody) having the desired specificity, affinity, and performance, such as a mouse, rat, or rabbit CDR. In some embodiments, residues in the human immunoglobulin Fv framework region are replaced by the corresponding non-human residues. A humanized antibody may further contain amino acid residues that do not belong to the recipient antibody and are not present in the introduced CDR or framework region sequences. Typically, a humanized antibody contains at least one, usually two, variable domains, in which 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.

[0273] Typically, a humanized antibody or humanized antibody portion contains one or more amino acid residues introduced from a non-human source. These non-human amino acid residues are typically referred to as "import" residues and typically are derived from the "import" variable domain. In some embodiments, humanization is performed substantially according to the following method of Winter and his 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)), replacing the corresponding sequences of the humanized antibody with rodent CDRs or CDR sequences. Thus, such "humanized" antibody portions (U.S. Patent No. 4,816,567) are less complete than substantially complete humanized antibodies, and their variable domains have been replaced with the corresponding sequences already derived from non-humanization. In practice, a humanized antibody portion is typically a human antibody portion in which some CDR residues and possibly some framework region residues have been replaced by residues from the analogous sites of the rodent antibody.

[0274] The production of human antibody portions is one alternative form of humanization. For example, currently, it is possible to generate normal genetically modified animals (e.g., mice) that can produce a complete fully human antibody library without producing endogenous immunoglobulins after immunization. For example, in chimeric and germline mutant mice, homozygous deletion of the antibody heavy chain joining region (JH) gene has been reported to completely inhibit the production of endogenous antibodies. Transferring the human germline immunoglobulin gene array into such germline mutant mice allows for the production of fully human antibodies under antigen stimulation. For example, Jakobovits et al., PNAS See USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immunol., 7:33 (1993); U.S. Patent Nos. 5,545,806, 5,569,825, 5,591,669, 5,545,807; and WO97 / 17852. Alternatively, fully human antibodies can be prepared by introducing human immunoglobulin loci into genetically modified animals (e.g., mice in which the endogenous immunoglobulin genes have been partially or completely silenced). Upon antigen stimulation, it has been found that the production of fully human antibodies is very similar to that in humans in all aspects, including gene rearrangement, assembly, and antibody libraries. This method is described, for example, in U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 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).

[0275] Human antibodies or human antibody portions can also be produced by activating B cells in vitro (see U.S. 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 such as those of Cole et al. and Boerner et al. are also used in the preparation of 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).

[0276] An anti-PcrV antibody, an anti-Psl antibody, or a variant of a bispecific antibody that specifically recognizes Pseudomonas PcrV and Pseudomonas Psl In some embodiments, the amino acid sequences of variants of the antigen-binding domains or antigen-binding proteins provided herein (e.g., antibody portions that specifically recognize PcrV or Psl, or bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl) are also included. For example, it may be desirable to improve the binding affinity and / or other biological activities of the antigen-binding domain or antigen-binding protein. The amino acid sequences of variants of the antigen-binding entity can be prepared by introducing appropriate modifications into the nucleotide sequences encoding the antigen-binding entity, 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 antigen-binding entity. The final construct can be completed by any combination of deletions, insertions, and substitutions of amino acid residues so as to have the desired properties such as antigen-binding.

[0277] In some embodiments, an antigen-binding domain or an antigen-binding protein variant having one or more amino acid substitutions is provided. The targeted sites of the substituted mutations include hypervariable regions (HVRs) and framework regions (FRs). Amino acid substitutions can be introduced into the target antibody, and products with a desired activity (e.g., improved biological activity, maintained / improved antigen-binding ability, reduced immunogenicity, reduced binding of pathogens to epithelial cells, or improved opsonophagocytic killing (OPK) of pathogenic bacteria such as Pseudomonas aeruginosa) can be screened. In some embodiments, the amino acid substitutions described herein are limited to the "exemplary substitutions" in Table 7 of the present invention. In some embodiments, the amino acid substitutions are limited to the "preferred substitutions" in Table 7 of the present invention.

[0278] Conservative substitutions are shown in Table 7 below.

[0279]

Table 7

[0280] a. Hydrophobic amino acids: Norleucine, Methionine (Met), Alanine (Ala), Valine (Val), Leucine (Leu), Isoleucine (Ile); b. Neutral hydrophilic amino acids: Cysteine (Cys), Serine (Ser), Threonine (Thr), Asparagine (Asn), Glutamine (Gln); c. Acidic amino acids: Aspartic acid (Asp), Glutamic acid (Glu); d. Basic amino acids: Histidine (His), Lysine (Lys), Arginine (Arg); e. Amino acids that affect the direction of the chain: Glycine (Gly), Proline (Pro); f. Aromatic amino acids: Tryptophan (Trp), Tyrosine (Tyr), Phenylalanine (Phe).

[0281] Substitutions of non-conserved amino acids include substitutions from one such type to another.

[0282] Exemplary substitution variants are, for example, affinity matured antibodies produced by affinity maturation techniques based on phage display. Briefly, one or more CDR residues are mutated, the variant antibody moiety is displayed on a phage, and variants having a particular biological activity (e.g., based on an RBC cell lysis inhibition assay or binding affinity) are screened. Alterations (e.g., substitutions) can be made in the HVRs region to obtain improved biological activity based on the RBC cell lysis inhibition assay or binding affinity. Such alterations are made in the "hot spot regions" of the HVR, i.e., residues encoded by codons that have undergone high frequency mutation during somatic maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and / or specificity determining residues (SDRs), and the binding affinities of the resulting variant V and V H and V L are measured. Affinity maturation by construction and rescreening of secondary libraries is described in several references. See, e.g., Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)).

[0283] In some examples of affinity maturation, diversity is introduced into the variable genes used for affinity maturation by any of a variety of methods (e.g., Error Prone PCR, chain shuffling or oligonucleotide-directed mutagenesis). Then, a secondary library is created. The library is screened to identify variants of the antibody with the desired affinity. Another method of introducing diversity involves the intervening of HVRs, where some HVR residues (e.g., 4 to 6 residues at a time) are randomized. The HVR residues involved in antigen binding are specifically identified and mutagenized or modeled, for example, by alanine scanning. Usually, the CDR-H3 and CDR-L3 regions are particularly important targets.

[0284] In some embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, provided that such changes do not substantially reduce the antibody's ability to bind to the antigen. For example, conservative changes (e.g., conservative substitutions as referred to herein) that do not substantially reduce the binding affinity are generated in the HVRs. These changes may occur outside the "hot spot regions" or SDR regions of the HVRs. In some embodiments, the variant V H and V L each HVR of the sequence does not change or contains one, two, or three or fewer amino acid substitutions.

[0285] As described in Cunningham and Wells (1989) Science, 244:1081-1085, a useful method for identifying amino acid residues or regions that can be targeted for mutagenesis from an antibody is called "alanine scanning mutagenesis." In this method, one or a group of target residues (e.g., charged residues such as arginine, aspartic acid, histidine, lysine, and glutamic acid) are substituted with neutral or negatively charged amino acids (e.g., alanine or glutamic acid) to determine whether the interaction between the antibody and the antigen is affected. Further substitutions can be introduced at the amino acid position to demonstrate that the position is functionally sensitive to the first substitution. Alternatively / Additionally, the crystal structure of the antigen-antibody complex can be used to identify the contact sites between the antibody and the antigen. Residues located at these contact sites and adjacent residues can be targeted or deleted as substitution candidates. Mutants are screened to determine whether they have the desired properties.

[0286] Insertion of an amino acid sequence includes fusing a polypeptide having a length of from 1 residue to over 100 residues at the amino terminus and / or carboxyl terminus, and further includes inserting one or more amino acid residues into the sequence. An example of a terminal insertion includes an antigen-binding portion having a methionyl residue at the N terminus. Insertion mutants of other antigen-binding portions include fusing an enzyme (e.g., ADEPT) at the N terminus or C terminus of the antigen-binding portion, or a polypeptide that increases the serum half-life of the antigen-binding portion.

[0287] Fc region mutants In some embodiments, one or more amino acid modifications are made to the antigen-binding portions described herein (e.g., full-length anti-PcrV antibodies, anti-Psl antibodies, Pseudomonas PcrV and It is introduced into the Fc region of a bispecific antibody that specifically recognizes Psl, an anti-PcrV antibody, an anti-Psl antibody, or a fusion protein of a bispecific molecule that specifically recognizes Pseudomonas PcrV and Psl, thereby generating an Fc region variant. In some embodiments, the Fc region variant has an enhanced ADCC effector function and is typically associated with receptors (FcRs) that bind to Fc. In some embodiments, the Fc region variant has a reduced ADCC effector function. There are many examples of changes or mutations in the Fc sequence that affect its effector function. For example, in WO00 / 42072 and Shields et al. J Biol. Chem. 9(2):6591-6604 (2001), variants of antibodies related to enhancing or reducing binding to FcRs were described. The content of this publication is incorporated herein by reference.

[0288] Antibody-dependent cell-mediated cytotoxicity (ADCC) is a mechanism of action of therapeutic antibodies against tumor cells. ADCC is a cell-mediated immune defense. When an antigen on the surface of a target cell membrane is bound by a specific antigen-binding moiety (e.g., an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl), effector cells of the immune system spontaneously lyse the target cell (e.g., an infected cell). Usually, the ADCC effect involves NK cells activated by the antibody. NK cells express the Fc receptor CD16. This receptor recognizes and binds to the Fc portion of the antibody molecule that binds to the surface of the target cell. The most common Fc receptor on the surface of NK cells is CD16 or FcγRIII. The binding of the Fc receptor to the Fc region of the antibody causes activation of NK cells, release of cell-lysing particles, and subsequent apoptosis of the target cell.

[0289] In some embodiments, the invention also provides a variant of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a variant of a full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl), wherein the variant comprises an Fc region having one or more effector functions, thereby making it an ideal candidate antibody. Although the half-life of an anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl in vivo is important, certain effector functions (such as CDC and ADCC) may not be necessary or may be harmful. By performing in vitro and / or in vivo cytotoxicity tests, a decrease / elimination of CDC and / or ADCC activity can be confirmed. For example, an Fc receptor (FcR) binding assay can be performed to confirm that the antibody lacks FcγR binding (and thus may lack ADCC activity) but retains FcRn binding ability. Among the main cells involved in ADCC, NK cells express only FcγRIII, and monocytes express FcγRI, FcγRII, and FcγRIII. Table 3 on page 464 of Ravetch and Kinet Annu. Rev. Immunol. 9:457-492 (1991) summarizes FcR expression in hematopoietic cells. Non-limiting examples of evaluating the ADCC activity of a target molecule in vitro are described in U.S. Pat. No. 5,500,362 (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); see U.S. Pat. No. 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Also, a non-radioactive detection method may be used (e.g., ACTI TMFlow cytometry non-radioactive cytotoxicity assay (CellTechnology, Inc. Mountain View, Calif.) and CYTOTOX 96 TM Non-radioactive cytotoxicity assays (Promega, Madison, Wis.). Effector cells used in these experiments were peripheral blood mononuclear cells (PBMCs) and naive cells. These include NK and NK cells. Also, the ADCC activity of the molecule of interest is tested in vivo, for example in an animal model, as described in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). At the same time, a C1q binding test can be carried out to confirm that the antibody cannot bind to C1q and lacks CDC activity. See, for example, the C1q and C3c binding ELISA in WO2006 / 029879 and WO2005 / 100402. To assess complement activation status, CDC measurements are performed (see, e.g., 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 measured using methods known in the art (see, e.g., Petkova, S. (See B. et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

[0290] Antibodies with reduced effector function include those with one or more residue substitutions at residue positions 238, 265, 269, 270, 297, 327 and 329 of the Fc region (US Pat. No. 6,737,056). These Fc variants include Fc variants with two or more residue substitutions at positions 265, 269, 270, 297 and 327, including an Fc variant designated "DANA" in which residues 265 and 297 are substituted with alanine (US Pat. No. 7,332,581).

[0291] Variants of antibodies related to an improvement or reduction in the ability to bind to FcRs have already been described (see, for example, U.S. Pat. No. 6,737,056; WO2004 / 056312 and Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001)).

[0292] In some embodiments, the modification of the Fc region results in a modification (i.e., improvement or reduction) of opsonization, see those described in Moore et al., MAbs. 2(2):181-189 (2010).

[0293] In some embodiments, provided are variants of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl), which variants include an Fc region variant having one or more amino acid substitutions and can extend the half-life or enhance the binding to the Fc receptor (FcRn). Antibodies that can extend the half-life or improve FcRn binding are described in US2005 / 0014934A1 (Hinton et al.). These antibody Fc regions have one or more amino acids substituted to enhance the binding between the Fc region and FcRn. These Fc variants include those in which residues at positions 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434 in the Fc region are substituted, for example, those in which the residue at position 434 of the Fc region is substituted (U.S. Pat. No. 7,371,826).

[0294] Also, see examples of other mutant Fc regions described in Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821 and WO94 / 29351.

[0295] The present invention relates to an anti- PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) is also included.

[0296] glycosylation variant In some embodiments, the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl provided herein (e.g., a full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) is modified to increase or decrease the degree of glycosylation of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl. One or more glycosylation sites are conveniently added or removed by changing the amino acid sequence of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl or its polypeptide portion to add or delete a glycosylation site on the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl.

[0297] Here, when the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl contains an Fc region, the carbohydrate linked thereto can be changed. Natural antibodies produced by mammalian cells usually contain branched biantennary oligosaccharides, which are usually linked to Asn297 in the CH2 domain of the Fc region through an N-linkage. See, for example, Wright et al., TIBTECH 15:26-32 (1997). The oligosaccharide can contain, for example, mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as various sugars such as trehalose linked to GlcNAc in the "stem" portion of the biantennary oligosaccharide structure. In some embodiments, the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody of the present invention that specifically recognizes Pseudomonas PcrV and Psl is oligosaccharide-modified to generate mutants of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl having certain improved properties.

[0298] 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. Usually, in human serum, a very small number of naturally occurring non-fucosylated IgGs are detected. N-glycosylation of the Fc region is important for the binding of the Fc region to FcγR, but non-fucosylated N-glycans enhance the binding ability of Fc to FcγRIIIa. Enhancement of the binding ability to FcγRIIIa enhances the ADCC effect, which is advantageous in certain antibody therapy applications that require cytotoxicity.

[0299] In some embodiments, enhanced effector function can be detrimental when Fc-mediated cytotoxicity is not required. In some embodiments, the Fc fragment or CH2 domain is non-glycosylated. In some embodiments, the N-glycosylation site in the CH2 domain is mutated to prevent its glycosylation.

[0300] In some embodiments, an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl Provided are variants of bispecific antibodies) that contain an Fc region, where the carbohydrate structure linked to the Fc region has reduced fucose or lacks fucose, which may enhance the function of ADCC. Specifically, this specification provides an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl, which has reduced fucose compared to the same anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl produced by wild-type CHO cells. That is, they are characterized by having a lower amount of fucose compared to antibodies produced by natural CHO cells (e.g., CHO cells that produce natural glycosylation forms, CHO cells containing the natural FUT8 gene). In some embodiments, the N-linked glycans of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl have less than 50%, 40%, 30%, 20%, 10%, or 5% fucose. For example, the fucose content of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl can be 1%-80%, 1%-65%, 5%-65%, or 20%-40%. In some embodiments, the N-linked glycans of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl do not contain fucose, i.e., the bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl does not completely contain fucose or has no fucose or is defucosylated. The fucose content is determined by calculating the average content of fucose in the sugar chain linked to Asn297 relative to the total amount of all sugar structures (e.g., complex, hybrid, or mannose structures) linked to Asn297, measured by MALDI-TOF mass spectrometry as described in WO2008 / 077546. Asn297 is the asparagine residue at position 297 of the Fc region (EU Fc region residue numbering system).However, due to minor changes in the antibody sequence, Asn297 may also be located approximately ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300. These fucosylation variants may have enhanced ADCC function. See, for example, US Patent Publication Nos. US2003 / 0157108 (Presta, L.), US2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to variants of "defucosylated" or "fucose-deficient" antibodies include US2003 / 0157108; WO2000 / 61739; WO2001 / 29246; US2003 / 0115614; US2002 / 0164328; US2004 / 0093621; US2004 / 0132140; US2004 / 0110704; US2004 / 0110282; US2004 / 0109865; WO2003 / 085119; WO2003 / 084570; WO2005 / 035586; WO2005 / 035778; WO2005 / 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 function (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US Pat Appl No US2003 / 0157108A1, Presta, L; and WO2004 / 056312A1, Adams et al., particularly Example 11), and knockout cell lines such as CHO cells in which the α-1,6-fucosyltransferase gene, the FUT8 gene, has been knocked out (Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107).

[0301] Variants of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) further provide a bisected oligosaccharide. For example, a bisected oligosaccharide linked to the Fc region of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl is evenly divided by GlcNAc. Such variants of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) may have reduced fucosylation and / or enhanced ADCC function. Examples of such antibody variants are described in WO2003 / 011878 (Jean-Mairet et al.); U.S. Pat. No. 6,602,684 (Umana et al.); US2005 / 0123546 (Umana et al.), and Ferrara et al., Biotechnology and Bioengineering, 93(5):851-861 (2006). The present invention also provides variants of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) that have at least one galactose residue in the oligosaccharide bound to the Fc region. Such variants of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl may have enhanced CDC function. Such variants are described, for example, in WO1997 / 30087 (Patel et al.); WO1998 / 58964 (Raju, S.); and WO1999 / 22764 (Raju, S.).

[0302] In some embodiments, variants of said anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) comprise an Fc region capable of binding to FcγRIII. In some embodiments, variants of said anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl and comprises an Fc region (e.g., full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) exhibit ADCC activity in the presence of human effector cells (e.g., T cells), or have enhanced ADCC activity in the presence of human effector cells as compared to other same anti-PcrV antibodies, anti-Psl antibodies, or bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl and have a human wild-type IgG1 Fc region (e.g., full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl).

[0303] Cysteine-engineered variant In some embodiments, the preparation of an anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl in which cysteine is engineered (e.g., full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl) is required, and in said antibody, one or more amino acid residues are substituted with cysteine residues. In some embodiments, the substituted residues appear at accessible sites of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl. By substituting these residues with cysteine, the active sulfhydryl groups are located at accessible sites of the anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl, and said anti-Pc Coupling of an rV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl with other moieties, such as a drug moiety or a linker-drug moiety, is used to prepare an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody immunocomplex that specifically recognizes Pseudomonas PcrV and Psl as further described herein. Anti-PcrV antibodies, anti-Psl antibodies, or bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl in which cysteine has been engineered (e.g., full-length anti-PcrV antibodies, anti-Psl antibodies, or bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl) can be prepared, for example, as described in U.S.Pat.No.7,521,541.

[0304] Derivative In some embodiments, bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl provided herein (e.g., full-length bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl) are known in the art and can be further modified to include other non-protein moieties that are readily available. Moieties suitable for derivatizing bispecific antibodies that specifically recognize Pseudomonas PcrV and Psl include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers), dextran or poly(n-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymers, propylene oxide / ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerin), polyvinyl alcohol, and mixtures thereof, but are not limited thereto. Polyethylene glycol propionaldehyde has advantages in manufacture because it is stable in water. The polymer can be of any molecular weight and can be branched or unbranched. The number of polymers that bind to the bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl may vary, and if multiple polymers are bound, they may be the same molecule or different molecules. Generally, the amount and / or type of polymer used for derivatization is determined by factors including, but not limited to, the property or function to be improved of the bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl, whether the bispecific antibody derivative that specifically recognizes Pseudomonas PcrV and Psl is to be used for treatment under specific conditions, etc.

[0305] Pharmaceutical composition This specification further provides a composition (e.g., a pharmaceutical composition, also referred to herein as a formulation) containing any one of an anti-PcrV antibody, an anti-Psl antibody, or a bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl (e.g., a full-length anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl), a nucleic acid encoding the antibody, a vector containing the nucleic acid encoding the antibody, or a host cell containing the nucleic acid or vector described herein. In some embodiments, a pharmaceutical composition containing any one of an anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl described herein, and a pharmaceutically acceptable vector is provided.

[0306] Mixing an anti-PcrV antibody, anti-Psl antibody, or bispecific antibody that specifically recognizes Pseudomonas PcrV and Psl of the required purity with any pharmaceutically acceptable vector, excipient, or stabilizer (Remington’s Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) results in an appropriate anti-PcrV antibody, anti-Psl antibody, or Pseudomonas P Obtain a formulation of a bispecific antibody that specifically recognizes crV and Psl, and prepare it in the form of a lyophilized formulation or a liquid formulation. Acceptable vectors, excipients, or stabilizers are non-toxic to the subject at the amounts and concentrations used, and these include buffers such as phosphates, citrates, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethylammonium chloride; benzalkonium chloride; benzethonium chloride; phenol; butanol or benzyl alcohol; alkyl p-hydroxybenzoates such as methyl p-hydroxybenzoate or propyl p-hydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); polypeptides of low molecular weight (less than about 10 residues); proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; saccharides such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., zinc-protein complexes); and / or nonionic surfactants such as TWEEN TM , PLURONICS TM or nonionic surfactants such as polyethylene glycol (PEG); exemplary formulations are described in WO98 / 56418, the contents of which are hereby expressly incorporated ...

Claims

1. A bispecific molecule, wherein the bispecific molecule comprises a first antigen-binding domain that specifically recognizes Pseudomonas PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas Psl, The first antigen-binding domain comprises a heavy-chain variable domain V Ｈ and a light-chain variable domain V Ｌ and the V Ｈ comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 6, and the V Ｌ comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 12, and, the second antigen-binding domain is, (i) V Ｈ and V Ｌ and the V Ｈ comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 48, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 52, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 57, and the V Ｌ comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 65, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 70, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 74, or, (ii) V Ｈ and V Ｌ and the V Ｈ comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 48, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 53, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 60, and the V Ｌ comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 65, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 70, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 74, or, (iii) V Ｈ and V Ｌ and the V Ｈ comprises an HC-CDR1 having an amino acid sequence of SEQ ID NO: 48, an HC-CDR2 having an amino acid sequence of SEQ ID NO: 52, and an HC-CDR3 having an amino acid sequence of SEQ ID NO: 59, Ｌ comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:65, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:70, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74; or (iv) V Ｈ and V Ｌ and V Ｈ comprises an HC-CDR1 having an amino acid sequence of SEQ ID NO: 48, an HC-CDR2 having an amino acid sequence of SEQ ID NO: 53, and an HC-CDR3 having an amino acid sequence of SEQ ID NO: 58, Ｌ LC-CDR1 comprises the amino acid sequence SEQ ID NO:66, and LC-CDR2 comprises the amino acid sequence SEQ ID NO:

70. and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:74; or (v) V Ｈ and V Ｌ and V Ｈ comprises an HC-CDR1 having an amino acid sequence of SEQ ID NO: 49, an HC-CDR2 having an amino acid sequence of SEQ ID NO: 54, and an HC-CDR3 having an amino acid sequence of SEQ ID NO: 61, Ｌ comprises an LC-CDR1 comprising the amino acid sequence SEQ ID NO:67, an LC-CDR2 comprising the amino acid sequence SEQ ID NO:71, and an LC-CDR3 comprising the amino acid sequence SEQ ID NO:75; or (vi) V Ｈ and V Ｌ and V Ｈ comprises an HC-CDR1 having an amino acid sequence of SEQ ID NO: 50, an HC-CDR2 having an amino acid sequence of SEQ ID NO: 55, and an HC-CDR3 having an amino acid sequence of SEQ ID NO: 62, Ｌ a LC-CDR1 comprising the amino acid sequence SEQ ID NO:68, a LC-CDR2 comprising the amino acid sequence SEQ ID NO:72, and a LC-CDR3 comprising the amino acid sequence SEQ ID NO:

76.

2. A bispecific molecule comprising a first antigen-binding domain that specifically recognizes Pseudomonas aeruginosa PcrV and a second antigen-binding domain that specifically recognizes Pseudomonas aeruginosa Psl, wherein the first antigen-binding domain (i) comprises V Ｈ and V Ｌ and the V Ｈ comprises the amino acid sequence of SEQ ID NO: 91 or 161, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91 or 161, and the V Ｌ comprises the amino acid sequence of SEQ ID NO: 112 or 182, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112 or 182, and, wherein the second antigen-binding domain (i) comprises V Ｈ and V Ｌ and the V Ｈ comprises the amino acid sequence of SEQ ID NO: 103 or 173, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 103 or 173, and the V Ｌ comprises the amino acid sequence of SEQ ID NO: 117 or 187, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117 or 187, or (ii) comprises V Ｈ and V Ｌ and the V Ｈ comprises the amino acid sequence of SEQ ID NO: 106 or 176, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 106 or 176, and the V Ｌ comprises the amino acid sequence of SEQ ID NO: 117 or 187, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117 or 187, or (iii) comprises V Ｈ and V Ｌ and the V Ｈcomprises the amino acid sequence of SEQ ID NO: 105 or 175, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 105 or 175, wherein said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117 or 187, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117 or 187, or (iv) V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 104 or 174, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 104 or 174, wherein said V Ｌ comprises the amino acid sequence of SEQ ID NO: 118 or 188, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 118 or 188, or (v) V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 107 or 177, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 107 or 177, wherein said V Ｌ comprises the amino acid sequence of SEQ ID NO: 119 or 189, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 119 or 189, or (vi) V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 108 or 178, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 108 or 178, wherein said V Ｌ comprises the amino acid sequence of SEQ ID NO: 119 is 189, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 119 or 189, or (vii) V Ｈ and V Ｌ comprising said V Ｈcomprises the amino acid sequence of SEQ ID NO: 109 or 179, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 109 or 179, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 120 or 190, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 120 or 190, the bispecific molecule according to claim 1. **Claim 3** a) the first antigen-binding domain is a Fab arm that specifically recognizes PcrV, the second antigen-binding domain is a single-chain variable region fragment (scFv) that specifically recognizes Psl, and the bispecific molecule further comprises an Fc region comprising CH2 and CH3 domains, the single-chain variable region fragment (scFv) is interconnected with the Fab arm via a first polypeptide linker (L1) and with the Fc region via a second polypeptide linker (L2), and the bispecific molecule is bivalent for binding to each of PcrV and Psl, or b) the first antigen-binding domain is a single-chain variable region fragment (scFv) that specifically recognizes PcrV, the second antigen-binding domain is a Fab arm that specifically recognizes Psl, and the bispecific molecule further comprises an Fc region comprising CH2 and CH3 domains, the single-chain variable region fragment (scFv) is interconnected with the Fab arm via a first polypeptide linker (L1) and with the Fc region via a second polypeptide linker (L2), and the bispecific molecule is bivalent for binding to each of PcrV and Psl, the bispecific molecule according to claim 1 or 2. **Claim 4** The bispecific molecule is a) the amino acid sequence of SEQ ID NO: 135, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 135, and The bispecific molecule according to claim 1 or 2, comprising an amino acid sequence of any one of SEQ ID NOs: 195-204, or a mutant sequence having at least 90% sequence identity with an amino acid sequence of any one of SEQ ID NOs: 195-204.

5. The bispecific molecule according to claim 3, wherein the Fc region is selected from the group consisting of the Fc regions of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE and IgD.

6. The bispecific molecule according to claim 3, wherein the Fc region comprises a variable Fc region.

7. The bispecific molecule according to claim 3, wherein the Fc region is glycosylated or the Fc region is deglycosylated.

8. The bispecific molecule according to claim 3, wherein the Fc region has reduced fucosylation or is afucosylated.

9. The bispecific molecule according to claim 6, wherein the variable Fc region comprises substitutions at one or more sites among sites 239, 282, 289, 297, 312, 324, 330, 335, 337, 339, 356, 359, 361, 383, 384, 398, 400, 440, 422 and 442 according to the EU index number.

10. The bispecific molecule according to claim 9, wherein the variable Fc region comprises a substitution at site 297.

11. The bispecific molecule according to claim 10, wherein the substitution at site 297 is 297Q.

12. The bispecific molecule is a) a chimeric heavy chain comprising an amino acid sequence of any one of SEQ ID NOs: 137-149, or a mutant sequence having at least 90% sequence identity with an amino acid sequence of any one of SEQ ID NOs: 137-149, and The bispecific molecule according to claim 1 or 2, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 135, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO:

135.

13. (i) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas aeruginosa PcrV, and (ii) an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas aeruginosa PcrV and / or an antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl, or (i) an antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl, and (ii) an antigen-binding protein that specifically recognizes a first epitope on Pseudomonas aeruginosa PcrV and / or an antigen-binding protein that specifically recognizes a second epitope on Pseudomonas aeruginosa PcrV, a pharmaceutical composition comprising The antigen-binding protein that specifically recognizes the first epitope or the second epitope on Pseudomonas aeruginosa PcrV is (i) V Ｈ and V Ｌ comprising, said V Ｈ comprises an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 2, 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: 7, said V Ｌ comprises an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 13, or (ii) V Ｈ and V Ｌ comprising, said V Ｈ 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: 4, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 6, said V Ｌ comprises an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12, or (iii) V Ｈ and V Ｌ comprising, said V Ｈ includes an HC - CDR1 having the amino acid sequence of SEQ ID NO: 22, an HC - CDR2 having the amino acid sequence of SEQ ID NO: 23, and an HC - CDR3 having the amino acid sequence of SEQ ID NO: 26, said V Ｌ includes an LC - CDR1 having the amino acid sequence of SEQ ID NO: 30, an LC - CDR2 having the amino acid sequence of SEQ ID NO: 33, and an LC - CDR3 having the amino acid sequence of SEQ ID NO: 35, or (iv) V Ｈ and V Ｌ comprising, said V Ｈ includes an HC - CDR1 having the amino acid sequence of SEQ ID NO: 22, an HC - CDR2 having the amino acid sequence of SEQ ID NO: 24, and an HC - CDR3 having the amino acid sequence of SEQ ID NO: 27, said V Ｌ includes an LC - CDR1 having the amino acid sequence of SEQ ID NO: 31, an LC - CDR2 having the amino acid sequence of SEQ ID NO: 34, and an LC - CDR3 having the amino acid sequence of SEQ ID NO: 36, and, said antigen - binding protein that specifically recognizes Pseudomonas Psl is (i) V Ｈ and V Ｌ comprising, said V Ｈ includes an HC - CDR1 having the amino acid sequence of SEQ ID NO: 48, an HC - CDR2 having the amino acid sequence of SEQ ID NO: 52, and an HC - CDR3 having the amino acid sequence of SEQ ID NO: 57, said V Ｌ includes an LC - CDR1 having the amino acid sequence of SEQ ID NO: 65, an LC - CDR2 having the amino acid sequence of SEQ ID NO: 70, and an LC - CDR3 having the amino acid sequence of SEQ ID NO: 74, or (ii) V Ｈ and V Ｌ comprising, said V Ｈ includes an HC - CDR1 having the amino acid sequence of SEQ ID NO: 48, an HC - CDR2 having the amino acid sequence of SEQ ID NO: 53, and an amino acid sequence of SEQ ID NO: 60 for HC - CDR3, said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 65, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 70, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 74, or (iii) V Ｈ and V Ｌ and said V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 49, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 54, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 61, and said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 67, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 71, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 75, or (iv) V Ｈ and V Ｌ and said V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 51, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 56, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 64, and said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 69, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 73, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 80, a pharmaceutical composition.

14. (a) The antigen - binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and said V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 1, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 4, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 6, and said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 12, and the antigen - binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ comprising the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 48, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 52, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 57, and the V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 65, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 70, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 74, or (b) the antigen - binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ comprising the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 1, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 4, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 6, and the V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 12, and the antigen - binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ comprising the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 49, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 54, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 61, and the V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 67, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 71, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 75, or (c) the antigen - binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ comprising the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 2, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 5, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 7, and the V Ｌ comprises an LC-CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 containing the amino acid sequence of SEQ ID NO: 13, and the antigen-binding protein that specifically recognizes Psl of the genus Pseudomonas is V Ｈ and V Ｌ and the V Ｈ comprises an HC-CDR1 containing the amino acid sequence of SEQ ID NO: 48, an HC-CDR2 containing the amino acid sequence of SEQ ID NO: 52, and an HC-CDR3 containing the amino acid sequence of SEQ ID NO: 57, and the V Ｌ comprises an LC-CDR1 containing the amino acid sequence of SEQ ID NO: 65, an LC-CDR2 containing the amino acid sequence of SEQ ID NO: 70, and an LC-CDR3 containing the amino acid sequence of SEQ ID NO: 74, or (d) the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and the V Ｈ comprises an HC-CDR1 containing the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 containing the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 containing the amino acid sequence of SEQ ID NO: 6, and the V Ｌ comprises an LC-CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 containing the amino acid sequence of SEQ ID NO: 12, and, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and the V Ｈ comprises an HC-CDR1 containing the amino acid sequence of SEQ ID NO: 22, an HC-CDR2 containing the amino acid sequence of SEQ ID NO: 24, and an HC-CDR3 containing the amino acid sequence of SEQ ID NO: 27, and the V Ｌ comprises an LC-CDR1 containing the amino acid sequence of SEQ ID NO: 31, an LC-CDR2 containing the amino acid sequence of SEQ ID NO: 34, and an LC-CDR3 containing the amino acid sequence of SEQ ID NO: 36, and The antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl is V Ｈ and V Ｌ and includes, the V Ｈ includes an HC-CDR1 having the amino acid sequence of SEQ ID NO: 48, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 52, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 57, and the V Ｌ includes an LC-CDR1 having the amino acid sequence of SEQ ID NO: 65, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 70, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 74, or (e) The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV is V Ｈ and V Ｌ and includes, the V Ｈ includes an HC-CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 4, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 6, and the V Ｌ includes an LC-CDR1 having the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 12, and The antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl is V Ｈ and V Ｌ and includes, the V Ｈ includes an HC-CDR1 having the amino acid sequence of SEQ ID NO: 51, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 56, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 64, and the V Ｌ includes an LC-CDR1 having the amino acid sequence of SEQ ID NO: 69, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 73, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 80, or (f) The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV is V Ｈ and V Ｌ and includes, the V Ｈcomprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 1, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 4, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 6, wherein said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 12, and said antigen - binding protein that specifically recognizes Psl of the genus Pseudomonas is V Ｈ and V Ｌ and comprises, wherein said V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 48, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 53, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 60, wherein said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 65, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 70, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 74, or (g) said antigen - binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and comprises, wherein said V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 1, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 4, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 6, wherein said V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 12, and said antigen - binding protein that specifically recognizes said second epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and comprises, wherein said V Ｈ comprises an H C - CDR1 containing the amino acid sequence of SEQ ID NO: 22, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 24, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 27, wherein said V Ｌcomprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 31, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 34, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 36, and, the antigen - binding protein that specifically recognizes Pseudomonas sp. Psl is V Ｈ and V Ｌ and the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 48, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 53, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 60, and the V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 65, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 70, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 74, or, (h) the antigen - binding protein that specifically recognizes the first epitope on Pseudomonas sp. PcrV is V Ｈ and V Ｌ and the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 2, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 5, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 7, and the V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 10, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 11, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 13, and, the antigen - binding protein that specifically recognizes the second epitope on Pseudomonas sp. PcrV is V Ｈ and V Ｌ and the V Ｈ comprises an HC - CDR1 containing the amino acid sequence of SEQ ID NO: 22, an HC - CDR2 containing the amino acid sequence of SEQ ID NO: 23, and an HC - CDR3 containing the amino acid sequence of SEQ ID NO: 26, and the V Ｌ comprises an LC - CDR1 containing the amino acid sequence of SEQ ID NO: 30, an LC - CDR2 containing the amino acid sequence of SEQ ID NO: 33, and an LC - CDR3 containing the amino acid sequence of SEQ ID NO: 35, or, (i) The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV is V Ｈ and V Ｌ and includes, wherein the V Ｈ includes an HC-CDR1 having the amino acid sequence of SEQ ID NO: 2, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 5, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 7, and the V Ｌ includes an LC-CDR1 having the amino acid sequence of SEQ ID NO: 10, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 11, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 13, and the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas aeruginosa PcrV is V Ｈ and V Ｌ and includes, wherein the V Ｈ includes an HC-CDR1 having the amino acid sequence of SEQ ID NO: 22, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 23, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 26, and the V Ｌ includes an LC-CDR1 having the amino acid sequence of SEQ ID NO: 30, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 33, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 35, and, the antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl is V Ｈ and V Ｌ and includes, wherein the V Ｈ includes an HC-CDR1 having the amino acid sequence of SEQ ID NO: 48, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 52, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 57, and the V Ｌ includes an LC-CDR1 having the amino acid sequence of SEQ ID NO: 65, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 70, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 74, The pharmaceutical composition according to claim 13.

15. The antigen-binding protein that specifically recognizes the first epitope or the second epitope on Pseudomonas aeruginosa PcrV is (i) V Ｈ and V Ｌ comprising V Ｈ comprises the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, or (ii) V Ｈ and V Ｌ comprising V Ｈ comprises the amino acid sequence of SEQ ID NO: 92, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 92 and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 113, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 113, or (iii) V Ｈ and V Ｌ comprising V Ｈ comprises the amino acid sequence of SEQ ID NO: 95, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 95, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 114, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 114, or (iv) V Ｈ and V Ｌ comprising V Ｈ comprises the amino acid sequence of SEQ ID NO: 96, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 96, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 115, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 115, and, the antigen-binding protein that specifically recognizes Pseudomonas Psl is (i) V Ｈ and V Ｌcomprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 103, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 103, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or (ii) V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 106, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 106, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or (iii) V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 107, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 107, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 119, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 119, or (iv) V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 111, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 111, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 124, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 124, The pharmaceutical composition according to claim 13.

16. (i) The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV is V Ｈ and V Ｌcomprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and said antigen-binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 103, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 103, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or (ii) said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 92, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 92, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 113, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 113, and said antigen-binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ comprising said V Ｈ comprises the amino acid sequence of SEQ ID NO: 103, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 103, said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or (iii) said antigen-binding protein that specifically recognizes said first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ comprises, wherein said V Ｈ comprises the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and said antigen-binding protein that specifically recognizes Pseudomonas sp. Psl is V Ｈ and V Ｌ comprises, wherein said V Ｈ comprises the amino acid sequence of SEQ ID NO: 107, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 107, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 119, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 119, or (iv) said antigen-binding protein that specifically recognizes the first epitope on Pseudomonas sp. PcrV is V Ｈ and V Ｌ comprises, wherein said V Ｈ comprises the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and, said antigen-binding protein that specifically recognizes the second epitope on Pseudomonas sp. PcrV is V Ｈ and V Ｌ comprises, wherein said V Ｈ comprises the amino acid sequence of SEQ ID NO: 96, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 96, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 115, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 115, and The antigen-binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ and includes, the V Ｈ includes the amino acid sequence of SEQ ID NO: 103, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 103, and the V Ｌ includes the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or (v) The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and includes, the V Ｈ includes the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, and the V Ｌ includes the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and The antigen-binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ and includes, the V Ｈ includes the amino acid sequence of SEQ ID NO: 111, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 111, and the V Ｌ includes the amino acid sequence of SEQ ID NO: 124, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 124, or (vi) The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and includes, the V Ｈ includes the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, and the V Ｌcomprises the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ and comprises, the V Ｈ is the amino acid sequence of SEQ ID NO: 106, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 106, the V comprises the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or Ｌ (vii) the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V and V Ｈ and comprises, the V Ｌ is the amino acid sequence of SEQ ID NO: 91, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 91, the V Ｈ comprises the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and Ｌ (ix) the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V and V Ｈ and comprises, the V Ｌ is the amino acid sequence of SEQ ID NO: 96, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 96, the V Ｈ comprises the amino acid sequence of SEQ ID NO: 115, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 115, and Ｌ (x) the antigen-binding protein that specifically recognizes Pseudomonas Psl is V and V Ｈ and comprises, the V Ｌ is the amino acid sequence of SEQ ID NO: 112, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 112, and Ｈcomprises the amino acid sequence of SEQ ID NO: 106, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 106, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, or (viii) the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and said V Ｈ comprises the amino acid sequence of SEQ ID NO: 92, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 92, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 113, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 113, and, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and said V Ｈ comprises the amino acid sequence of SEQ ID NO: 95, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 95, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 114, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 114, or (ix) the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is V Ｈ and V Ｌ and said V Ｈ comprises the amino acid sequence of SEQ ID NO: 92, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 92, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 113, or a variant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 113, and, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is V Ｈ and V Ｌ comprises, wherein said V Ｈ comprises the amino acid sequence of SEQ ID NO: 95, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 95, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 114, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 114, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is V Ｈ and V Ｌ comprises, wherein said V Ｈ comprises the amino acid sequence of SEQ ID NO: 103, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 103, and said V Ｌ comprises the amino acid sequence of SEQ ID NO: 117, or a mutant sequence having at least 90% sequence identity with the amino acid sequence of SEQ ID NO: 117, the pharmaceutical composition according to any one of claims 13 to 15.

17. (i) the molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:5, or (ii) the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:5, or (iii) the molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV, and the antigen-binding protein that specifically recognizes Pseudomonas Psl is 1:1:1 or 1:1:2, the pharmaceutical composition according to any one of claims 13 to 16. ​

18. (i) The molar ratio of the antigen-binding protein that specifically recognizes Pseudomonas Psl to the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV is 2:1 or 1:1, or (ii) The molar ratio of the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV to the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV is 2:1 or 1:

1. The pharmaceutical composition according to claim 17.

19. A pharmaceutical composition comprising the bispecific molecule according to any one of claims 1 to 12.

20. A pharmaceutical composition according to any one of claims 13 to 19 for treating and / or preventing a disease or medical condition, wherein the disease or medical condition is a pathogenic infection, the infection is a Gram-negative bacterial infection, and the Gram-negative bacterium is Pseudomonas aeruginosa, and / or wherein the disease or medical condition comprises one or more of the symptoms caused by Pseudomonas aeruginosa infection.

21. The symptoms include one or more of fever, chills, fatigue, muscle and joint pain, joint swelling, headache, diarrhea, skin rash, wound pus, bacteremia, acute pneumonia, intra-abdominal infection, respiratory tract infection, septic shock, suppurative arthritis, enteritis, skin and soft tissue infection, urinary tract infection, intestinal infection, ulcerative keratitis, chronic suppurative otitis media, mastoiditis, sinusitis, and endocarditis. The pharmaceutical composition according to claim 20.

22. The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas PcrV and the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas PcrV and / or the antigen-binding protein that specifically recognizes Pseudomonas Psl are administered simultaneously, or The antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV, the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas aeruginosa PcrV, and / or the antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl are administered sequentially, or, The antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV, and / or the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas aeruginosa PcrV are administered simultaneously, or, The antigen-binding protein that specifically recognizes Pseudomonas aeruginosa Psl, the antigen-binding protein that specifically recognizes the first epitope on Pseudomonas aeruginosa PcrV, and / or the antigen-binding protein that specifically recognizes the second epitope on Pseudomonas aeruginosa PcrV are administered sequentially, the pharmaceutical composition according to claim 13.

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