Antibodies against multi-drug resistant klebsiella pneumoniae

Abstract

The present invention relates to a monoclonal antibody or an antigen-binding fragment thereof having potent antibacterial activity against klebsiella pneumoniae, in particular against multi-drug resistant klebsiella pneumoniae strains. The invention also relates to application of the monoclonal antibody or the antigen binding fragment thereof in treatment, prevention and diagnosis of Klebsiella pneumoniae infection, especially Klebsiella pneumoniae related diseases.

Description

TECHNICAL FIELD

[0001] The present invention relates to monoclonal antibodies or antigen-binding portions thereof having potent bactericidal activity against multi-drug resistant Klebsiella pneumoniae, in particular against Klebsiella pneumoniae strains producing New Delhi Metallo-β-lactamases (NDM). The present invention further relates to the use of such monoclonal antibodies or antigen-binding portions thereof in the treatment, prevention and diagnosis of Klebsiella pneumoniae infections, in particular Klebsiella pneumoniae dependent diseases. BACKGROUND

[0002] Antimicrobial resistance (AMR) has been ranked by the World Health Organization and the European Medicines Agency as one of the top ten global health priorities due to its impact on global human health and socio-economic well-being. Among human pathogens, an increasing number of pathogens have developed resistance to antibiotics (this phenomenon is known as multi-drug resistance (MDR)), and Klebsiella pneumoniae (herein also referred to as “Kp”) is undoubtedly the most common MDR pathogen. Kp can cause both nosocomial infections (i.e. urinary tract infections, pneumonia, wound and surgical site infections, sepsis) and invasive community-acquired diseases such as pyogenic liver abscess, endophthalmitis and meningitis. Treatment is often limited due to the increasing frequency of emergence of carbapenemase-producing MDR Kp isolates, carbapenemases being a class of enzymes that can lead to resistance to carbapenems, a subclass of β-lactam antibiotics. Among carbapenemases, New Delhi Metallo-β-lactamases (NDMs) pose the greatest threat, as this class of enzymes also leads to resistance to novel β-lactamase inhibitor combination formulations (i.e. ceftazidime-avibactam, imipenem-relebactam, and meropenem-vaborbactam), which are considered to be the last line of defense. NDMs are encoded by the blaNDMgene located on large multi-drug resistant plasmids that have spread widely among high-risk MDR Kp strains on all continents. NDM

[0003] ​NDM-producing strains are of concern because their emergence is associated with specific genetic features that endow them with high virulence traits, increasing the probability of Kp spread. It is worth noting that since 2018, Tuscany, Italy, has been experiencing a nosocomial epidemic caused by NDM-1 -positive ST147 clonal lineages. Overall, ST147 is widespread in India and Southeast Asia and has recently been classified as a pan-drug resistant strain. Genomic surveillance data show that ST147 isolates from Tuscany carry highly diverse plasmid content, encoding both MDR and high virulence genes. Similar genetic rearrangements have also been observed in Russia (2017), the United Kingdom (2018-2019), and Egypt (2019), where different Kp ST (ST15, ST147, ST395, and ST874) carrying hybrid plasmids containing both virulence genes and bla NDM genes have been reported. Similar scenarios have also been observed in the 2019 German epidemic, where a hybrid element was detected in a hypervirulent ST307 isolate carrying bla NDM-1 / bla OXA-48 . Thus, in these epidemic clonal lineages, there is a convergence of genetic elements conferring MDR and high virulence traits, a worrying evolutionary development that not only highlights the possibility of similar clonal lineages triggering larger epidemics, but also urgently calls for alternative therapeutic strategies.

[0004] Human monoclonal antibodies (mAbs) can represent a new powerful tool that can be rapidly translated into innovative prevention or treatment strategies against AMR. The uniqueness of mAbs is that they can target a virtually unlimited repertoire of antigens specific to a given pathogen; they are inherently safe and avoid damaging the host microbiota.

[0005] However, there is an urgent need for potent, broad-spectrum antibody therapies for the treatment, prevention, and diagnosis of Kp infections, particularly MDR Kp-related diseases. SUMMARY

[0006] To identify potent human mAbs against Kp, particularly against multi-drug resistant strains of Kp, the inventors developed a powerful unbiased mAb selection method that does not bind to specific predetermined antigens, allowed the isolation of human mAbs from patients who experienced Kp blood infections during the 2018 Tuscany epidemic, and performed a functional screening.

[0007] This strategy, detailed in the examples, allowed the identification of Kp-specific mAbs with potent in vitro bactericidal activity in the picomolar range, with the best drug candidates having a protective effect against multidrug-resistant Kp in an in vivo bacteremia model.

[0008] Notably, as will be clear from the examples, the authors of the present application found that the selected ST147 NDM-1 Capsular mAbs were able to promote bacterial uptake in phagocytosis assays and to trigger enchained bacterial growth, which correlated with their ability to protect against bloodstream infection in vivo.

[0009] In certain aspects, the present application thus provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of Kp.

[0010] In certain aspects, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of a drug-resistant or multidrug-resistant strain of Kp, in particular a Kp strain producing New Delhi metallo-beta-lactamase (NDM).

[0011] In certain aspects, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of at least one K. pneumoniae capsular type K64 strain.

[0012] In certain aspects, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of K. pneumoniae sequence type 147 (ST147). In certain aspects, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a capsular antigen (also referred to herein as K antigen) or O antigen of at least one Kp strain.

[0013] In certain aspects, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a capsular antigen of at least one of the following strains and is bactericidal against it: a NDM-1 positive K. pneumoniae sequence type 147 (ST147) strain and a NDM-9 positive K. pneumoniae sequence type 147 (ST147) strain.

[0014] In certain aspects, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a capsular antigen of a NDM-1 positive K. pneumoniae sequence type 147 (ST147) strain and is bactericidal against it.

[0015] In certain aspects, the application provides a human monoclonal antibody or antigen binding portion thereof that specifically binds to a surface antigen of Klebsiella pneumoniae, comprising a light chain variable domain (VL) and a heavy chain variable domain (VH) of a monoclonal antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13.

[0016] In certain aspects, the application provides a human monoclonal antibody or antigen binding portion thereof that specifically binds to a region of a surface antigen of Klebsiella pneumoniae, comprising a CDR of a monoclonal antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13.

[0017] In certain aspects, the present application provides a human monoclonal antibody or antigen binding portion thereof, which specifically binds to a region of a surface antigen of Kp, comprising VL and VH domains that are at least 85%, 90%, 95%, 97%, 98%, or 99% identical in amino acid sequence to the VL and VH domains, respectively, of a monoclonal antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13.

[0018] In certain aspects, the present application provides a human monoclonal antibody or antigen binding portion thereof, which competes with any of the antibodies disclosed herein for a surface antigen of Kp.

[0019] In certain aspects, the present application provides a human monoclonal antibody or antigen binding portion according to any of the embodiments disclosed herein for use in the prophylactic or therapeutic treatment of an infection with Kp, preferably an infection with the Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, or a condition or disease caused by such an infection.

[0020] In certain aspects, the present application provides a human monoclonal antibody or antigen binding portion according to any of the embodiments disclosed herein for use in the prophylactic or therapeutic treatment of an infection with Kp, preferably an infection with the Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, or a condition or disease caused by such an infection.

[0021] In certain aspects, the present application provides a method of preventing or treating an infection with Kp, or a condition or disease caused by such an infection, comprising administering to a subject in need thereof a human monoclonal antibody or antigen binding portion according to any of the embodiments disclosed herein.

[0022] The present application further provides a human monoclonal antibody or antigen binding portion according to any of the embodiments disclosed herein for use in the diagnosis, prevention and / or treatment of a subject suffering from or at risk of developing an infection with Kp, preferably an infection with the Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein. Furthermore, the present application relates to the use of the human binding molecules and / or nucleic acid molecules of the present application in the diagnosis / detection of such an infection.

[0023] In certain aspects, the present application provides a pharmaceutical composition comprising at least one or more human monoclonal antibodies or antigen binding portions thereof according to any of the embodiments disclosed herein and a pharmaceutically acceptable carrier, and use thereof in the prevention and / or treatment of Kp infection, preferably of infection by the K. pneumoniae capsular type K64 strain according to any of the variants disclosed herein, or a condition or disease caused by such infection.

[0024] In certain aspects, the present application provides an isolated cell line producing an antibody or antigen binding portion thereof according to any of the embodiments disclosed herein.

[0025] In certain aspects, the present application provides an isolated nucleic acid molecule comprising a nucleotide sequence encoding an antibody or antigen binding portion thereof according to any of the embodiments disclosed herein.

[0026] In certain aspects, the present application provides a vector comprising a nucleic acid molecule encoding an antibody or antigen binding portion thereof according to any of the embodiments disclosed herein, wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule.

[0027] In certain aspects, the present application provides a non-human transgenic animal or transgenic plant comprising a nucleic acid according to any of the preceding embodiments, wherein the non-human transgenic animal or transgenic plant expresses the nucleic acid. In certain embodiments, the non-human transgenic animal is a mammal.

[0028] In certain aspects, the present application provides the use of a human monoclonal antibody or antigen binding portion thereof according to any of the embodiments disclosed herein for the diagnosis of Kp infection.

[0029] In certain aspects, the present application provides an in vitro method for revealing the presence of K. pneumoniae in a sample, comprising the steps of:

[0030] i) contacting an antibody or antigen binding portion thereof according to any of the embodiments disclosed herein;

[0031] ii) detecting the binding of said antibody or antigen binding portion thereof to a surface antigen of Kp, preferably to a K. pneumoniae capsular antigen, more preferably to a K64 type capsular antigen.

[0032] In certain aspects, the present application provides an in vitro method for diagnosing Kp infection in a subject, comprising the steps of:

[0033] i) contacting an antibody or antigen binding portion thereof according to any of the embodiments disclosed herein with a biological sample of said subject;

[0034] ii) detecting binding of said antibody, or antigen binding portion thereof, to a surface antigen of Kp, preferably detecting binding of said antibody, or antigen binding portion thereof, to a Klebsiella pneumoniae capsular antigen, more preferably a K64 type capsular antigen.

[0035] The present application contemplates any combination of the foregoing aspects and embodiments of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 . Against ST147 NDM-1 Isolation and selection of Kp-specific mAbs of the Toscana outbreak strain. A) Scheme of the anti-Kp antibody isolation procedure. B) ELISA screening of 18'390 single cell sorted MBC supernatants isolated from 7 convalescent patients. This assay detected IgG / IgA against ST147 NDM-1 c.i.1 and ST147 NDM-1 c.i.2 surface antigens. The signal of 214 supernatants was at least 2-fold higher than the blank OD 405 and were selected for further analysis. C) L-SBA of recombinant mAbs against ST147 NDM-1 c.i.1. 20 bactericidal mAbs were selected with a 30% reduction of bacterial viability as cut-off.

[0037] Figure 2 . Binding properties of the 20 bactericidal ST147 NDM-1 specific mAbs. A) Heatmap representing high-throughput flow cytometry screening of 20 functional mAbs against a genetically diverse panel of Kp. MFI has been normalized against the control and converted into log scale. B) ELISA heatmap of purified K- (capsular) and O- antigens. ELISA plates were coated with 100 μg / ml of different Kp-specific O-antigens (i.e. LPS O2a, internally purified O2a from ST147 NDM-1 LPS O2afg, internally purified O1v2 (v2: O2afg) from ST147 OXA-48 LPS O2afg, internally purified O1v2 (v2: O2afg) from ST147 NDM-1 LPS O2afg, internally purified O1v2 (v2: O2afg) from ST147 405A value at least 3 times higher is considered positive. CE) shows representative immunoblot patterns of selected anti-Kp mAbs detected by total sugar extracts of the strains shown. Results 08O09 (Figure C) show diffuse high-molecular-weight bands in the presence of total sugar extracts of ST147 Kp, indicating recognition of the KL64 capsule. Results 05N02 (Figure D) show broad ladder patterns for a wide range of Kp STs carrying both O2 and O1 antigens. Results 05D08 (Figure E) show medium-molecular-weight ladder signals for Kp strains carrying O2.

[0038] Figure 3 Anti-Kp mAb and ST147 NDM1 Microscopic characterization of strain binding. A) Image shows ST147 expressing sfmCherry stained with 08O09, 05D08, and 05N02 mAb (secondary antibody labeled with anti-human A488 conjugated, green). NDM1 B) Bacterial DNA was stained with dapi (blue). Scale bar 2 mm. B) Scatter plot shows the quantification of A488 intensity at the single bacterial level (n = ... - ... analytes). C) Scatter plot shows the area measurement of a single A488 point. D) Representative image showing ST147 stained with mAbs conjugated with 08O09-A488, 05N02-A555, and 05D08-A647. NDM1 Scale bar 2mm.

[0039] Figure 4 20 targeted ST147 NDM-1 In vitro functional characterization of bactericidal mAbs. A) The heatmap shows the ICs obtained from F-SBA screening of 20 anti-Kp mAbs against the indicated bacterial strain group. 50 A) Bacterial uptake fold of THP1 macrophages measured in a phagocytosis assay with anti-Kp mAb from cluster 1. B) Bacterial uptake fold of THP1 cells measured in a sample treated with anti-Kp mAb from cluster 2. C) The figure shows a time-lapse still frame of bacteria expressing sfmCherry grown in 100 mg / mL of the mAb shown. Scale bar 5 mm.

[0040] Figure 5 In immune-normal ST147 NDM-1Evaluation of 08O09, 05D08 and 05N02 mAbs for in vivo protective properties in bacteremia model. A) Scheme of the prophylactic (PRO) study. Survival analysis of 08O09 (B), 05N02 (C) and 05D08 (D) PRO schemes and sham-operated control group (n=10 mice per group). (E) Survival analysis of 08O09 PRO scheme at 5 mg / kg single dose (n=10 mice per group). (F) Scheme of the therapeutic (THR) study. (G) Survival analysis of 08O09 THR scheme and sham-operated control group (n=10 mice per group). (H) Scheme of the prophylactic plus therapeutic (PRO+THR). (I) Survival analysis of 08O09 PRO scheme and PRO+THR scheme at 1 mg / kg and 5 mg / kg single dose (n=10 animals per group).

[0041] Figure 6 . Gating strategy for total memory B cell single cell sorting. Flow cytometry plots report the gating strategy to identify the following cells: live cells, lymphocyte population, single cells, CD19 positive cells (B cells), CD19 + CD27 + IgD- cells (memory B cells), CD19 + CD27 + IgD - IgM - B cells (memory B cells expressing IgG, IgA or IgE).

[0042] Figure 7 . 134 recombinant mAbs against ST147 NDM-1 ELISA results heat map of clinical isolates. ELISA was performed against two different ST147 NDM-1 clinical isolates to confirm the binding of mAbs to Kp surface. Values at least 3-fold higher than the blank OD 405 were considered as positive results.

[0043] Figure 8 . Heat map showing the summary of the high-throughput flow cytometry screening of 20 functional anti-Kp mAbs against a selected group of different Klebsiella species and commensal bacteria. MFI was normalized to the control and converted to log scale.

[0044] Figure 9 . Purification of capsular KL64. A) SEC-HPLC profile of purified KL64 capsule from ST147 NDM-1 . Three arrows indicate the 410, 80 and 12 kDa molecular weight standards. B) Theoretical structure of capsular KL64. NDM-1 Capsular polysaccharides have been purified as reported in the Materials and Methods section.

[0045] Figure 10 Purification of different O antigen subtypes. SEC-HPLC profile of purified O antigens, (A) from ST147 NDM-1 O2v1 version; and (B) from ST13- OXA48 Olv2 version. Three arrows indicate the 410, 80 and 12 kDa molecular weight standards. C) Molecular structure of O antigens of O1, O2a and O2a fg types.

[0046] Figure 11 O antigen-deficient strain ST147 NDM-9 Polysaccharide characterization. A) SEC-HPLC analysis of total sugar content after acetic acid hydrolysis. Three peaks correspond to the capsule, O antigen and core. Three arrows indicate the 410, 80 and 12 kDa molecular weight standards. B) Silver staining analysis of total sugar extracts from Kp strains used in this study. ST147 NDM-9 Lacks LPS ladder signal (25-50 kDa range).

[0047] Figure 12 ROI definition and spot detection for mAb binding characterization. Left panel shows the binding pattern of A488-labeled anti-Kp mAbs on ST147 NDM-1 Kp, middle panel shows the ROIs where A488 signal was detected, right panel shows the morphology of A488 spots. Scale bar is 2 pm.

[0048] Figure 13. Sequence analysis of 20 selected mAbs. A) IGHV and IGHJ gene usage frequency (left), IGKV and IGKJ gene usage frequency (right). B) IGHV and IGHJ gene pairing heat map (left), IGKV and IGKJ gene pairing heat map (right). C) IGHV and IGKV gene pairing heat map (left), IGHJ and IGKJ gene pairing heat map (right). D) CDR3 length distribution, heavy chain (left) and light chain (right). E) Percentage of variable chain identity relative to inferred germline.

[0049] Figure 14 F-SBA analysis of 20 functional mAbs against complement-sensitive pathogenic Kp strains. 20 bactericidal anti-Kp mAbs against ST147 NDM-1 c.i.1 (A), ST147 NDM-1 c.i.2 (B), ST147 NDM-9 (C) and ST307 NDM-5(D) F-SBA. Individual experiments were normalized relative to the negative control (no mAb). For each mAb, bactericidal curves and corresponding IC50s were obtained using [Inhibitor] vs. normalized response, variable slope analysis on GraphPad Prism. 50 Value report at Figure 4 In A.

[0050] Figure 15 Complement-dependent killing efficacy of 20 functional mAbs against four pathogenic Kp strains. Thermographs showing the percentage reduction in rezamidium fluorescence were used as readings of bacterial viability and normalized relative to the control group (no mAb). Results were extrapolated from F-SBA of individual mAbs.

[0051] Figure 16 In immune-normal ST147 NDM-1 Evaluation of the in vivo protective properties of 08O09, 05D08, and 05N02 mAbs in a bacteremia model. (A) Survival analysis of mice infected with ST147 isolate 30 under four test inoculums (logarithmic value). 10 (CFU / mL). (B) Compared with the control group, the 08O09 prevention (PRO) regimen resulted in a lower CFU / mL ratio per spleen at the endpoint. 10 CFU. (C) Compared with the control group, the 08O09 treatment (THR) regimen resulted in a lower rate of CFU per spleen at the endpoint. 10 CFU. (D) Comparison of 96-hour survival time in mice infected and treated with a single dose of mAb 08O09 at 1 mg / kg (E) and 5 mg / kg (PRO+THR). Each curve tested 10 animals.

[0052] Figure 17 Pharmacokinetic experiments in the THR and PRO studies. The concentration of human mAb in mouse plasma was measured by quantitative ELISA, administered via single IV or IP dose of 5 mg / kg. Absorbance was measured at 405 nm for 20 minutes, and the values ​​were extrapolated using GraphPad in the presence of mouse plasma based on an S-shaped curve. Detailed Implementation

[0053] Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Generally, nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art.

[0054] The methods and techniques of the present application are generally performed according to conventional methods well-known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. See e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989); and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates (1992); and Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1990), all of which are incorporated herein by reference.

[0055] The following terms, unless otherwise indicated, shall have the following meanings:

[0056] The term "polypeptide" encompasses proteins, protein fragments, and polypeptide analogs of protein sequences, either naturally occurring or artificially synthesized. A polypeptide can be monomeric or polymeric. The term "isolated protein," "isolated polypeptide," or "isolated antibody" refers to a protein, polypeptide, or antibody that (1) is free from the natural associating components that accompany the protein in its native state, (2) is free from other proteins with which it is associated in its natural state, (3) is expressed by a cell in a different species, or (4) does not occur in nature. Thus, a polypeptide that is chemically synthesized or synthesized in a cell system other than the cell from which it naturally originates will be "isolated" from its natural associating components. An isolated protein can also be produced by isolation with the use of protein purification techniques well known in the art, such that the protein is substantially free of naturally-occurring components. Examples of isolated antibodies include anti-Kp antibodies that have been affinity purified using Kp whole bacteria or portions thereof, particularly surface antigens, anti-Kp antibodies synthesized in vitro by hybridomas or other cell lines, and human anti-Kp antibodies derived from transgenic animals. A protein or polypeptide is "substantially pure," "substantially homogeneous," or "substantially purified" when at least about 60% to 75% of the sample presents a single polypeptide. A polypeptide or protein can be monomeric or polymeric. A substantially pure polypeptide or protein typically comprises at least about 50%, 60%, 70%, 80%, or 90% (by weight) of the protein sample, more typically about 95%, and preferably more than 99% pure. Protein purity or homogeneity can be indicated by a number of methods well known in the art, such as by polyacrylamide gel electrophoresis using a dye that is well known in the art to visualize the individual polypeptide bands. For even greater resolution, HPLC or other well-known purification methods can be used to provide even greater resolution for certain purposes. The term "polypeptide fragment" as used herein refers to a polypeptide that has an amino-terminal and / or carboxy-terminal deletion, but otherwise has the same amino acid sequence as the corresponding position in the native sequence. In some embodiments, the fragment is at least 5, 6, 8, or 10 amino acids in length. In other embodiments, the fragment is at least 14, at least 20, at least 50, or at least 70, 80, 90, 100, 150, or 200 amino acids in length.

[0057] The term "polypeptide analog" as used herein refers to a polypeptide comprising a segment that is substantially identical to a portion of an amino acid sequence and which segment has at least one of the following properties: (1) is capable of specifically binding to a Kp surface antigen under suitable binding conditions according to any of the embodiments disclosed herein; (2) has bactericidal activity against Kp. Typically, the polypeptide analog comprises conservative amino acid substitutions (or insertions or deletions) relative to the native sequence. The analog is typically at least 20 or 25 amino acids long, preferably at least 50, 60, 70, 80, 90, 100, 150, or 200 amino acids long or longer, and can be as long as the full-length polypeptide. Some embodiments of the application include polypeptide fragment or polypeptide analog antibodies having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 substitutions between the germline amino acid sequence. In certain embodiments, the amino acid substitutions of the anti-Kp antibody, or antigen binding portion thereof, are amino acid substitutions that: (1) decrease susceptibility to proteolysis; (2) decrease susceptibility to oxidation; (3) alter binding affinity for forming protein complexes; and (4) impart or modify other physicochemical or functional properties of such analogs, but still retain specific binding to Kp surface antigens. The analogs can include a variety of muteins, whose sequences differ from the normal peptide sequence. For example, single or multiple amino acid substitutions, preferably conservative amino acid substitutions, can be made in the normally occurring sequence, preferably in the portion of the polypeptide outside of the one or more domains that form intermolecular contacts. Conservative amino acid substitutions should not significantly alter the structural characteristics of the parent sequence, e.g., the replacement amino acid should not alter the reverse-parallel beta sheet that constitutes the immunoglobulin binding domain of the parent sequence, nor should it disrupt other types of secondary structure unique to the parent sequence. In general, glycine and proline are not used in reverse-parallel beta sheets. Examples of polypeptide secondary and tertiary structures that are well-recognized in the art are described in Proteins, Structures and Molecular Principles (Creighton, ed., W. H. Freeman and Company, New York (1984)); Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); and Thornton et al., Nature 354:105 (1991), which are incorporated herein by reference.

[0058] The term "Klebsiella pneumoniae", also abbreviated "Kp" in the instant specification, as used herein, refers to a gram-negative, non-motile, encapsulated, lactose-fermenting, facultative anaerobic bacillus. It is mucoid on MacConkey agar and ferments lactose. In a clinical setting, it is the most important member of the genus Klebsiella in the family Enterobacteriaceae. In recent years, Klebsiella has emerged as an important pathogen of nosocomial infections.

[0059] Kp is present in the normal flora of the mouth, skin, and intestinal tract, but if inhaled, it causes destructive changes to the lungs, particularly the alveoli, of humans and animals, resulting in sputum that is blood, brown, or yellowish jelly. Notably, Kp can cause both nosocomial infections (i.e., urinary tract infections, pneumonia, wound and surgical site infections, septicemia) and invasive community-acquired diseases, such as pyogenic liver abscesses, endophthalmitis, and meningitis.

[0060] Kp also occurs naturally in soil, and about 30% of strains are capable of nitrogen fixation under anaerobic conditions. As an independent living nitrogen-fixing bacterium, its nitrogen fixation system has been extensively studied and has agricultural application value, as Kp has been shown to be able to increase crop yield under agricultural conditions.

[0061] When reference is made herein to "antibodies" associated with the present application, it is understood that antigen-binding portions thereof can also be used. Antigen-binding portions compete with full antibody for specific binding. See Fundamental Immunology, Chapter 7 (Paul, W., ed., 2nded. Raven Press, N.Y. (1989)) (incorporated by reference in its entirety for all purposes). Antigen-binding portions can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies.

[0062] In some embodiments, antigen-binding portions include Fab, Fab', F(ab')2, Fd, Fv, dAb, and complementarity determining region (CDR) fragments, single-chain antibodies (scFv), chimeric antibodies, diabodies, nanobodies, and any polypeptide that contains at least a portion of an antibody that is sufficient to confer specific antigen-binding to the polypeptide.

[0063] From N- to C-terminus, both mature light and heavy chain variable domains comprise FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 regions. The assignment of amino acids in each domain herein follows the definition of the IMGT convention described in Lefranc et al. (2003) Developmental & Comparative Immunology 27.1 (2003): 55-77.

[0064] In the present document, the antibodies referred to by number are identical to the monoclonal antibodies obtained from human peripheral blood mononuclear cells (PBMCs) isolated from the same number donor. For example, the monoclonal antibody SBJ08-O09 (also referred to herein as J08O09) is identical to the antibody obtained from PBMCs isolated from the subject identified by code 08 or a subclone thereof. In the present document, an antibody or antigen-binding portion thereof that specifically binds to a Kp surface antigen is also referred to as an anti-Kp antibody or antigen-binding portion thereof, in accordance with any of the embodiments disclosed in the specification and claims.

[0065] As used herein, an Fd fragment refers to an antibody fragment consisting of the VH and CHI domains; an Fv fragment consists of the VL and VH domains of a single arm of an antibody; and a dAb fragment (Ward et al., Nature 341 :544-546 (1989)) consists of a VH domain.

[0066] In some embodiments, the antibody is a single chain antibody (scFv), in which the VL and VH domains are paired by a synthetic linker that enables them to be made as a single protein chain. (Bird et al., Science 242:423-426 (1988) and Huston et al., Proc. Natl Acad. Sci. USA 85:5879-5883 (1988)). In some embodiments, the antibody is a diabody, i.e., a bivalent antibody in which the VH and VL domains are expressed on a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain, thereby forcing the domains to pair with the complementary domains of another chain and creating two antigen binding sites. (See, e.g., Holliger P. et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993), and Poljak R.J. et al., Structure 2: 1121-1123 (1994)). In such embodiments, one or more CDRs can be incorporated as part of a larger polypeptide chain, can be covalently linked to another polypeptide chain, or can be incorporated non-covalently. In embodiments having one or more binding sites, the binding sites can be identical or different from one another.

[0067] The term "human antibody", as used herein, refers to any antibody whose variable and constant domain sequences are of human sequence, or whose variable domain sequence's any CDRs are of human sequence. The term encompasses antibodies whose sequences are derived from human genes but have been altered, e.g., to reduce possible immunogenicity, to increase affinity, to eliminate a cysteine that can lead to undesirable folding, etc. The term encompasses such antibodies that are recombinantly produced in non-human cells, which can produce atypical glycosylation in human cells. The term "chimeric antibody", as used herein, refers to an antibody that comprises regions from two or more different antibodies.

[0068] The term "epitope" includes any protein determinant capable of specific binding to an immunoglobulin or T-cell receptor, or to other molecules, such as glycosylation of molecules. Epitopes or antigenic determinants are generally formed both by chemically active surface groups of molecules, such as amino acids or carbohydrate or sugar side chains, and by regions of specific three-dimensional structural characterization, as well as specific charge characteristics. Epitopes can be "linear" or "conformational". In a linear epitope, all of the interaction points between the protein and the interacting molecule (e.g., an antibody) occur linearly along the primary amino acid sequence of the protein. In a conformational epitope, the interaction points are between amino acid residues that are separated from one another on the protein.

[0069] An antibody having "bactericidal activity" or "antibacterial activity", as used herein, refers to an antibody that exhibits the ability to destroy and / or inhibit the action of its target, i.e., one or more Kp strains according to any embodiment disclosed herein, or that exhibits the ability to kill one or more Kp strains according to any embodiment disclosed herein.

[0070] In particular, an antibody or antigen-binding portion thereof according to the application is said to exhibit "functional activity" against Kp organisms when it is determined to exhibit complement-mediated bactericidal activity against Kp using any of the standard assays described herein (e.g., a fluorescent serum bactericidal assay using resazurin staining as a readout of cell viability).

[0071] An antibody having "bactericidal activity" or "antibacterial activity", as used herein, in particular complement-mediated bactericidal activity or antibacterial activity, refers to an antibody or antigen-binding portion thereof that exhibits a 50% inhibitory concentration (IC50) of less than 100 ng / ml, preferably less than 10 ng / ml, more preferably less than 5 ng / ml, for example, when tested by an in vitro fluorescent serum bactericidal assay (F-SBA) against Kp, e.g., as disclosed in the specification and examples.

[0072] The term "polynucleotide" as referred to herein means a polymer of nucleotides of at least 10 bases in length, which can be ribonucleotides, deoxyribonucleotides or a modified form of either type of nucleotide. The term includes single and double stranded forms.

[0073] The term "isolated polynucleotide" as used herein refers to a polynucleotide of genomic, cDNA, or synthetic origin, or some combination thereof, which, in the sole or principal sense, is (1) not associated with all or a portion of the polynucleotide with which the "isolated polynucleotide" is associated in nature, (2) operably linked to a polynucleotide to which it is not linked in nature, or (3) does not occur in nature as part of a larger sequence.

[0074] The term "naturally occurring nucleotide" as used herein includes deoxyribonucleotides and ribonucleotides. The term "modified nucleotide" as used herein includes nucleotides having modified or substituted sugar groups and the like. The term "oligonucleotide linkage" as referred to herein includes oligonucleotide linkages such as phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phoshoraniladate, phosphoroamidate and the like. See, e.g., LaPlanche et al., Nucl. Acids Res. 14:9081 (1986); Stec et al., J. Am. Chem. Soc. 106:6077 (1984); Stein et al., Nucl. Acids Res. 16:3209 (1988); Zon et al., Anti-Cancer Drug Design 6:539 (1991); Zon et al., Oligonucleotides and Analogues: A Practical Approach, pp. 87-108 (F. Eckstein, ed., Oxford University Press, Oxford England (1991)); U.S. Patent No. 5,151,510; Uhlmann and Peyman, Chemical Reviews 90:543 (1990), the disclosures of which are incorporated herein by reference. If desired, the oligonucleotide can comprise a label for detection. "Operably linked" sequences include both expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or at a distance to control the expression of the gene of interest. The term "expression control sequence" as used herein refers to polynucleotide sequences that are necessary to affect the expression and processing of coding sequences to which they are ligated. Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequence); sequences that enhance protein stability; and when desired, sequences that enhance protein secretion. The nature of such control sequences will vary depending on the host organism in which the gene is to be expressed; in prokaryotes, such control sequences generally include promoter, ribosomal binding site and termination sequences; in eukaryotes, such control sequences generally include promoters and termination sequences. The term "control sequences" is intended to include, at a minimum, all components whose presence is essential for expression and processing, and can also include additional components such as leader sequences and fusion partner sequences.The term "vector," as used herein, refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. In some embodiments, a vector is a plasmid, i.e., a circular double stranded DNA piece into which additional DNA segments can be ligated. In some embodiments, a vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. In some embodiments, a vector is capable of autonomous replication in a host cell into which it is introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). In other embodiments, a vector (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby be replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply, "expression vectors").

[0075] The term "recombinant host cell" (or simply "host cell"), as used herein, refers to a cell into which a recombinant expression vector has been introduced. It should be understood that "recombinant host cell" and "host cell" refer not only to the particular subject cell but to the progeny of the cell. Because certain modifications can occur in succeeding generations due to either mutation or environmental influences, such progeny can not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein.

[0076] In the context of nucleotide or amino acid sequences, the term "percent sequence identity" means the percentage of residues in the two sequences that are the same when aligned for maximum correspondence. The length of sequence identity comparisons can cover at least about 9 nucleotides, usually at least about 18 nucleotides, more usually at least about 24 nucleotides, typically at least about 28 nucleotides, more typically at least about 32 nucleotides, and preferably at least about 36, 48 or more nucleotides, of a segment. A number of different algorithms known in the art can be used to measure the nucleotide sequence identity. For example, polynucleotide sequences can be compared using FASTA, Gap, or Bestfit, which are programs contained in the ACCES / SWISS-PROT search programs, which provide alignments and percent sequence identity of the two sequences being compared (Pearson, Methods Enzymol. 183:63-98 (1990); Pearson, Methods MoI. Biol. 132:185-219 (2000); Pearson, Methods Enzymol. 266:227-258 (1996); Pearson, J MoI. Biol 276:71-84 (1998); which are incorporated herein by reference). When referring to nucleic acids or fragments thereof or to amino acids, the term "substantially similar" or "substantially identical" means that a nucleotide sequence will have at least about 85%, preferably at least about 90%, more preferably at least about 95%, 96%, 97%, 98%, or 99% nucleotide sequence identity when compared to another nucleic acid (or its complementary strand) using a suitable nucleotide insertion or deletion, as measured by any of the well-known sequence identity algorithms (e.g., FASTA, BLAST, or Gap as described above). The term "substantially the same" or "substantial identity" when applied to polypeptides means that two peptide sequences, when optimally aligned, will have at least 70%, 75%, or 80% sequence identity, preferably at least 90% or 95% sequence identity, more preferably at least 97%, 98%, or 99% sequence identity. In certain embodiments, changes in residue different positions are conservative amino acid substitutions. A "conservative amino acid substitution" means the substitution of one amino acid residue for another residue with a side chain R group of similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not significantly change the functional properties of a protein. If two or more amino acid sequences differ in residue positions due to conservative substitutions, then the percent sequence identity can be adjusted upwards to account for the conservative nature of the substitutions. Means for making this adjustment are well known to those of ordinary skill in the art. See, e.g., Pearson, Methods MoI. Biol. 243:307-31 (1994).Examples of groups of amino acids with chemically similar side chains include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic- hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine and methionine. Conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid- aspartic acid, and asparagine-glutamine. Alternatively, conservative replacements are ones in which the change is to an amino acid with a structurally similar side chain. For example, amino acids are divided into groups based on the nature of their side chains: (1) hydrophobic: norleucine, alanine, valine, leucine, isoleucine, methionine, proline; (2) neutral hydrophilic: glycine, serine, threonine, cysteine, tyrosine; (3) acidic: aspartic acid, glutamic acid; (4) basic: histidine, lysine, arginine; (5) residues that affect chain absorption: norleucine, tyrosine; (6) aromatic: phenylalanine, tyrosine, tryptophan; and (7) small, nonpolar: alanine, cysteine, valine, leucine, isoleucine, methionine, proline. Examples of conservative changes include: substitution of one hydrophobic residue such as isoleucine, valine, leucine or methionine for another; substitution of one polar residue for another, such as between arginine and lysine, between glutamic and aspartic acid, and between phenylalanine and tyrosine; and substitution of one small, nonpolar residue, such as alanine or serine, for another. Alternatively, conservative replacement of an amino acid can be determined based on the hydropathic index of amino acids, as described by Kyte and Doolittle, 1982, J. Mol. Biol. 157: 105-132, which is incorporated herein by reference. In making such determinations, replacement amino acids have hydropathic indices within 2 units of the reference amino acid. The hydropathic index of an amino acid is based on the difference in free energy between the amino acid in a polar and nonpolar environment. The hydropathic index of amino acids is as follows: isoleucine (+5.4); valine (+5.2); leucine (+3.8); phenylalanine (+2.7); cysteine / cysteine (+2.5); methionine (+1.9); proline (-0.5); histidine (-0.5); lysine (-3.9); tyrosine (-2.3); tryptophan (-4.5); serine (-0.8); threonine (-0.7); asparagine (-6.4); glutamine (-6.3); glutamic acid (-8.5); aspartic acid (-8.7); arginine (-11.5); and

[0077] The term "labeled" or "labeled" as used herein refers to the incorporation of another molecule into an antibody. In one embodiment, the label is a detectable marker, such as incorporation of a radiolabeled amino acid or attachment of a biotinylated moiety that can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected by optical or colorimetric methods). In another embodiment, the label or marker can be therapeutic, such as a drug conjugate or toxin. Methods for labeling polypeptides and glycoproteins are known in the art and can be used.

[0078] The expressions "drug resistant" and "multidrug resistant" (abbreviated as MDR) when used herein in relation to bacteria define bacteria that are resistant to at least one class of drugs or to two or more classes of drugs, respectively. In particular, the expression "multidrug resistant" is used to define bacteria that are resistant to two or more classes of antibiotics, including but not limited to beta-lactam antibiotics (also referred to herein as beta-lactam antibiotics), in particular carbapenems.

[0079] The term "capsular antigen" or "K antigen" used synonymously in the present specification refers to one or more antigens belonging to the bacterial capsular polysaccharide (CPS) of Kp according to any of the variants disclosed herein. The term "capsular antigen" or "K antigen" used herein encompasses the capsular polysaccharide of said Kp or any portion or fragment thereof, unless otherwise defined. The term "O antigen" used herein refers to a somatic antigen on the surface of Kp cells. The O antigen is the most surface-exposed part of the bacterial lipopolysaccharide. It is composed of oligosaccharide repeat units of varying lengths, each unit usually containing 2 to 7 sugar moieties.

[0080] The expression "carbapenemase-producing strain" used herein refers to any Kp strain producing carbapenemases, which are a class of beta-lactamases that can lead to resistance to carbapenems, a subclass of beta-lactam antibiotics. The structure of carbapenems is very similar to that of penicillins (penams), but the sulfur atom at position 1 is replaced by a carbon atom and an unsaturated bond is introduced - hence the name carbapenems for this class of antibiotics.

[0081] Carbapenemases give Kp strains a broad antibiotic resistance profile, as these enzymes can not only hydrolyze and inactivate carbapenem antibiotics, but also a broad spectrum of penicillins, oxyimino-cephalosporins and cephamycins.

[0082] The expression "New Delhi metallo-beta-lactamase (NDM)-producing strain" used herein refers to any Kp strain producing New Delhi metallo-beta-lactamase (NDM, also referred to herein as New Delhi metallo-beta-lactamase), which is a specific member of a large family of genes encoding beta-lactamases known as carbapenemases. NDM is not only able to confer resistance to a broad range of beta-lactam antibiotics, but also to novel beta-lactamase inhibitor combinations, including but not limited to the combinations of ceftazidime-avibactam, imipenem-relebactam and meropenem-vaborbactam.

[0083] NDM is encoded by the bla NDM gene located on large multi-drug resistant plasmids that have spread among high-risk MDR Kp strains on all continents (di Pilato V. et al., 2022 Mar 1; 3(3): e224-34; Martin MJ et al., PNAS, 2021; 118: 1-8). NDM comprises several variants encoded by the corresponding bla NDM genes, which have different properties.

[0084] As used herein, the expression "NDM-1 positive strain", "NDM-5 positive strain" or "NDM-9 positive strain" (herein also abbreviated as NDM-1 + , NDM-5 + and NDM-9 + , respectively, refers to any Kp strain capable of producing the following enzymes: New Delhi metallo-β-lactamase 1, New Delhi metallo-β-lactamase 5 and New Delhi metallo-β-lactamase 9, each of which is encoded by a respective gene, namely bla NDM-1 , bla NDM-5 and bla NDM-9 , respectively.

[0085] The NDM-1 enzyme was originally named after the Indian capital New Delhi, as it was first described by Yong et al. in December 2009 in a Swedish citizen who had become ill in India with an antibiotic-resistant bacterium (Yong D et al., 2009, Antimicrob. Agents Chemother. 53(12): 5046-5054). The infection was identified as a carbapenem-resistant Klebsiella pneumoniae strain carrying a novel gene, bla NDM-1 .

[0086] In the context of the present specification, an NDM-producing Kp strain belonging to a particular sequence type (ST) can be denoted by the following nomenclature: "STX NDMY " or "STX NDM-Y " or "STX NDMY+ ", wherein X stands for the particular ST number of the Kp strain and Y stands for the particular NDM variant produced by said strain.

[0087] As used herein, the expression "Klebsiella pneumoniae capsular type K64 strain" refers to any Kp strain carrying one or more genes from the capsular type 64 biosynthetic locus, i.e. one or more genes encoding one or more enzymes responsible for or involved in the biosynthesis of K64-type capsular polysaccharide (CPS).

[0088] In other words, as used herein, the expression "Klebsiella pneumoniae capsular type K64 strain" refers to any strain or clonal lineage of Klebsiella pneumoniae carrying K64-type capsular polysaccharide (CPS).

[0089] In the context of the present specification, the terms "K64", "KL64", "K-64", "KL-64" all refer to Klebsiella pneumoniae of capsular type 64 and are used interchangeably.

[0090] As used herein, the term "IC 50"IC50" refers to the concentration of antibody that results in a 50% reduction in Kp cell viability as measured by, for example, a fluorescent serum bactericidal assay, in particular, the resazurin-based serum bactericidal assay disclosed herein.

[0091] Human anti-Kp antibodies and their characterization

[0092] In one embodiment, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of Kp.

[0093] Preferably, in one embodiment, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of at least one drug resistant or multidrug resistant Kp strain, more preferably at least one carbapenemase-producing Kp strain, even more preferably at least one NDM-producing Kp strain.

[0094] Preferably, in one embodiment, the present application provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of at least one capsular type K64 Klebsiella pneumoniae strain.

[0095] In a preferred embodiment, the Kp strain comprises one or more Kp strains selected from the group consisting of NDM-1 positive strains, NDM-9 positive strains, and NDM-5 positive strains.

[0096] In a preferred embodiment, the Kp strain according to any of the variants disclosed herein comprises one or more Kp strains or clinical isolates selected from the group consisting of Sequence Type 147 (ST147), Sequence Type 258 (ST258), Sequence Type 493 (ST493), Sequence Type 307 (ST307), Sequence Type 13 (ST13), and Sequence Type 512 (ST512), in particular strains from ST147, ST258 or clonal lineages defined thereby.

[0097] In one embodiment, the present application specifically provides a human monoclonal antibody or an antigen binding portion thereof that specifically binds to a surface antigen of at least one NDM-producing Kp strain selected from the group consisting of ST147, ST258, ST493, ST307, and ST13, in particular ST147, ST258, ST307, and ST493.

[0098] Preferably, the Kp comprises at least one NDM-producing Kp strain selected from the group consisting of ST147 NDM-1 , ST147 NDM-9 , and ST307 NDM-5 .

[0099] In some aspects, the human monoclonal antibody or antigen binding portion thereof according to any of the embodiments disclosed in the present specification and claims specifically binds to a surface antigen of at least one strain, at least two strains, at least three strains, at least four strains, or at least five strains of Kp according to any of the variants disclosed in the present specification and claims.

[0100] According to a more preferred embodiment, the present application provides a human monoclonal antibody or antigen binding portion thereof, which specifically binds to a surface antigen of ST147 NDM-1 of Kp, which strain has been causing nosocomial outbreaks in Tuscany, Italy since 2018. Overall, ST147 is widely spread in India and Southeast Asia and has recently been classified as a pandrug-resistant strain.

[0101] In a preferred embodiment, the Kp is ST147 NDM-1 and one, more or all of the ST147 NDM-9 strains, more preferably ST147 NDM-1 and one or all of the ST147 NDM-9 strains, and the human monoclonal antibody or antigen binding portion thereof is selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ08-H10, SBJ08-N23, SBJ08-K19, SBJ08-M13, SBJ03-L02, SBJ08-J10, SBJ10-H18, SBJ08-O03, SBJ05-K07, SBJ09-I10 and SBJ11-C06.

[0102] In another preferred embodiment, the Kp is one or more of the strains ST147 NDM-1 , ST307 NDM-5 , ST258 and ST493, preferably comprising all strains, and the human monoclonal antibody or antigen binding portion thereof is selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, SBJ05-M13, SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04 and SBJ05-B17.

[0103] In another preferred embodiment, the Kp is one or more of the strains ST147 NDM-1 and ST307 NDM-5 , preferably comprising all strains, and the human monoclonal antibody or antigen binding portion thereof is selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13 and SBJ05-M13.

[0104] In another preferred embodiment, the Kp is one or more of the strains ST147 NDM-1 , ST307 NDM-5 , and ST493, preferably comprising all strains, more preferably the ST493 strain, and the human monoclonal antibody or antigen binding portion thereof is selected from the group consisting of SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04, and SBJ05-B17.

[0105] As used in the present description and claims, the term "surface antigen" encompasses any antigen displayed on the surface of a Kp bacterium, in particular of a Kp strain according to any of the variants disclosed herein.

[0106] In one embodiment, the surface antigen is selected from the group consisting of (i) a capsular antigen (K antigen) and / or (ii) an O antigen of at least one Kp strain according to any of the variants disclosed herein.

[0107] According to a more preferred embodiment, the present application provides a human monoclonal antibody or antigen binding portion thereof, which specifically binds to and has a bactericidal effect on the capsular antigen of at least one of the following strains: a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain and a NDM-9 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

[0108] In particular, the present application provides a human monoclonal antibody or antigen binding portion thereof, which specifically binds to and has a bactericidal effect on the capsular antigen of a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

[0109] In a preferred embodiment, the capsular antigen is a capsular polysaccharide, more preferably a K64 type capsular polysaccharide.

[0110] In one embodiment, the O antigen is at least one O1 type antigen and / or O2 type antigen of a Kp strain according to any of the variants disclosed herein.

[0111] In another embodiment, the O2 type antigen is selected from the group consisting of O2a and O2afg antigens.

[0112] In a preferred embodiment, the surface antigen is a capsular antigen and the human monoclonal antibody or antigen binding portion thereof is selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ08-H10, SBJ08-N23, SBJ08-K19, SBJ08-M13, SBJ03-L02, SBJ08-J10, SBJ10-H18, SBJ08-O03, SBJ05-K07, SBJ09-I10, and SBJ11-C06, preferably SBJ08-O09.

[0113] Preferably, when the surface antigen is a capsular antigen, in particular a capsular antigen from KL64 type capsule, the Kp is one or more of, preferably comprising all of, strains ST147 NDM-1 and ST147 NDM-9 , preferably comprising all of strains ST147 NDM-1 and ST147 NDM-9 .

[0114] In another preferred embodiment, the surface antigen is an O antigen, and the human monoclonal antibody or antigen-binding portion thereof is selected from SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, SBJ05-M13, SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04, and SBJ05-B17.

[0115] Preferably, when the surface antigen is an O antigen, the Kp is one or more of, preferably comprising all of, strains ST147 NDM-1 , ST307 NDM-5 , ST258, and ST493. In another preferred embodiment, the surface antigen is an O2 type antigen, and the human monoclonal antibody or antigen-binding portion thereof is selected from SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, and SBJ05-M13.

[0116] Preferably, when the surface antigen is an O2 type antigen, the Kp is one or more of, preferably comprising all of, strains ST147 NDM-1 and ST307 NDM-5 .

[0117] In another preferred embodiment, the surface antigen is an epitope common to O1 and O2 type O antigens, and the human monoclonal antibody or antigen-binding portion thereof is selected from SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04, and SBJ05-B17.

[0118] Preferably, when the surface antigen is an O1 type antigen, the Kp is the ST493 strain.

[0119] In certain aspects, the human monoclonal antibody or antigen-binding portion thereof that specifically binds to a Kp surface antigen, according to any of the embodiments disclosed herein, exhibits bactericidal or antibacterial activity, in particular complement-mediated bactericidal or antibacterial activity.

[0120] The complement-mediated bactericidal activity of the antibodies can be assessed using standard assays, such as luminescent or fluorescent serum bactericidal assays known to the person skilled in the art. By way of example only, in these standard assays, Kp is not only reacted with the antibody to be tested, but also with a source of complement. Bacterial counts are determined at different sampling times. Antibodies with complement-mediated bactericidal activity are those for which the viable bacterial cell count is reduced in a dose-dependent manner after co-incubation with the antibody and complement, and to a minimum of 50% compared to the colony count at time zero, such antibodies are considered to have bactericidal activity for the purposes of the present application and are suitable for further use.

[0121] In one embodiment, the present application provides a human monoclonal antibody or antigen binding portion that specifically binds to a surface antigen of Kp according to any of the embodiments disclosed herein, wherein the antibody or antigen binding portion thereof provides a percentage of bacterial killing, in particular a percentage of complement-dependent bacterial killing, of greater than 30%, preferably greater than 60%, preferably greater than 70%, more preferably greater than 80%, even more preferably greater than 85%, 90%, 95%, 98% or 99%, as measured by, for example, an in vitro luminescent serum bactericidal assay (L-SBA) and / or by an in vitro fluorescent serum bactericidal assay (F-SBA) as disclosed in the Examples.

[0122] In a preferred embodiment, bacterial killing is measured by L-SBA assay by incubating at least one Kp strain according to any of the variants disclosed herein with at least four serial dilutions of mAb in the presence of an appropriate amount of baby rabbit complement (BRC). After the addition of luciferase, the luminescence signal generated by the luciferin turnover in the presence of bacterial ATP is measured as an indicator of Kp viability, allowing the rapid and sensitive quantification of bacterial survival. The luminescence values obtained from all mAbs in each experiment can be used to calculate the median value for each dilution tested. The difference between the luminescence value of each single mAb and the median value can be used as a readout of bacterial viability. The appropriate amount or concentration of BRC used in the assay can be optimized by performing a complement sensitivity screen on selected Kp strains, for example as disclosed in the Examples.

[0123] According to a preferred aspect, the human monoclonal antibody of the application or antigen binding portion thereof displays a 50% inhibitory concentration (IC50) of less than 100 ng / ml, preferably less than 50, 25, 20, 10, 8, 6, 5, 4, 3, 2 or 1 ng / ml when tested by in vitro fluorescent serum bactericidal assay (F-SBA) against Kp, for example against Kp ST147 NDM-1 strain, ST147 NDM-9 strain and / or ST307 NDM-5 strain, for example as disclosed in the Examples.

[0124] In preferred embodiments, the F-SBA assay is performed by incubating at least one Kp strain according to any of the variants disclosed herein with at least three serial dilutions of the mAb in the presence of an appropriate amount of baby rabbit complement (BRC), e.g. as disclosed in the examples, in particular using resazurin staining as a fluorescent readout for bacterial viability. If resazurin is used, the F-SBA assay is also abbreviated as "R-SBA" herein.

[0125] As disclosed in more detail in the examples, the inventors found that the most potent bactericidal antibodies SBJ08-O09, SBJ08-D18 and SBJ03-F18 surprisingly exhibit bactericidal activity against all ST147 NDM-1 , ST147 NDM-9 strains.

[0126] In certain aspects, the present application provides a human monoclonal antibody, or antigen binding portion thereof, that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, particularly at least one surface antigen of a Kp strain according to any of the variants disclosed herein, comprising: (a) a heavy chain variable domain amino acid sequence comprising the amino acid sequence of the heavy chain variable domain of an antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13; (b) a light chain variable domain amino acid sequence comprising the amino acid sequence of the light chain variable domain of an antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13; (c) the heavy chain variable domain of (a) and the light chain variable domain of (b); or (d) heavy and light chain variable domain amino acid sequences comprising the heavy and light chain variable domain amino acid sequences, respectively, from the same antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.

[0127] Nucleic acids encoding the full length or a portion comprising the variable domain of the heavy and light chains of the antibodies of the application as well as the corresponding deduced amino acid sequences can be found in the sequence listing attached herewith.

[0128] In certain aspects, the present application provides a monoclonal antibody or antigen binding portion thereof that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, in particular a surface antigen of at least one Kp strain according to any of the variants disclosed herein, comprising: (a) a heavy chain variable domain amino acid sequence comprising the heavy chain CDR1, CDR2, and CDR3 amino acid sequences of an antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13; (b) a light chain variable domain amino acid sequence comprising the light chain CDR1, CDR2, and CDR3 amino acid sequences of an antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13; (c) the heavy chain variable domain of (a) and the light chain variable domain of (b); or (d) the heavy chain variable domain and the light chain variable domain of (c) comprising the heavy chain and light chain CDR amino acid sequences from the same antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.

[0129] In certain aspects, the present application provides a monoclonal antibody that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, in particular to a surface antigen of at least one Kp strain according to any of the variants disclosed herein, wherein the antibody comprises a heavy chain of an antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13. In certain aspects, the present application provides a monoclonal antibody that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, in particular to a surface antigen of at least one Kp strain according to any of the variants disclosed herein, wherein the antibody comprises a light chain of an antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.In certain aspects, the present application provides a monoclonal antibody that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, in particular to a surface antigen of at least one Kp strain according to any of the variants disclosed herein, wherein the antibody comprises the heavy and light chain of the same antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.

[0130] In certain aspects, the present application provides a human monoclonal antibody or antigen binding portion thereof that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, in particular to a surface antigen of at least one Kp strain according to any of the variants disclosed herein, comprising VL and VH domains, wherein the VL and VH domains are at least 85%, 90%, 95%, 97%, 98%, or 99% identical in amino acid sequence to the VL and VH domains, respectively, of a monoclonal antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.

[0131] In certain aspects, the present application provides a human monoclonal antibody, or antigen binding portion thereof, that specifically binds to a surface antigen of a Kp according to any of the embodiments disclosed herein, particularly at least one surface antigen of a Kp strain according to any of the variants disclosed herein, comprising a light chain and a heavy chain, wherein the light chain and the heavy chain are at least 85%, 90%, 95%, 97%, 98%, or 99% identical in amino acid sequence to the light chain and the heavy chain, respectively, of a monoclonal antibody selected from the group consisting of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.

[0132] One amino acid substitution that can be made is to change one or more cysteines in the antibody that can be chemically reactive to another residue, such as but not limited to alanine or serine. In one embodiment, there is a substitution of a non-canonical cysteine. The substitution can be made in the variable domain CDRs or framework regions of the antibody, or in the constant domain of the antibody. In some embodiments, the cysteine is canonical.

[0133] Another amino acid substitution that can be made is to change any potential proteolytic site in the antibody. Such sites can be in the variable domain CDRs or framework regions of the antibody, or in the constant domain of the antibody. Replacing the cysteine residue and removing the proteolytic site can reduce any risk of heterogeneity in the antibody product, thereby increasing its homogeneity. Another amino acid substitution is to eliminate asparagine-glycine pairs that form potential deamidation sites, by changing one or both of the residues. In some embodiments, the C-terminal lysine of the anti-Kp antibody heavy chain of the present application is cleaved. In various embodiments of the present application, the heavy chain and light chain of the anti-Kp antibody can optionally comprise a signal sequence.

[0134] The class and subclass of an anti-Kp antibody can be determined by any method known in the art. Generally, antibodies specific for the particular class and subclass of antibodies can be used to determine the class and subclass of an antibody. Such antibodies are commercially available. The class and subclass can be determined by ELISA or Western blot (immunoblot) as well as other techniques. Alternatively, the class and subclass can be determined by sequencing all or a portion of the antibody heavy and / or light chain constant domains, comparing their amino acid sequences to the known amino acid sequences of various class and subclass immunoglobulins, and determining the class and subclass of the antibody.

[0135] In some embodiments, the human monoclonal antibodies of the application are IgG, IgM, IgE, IgA, or IgD molecules. In one embodiment, the human monoclonal antibodies are IgG, and are of the IgGl, IgG2, IgG3, IgG4 subclass. In another embodiment, the human antibody subclass is IgGl.

[0136] Binding affinity of anti-Kp antibodies to Kp surface antigens.

[0137] In some embodiments of the application, the anti-Kp antibodies bind with high affinity to a surface antigen of a Kp according to any of the embodiments disclosed herein. In some embodiments, the anti-Kp antibodies bind with high affinity to at least one of the capsular antigens or polysaccharides of a Kp strain according to any of the variants disclosed herein, preferably to the K64 type capsular antigen. In some embodiments, the anti-Kp antibodies bind to the O antigen of at least one of the Kp strains according to any of the variants disclosed herein, in particular to the O1 antigen and / or the O2 antigen. The binding affinity and off-rate of an anti-Kp antibody to a Kp surface antigen according to any of the embodiments disclosed herein can be determined by methods known in the art. The binding affinity can be measured by ELISA, RIA, flow cytometry, surface plasmon resonance (e.g. BIACORE(TM)). The off-rate can be measured by surface plasmon resonance. Preferably, the binding affinity and off-rate are measured by surface plasmon resonance. More preferably, the binding affinity and off-rate are measured using BIACORE(TM). Whether an antibody has substantially the same KD as an anti-Kp antibody can be determined using methods known in the art.

[0138] The present invention provides human anti-Kp monoclonal antibodies that bind to a surface antigen of Kp according to any of the embodiments disclosed herein, in particular to a surface antigen of at least one Kp strain according to any of the variants disclosed herein, and compete or cross-compete with and / or bind to the same epitope as an antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13. Two antibodies are said to cross-compete if they compete with each other for binding to a Kp surface antigen.

[0139] Whether an antibody binds to the same epitope or cross-competes for binding with an anti-Kp antibody can be determined using methods known in the art. In one embodiment, an anti-Kp antibody of the present invention is allowed to bind to a Kp surface antigen under saturating conditions, and then the ability of a test antibody to bind to the Kp surface antigen is measured. If the test antibody is able to bind to the Kp surface antigen at the same time as the anti-Kp antibody, then the test antibody binds to a different epitope than the anti-Kp antibody. However, if the test antibody is not able to bind to the Kp surface antigen at the same time, then the test antibody binds to the same epitope, to an overlapping epitope, or to an epitope in close proximity to the epitope bound by the human anti-Kp antibody, or the binding by the human anti-Kp antibody can induce a conformational change in the surface antigen that prevents or reduces binding by the test antibody. This experiment can be performed using ELISA, RIA, BIACORE(TM), flow cytometry, or other methods known in the art.

[0140] To test whether an anti-Kp antibody cross-competes with another anti-Kp antibody, the competition method described above can be used from both directions, i.e., to determine whether the reference antibody blocks the test antibody, and vice versa. In one embodiment, the experiment is performed using ELISA. Methods for determining KD will be discussed further below.

[0141] In another embodiment, the present invention provides an anti-Kp antibody that disrupts, inhibits, blocks, neutralizes, or reduces a Kp action or function, in particular a bactericidal or Kp cell-killing action.

[0142] In another embodiment, the present invention provides an anti-Kp antibody that is capable of killing Kp cells, in particular cells of at least one Kp strain according to any of the variants disclosed herein, in a complement-dependent manner.

[0143] In certain embodiments, the present application provides anti-Kp antibodies that inhibit, block, or reduce by 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% the symptoms or conditions caused by Kp infection, particularly infection by MDR Kp strains, particularly infection by one or more NDM-producing strains according to any of the variants disclosed herein, for a day, a week, a month, six months, a year, or the rest of the subject's life. In certain embodiments, the present application provides anti-Kp antibodies that can achieve any combination of the foregoing embodiments.

[0144] In certain embodiments, to increase complement-dependent Kp killing, mutations that enhance the process of natural hexamerization that occurs between the Fc regions of antibodies after opsonization will be introduced into the Fc portion of the IgGl antibody as previously described, preferably the E430G mutation as described in de Jong RN et al. "A Novel Platform for the Potentiation of Therapeutic Antibodies Based on Antigen-Dependent Formation of IgG Hexamers at the Cell Surface." PLoS Biol. 2016 Jan 6; 14(1).

[0145] All these modifications can be made by way of site-directed mutagenesis, for example using the Agilent Quick-Change II Site-Directed Mutagenesis Kit according to the manufacturer's recommendations.

[0146] In certain embodiments, the antibody comprises a variable region of an antibody selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13, and a mutated IgGl constant region backbone containing an E430G mutation in the Fc domain (as described in de Jong RN et al. PLoS Biol. 2016 Jan 6;14(l)).

[0147] Nucleic acids, vectors, host cells, and recombinant methods of making antibody nucleic acids

[0148] The present application also encompasses nucleic acid molecules encoding an anti-Kp antibody or antigen binding portion thereof. In some embodiments, different nucleic acid molecules encode the heavy and light chains of an anti-Kp immunoglobulin. In other embodiments, the same nucleic acid molecule encodes both the heavy and light chains of an anti-Kp immunoglobulin. In one embodiment, the nucleic acid molecule encodes a Kp antibody or antigen binding portion thereof of the present application. In some embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a VL amino acid sequence comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservative amino acid substitutions and / or 1, 2, or 3 non-conservative substitutions compared to the germline. The substitutions can occur in the CDR regions, framework regions, or constant regions. In some embodiments, the nucleic acid molecule encodes a VL amino acid sequence comprising one or more variants identical to the changes found in the VL of one of the antibodies selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13 compared to the germline sequence.

[0149] In some embodiments, the nucleic acid molecule encodes a sequence of at least three amino acid substitutions compared to the germline sequence found in the VL of one of the antibodies selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, and SBJ11-J13.

[0150] In some embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a VL amino acid sequence of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, or a variant or portion thereof. In some embodiments, the nucleic acid encodes an amino acid sequence comprising the light chain CDRs of one of the above antibodies. In some embodiments, the portion is a contiguous portion comprising CDR1-CDR3. In some embodiments, the nucleic acid encodes an amino acid sequence of the light chain CDRs of the antibody. In some embodiments, the portion encodes a contiguous region of the light chain CDR1-CDR3 of an anti-Kp antibody.

[0151] In some embodiments, the nucleic acid molecule encodes a VL amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% identical to the VL amino acid sequence of the VL region of any of the following antibodies: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13. Nucleic acid molecules of the application include nucleic acids that hybridize to a nucleotide sequence encoding a VL region amino acid sequence under highly stringent conditions (e.g., the conditions described above).

[0152] In another embodiment, the nucleic acid encodes a full length light chain of an antibody selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, or a light chain comprising a mutation, e.g., a mutation disclosed herein.

[0153] In yet another embodiment, the nucleic acid molecule encodes a heavy chain (VH) variable domain comprising a human VH1, VH3, or VH4 family gene sequence or a sequence derived therefrom. In some embodiments, the nucleic acid molecule encodes a sequence of one or more amino acid mutations identical to those found in the VH of one of the following monoclonal antibodies: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, compared to the germline sequence.

[0154] In some embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding at least a portion of a VL amino acid sequence of a monoclonal antibody selected from the group consisting of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, all three CDR regions, contiguous portions including CDR1-CDR3, or the entire VL region (with or without signal sequence). In some embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding an amino acid sequence of one of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, or said sequence lacking the signal sequence. In some preferred embodiments, the nucleic acid molecule comprises at least a portion of a nucleotide sequence of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, or said sequence lacking the signal sequence. In some embodiments, the portion encodes a VL region (with or without signal sequence), a CDR3 region, all three CDR regions, or a contiguous region including CDR1-CDR3.

[0155] In some embodiments, the nucleic acid molecule encodes a VH amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% identical to the VH amino acid sequence of any of the following antibodies: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13.

[0156] The nucleic acid molecules of the application include nucleic acids that hybridize under high stringency conditions (e.g., the conditions described above) to a nucleotide sequence encoding the following amino acid sequences or those encoding the VH region thereof: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13.

[0157] In another embodiment, the nucleic acid encodes a full length heavy chain of an antibody selected from the group consisting of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, or a heavy chain having the amino acid sequence of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, with or without the signal sequence, or a heavy chain comprising a mutation, such as one of the variants discussed herein. Further, the nucleic acid can comprise the nucleotide sequence of SBJ08-009, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, with or without the signal sequence, or a nucleic acid molecule encoding a heavy chain comprising a mutation, such as one of the variants discussed herein.

[0158] A nucleic acid molecule encoding an anti-Kp antibody heavy chain or light chain, or portion thereof, can be isolated from any source that produces such an antibody. In various embodiments, the nucleic acid molecule is isolated from a B cell isolated from an animal immunized with a surface antigen, or from a such a B cell-derived immortalized cell expressing or encoding an anti-Kp antibody. Methods of isolating mRNA encoding an antibody are well known in the art. See, e.g., Sambrook et al. The mRNA can be used to generate cDNA, for polymerase chain reaction (PCR) or cDNA cloning of antibody genes. In one embodiment, the nucleic acid molecule is isolated from a hybridoma having as one of the fusion partners a human immunoglobulin-producing cell from a non-human transgenic animal. In an even more preferred embodiment, the human immunoglobulin-producing cell is isolated from a XENOMOUSE animal. In another embodiment, the human immunoglobulin-producing cell is from a non-human, non-mouse transgenic animal as described above. In another embodiment, the nucleic acid is isolated from a non-human, non-transgenic animal. Nucleic acid molecules isolated from a non-human, non-transgenic animal can be used, e.g., in humanized antibodies. In some embodiments, a nucleic acid encoding a heavy chain of an anti-Kp antibody of the application can comprise a nucleotide sequence encoding a VH domain of the application linked in-frame to a nucleotide sequence encoding a heavy chain constant domain of any origin. Similarly, a nucleic acid molecule encoding a light chain of an anti-Kp antibody of the application can comprise a nucleotide sequence encoding a VL domain of the application linked in-frame to a nucleotide sequence encoding a light chain constant domain of any origin. In another aspect of the application, a nucleic acid molecule encoding a heavy chain (VH) and / or light chain (VL) variable domain is "converted" into a full-length antibody gene. In one embodiment, a nucleic acid molecule encoding a VH or VL domain is converted into a full-length antibody gene by operably linking the VH segment to one or more CH segments within an expression vector that has already been encoding a heavy chain constant (CH) or light chain constant (CL) domain, respectively, and / or by operably linking the VL segment to a CL segment within an expression vector. In another embodiment, a nucleic acid molecule encoding a VH and / or VL domain is converted into a full-length antibody gene by joining (e.g., ligating) a nucleic acid molecule encoding a CH and / or CL domain to a nucleic acid molecule encoding a VH and / or VL domain using standard molecular biology techniques. The nucleotide sequences of human heavy and light chain immunoglobulin constant domain genes are known in the art. See, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. NIH Publ. No. 91-3242, 1991. The nucleic acid molecule encoding the full-length heavy and / or light chain can then be expressed from a cell into which it has been introduced, and the anti-Kp antibody isolated.

[0159] The nucleic acid molecules can be used to recombinantly express large amounts of anti-Kp antibodies. The nucleic acid molecules can also be used to produce chimeric antibodies, bispecific antibodies, single chain antibodies, immunoadhesins, diabodies, mutant antibodies, and antibody derivatives, as described below. If the nucleic acid molecules are derived from a non-human, non-transgenic animal, the nucleic acid molecules can be used for antibody humanization, as described below.

[0160] In another embodiment, the nucleic acid molecules of the application are used as probes or PCR primers for specific antibody sequences. For example, the nucleic acids can be used as probes in diagnostic methods, or as PCR primers to amplify DNA regions that are particularly useful for isolating nucleic acid molecules encoding additional anti-Kp antibody variable domains. In some embodiments, the nucleic acid molecules are oligonucleotides. In some embodiments, the oligonucleotides are from the hypervariable domains of the heavy and light chains of the antibodies of interest. In some embodiments, the oligonucleotides encode all or part of one or more CDRs of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, or a variant thereof as described herein. The nucleic acid molecules disclosed herein can be DNA or RNA molecules.

[0161] Vectors

[0162] The present application provides vectors comprising nucleic acid molecules encoding the heavy chain of an anti-Kp antibody or antigen binding portion thereof of the present application. The present application also provides vectors comprising nucleic acid molecules encoding the light chain of such an antibody or antigen binding portion thereof. The present application further provides vectors comprising nucleic acid molecules encoding fusion proteins, modified antibodies, antibody fragments, and probes thereof. In some embodiments, expression of an anti-Kp antibody or antigen binding portion of the present application is carried out by inserting DNA encoding the partial or full light and heavy chains, obtained as described above, into expression vectors such that the genes are operably linked to the necessary expression control sequences (e.g., transcription and translation control sequences). Expression vectors include plasmids, retroviruses, adenoviruses, adeno-associated viruses (AAV), plant viruses (e.g., cauliflower mosaic virus, tobacco mosaic virus), cosmids, YACs, EBV-derived episomes, and the like. The antibody genes are ligated into the vectors such that the transcriptional and translational control sequences within the vector function to produce the desired transcription and translation of the antibody genes. The choice of expression vector and expression control sequences will depend on the host cell used. The antibody light chain gene and the antibody heavy chain gene can be inserted into separate vectors or, more typically, into the same vector. In one embodiment, both genes are inserted into the same expression vector. The antibody genes are inserted into the expression vector by standard methods (e.g., ligation of adaptors to the ends of the antibody gene fragments and complementary restriction sites on the vector, or blunt-end ligation if no restriction sites are present). One convenient vector is a vector encoding a functionally complete human CH or CL immunoglobulin sequence, which has been engineered to have appropriate restriction sites so that any VHand VLsequences can be readily inserted and expressed, as described above. In such a vector, splicing usually occurs at the splice donor site at the junction of the inserted J region and the splice acceptor site preceding the human C domain, as well as at the splice regions within the human CH exons. Polyadenylation and transcription termination occur at the natural chromosomal site downstream of the coding region. The recombinant expression vectors can also encode a signal peptide that facilitates secretion of the antibody chain(s) from a host cell. The antibody chain genes can be cloned into the vector such that the signal peptide is linked in-frame to the amino terminus of the immunoglobulin chain. The signal peptide can be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., from a non-immunoglobulin protein). In addition to the antibody chain genes, the recombinant expression vectors of the present application carry regulatory sequences that control the expression of the antibody chain genes in a host cell. One of skill will appreciate that the design of the expression vector, including the selection of regulatory sequences, mayPreferred regulatory sequences for mammalian host cell expression include viral elements that direct high levels of protein expression in mammalian cells, such as those derived from retroviral LTRs, cytomegalovirus (CMV), (such as the CMV promoter / enhancer), Simian Virus 40 (SV40), (such as the SV40 promoter / enhancer), adenovirus, (such as the adenovirus major late promoter (AdMLP)), poliomavirus, and / or the strong promoters and enhancers of the long terminal repeat of human T cell leukemia virus, herpes simplex virus, and human immunodeficiency virus. For a description of viral regulatory elements and sequences, see, e.g., U.S. Pat. No. 5, 168,062, U.S. Pat. No. 4,510,245, and U.S. Pat. No. 4,968,615. Methods of expressing antibodies in plants, including descriptions of promoters and vectors, and transformation of plants are known in the art. See, e.g., U.S. Pat. No. 6,517,529, incorporated herein by reference. Methods of expressing polypeptides in bacterial cells or fungal cells (e.g., yeast cells) are also well known in the art. In addition to antibody chain genes and regulatory sequences, a recombinant expression vector of the application can carry other sequences, such as a sequence enabling the vector to replicate in the host cell in question (e.g., an origin of replication) and a selectable marker gene. The selectable marker gene facilitates selection of host cells into which the vector has been introduced (see e.g., U.S. Pat. Nos. 4,399,216, 4,634,665 and 5,179,017, incorporated herein by reference). For example, a selectable marker gene can confer resistance to drugs (e.g., G418, hygromycin or methotrexate) on host cells into which the vector has been introduced. Preferred selectable marker genes include dihydrofolate reductase (DHFR) genes (for use in dhfr host cells with methotrexate selection / amplification), the neo gene (for G418 selection), and glutamine synthetase genes.

[0163] In some embodiments, the vector is selected from the group consisting of an RNA viral vector, a DNA viral vector, a plasmid viral vector, an adenoviral vector, an adeno-associated viral vector, a herpes viral vector, and a retroviral vector.

[0164] In some aspects, the present application provides compositions (e.g., pharmaceutical compositions), methods, kits, and reagents comprising one or more isolated nucleic acid molecules or vectors according to any of the embodiments disclosed herein for use in the prevention and / or treatment of Kp infection, particularly in humans and other mammals. In some embodiments, such nucleic acid molecules and vectors are formulated in nanoparticles, such as lipid nanoparticles, cationic lipid nanoparticles, examples of such formulations can be found in US2020197510, incorporated herein by reference.

[0165] Non-hybridoma host cells and methods of recombinantly producing proteins

[0166] Nucleic acid molecules encoding anti-Kp antibodies and vectors comprising these nucleic acid molecules can be used to transfect or transform suitable mammalian, plant, bacterial, or yeast host cells. Transfection or transformation can be performed by any known method of introducing polynucleotides into a host cell. Methods of introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide in liposomes, and direct microinjection of the DNA into the nucleus of the cell. In addition, nucleic acid molecules can be introduced into mammalian cells by viral vectors. Methods of transforming cells are well known in the art (see, e.g., U.S. Patent Nos. 4,399,216, 4,912,040, 4,740,461, and 4,959,455, which are incorporated herein by reference). Methods of transforming plant cells are well known in the art and include, for example, Agrobacterium-mediated transformation, biolistic transformation, direct injection, electroporation, and viral transformation. Methods of transforming bacterial and yeast cells are also well known in the art. Mammalian cell lines useful as expression hosts are well known in the art and include a number of immortalized cell lines available from the American Type Culture Collection (ATCC). These cell lines include, but are not limited to, Chinese hamster ovary (CHO) cells, N50 cells, SP2 cells, HEK-293T cells, NIH-3T3 cells, HeLa cells, baby hamster kidney (BHK) cells, African green monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, and many others. Particularly preferred cell lines are selected by determining which cell lines have high expression levels. Other useful cell lines are insect cell lines, such as Sf9 or Sf21 cells. When recombinant expression vectors encoding antibody genes are introduced into mammalian host cells, the ability to produce the antibody is determined by culturing the host cells in a culture medium under conditions suitable for expression of the antibody. More preferably, the antibody is secreted into the culture medium from which it is recovered. Standard protein purification methods can be employed. Plant host cells include, for example, Nicotiana, Arabidopsis, duckweed, corn, wheat, potato, and the like. Bacterial host cells include E. coli and Streptomyces. Yeast host cells include Schizosaccharomyces pombe, Saccharomyces cerevisiae, and Pichia pastoris. In addition, various known techniques can be used to enhance expression of the antibodies of the application from the production cell line. For example, the glutamine synthetase gene expression system (GS system) is a common method used to enhance expression under certain conditions.European Patent Nos. 0 216 846, 0 256 055, 0 323 997, and 0 338 841 have all or in part discussed GS systems. Antibodies expressed by different cell lines or transgenic animals can have different glycosylation from one another. However, all antibodies encoded by the nucleic acid molecules provided herein, or comprising the amino acid sequences provided herein, are part of the present application regardless of their glycosylation.

[0167] Transgenic animals and plants

[0168] Anti-Kp antibodies of the present application can also be produced transgenically, i.e., by generating transgenic mammals or plants that carry the immunoglobulin heavy and light chain sequences of interest and producing the antibodies in recoverable form therefrom. With respect to transgenic production in mammals, anti-Kp antibodies can be produced in and recovered from the milk of goats, cows, or other mammals. See, e.g., U.S. Patent Nos. 5,827,690, 5,756,687, 5,750,172, and 5,741,957, which are incorporated herein by reference. In some embodiments, a non-human transgenic animal comprising a human immunoglobulin locus is immunized with Kp whole cells or immunogenic portions thereof (e.g., one or more surface antigens as described above). Methods of making antibodies in plants are described in, e.g., U.S. Patent Nos. 6,046,037 and 5,959,177, which are incorporated herein by reference.

[0169] In some embodiments, the non-human transgenic animal or plant is produced by standard transgenic techniques by introducing one or more nucleic acid molecules encoding an anti-Kp antibody of the application into the animal or plant. See Hogan and U.S. Patent 6,417,429, supra. The transgenic cell used to make the transgenic animal can be an embryonic stem cell or a somatic cell or a zygote. The transgenic non-human organism can be a chimera, a non-chimeric heterozygote, and a non-chimeric homozygote. See, e.g., Hogan et al. Manipulating the Mouse Embryo: A Laboratory Manual Second Edition, Cold Spring Harbor Press (1999); Jackson et al. Mouse Genetics and Transgenics: A Practical Approach, Oxford University Press (2000); and Pinkert, Transgenic Animal Technology: A Laboratory Handbook, Academic Press (1999), which are incorporated herein by reference. In some embodiments, the transgenic non-human animal is targeted for disruption and replacement by a targeted construct encoding a heavy chain and / or light chain of interest. In one embodiment, the transgenic animal comprises and expresses nucleic acid molecules encoding heavy and light chains that specifically bind to Kp, preferably to (i) the capsular polysaccharide of a Kp strain according to any of the embodiments disclosed herein or (ii) the O-antigen; or (iii) both (i) and (ii). In one embodiment, the transgenic animal comprises and expresses nucleic acid molecules encoding heavy and light chains that specifically bind to the surface antigen of at least one NDM-producing Kp strain according to any of the variants disclosed herein, preferably to the surface antigen of at least two, at least three, at least four, or at least five strains. In some embodiments, the transgenic animal comprises nucleic acid molecules encoding a modified antibody (e.g., a single chain antibody, a chimeric antibody, or a humanized antibody). The anti-Kp antibody can be made in any transgenic animal. In one embodiment, the non-human animal is a mouse, a rat, a sheep, a pig, a goat, a cow, or a horse. The non-human transgenic animal expresses the encoded polypeptides in blood, milk, urine, saliva, tears, mucus, and other bodily fluids.

[0170] Class switching

[0171] Another aspect of the application provides a method of class or subclass switching an anti-Kp antibody to another class or subclass. In some embodiments, a nucleic acid molecule encoding a VL or VH (not including sequences encoding a CL or CH) is isolated using methods well known in the art. The nucleic acid molecule is then operably linked to a nucleotide sequence encoding a CL or CH from the desired immunoglobulin class or subclass. This can be accomplished using a vector or nucleic acid molecule comprising a CL or CH chain, as described above. For example, an anti-Kp antibody that is initially IgM can be class switched to IgG. Further, class switching can be used to switch one IgG subclass to another, for example from IgGl to IgG2. Another method for producing an antibody of the application comprising a desired isotype includes the steps of isolating a nucleic acid encoding a heavy chain of an anti-Kp antibody and a nucleic acid encoding a light chain of an anti-Kp antibody, isolating a sequence encoding a VH region, linking the VH sequence to a sequence encoding a heavy chain constant domain of a desired isotype, expressing the light chain gene and the heavy chain construct in a cell, and collecting the anti-Kp antibody having the desired isotype.

[0172] Modifying antibodies

[0173] According to one embodiment, the application provides an immunospecific polypeptide comprising at least one variable domain, in particular at least one heavy chain variable domain (VH) and light chain variable domain (VL) of a human monoclonal antibody as defined in any of the embodiments disclosed herein, or CDRs as defined in any of the embodiments disclosed herein, wherein the immunospecific polypeptide is, for example, a multispecific antibody, a bispecific antibody, a trispecific antibody, a diabody, a triabody, a tetrabody, a minibody, a linear antibody, a monoclonal antibody, a chelating recombinant antibody, a triabody, a bibody, an intrabody, a nanobody, a binding domain immunoglobulin fusion protein, a fusion antibody, an immunoadhesin, or an antigen binding portion thereof.

[0174] In a particular embodiment, the antibody or antigen binding fragment is a Fab, F(ab')2, Fv or single chain antibody fragment.

[0175] In certain embodiments, the antibody or antigen-binding fragment thereof in any of the embodiments disclosed herein is bispecific, trispecific, or multispecific. Bispecific, trispecific, or multispecific antibodies of the application can be formed using methods well known in the art, e.g., chemically coupling one or more antibodies or antigen-binding variable domains disclosed herein to each other and / or to a different epitope binding polypeptide, wherein the binding domains of the bispecific, trispecific, or multispecific molecule exhibit affinity for at least two different antigens. For example, an antibody according to the application can comprise a first and a second VL domain, or a first and a second VH domain, wherein the first and second domains have different binding specificities (i.e., bind different antigens).

[0176] According to one preferred embodiment, the antibody or antigen-binding fragment thereof according to the application comprises a first and a second VL domain, or a first and a second VH domain, according to any of the embodiments disclosed in the present specification and claims, wherein the first and second domains have different binding specificities for Kp (i.e., bind different antigens).

[0177] According to one object of the application, the immunospecific polypeptide of the application comprises a VL domain or a VH domain according to any of the embodiments disclosed in the present specification and claims, and an antigen binding polypeptide, wherein the VL domain or VH domain exhibits a different binding specificity than the polypeptide. In certain embodiments of the application, at least one antigen binding domain of the bispecific, trispecific, or multispecific antibody or antigen-binding fragment of the application immunospecifically binds Kp. In other embodiments of the application, at least one antigen binding domain of the bispecific, trispecific, or multispecific molecule of the application immunospecifically binds a surface antigen of Kp.

[0178] In another embodiment, the present application provides a fusion antibody or immunoadhesin prepared to comprise all or a portion of at least one anti-Kp antibody of the present application according to any of the embodiments disclosed herein linked to another polypeptide. In one embodiment, only the variable domains of at least one, at least two, or at least three anti-Kp antibodies according to any of the embodiments disclosed herein are linked to the polypeptide. In another embodiment, the VH domain of an anti-Kp antibody is linked to a first polypeptide, while the VL domain of an anti-Kp antibody is linked to a second polypeptide, which is linked to the first polypeptide such that the VH and VL domains can interact with one another to form an antigen binding site. In another embodiment, the VH domain is separated from the VL domain by a linker such that the VH and VL domains can interact with one another (see the section on "Single Chain Antibodies" below). The VH-linker-VL antibody is then linked to a polypeptide of interest. The fusion antibody can be used to direct the polypeptide of interest to cells or tissues expressing the Kp surface antigen. The polypeptide of interest can be a therapeutic agent, such as a toxin, a chemokine, or other modulating protein, or it can be a diagnostic agent, such as an enzyme that is easily visualized, e.g., horseradish peroxidase. In addition, fusion antibodies can be constructed in which two (or more) single chain antibodies are linked to one another.

[0179] This is useful if one wants to create a bivalent or multivalent antibody on a single polypeptide chain, or if one wants to create a bispecific antibody or nanobody.

[0180] According to a preferred embodiment, the antibody or antigen binding fragment is a single chain antibody (scFv). To construct a single chain antibody (scFv), the VH and VL encoding DNA fragments according to any of the embodiments disclosed herein are operatively linked to another fragment encoding a flexible linker (e.g., encoding the amino acid sequence (Gly4-Ser)3) such that the VH and VL sequences can be expressed as a contiguous single chain protein in which the VL and VH domains are linked by the flexible linker. See, e.g., Bird et al., Science 242:423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); McCafferty et al., Nature 348:552-554 (1990).

[0181] If only a single VH and VL are used, the single chain antibody can be monovalent; if two VH and VL are used, the single chain antibody can be bivalent; if more than two VH and VL are used, the single chain antibody can be multivalent.

[0182] Bispecific or multivalent antibodies that specifically bind to a Kp surface antigen and another molecule can be generated. Bispecific antibodies or antigen binding fragments can be produced by a variety of methods including hybridoma fusion or linking of Fab’ fragments. See, e.g., Songsivilai and Lachmann, Clin. Exp. Immunol. 79:315-321 (1990), Kostelny et al., J. Immunol. 148: 1547-1553 (1992).

[0183] In addition, bispecific antibodies can be formed as “diabodies” or “Janusins.” In some embodiments, the bispecific antibody binds to two different epitopes of a Kp surface antigen according to any of the variants disclosed herein. In some embodiments, the bispecific antibody has a first heavy chain and a first light chain from the following monoclonal antibodies: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13, and additional antibody heavy and light chains. In some embodiments, the additional light and heavy chains are also from one of the monoclonal antibodies identified above, but are different from the first heavy and light chains. In some embodiments, the modified antibodies described above are made using one or more variable domains or CDR regions of the human anti-Kp monoclonal antibodies provided herein. The production of bispecific, trispecific, or multispecific antibodies or nanobodies according to any of the variants disclosed herein using one or more antigen binding regions or variable domains of a human monoclonal antibody according to any of the embodiments disclosed in the specification and claims also forms part of the present application.

[0184] The immunospecific polypeptides, such as multispecific antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies, chelating recombinant antibodies, triabodies, diabodies, monoclonal antibodies, intrabodies, nanobodies, binding-domain immunoglobulin fusion proteins, fusion antibodies, immunoadhesins, or antigen-binding fragments thereof according to any of the embodiments disclosed in the present specification and claims, can be used for prophylactic or therapeutic treatment of a Klebsiella pneumoniae infection or a condition or disease resulting from such an infection, in particular for prophylaxis and / or treatment of an infection with a drug-resistant or multi-drug resistant Klebsiella pneumoniae strain, in particular an infection with a drug-resistant or multi-drug resistant Klebsiella pneumoniae strain according to any of the embodiments disclosed herein.

[0185] Accordingly, the present application also provides a pharmaceutical composition comprising an immunospecific polypeptide, such as one or more multispecific antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies, monoclonal antibodies, chelating recombinant antibodies, triabodies, diabodies, intrabodies, nanobodies, binding-domain immunoglobulin fusion proteins, fusion antibodies, immunoadhesins, or antigen-binding fragments thereof according to any of the embodiments disclosed in the present specification and claims, and at least one pharmaceutically acceptable carrier. The composition is used for prophylaxis and / or treatment of a Kp infection according to any of the embodiments disclosed herein, preferably for prophylaxis and / or treatment of an infection with the Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein.

[0186] The multispecific antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies, chelating recombinant antibodies, triabodies, diabodies, intrabodies, nanobodies, binding-domain immunoglobulin fusion proteins, fusion antibodies, immunoadhesins, or antigen-binding fragments thereof according to any of the embodiments disclosed in the present specification and claims can also be used in a method for in vitro diagnosis of Kp, or for designing a vaccine against Kp according to any of the variants disclosed in the present specification and claims, preferably a vaccine against the Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein.

[0187] Derivatizing and labeling antibodies

[0188] An anti-Kp antibody or antigen-binding portion of the present application can be derivatized or linked to another molecule (e.g., another peptide or protein). Typically, the antibody or portion thereof is derivatized so that binding to Kp surface antigens according to any of the embodiments disclosed herein is not adversely affected by the derivatization or label. Thus, antibodies and antibody portions of the present application are intended to include both full-length and modified forms of the human anti-Kp antibodies described herein. For example, an antibody or antibody portion of the present application can be functionally linked (by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or diabody), a detector molecule, a cytotoxic agent, a pharmaceutical agent, and / or a protein or peptide that can mediate association of the antibody or antibody portion with another molecule (e.g., a streptavidin core region or a polyhistidine tag). One type of derivatized antibody is produced by cross-linking two or more antibodies (of the same or different type, e.g., to produce bispecific antibodies). Suitable cross-linking agents include heterobifunctional (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide ester) or homobifunctional (e.g., disuccinimidyl suberate) cross-linking reagents. Such cross-linking agents are available from Pierce Chemical Company, Rockford, IL.

[0179] Another derivatized antibody is a labeled antibody. Detecting agents that can be used to derivatize an antibody or antigen-binding portion of the present application include fluorescent compounds, including fluorescein, fluorescein isothiocyanate, rhodamine, phycoerythrin, 5-dimethylamine-l-napthalenesulfonyl chloride, lanthanide phosphors, and the like. Antibodies can also be labeled with enzymes that can be used for detection, e.g., horseradish peroxidase, beta-galactosidase, luciferase, alkaline phosphatase, glucose oxidase, and the like. When antibodies are labeled with a detectable enzyme, the presence of the enzyme can be detected by adding additional reagents that are substrates for the enzyme, which produce a recognizable reaction product. For example, when horseradish peroxidase is present, the addition of hydrogen peroxide and diaminobenzidine results in a detectable colored reaction product. Antibodies can also be labeled with biotin, and detected through indirect measurement of avidin or streptavidin binding. Antibodies can also be labeled with a predetermined polypeptide epitope (e.g., a leucine zipper pair sequence, a tag for epitope tags, a metal binding domain, an epitope tag) that is recognized by a secondary reporter molecule. In some embodiments, the label is attached by way of a spacer arm to reduce potential steric hindrance. Anti-Kp antibodies can also be labeled with radiolabeled amino acids. The radiolabel can be used for both diagnostic and therapeutic purposes. In addition, the radiolabel can serve as a toxin for cancer cells or tumors for treatment. In some embodiments, the anti-Kp antibodies can be labeled with paramagnetic, radioactive, or florigenic ions that can be detected by imaging.In some embodiments, the paramagnetic ion is chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III), or erbium (III). In other embodiments, the radioactive ion is iodine 123, technetium 99, indium 111, rhenium 188, rhenium 186, copper 67, iodine 131, yttrium 90, iodine 125, astatine 211, and gallium 67. In other embodiments, the anti-Kp antibody is labeled with an X-ray imaging agent, such as lanthanum (III), gold (III), lead (II), and bismuth (III).

[0189] Compositions and kits

[0190] The present application relates to a composition comprising a human anti-Kp antibody of the present application and one or more pharmaceutically acceptable excipients and / or carriers.

[0191] In certain embodiments, the composition can comprise one or more antibodies or binding portions thereof of any of the foregoing embodiments.

[0192] In a preferred embodiment, the composition comprises one or more antibodies selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ08-H10, SBJ08-N23, SBJ08-K19, SBJ08-M13, SBJ03-L02, SBJ08-J10, SBJ10-H18, SBJ08-O03, SBJ05-K07, SBJ09-I10, and SBJ11-C06, preferably comprising all of these antibodies.

[0193] In another preferred embodiment, the composition comprises one or more antibodies selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, SBJ05-M13, SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04, and SBJ05-B17, preferably comprising all of these antibodies.

[0194] In another preferred embodiment, the composition comprises one or more antibodies selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, and SBJ05-M13, preferably comprising all of these antibodies.

[0195] In another preferred embodiment, the composition comprises one or more antibodies selected from the group consisting of SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04 and SBJ05-B17, preferably comprising all of these antibodies.

[0196] In one embodiment, the composition can further comprise one or more multispecific antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies, chelating recombinant antibodies, triabodies, bibodies, intrabodies, nanobodies, binding domain immunoglobulin fusion proteins, fusion antibodies, immunoadhesins, or antigen-binding fragments thereof according to any of the embodiments disclosed in the specification and claims.

[0197] The compositions according to any of the embodiments disclosed herein are particularly useful for preventing and / or treating Kp infections. In some embodiments, the subject being treated is a human. In other embodiments, the subject is a veterinary subject. In some embodiments, the antagonistic anti-Kp antibodies that bind to the capsular polysaccharide and the antagonistic anti-Kp antibodies that bind to the O antigen, or antigen-binding portions of either or both, are co-administered to the subject together or separately. In certain embodiments, the antibodies are present in a composition comprising a pharmaceutically acceptable carrier. In another embodiment, one or more antagonistic Kp antibodies of the present application are administered in combination with one or more additional antagonistic antibodies that bind to a different surface antigen of Kp or to a different epitope on a Kp surface antigen, and / or that bind to a surface antigen from a different Kp isolate.

[0198] As used herein, "pharmaceutically acceptable carrier" means any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Some examples of pharmaceutically acceptable carriers include water, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like, as well as combinations thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Further examples of pharmaceutically acceptable substances include wetting agents or minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the antibody. The compositions of the present application can be in unit- or multi-dose form, for example, as a liquid, semi-solid or solid dosage form, such as liquid solutions (e.g., injectable solutions and infusions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories, for example. The preferred form depends on the intended mode of administration and therapeutic application. Typical preferred compositions are in the form of injectable solutions or infusions, for example, similar to compositions used for passive immunization of humans. The preferred mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In one embodiment, the antibody is administered by intravenous infusion or injection. In another embodiment, the antibody is administered by intramuscular or subcutaneous injection. Therapeutic compositions typically are sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable for high drug concentration. Sterile injectable solutions can be prepared by incorporating the anti-Kp antibody in the required amount in the appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. Proper fluidity can be maintained, for example, by the use of coating such as lecithin, by the maintenance of required particle size in the case of dispersion and by the use of surfactants. Prolonged absorption of injectable compositions can be brought about by the inclusion of agents that delay absorption, for example, monostearate salts and gelatin. The antibodies of the present application can be administered by a variety of methods known in the art, although for many therapeutic applications, the preferred route / mode of administration is subcutaneous, intramuscular or intravenous infusion. Those of skill in the art will appreciate that the route and / or mode of administration will vary depending upon the desired results and the condition of the subject. Other modes of administration include intraperitoneal, intrabronchial, transmucosal, intraspinal, intrasynovial, intra-aortic, intranasal, ocular, otic, topical, and buccal. In certain embodiments, the active compound of the antibody composition can be prepared with a carrier that will protect the antibody against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many of these formulations are known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug Delivery Systems (J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978). The present application also provides compositions suitable for inhaled delivery of an anti-Kp antibody described herein. The anti-Kp antibody can be conveniently delivered to the subject in the form of an aerosol spray from pressurized packs or a nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges (e.g., gelatin) for use in an inhaler or insufflator can be formulated to contain a powder mix of the compound and a suitable powder base such as lactose or starch. Formulations for pulmonary delivery of antibodies are described in Dellamary et al. (2004) J Control Release. ; 95(3):489-500. The present application also provides compositions suitable for administration via the oral mucosa comprising an anti-Kp antibody described herein. Transmucosal delivery refers to delivery of a delivery vehicle across the mucosa of the oral cavity, pharynx, or esophagus, which is distinct from, e.g., traditional oral delivery, in which absorption of the drug occurs in the gut. Thus, administration routes in which an anti-Kp antibody is absorbed through the buccal, sublingual, gingival, pharyngeal, and / or esophageal mucosa are all encompassed by the term "transmucosal delivery" as used herein. To administer the anti-Kp antibody transmucosally, the anti-Kp antibody can be formulated, e.g., as a suckable pastille (see U.S. Patent No. 5,711,961) or a buccal patch (see, e.g., U.S. Patent No. 5,298,256). The present application also provides compositions suitable for administration via the vaginal mucosa comprising an anti-Kp antibody described herein. The anti-Kp antibody of the present application can be formulated as a vaginal suppository, foam, cream, tablet, capsule, ointment, or gel. In certain embodiments, the composition comprising the anti-Kp antibody is formulated with a permeant suitable for the transmucosal barrier to be penetrated. Such permeants are generally known in the art, including, e.g., bile salts and fusidic acid derivatives for transmucosal administration. In certain embodiments, the anti-Kp antibody of the present application can be administered orally, e.g., with an inert diluent or an assimilable edible carrier. The compound (and other ingredients if desired) can also be enclosed in a hard or soft shell gelatin capsule, compressed into tablets, or added to the subject's diet.For oral therapeutic administration, the anti-Kp antibody can be mixed with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. For administration by modes of use other than parenteral, it can be necessary to coat the compound or to co-administer the compound with a material to prevent its inactivation. Additional active compounds can also be added to the composition. In certain embodiments, the inhibitory anti-Kp antibody of the present application is co-formulated and / or co-administered with one or more additional therapeutic agents, particularly antimicrobial agents, more particularly one or more beta-lactamase inhibitors.

[0199] These therapeutic agents include, but are not limited to, antibodies that bind other targets, photosensitizers, androgens, estrogens, non-steroidal anti-inflammatory agents, anti-hypertensive agents, analgesic agents, anti-depressants, antibiotics, anti-cancer agents, anesthetics, anti-emetics, anti-infective agents, contraceptives, anti-diabetic agents, steroids, anti-allergic agents, chemotherapeutic agents, anti-migraine agents, anti-smoking agents, anti-viral agents, immunosuppressive agents, thrombolytic agents, cholesterol-lowering agents, and anti-obesity agents. Therapeutic agents also include peptide analogs that inhibit or kill Kp cells, as well as agents that inhibit Kp function or activity. In one embodiment, other inhibitory agents are also known in the art.

[0200] Such combination therapy can require lower dosages of the inhibitory anti-Kp antibody and the co-administered agent, thereby avoiding toxicity or complications associated with each therapy when used alone. In certain embodiments, the one or more therapeutic agents co-formulated and / or co-administered with the inhibitory anti-Kp antibody of the present application is an antibacterial agent. Antibacterial agents include antibiotics (e.g., antibacterial agents), antiviral agents, antifungal agents, and antiprotozoal agents. Non-limiting examples of antibacterial agents are sulfonamides, trimethoprim-sulfamethoxazole, quinolones, penicillins, and cephalosporins.

[0201] The compositions of the present application can comprise a "therapeutically effective amount" or a "prophylactically effective amount" of the antibody or antigen-binding portion of the present application. A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the antibody or antibody portion can vary depending upon factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody portion to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at the early stages of disease, the prophylactically effective amount can be less than the therapeutically effective amount.

[0202] The dosage regimen can be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response). For example, a single bolus can be administered, several divided doses can be administered over time or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the application are dictated by and directly dependent on (a) the unique characteristics of the anti-Kp antibody or portion thereof and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of compounding such antibodies for the treatment of individuals with allergies.

[0203] In a preferred embodiment, the pharmaceutical composition comprises a human monoclonal antibody or antigen binding portion thereof equal to or less than 400 mg per dosage unit, more preferably less than 400, 350, 300, 250, 200, 150, 100, 50, 25, 10 mg per dosage unit.

[0204] The pharmaceutical composition comprising such dosage unit is preferably for parenteral administration, e.g. for intravenous, subcutaneous, intraperitoneal or intramuscular administration. In a preferred embodiment, the pharmaceutical composition is in liquid form, at a concentration of 20 to 200 mg / ml, more preferably 40 to 80 mg / ml.

[0205] An exemplary, non-limiting range of a therapeutically or prophylactically effective amount of the antibody or antibody portion of the application is 0.025 to 50 mg / kg, more preferably 0.1 to 5 mg / kg, more preferably 0.1 to 5, 0.1 to 4 or 0.25 to 3 mg / kg.

[0206] In one embodiment, the present application provides a human monoclonal antibody or antigen binding portion according to any of the embodiments disclosed herein for use in a method of prophylactic or therapeutic treatment of a Kp infection or a condition or disease caused by such infection, in particular an infection caused by at least one multi-drug resistant strain of Kp, preferably an infection caused by a Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, more preferably an infection caused by at least one NDM-producing Kp strain according to any of the variants disclosed herein, wherein the method comprises the step of administering to the patient a therapeutically effective amount of said antibody or antigen binding portion thereof, preferably 0.025 to 50 mg / kg, more preferably 0.1 to 5 mg / kg, more preferably 0.1 to 5, 0.1 to 4 or 0.25 to 3 mg / kg, once a day, for example for at least one, two, three, four, five, six, seven, eight, nine, ten, eleven days. The patient is a mammal, preferably a human.

[0207] It is noted that dosage values can vary with the type and severity of the condition to be alleviated. It is to be understood that the specific treatment regime for any particular patient will depend on a variety of factors, including the patient's general health status, the specific composition to be administered, and the judgment of the treating physician. The dosage range disclosed herein is exemplary only, and is not intended to suggest that the scope or practice of the claimed compositions is limited to such exemplary dose ranges. Another aspect of the application provides a kit comprising an anti-Kp or antigen binding portion of the application, or a composition comprising such an antibody or antigen binding fragment. In addition to the antibody or composition, the kit can comprise a diagnostic or therapeutic agent. The kit can also comprise instructions for a diagnostic or therapeutic method, as well as packaging material, such as, but not limited to, ice packs, dry ice, Styrofoam, bubble wrap, plastic, cellophane, shrink wrap, bubble wrap, cardboard, and starch peanut. In one embodiment, the kit comprises an antibody or a composition comprising the antibody, and a diagnostic agent useful in the methods described below. In yet another embodiment, the kit comprises an antibody or a composition comprising the antibody, and one or more therapeutic agents useful in the methods described below.

[0208] In one embodiment, the antibody or binding portion thereof according to any of the embodiments disclosed herein, or a composition comprising such an antibody, is used to prevent or treat a patient infected with Kp, in particular a patient infected with at least one drug resistant or multi-drug resistant Kp strain, preferably a K. pneumoniae capsular type K64 strain according to any of the variants disclosed herein, for example a NDM-producing Kp strain, in particular a NDM-1 positive Kp strain, or any other strain disclosed in the specification and claims.

[0209] The use of such antibodies and compositions of Kp-specific mAbs includes, but is not limited to, passive immunization of at-risk populations (e.g., occupational exposure populations, populations living in endemic areas), as well as treatment of acute cases (whether or not hospitalized). The present application also relates to a composition for inhibiting Kp infection in a mammal, in particular infection with at least one drug resistant or multi-drug resistant Kp strain, more particularly a NDM-1 -producing Kp strain (e.g., ST147 NDM-1 strains), said composition comprising an amount of an antibody of the application, and optionally in combination with an amount of a beta-lactamase inhibitor, wherein the doses of the anti-Kp antibody and the inhibitor are jointly effective to kill the bacteria.

[0210] Diagnostic method uses

[0211] The antibodies according to the present application can also be used as diagnostic tools for rapid detection of Kp infection, in particular of infection with at least one drug resistant or multi-drug resistant Kp strain, preferably of infection with a Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, more particularly of infection with a NDM-producing Kp strain, or of infection with any other Kp strain according to any of the variants disclosed in the present specification. In another aspect, the present application provides diagnostic methods. Anti-Kp antibodies can be used to detect Kp in a biological sample, either in vitro or in vivo. In one embodiment, the present application provides a method for diagnosing the presence or location of Kp cells in a subject in need thereof. Anti-Kp antibodies can be used in conventional immunoassays, including but not limited to ELISA, RIA, flow cytometry, tissue immunohistochemistry, Western blotting (immunoblotting), or immunoprecipitation. Anti-Kp antibodies of the present application can be used to detect Kp in humans. The present application provides a method of detecting Kp cells in a biological sample, the method comprising contacting the biological sample with an anti-Kp antibody of the present application, and detecting the bound antibody. In one embodiment, the anti-Kp antibody is directly labeled with a detectable marker. In another embodiment, the anti-Kp antibody (primary antibody) is unlabeled, while a secondary antibody or other molecule that can bind to the anti-Kp antibody is labeled. It is well known to those skilled in the art that a secondary antibody should be chosen that is capable of specifically binding to the particular species and to the particular primary antibody type. For example, if the anti-Kp antibody is a human IgG, then the secondary antibody can be an anti-human IgG. Other molecules that can bind to antibodies include, but are not limited to, protein A and protein G, both of which are commercially available, for example, from Pierce Chemical Co. Examples of biological samples for use in the diagnostic methods disclosed herein are urine, feces, blood, saliva, biopsy samples, cerebrospinal fluid, nasopharyngeal and oropharyngeal washes, sputum, endotracheal aspirates, bronchoalveolar lavage fluid, or other biological samples obtainable from a human subject.

[0212] Suitable labels for the antibody or secondary antibody are disclosed above, including various enzymes, prosthetic groups, fluorescent, luminescent, and radiative moieties. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; examples of luminescent materials include luminol. In other embodiments, Kp or Kp surface antigen standards labeled with a detectable moiety and unlabeled anti-Kp antibody can be used in a competitive immunoassay to determine Kp cells in a biological sample. In this assay, the biological sample, labeled Kp or Kp surface antigen standard, and anti-Kp antibody are combined and the amount of labeled Kp or Kp antigen standard bound to the unlabeled antibody is determined. The amount of Kp cells or Kp surface antigen in the biological sample is inversely proportional to the amount of labeled Kp or Kp antigen standard bound to the anti-Kp antibody. The above-disclosed immunoassays can be used for a variety of purposes. For example, anti-Kp antibodies can be used to detect Kp cells in a sample of cultured cells, or in a diagnostic assay for a subject sample. Following a diagnostic method according to any of the embodiments disclosed herein, a further step of administering an anti-Kp drug to a positive subject can be performed, e.g., according to any of the treatment methods disclosed herein.

[0213] Therapeutic method uses

[0214] In another embodiment, the present application provides a method of neutralizing or inhibiting or killing Kp cells by administering to a patient in need thereof one or more anti-Kp antibodies according to any of the embodiments disclosed herein. Any of the types of antibodies described herein can be used for treatment. In various embodiments, the anti-Kp antibody is a human antibody. In some embodiments, the antibody or antigen binding portion thereof binds to at least one capsular antigen or polysaccharide and / or O antigen of a Kp strain according to any of the variants disclosed in the specification. In some embodiments, the patient is a human patient. Alternatively, the patient can be a mammal infected with Kp. In one embodiment, the present application provides a method of treating, adjunct treating, preventing or adjunct preventing a Kp infection in a subject, in particular an infection caused by at least one drug resistant or multi-drug resistant Kp strain, preferably an infection caused by Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, more particularly an infection caused by a NDM-producing Kp strain or any other Kp strain according to any of the variants disclosed in the specification, and a condition or disease resulting from such infection, by administering to the subject a therapeutically effective amount or a prophylactically effective amount of one or more anti-Kp antibodies of the present application. Antibodies and antigen binding fragments thereof that are antagonists of Kp or Kp surface antigens are useful as therapeutic agents for Kp infection. The antibodies can be administered locally or systemically. Therapeutic compositions comprising the anti-Kp antibodies can be administered to the subject in a variety of pharmaceutically acceptable dosage forms familiar to those skilled in the art, for example, by oral, nasal, vaginal, buccal, rectal, ocular, or pulmonary routes. For example, prior to administration, the formulation containing the anti-Kp antibodies can be conveniently packaged in a dual chamber unit dosage container, one chamber containing a lyophilized anti-Kp antibody formulation and the other chamber containing physiological saline. Serum concentrations of the antibodies can be measured by any method known in the art.

[0215] In some aspects, the method according to any of the embodiments disclosed herein comprises administering one or more antibodies selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ08-H10, SBJ08-N23, SBJ08-K19, SBJ08-M13, SBJ03-L02, SBJ08-J10, SBJ10-H18, SBJ08-O03, SBJ05-K07, SBJ09-I10, and SBJ11-C06.

[0216] In another preferred embodiment, the method comprises administering one or more antibodies selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, SBJ05-M13, SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04, and SBJ05-B17.

[0217] In another preferred embodiment, the method comprises administering one or more antibodies selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, and SBJ05-M13.

[0218] In another preferred embodiment, the method comprises administering one or more antibodies selected from the group consisting of SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04, and SBJ05-B17.

[0219] In another embodiment, the antibody of the present application is administered to a subject in combination with another therapeutic agent. In one embodiment, the additional therapeutic agent can treat the symptoms of a Kp infection alone, and can optionally act synergistically with the antibody. One of skill in the art can select an additional therapeutic agent for administration to treat an infection. Co-administration of the antibody with the additional therapeutic agent (combination therapy) encompasses administration of a composition comprising both the anti-Kp antibody and the additional therapeutic agent, as well as administration of two or more separate compositions, one comprising the anti-Kp antibody and the other comprising the additional therapeutic agent. Further, while co-administration or combination therapy generally means that the antibody and the additional therapeutic agent are administered at the same time, it also encompasses situations where the antibody and the additional therapeutic agent are administered at different times. For example, the antibody can be administered once every three days, while the additional therapeutic agent is administered once a day. Alternatively, the antibody can be administered prior to or after treatment with the additional therapeutic agent, for example, after a patient has failed treatment with the other agent. Similarly, administration of the anti-Kp antibody can also precede or follow other treatments.

[0220] The antibody and the one or more additional therapeutic agents (combination therapy) can be administered once, twice, or for a period of time until the condition is treated, alleviated, or cured. Preferably, the combination therapy is administered in multiple doses. The combination therapy is administered in doses ranging from three times a day to once every six months. The schedule of administration can be set, for example, at three times a day, twice a day, once a day, once every two days, once every three days, once a week, once every two weeks, once a month, once every two months, once every three months, and once every six months, or can be administered continuously by a minipump. The combination therapy can be administered orally, mucosally, buccally, intranasally, by inhalation, intravenously, subcutaneously, intramuscularly, or parenterally.

[0221] In certain aspects, the present application provides methods for treating, preventing or ameliorating the symptoms of a Kp-mediated disease, particularly an infection with at least one drug resistant or multi-drug resistant Kp strain, preferably an infection with a Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, more particularly an infection with an NDM-producing Kp strain, or an infection with any other Kp strain according to any of the variants disclosed in the present specification, in a subject in need thereof, comprising the step of administering to said subject an antibody or antigen binding portion according to any of the foregoing embodiments, further comprising administering at least one additional therapeutic agent selected from the group consisting of: (a) one or more antibodies selected from the group consisting of: SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13;

[0222] In particular,

[0223] (b) one or more antibodies that specifically bind to multiple Kp strain surface antigens; and / or

[0224] (c) one or more neutralizing antibodies that do not bind to Kp surface antigens; and / or

[0225] (d) one or more antibacterial agents; and / or

[0226] (e) one or more beta-lactamase inhibitors.

[0227] In certain aspects, the present application provides a kit for treating, preventing or alleviating symptoms of a Kp-mediated disease, in particular an infection with at least one drug resistant or multi-drug resistant Kp strain, preferably an infection with a Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, more particularly an infection with a NDM-producing Kp strain, or an infection with any other Kp strain according to any of the variants disclosed in the present specification, in a subject in need thereof, the kit comprising a) one or more antibodies selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ03-L02, SBJ05-B17, SBJ05-C11, SBJ05-D08, SBJ05-D14, SBJ05-K07, SBJ05-M13, SBJ05-N02, SBJ08-F04, SBJ08-H10, SBJ08-J10, SBJ08-K19, SBJ08-M13, SBJ08-N23, SBJ08-O03, SBJ09-G14, SBJ09-I10, SBJ09-M06, SBJ10-H18, SBJ11-C06, SBJ11-J13; in particular,

[0228] (b) one or more of these antibodies that specifically bind to surface antigens of a plurality of Kp strains; and / or

[0229] (c) one or more neutralizing antibodies that do not bind to Kp surface antigens; and / or

[0230] (d) one or more antibacterial agents; and / or

[0231] (e) one or more beta-lactamase inhibitors.

[0232] In a preferred embodiment, the kit comprises or consists of one or more antibodies selected from the group consisting of SBJ08-O09, SBJ08-D18, SBJ03-F18, SBJ08-H10, SBJ08-N23, SBJ08-K19, SBJ08-M13, SBJ03-L02, SBJ08-J10, SBJ10-H18, SBJ08-O03, SBJ05-K07, SBJ09-I10 and SBJ11-C06, preferably all of these antibodies.

[0233] In another preferred embodiment, the kit comprises or consists of one or more antibodies selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13, SBJ05-M13, SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04 and SBJ05-B17, preferably comprises all of these antibodies or consists thereof.

[0234] In another preferred embodiment, the kit comprises or consists of one or more antibodies selected from the group consisting of SBJ05-D08, SBJ05-C11, SBJ05-D14, SBJ11-J13 and SBJ05-M13, preferably comprises all of these antibodies or consists thereof.

[0235] In another preferred embodiment, the kit comprises or consists of one or more antibodies selected from the group consisting of SBJ09-G14, SBJ05-N02, SBJ09-M06, SBJ08-F04 and SBJ05-B17, preferably comprises all of these antibodies or consists thereof.

[0236] The human monoclonal antibodies or antigen-binding portions thereof disclosed herein can also be advantageously used as diagnostic reagents in in vitro methods for detecting anti-Kp antibodies in a biological sample previously obtained from a patient, e.g. a serum, plasma, blood sample or any other suitable biological material obtained from a patient, preferably a human. These antibodies can be present in a biological sample obtained from a patient, e.g. due to a previous contact with the virus, or because a monoclonal antibody of the application has been previously administered to the patient for therapeutic, prophylactic or research purposes. Therefore, diagnostic kits comprising a human monoclonal antibody of the application or an antigen-binding portion thereof as disclosed herein as a specific reagent, specifically designed for detecting and / or quantifying anti-Kp antibodies in a biological sample previously obtained from a patient, also belong to the scope of the present application.

[0237] According to one embodiment, the kit can further comprise one or more multispecific antibodies, diabodies, triabodies, tetrabodies, minibodies, linear antibodies, chelating recombinant antibodies, triabodies, diabodies, intrabodies, nanobodies, binding-domain immunoglobulin fusion proteins, fusion antibodies, immunoadhesins, or antigen-binding fragments thereof according to any of the embodiments disclosed in the present specification and claims.

[0238] The human monoclonal antibodies or antigen binding portions thereof disclosed herein can also be advantageously used for the design of vaccines against Kp, in particular against at least one drug resistant or multi-drug resistant Kp strain, preferably against a Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein, more particularly against at least one NDM-producing Kp strain or against a Kp strain according to any of the variants disclosed in the specification and claims. As disclosed in Rappuoli, Rino, et al. "Reverse vaccinology 2.0: Human immunology instructs vaccine antigen design." The Journal of experimental medicine vol. 213, 4 (2016): 469-81. doi:10.1084 / jem.20151960, human mAbs can be used to identify protective antigens / epitopes. Structural characterization of Ab-antigen complexes can be used to guide antigen design. Thus, the use of the human monoclonal antibodies or antigen binding portions thereof disclosed herein for the design of a vaccine against Kp, in particular against NDM-producing Kp strains, also falls within the scope of the present application.

[0239] The human monoclonal antibodies or antigen binding portions thereof disclosed herein can be used for the preparation of mimotopes, such as anti-idiotype antibodies, peptides, S-protein truncations or artificial forms or other mimotopes capable of eliciting the antibodies disclosed herein. Among these, anti-idiotype antibodies are preferred. Anti-idiotype antibodies are antibodies specific for the idiotype of the neutralizing antibodies used for their preparation and are thus able to mimic the key epitopes they recognize. The preparation of anti-idiotype antibodies employs methods known per se and need not to be explained in further detail here. Thus, mimotopes for the antibodies of the present application, preferably anti-idiotype antibodies, also fall within the scope of the present application. The human monoclonal antibodies or antigen binding portions thereof disclosed herein can be used for the preparation of anti-idiotype antibodies according to methods known per se. Anti-idiotype antibodies are antibodies specific for the idiotype of the broad-spectrum neutralizing antibodies used for their preparation and are thus able to mimic the key epitopes they recognize. Thus, anti-idiotype antibodies against the monoclonal antibodies of the present application are also comprised within the scope of the present application.

[0240] Preferred embodiments of the invention

[0241] Embodiment 1. The first embodiment of the present application relates to a human monoclonal antibody or antigen binding portion thereof that specifically binds to a surface antigen of Klebsiella pneumoniae.

[0242] Embodiment 2. There is also provided herein a human monoclonal antibody or antigen binding portion thereof according to embodiment 1 that specifically binds to a surface antigen of a drug resistant or multi-drug resistant strain of Klebsiella pneumoniae.

[0243] Embodiment 3. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to embodiment 1 or 2, which specifically binds to a surface antigen of carbapenemase-producing Klebsiella pneumoniae strains, preferably New Delhi metallo-beta-lactamase (NDM)-producing strains, more preferably at least the Klebsiella pneumoniae capsular type K64 strain according to any of the variants disclosed herein.

[0244] Embodiment 4. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to embodiment 3, wherein the strain is selected from the group consisting of NDM-1 positive strains, NDM-9 positive strains and NDM-5 positive strains.

[0245] Embodiment 5. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to embodiment 3 or 4, wherein the strain is selected from the group consisting of ST147, ST258, ST493, ST307 and ST13.

[0246] Embodiment 6. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 5, wherein the surface antigen is selected from the group consisting of capsular polysaccharide or O antigen.

[0247] Embodiment 7. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to embodiment 6, wherein the O antigen is an O1 antigen or an O2 antigen.

[0248] Embodiment 8. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 7, wherein the antibody or antigen binding portion thereof provides a bacterial killing percentage of greater than 30%, preferably greater than 60%, more preferably greater than 80% as measured by an in vitro fluorescent serum bactericidal assay (F-SBA).

[0249] Embodiment 9. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 8, wherein the antibody or antigen binding portion thereof displays an 50% inhibition concentration (IC50) of less than 100 ng / ml, preferably less than 10 ng / ml, more preferably less than 5 ng / ml when detected by an in vitro fluorescent serum bactericidal assay (F-SBA).

[0250] Embodiment 10. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to embodiment 9, wherein the antibody or antigen binding portion is detected in an in vitro F-SBA assay against the ST147 NDM-1 strain of Klebsiella pneumoniae, the ST147 NDM-9 strain of Klebsiella pneumoniae and / or the ST307 NDM-5 strain of Klebsiella pneumoniae.

[0251] Embodiment 11. Also provided herein are human monoclonal antibodies or antigen binding portions thereof according to any of embodiments 1 to 10, comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH and VL comprise the following complementarity determining regions (CDRs):

[0252] - a VH CDR1 having SEQ ID NO: 1,

[0253] - a VH CDR2 having SEQ ID NO: 2,

[0254] - a VH CDR3 having SEQ ID NO: 3,

[0255] - a VL CDR1 having SEQ ID NO: 4,

[0256] - a VL CDR2 having the sequence AAS (Ala-Ala-Ser), and

[0257] - a VL CDR3 having SEQ ID NO: 6;

[0258] or

[0259] - a VH CDR1 having SEQ ID NO: 19,

[0260] - a VH CDR2 having SEQ ID NO: 20,

[0261] - a VH CDR3 having SEQ ID NO: 21,

[0262] - a VL CDR1 having SEQ ID NO: 22,

[0263] - a VL CDR2 having the sequence GAS (Gly-Ala-Ser), and

[0264] - a VL CDR3 having SEQ ID NO: 24;

[0265] or

[0266] - a VH CDR1 having SEQ ID NO: 37,

[0267] - a VH CDR2 having SEQ ID NO: 38,

[0268] - a VH CDR3 having SEQ ID NO: 39,

[0269] - a VL CDR1 having SEQ ID NO: 40,

[0270] - VL CDR2 having the sequence AAS (Ala-Ala-Ser), and

[0271] - VL CDR3 having SEQ ID NO: 42,

[0272] or

[0273] - VH CDR1 having SEQ ID NO: 55,

[0274] - VH CDR2 having SEQ ID NO: 56,

[0275] - VH CDR3 having SEQ ID NO: 57,

[0276] - VL CDR1 having SEQ ID NO: 58,

[0277] - VL CDR2 having the sequence DDY (Asp-Asp-Tyr), and

[0278] - VL CDR3 having SEQ ID NO: 60

[0279] or

[0280] - VH CDR1 having SEQ ID NO: 73,

[0281] - VH CDR2 having SEQ ID NO: 74,

[0282] - VH CDR3 having SEQ ID NO: 75,

[0283] - VL CDR1 having SEQ ID NO: 76,

[0284] - VL CDR2 having the sequence DVS (Asp-Val-Ser), and

[0285] - VL CDR3 having SEQ ID NO: 78

[0286] or

[0287] - VH CDR1 having SEQ ID NO: 91,

[0288] - VH CDR2 having SEQ ID NO: 92,

[0289] - VH CDR3 having SEQ ID NO: 93,

[0290] - VL CDR1 having SEQ ID NO: 94,

[0291] - VL CDR2 having the sequence AAS (Ala-Ala-Ser), and

[0292] - VL CDR3 having SEQ ID NO: 132

[0293] or

[0294] - VH CDR1 having SEQ ID NO: 145,

[0295] - VH CDR2 having SEQ ID NO: 146,

[0296] - VH CDR3 having SEQ ID NO: 147,

[0297] - VL CDR1 having SEQ ID NO: 148,

[0298] - VL CDR2 having the sequence EIS (Glu-Ile-Ser), and

[0299] - VL CDR3 having SEQ ID NO: 150

[0300] or

[0301] - VH CDR1 having SEQ ID NO: 163,

[0302] - VH CDR2 having SEQ ID NO: 164,

[0303] - VH CDR3 having SEQ ID NO: 165,

[0304] - VL CDR1 having SEQ ID NO: 166,

[0305] - VL CDR2 having the sequence EIS (Glu-Ile-Ser), and

[0306] - VL CDR3 having SEQ ID NO: 150

[0307] or

[0308] - VH CDR1 having SEQ ID NO: 163,

[0309] - VH CDR2 having SEQ ID NO: 164,

[0310] - VH CDR3 having SEQ ID NO: 165,

[0311] - VL CDR1 having SEQ ID NO: 166,

[0312] - VL CDR2 having the sequence AAS (Ala-Ala-Ser), and

[0313] - VL CDR3 having SEQ ID NO: 186

[0314] or

[0315] - VH CDR1 having SEQ ID NO: 181,

[0316] - VH CDR2 having SEQ ID NO: 182,

[0317] - VH CDR3 having SEQ ID NO: 183,

[0318] - VL CDR1 having SEQ ID NO: 184,

[0319] - VL CDR2 having the sequence AAS (Ala-Ala-Ser), and

[0320] - VL CDR3 having SEQ ID NO: 186

[0321] or

[0322] - VH CDR1 having SEQ ID NO: 199,

[0323] - VH CDR2 having SEQ ID NO: 200,

[0324] - VH CDR3 having SEQ ID NO: 201,

[0325] - VL CDR1 having SEQ ID NO: 202,

[0326] - VL CDR2 having the sequence DAS (Asp-Ala-Ser), and

[0327] - VL CDR3 having SEQ ID NO: 204

[0328] or

[0329] - VH CDR1 having SEQ ID NO: 217,

[0330] - VH CDR2 having SEQ ID NO: 218,

[0331] - VH CDR3 having SEQ ID NO: 219,

[0332] - a VL CDR1 having the sequence GTS(Gly-Thr-Ser),

[0333] - a VL CDR2 having the sequence QIS(Gln-Ile-Ser), and

[0334] - a VL CDR3 having the sequence DAS(Asp-Ala-Ser)

[0335] or

[0336] - a VH CDR1 having the sequence GFT(Gly-Phe-Thr),

[0337] - a VH CDR2 having the sequence IIS(Ile-Ile-Ser),

[0338] - a VH CDR3 having the sequence DAS(Asp-Ala-Ser)

[0339] - a VL CDR1 having the sequence GTS(Gly-Thr-Ser),

[0340] - a VL CDR2 having the sequence QIS(Gln-Ile-Ser), and

[0341] - a VL CDR3 having the sequence DAS(Asp-Ala-Ser)

[0342] or

[0343] - a VH CDR1 having the sequence GFT(Gly-Phe-Thr),

[0344] - a VH CDR2 having the sequence IIS(Ile-Ile-Ser),

[0345] - a VH CDR3 having the sequence DAS(Asp-Ala-Ser)

[0346] - a VL CDR1 having the sequence GTS(Gly-Thr-Ser),

[0347] - a VL CDR2 having the sequence QIS(Gln-Ile-Ser), and

[0348] - a VL CDR3 having the sequence DAS(Asp-Ala-Ser)

[0349] or

[0350] - a VH CDR1 having the sequence GFT(Gly-Phe-Thr),

[0351] - a VH CDR2 having the sequence IIS(Ile-Ile-Ser),

[0352] - a VH CDR3 having the sequence DAS(Asp-Ala-Ser)

[0353] - a VL CDR1 having the sequence AAS (Ala-Ala-Ser),

[0354] - a VL CDR2 having the sequence KVS (Lys-Val-Ser), and

[0355] - a VL CDR3 having SEQ ID NO: 294

[0356] or

[0357] - a VH CDR1 having SEQ ID NO: 289,

[0358] - a VH CDR2 having SEQ ID NO: 290,

[0359] - a VH CDR3 having SEQ ID NO: 291,

[0360] - a VL CDR1 having SEQ ID NO: 292,

[0361] - a VL CDR2 having the sequence G AS (Gly-Ala-Ser), and

[0362] - a VL CDR3 having SEQ ID NO: 312

[0363] or

[0364] - a VH CDR1 having SEQ ID NO: 307,

[0365] - a VH CDR2 having SEQ ID NO: 308,

[0366] - a VH CDR3 having SEQ ID NO: 309,

[0367] - a VL CDR1 having SEQ ID NO: 310,

[0368] - a VL CDR2 having the sequence G AS (Gly-Ala-Ser), and

[0369] - a VL CDR3 having SEQ ID NO: 312

[0370] or

[0371] - a VH CDR1 having SEQ ID NO: 325,

[0372] - a VH CDR2 having SEQ ID NO: 326,

[0373] - a VH CDR3 having the sequence of SEQ ID NO: 330,

[0374] - a VL CDR1 having the sequence of SEQ ID NO: 328,

[0375] - a VL CDR2 having the sequence of DAS (Asp-Ala-Ser), and

[0376] - a VL CDR3 having the sequence of SEQ ID NO: 329

[0377] or

[0378] - a VH CDR1 having the sequence of SEQ ID NO: 343,

[0379] - a VH CDR2 having the sequence of SEQ ID NO: 344,

[0380] - a VH CDR3 having the sequence of SEQ ID NO: 345,

[0381] - a VL CDR1 having the sequence of SEQ ID NO: 346,

[0382] - a VL CDR2 having the sequence of EVS (Glu-Val-Ser), and

[0383] - a VL CDR3 having the sequence of SEQ ID NO: 348

[0384] or

[0385] - a VH CDR1 having the sequence of SEQ ID NO: 361,

[0386] - a VH CDR2 having the sequence of SEQ ID NO: 362,

[0387] - a VH CDR3 having the sequence of SEQ ID NO: 363,

[0388] - a VL CDR1 having the sequence of SEQ ID NO: 364,

[0389] - a VL CDR2 having the sequence of GAS (Gly-Ala-Ser), and

[0390] - a VL CDR3 having the sequence of SEQ ID NO: 366

[0391] or

[0392] - a VH CDR1 having the sequence of SEQ ID NO: 379,

[0393] - a VH CDR2 having the sequence of SEQ ID NO: 380,

[0394] - a VH CDR3 having SEQ ID NO: 381,

[0395] - a VL CDR1 having SEQ ID NO: 382,

[0396] - a VL CDR2 having the sequence EVS (Glu-Val-Ser), and

[0397] - a VL CDR3 having SEQ ID NO: 383

[0398] or

[0399] - a VH CDR1 having SEQ ID NO: 397,

[0400] - a VH CDR2 having SEQ ID NO: 398,

[0401] - a VH CDR3 having SEQ ID NO: 399,

[0402] - a VL CDR1 having SEQ ID NO: 400,

[0403] - a VL CDR2 having the sequence KVS (Lys-Val-Ser), and

[0404] - a VL CDR3 having SEQ ID NO: 402

[0405] or

[0406] - a VH CDR1 having SEQ ID NO: 415,

[0407] - a VH CDR2 having SEQ ID NO: 416,

[0408] - a VH CDR3 having SEQ ID NO: 417,

[0409] - a VL CDR1 having SEQ ID NO: 418,

[0410] - a VL CDR2 having the sequence GAS (Gly-Ala-Ser), and

[0411] - a VL CDR3 having SEQ ID NO: 420

[0412] or

[0413] - a VH CDR1 having SEQ ID NO: 433,

[0414] - a VH CDR2 having the sequence RIS (Arg-Ile-Ser),

[0415] - a VH CDR3 having the sequence RDS (Arg-Asp-Ser),

[0416] - a VL CDR1 having the sequence RVS (Arg-Val-Ser),

[0417] - a VL CDR2 having the sequence RVS (Arg-Val-Ser), and

[0418] - a VL CDR3 having the sequence RDS (Arg-Asp-Ser).

[0419] Embodiment 12. Also provided herein are human monoclonal antibodies or antigen binding portions thereof according to any of embodiments 1 to 11, comprising a heavy chain variable domain (VH) and a light chain variable domain (VL),

[0420] wherein the VH has SEQ ID NO: 7 and the VL has SEQ ID NO: 8; or

[0421] wherein the VH has SEQ ID NO: 25 and the VL has SEQ ID NO: 26; or

[0422] wherein the VH has SEQ ID NO: 43 and the VL has SEQ ID NO: 44 or

[0423] wherein the VH has SEQ ID NO: 61 and the VL has SEQ ID NO: 62 or

[0424] wherein the VH has SEQ ID NO: 79 and the VL has SEQ ID NO: 80 or

[0425] wherein the VH has SEQ ID NO: 97 and the VL has SEQ ID NO: 98 or

[0426] wherein the VH has SEQ ID NO: 133 and the VL has SEQ ID NO: 134 or

[0427] wherein the VH has SEQ ID NO: 151 and the VL has SEQ ID NO: 152 or

[0428] wherein the VH has SEQ ID NO: 169 and the VL has SEQ ID NO: 170 or

[0429] wherein the VH has SEQ ID NO: 187 and the VL has SEQ ID NO: 188 or

[0430] wherein the VH has SEQ ID NO: 205 and the VL has SEQ ID NO: 206 or

[0431] wherein the VH has SEQ ID NO: 223 and the VL has SEQ ID NO: 224 or

[0432] wherein the VH has SEQ ID NO: 241 and the VL has SEQ ID NO: 242 or

[0433] wherein the VH has SEQ ID NO: 259 and the VL has SEQ ID NO: 260 or

[0434] wherein the VH has SEQ ID NO: 277 and the VL has SEQ ID NO: 278 or

[0435] wherein the VH has SEQ ID NO: 295 and the VL has SEQ ID NO: 296 or

[0436] wherein the VH has SEQ ID NO: 313 and the VL has SEQ ID NO: 314 or

[0437] wherein the VH has SEQ ID NO: 331 and the VL has SEQ ID NO: 332 or

[0438] wherein the VH has SEQ ID NO: 349 and the VL has SEQ ID NO: 350 or

[0439] wherein the VH has SEQ ID NO: 367 and the VL has SEQ ID NO: 368 or

[0440] wherein the VH has SEQ ID NO: 385 and the VL has SEQ ID NO: 386 or

[0441] wherein the VH has SEQ ID NO: 403 and the VL has SEQ ID NO: 404 or

[0442] wherein the VH has SEQ ID NO: 421 and the VL has SEQ ID NO: 422 or

[0443] wherein the VH has SEQ ID NO: 439 and the VL has SEQ ID NO: 440.

[0444] Embodiment 13. Also provided herein are human monoclonal antibodies or antigen binding portions thereof according to any of embodiments 1 to 12, wherein the VL and the VH are at least 85%, preferably at least 95%, more preferably at least 99% identical to the amino acid sequences of the following VH and VL:

[0445] a VH having SEQ ID NO: 7 and the VL having SEQ ID NO: 8; or

[0446] a VH having SEQ ID NO: 25 and the VL having SEQ ID NO: 26; or

[0447] a VH having SEQ ID NO: 43 and the VL having SEQ ID NO: 44, or

[0448] a VH having SEQ ID NO: 61 and the VL having SEQ ID NO: 62, or

[0449] a VH having SEQ ID NO: 79 and the VL having SEQ ID NO: 80, or

[0450] a VH having SEQ ID NO: 97 and the VL having SEQ ID NO: 98, or

[0451] a VH having SEQ ID NO: 133 and the VL having SEQ ID NO: 134, or

[0452] a VH having SEQ ID NO: 151 and the VL having SEQ ID NO: 152, or

[0453] a VH having SEQ ID NO: 169 and the VL having SEQ ID NO: 170, or

[0454] a VH having SEQ ID NO: 187 and the VL having SEQ ID NO: 188, or

[0455] a VH having SEQ ID NO: 205 and the VL having SEQ ID NO: 206, or

[0456] a VH having SEQ ID NO: 223 and the VL having SEQ ID NO: 224, or

[0457] a VH having SEQ ID NO: 241 and a VL having SEQ ID NO: 242, or

[0458] a VH having SEQ ID NO: 259 and a VL having SEQ ID NO: 260, or

[0459] a VH having SEQ ID NO: 277 and a VL having SEQ ID NO: 278, or

[0460] a VH having SEQ ID NO: 295 and a VL having SEQ ID NO: 296, or

[0461] a VH having SEQ ID NO: 313 and a VL having SEQ ID NO: 314, or

[0462] a VH having SEQ ID NO: 331 and a VL having SEQ ID NO: 332, or

[0463] a VH having SEQ ID NO: 349 and a VL having SEQ ID NO: 350, or

[0464] a VH having SEQ ID NO: 367 and a VL having SEQ ID NO: 368, or

[0465] a VH having SEQ ID NO: 385 and a VL having SEQ ID NO: 386, or

[0466] a VH having SEQ ID NO: 403 and a VL having SEQ ID NO: 404, or

[0467] a VH having SEQ ID NO: 421 and a VL having SEQ ID NO: 422, or

[0468] a VH having SEQ ID NO: 439 and a VL having SEQ ID NO: 440.

[0469] In some embodiments, the human monoclonal antibody has a heavy chain having SEQ ID NO: 9 and a light chain having SEQ ID NO: 10, or

[0470] a heavy chain having SEQ ID NO: 27 and a light chain having SEQ ID NO: 28, or

[0471] the heavy chain of the antibody has SEQ ID NO: 45 and the light chain of the antibody has SEQ ID NO: 46, or

[0472] the heavy chain of the antibody has SEQ ID NO: 63 and the light chain of the antibody has SEQ ID NO: 64,

[0473] the heavy chain of the antibody has SEQ ID NO: 81 and the light chain of the antibody has SEQ ID NO: 82,

[0474] the heavy chain of the antibody has SEQ ID NO: 99 and the light chain of the antibody has SEQ ID NO: 100, or

[0475] the heavy chain of the antibody has SEQ ID NO: 135 and the light chain of the antibody has SEQ ID NO: 136, or

[0476] the heavy chain of the antibody has SEQ ID NO: 153 and the light chain of the antibody has SEQ ID NO: 154, or

[0477] the heavy chain of the antibody has SEQ ID NO: 171 and the light chain of the antibody has SEQ ID NO: 172, or

[0478] the heavy chain of the antibody has SEQ ID NO: 189 and the light chain of the antibody has SEQ ID NO: 190, or

[0479] the heavy chain of the antibody has SEQ ID NO: 207 and the light chain of the antibody has SEQ ID NO: 208, or

[0480] the heavy chain of the antibody has SEQ ID NO: 225 and the light chain of the antibody has SEQ ID NO: 226, or

[0481] the heavy chain of the antibody has SEQ ID NO: 243 and the light chain of the antibody has SEQ ID NO: 244, or

[0482] the heavy chain of the antibody has SEQ ID NO: 261 and the light chain of the antibody has SEQ ID NO: 262, or

[0483] the heavy chain of the antibody has SEQ ID NO: 279 and the light chain of the antibody has SEQ ID NO: 280, or

[0484] the heavy chain of the antibody has SEQ ID NO: 297 and the light chain of the antibody has SEQ ID NO: 298, or

[0485] the heavy chain of the antibody has SEQ ID NO: 315 and the light chain of the antibody has SEQ ID NO: 316, or

[0486] the heavy chain of the antibody has SEQ ID NO: 333 and the light chain of the antibody has SEQ ID NO: 334, or

[0487] the heavy chain of the antibody has SEQ ID NO: 351 and the light chain of the antibody has SEQ ID NO: 352, or

[0488] the heavy chain of the antibody has SEQ ID NO: 369 and the light chain of the antibody has SEQ ID NO: 370, or

[0489] the heavy chain of the antibody has SEQ ID NO: 387 and the light chain of the antibody has SEQ ID NO: 388, or

[0490] the heavy chain of the antibody has SEQ ID NO: 405 and the light chain of the antibody has SEQ ID NO: 406, or

[0491] the heavy chain of the antibody has SEQ ID NO: 423 and the light chain of the antibody has SEQ ID NO: 424, or

[0492] the heavy chain of the antibody has SEQ ID NO: 441 and the light chain of the antibody has SEQ ID NO: 442.

[0493] Embodiment 15. There is also provided herein a human monoclonal antibody according to any one of embodiments 1 to 14, wherein the fragment crystallizable (Fc) region of the IgGl backbone contains the following mutation E430G.

[0494] Embodiment 16. There is also provided herein a human monoclonal antibody or an antigen binding portion thereof which competes for binding to a surface antigen of a K. pneumoniae strain with any of the antibodies or antigen binding portions according to any one of embodiments 1 to 15.

[0495] Embodiment 17. There is also provided herein a human monoclonal antibody or an antigen binding portion thereof according to any one of embodiments 1 to 16 for use in the prophylactic or therapeutic treatment of a K. pneumoniae infection or a condition or disease resulting from such infection, in particular for use in the prophylaxis and / or treatment of an infection with a drug resistant or multi-drug resistant K. pneumoniae strain.

[0496] Embodiment 18. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 16 for use in the prophylactic or therapeutic treatment of an infection with NDM-producing K. pneumoniae strains, in particular NDM-1, NDM-5 and / or NDM-9 positive strains, or a condition or disease resulting from such an infection.

[0497] Embodiment 19. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 16 for use in the prophylactic or therapeutic treatment of an infection with the NDM-1 positive ST147 strain of K. pneumoniae, or a condition or disease resulting from such an infection.

[0498] Embodiment 20. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 16 for use in a method of prophylactic or therapeutic treatment of a K. pneumoniae infection, or a condition or disease resulting from such an infection, wherein the method comprises the step of administering to the patient 0.025 to 5 mg / kg of the human monoclonal antibody or antigen binding portion thereof.

[0499] Embodiment 21. Also provided herein is a human monoclonal antibody or antigen binding portion thereof according to any one of embodiments 1 to 16 for use in a method of prophylactic or therapeutic treatment of a K. pneumoniae infection, or a condition or disease resulting from such an infection, wherein the method comprises the step of administering to the patient 0.1 to 3 mg / kg, preferably 0.25 mg / kg, of the human monoclonal antibody or antigen binding portion.

[0500] Embodiment 22. Also provided herein is a human monoclonal antibody or antigen binding portion thereof for use according to any one of embodiments 17 to 21, wherein the human monoclonal antibody or antigen binding portion thereof is administered by intravenous, subcutaneous, intraperitoneal or intramuscular route.

[0501] Embodiment 23. Also provided herein is a human monoclonal antibody or antigen binding portion thereof for use according to any one of embodiments 17 to 21, wherein the method comprises the step of administering to the patient the human monoclonal antibody or antigen binding portion once a day for at least 3 days.

[0502] Embodiment 24. Also provided herein is a pharmaceutical composition comprising one or more human monoclonal antibodies or antigen binding portions thereof according to any one of embodiments 1 to 16 and a pharmaceutically acceptable carrier.

[0503] Embodiment 25. Also provided herein is a pharmaceutical composition according to embodiment 24, comprising a unit dose of the human monoclonal antibody or antigen binding portion thereof equal to or less than 400 mg.

[0504] Embodiment 26. Also provided herein is the pharmaceutical composition according to Embodiment 24, comprising a unit dose equal to or less than 100 mg of the human monoclonal antibody or antigen-binding portion thereof.

[0505] Embodiment 27. Also provided herein is the pharmaceutical composition according to any one of Embodiments 24 to 26, for intravenous, subcutaneous, intraperitoneal, or intramuscular administration.

[0506] Embodiment 28. Also provided herein is the composition according to any one of Embodiments 24 to 27, for use in the prevention and / or treatment of a Klebsiella pneumoniae infection.

[0507] Embodiment 29. Also provided herein is an isolated cell line producing the antibody or antigen-binding portion thereof according to any one of Embodiments 1 to 16.

[0508] Embodiment 30. Also provided herein is an isolated nucleic acid molecule comprising a nucleotide sequence encoding the antibody or antigen-binding portion thereof according to any one of Embodiments 1 to 16.

[0509] Embodiment 31. Also provided herein is a vector comprising the nucleic acid molecule according to Embodiment 30, wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule.

[0510] Embodiment 32. Also provided herein is the vector according to Embodiment 31, wherein the vector is selected from the group consisting of an RNA viral vector, a DNA viral vector, a plasmid viral vector, an adenoviral vector, an adeno-associated viral vector, a herpes viral vector, and a retroviral vector.

[0511] Embodiment 33. Also provided herein is a composition comprising the isolated nucleic acid molecule according to Embodiment 30 or the vector according to Embodiment 31 or 32, for use in the prevention and / or treatment of a Klebsiella pneumoniae infection.

[0512] Embodiment 34. Also provided herein is the composition according to Embodiment 33, wherein the nucleic acid molecule or the vector is formulated in a lipid nanoparticle.

[0513] Embodiment 35. Also provided herein is a host cell comprising the vector according to Embodiment 29 or the nucleic acid molecule according to Embodiment 30.

[0514] Embodiment 36. Also provided herein is a non-human transgenic animal or transgenic plant comprising the nucleic acid according to Embodiment 30, wherein the non-human transgenic animal or transgenic plant expresses the nucleic acid.

[0515] Embodiment 37. Also provided herein is the use of the human monoclonal antibody or antigen-binding portion thereof according to any one of Embodiments 1 to 16 in the diagnosis of a Klebsiella pneumoniae infection.

[0516] Embodiment 38. There is also provided herein an in vitro method for revealing the presence of a K. pneumoniae strain in a sample, comprising the steps of:

[0517] i) contacting an antibody, or an antigen-binding portion thereof, according to any one of embodiments 1 to 16, with said sample;

[0518] ii) detecting the binding of said antibody, or antigen-binding portion thereof, to a surface antigen of K. pneumoniae.

[0519] Embodiment 39. There is also provided herein an in vitro method for diagnosing a K. pneumoniae infection in a subject, comprising the steps of:

[0520] i) contacting an antibody, or an antigen-binding portion thereof, according to any one of embodiments 1 to 16, with a biological sample of said subject;

[0521] ii) detecting the binding of said antibody, or antigen-binding portion thereof, to a surface antigen of K. pneumoniae.

[0522] Embodiment 40. There is also provided herein an in vitro method according to embodiment 38 or 39, wherein said method is an immunoassay selected from the group consisting of ELISA, RIA, flow cytometry, tissue immunohistochemistry, Western blot (immunoblot), immunoprecipitation or any other equivalent assay.

[0523] Embodiment 41. There is also provided herein an in vitro method according to embodiments 38 to 40, wherein said antibody, or antigen-binding portion thereof, is directly labeled with a detectable label, or wherein said antibody, or antigen-binding portion thereof, (first antibody) is bound to a second labeled antibody or another labeled molecule.

[0524] Embodiment 42. There is also provided herein a diagnostic kit comprising an antibody, or antigen-binding portion thereof, according to any one of embodiments 1 to 16, as a specific reagent, said kit being intended in particular for use in a method for detecting or quantifying anti-K. pneumoniae antibodies, in particular anti-NMD-producing K. pneumoniae antibodies, in a patient biological sample.

[0525] Embodiment 43. There is also provided herein the use of an antibody, or antigen-binding portion thereof, according to any one of embodiments 1 to 16, for the design of a vaccine against K. pneumoniae.

[0526] Embodiment 44. There is also provided herein the use of an antibody, or antigen-binding portion thereof, according to any one of embodiments 1 to 16, for the design of a vaccine against NMD-producing K. pneumoniae strains.

[0527] Embodiment 45. Also provided herein are mimotopes specific for the idiotype of an antibody, or antigen binding portion thereof, according to any one of embodiments 1 to 16.

[0528] Embodiment 46. Also provided herein are anti-idiotype antibodies specific for the idiotype of an antibody, or antigen binding portion thereof, according to any one of embodiments 1 to 16.

[0529] Embodiment 47. Also provided herein are immunospecific polypeptides comprising at least a variable domain, in particular at least a heavy chain variable domain (VH) and a light chain variable domain (VL) as defined in any of the embodiments disclosed herein, or any of the CDRs as defined herein, in particular, the immunospecific polypeptides are multispecific antibodies, bispecific antibodies, trispecific antibodies, monoclonal antibodies, scFV, diabodies, triabodies, tetrabodies, minibodies, linear antibodies, chelating recombinant antibodies, triabodies, diabodies, intrabodies, nanobodies, binding-domain immunoglobulin fusion proteins, fusion antibodies, immunoadhesins, or antigen binding fragments thereof.

[0530] ***

[0531] The following experimental section is provided for illustrative purposes only and is not intended to limit the scope of the application as defined in the appended claims. The claims are an integral part of this description.

[0532] Examples

[0533] 1. Results

[0534] 1.1. Isolation of monoclonal antibodies recognizing epidemic strain ST147 NDM-1

[0535] To isolate K. pneumoniae-specific mAbs, which represent the breadth of the Ab repertoire induced by natural Kp infection, we employed an antigen-independent approach. Peripheral blood mononuclear cells (PBMCs) were collected from seven convalescent patients who recovered from Kp ST147 NDM-1 bloodstream infection during the Toscani outbreak and from one patient who had ST147 NDM-1 intestinal colonization during the Toscani outbreak (Table 1).

[0536] Table 1 - Information of patients from the Toscani outbreak participating in the study.

[0537]

[0538] A total of 18’390 CD19 + CD27 + IgM - IgD​- Memory B cells (MBCs) (IgG + or IgA + positive) were single cell sorted and plated with feeder cells expressing CD40L, IL-2 and IL-21 to promote MBC activation, proliferation and antibody secretion Figure 1 A; Figure 6 ). MBC culture supernatants were screened by enzyme-linked immunosorbent assay (ELISA) against immobilized whole ST147 NDM-1 bacteria to select Kp-reactive mAbs.

[0539] MBC culture supernatants were screened by ELISA against immobilized whole ST147 NDM-1 bacteria to identify 214 mAbs specific for antigens displayed on the bacterial surface (Table 2, B). Figure 1

[0540] Table 2. Kp isolates used in the original work.

[0541]

[0542] After molecular cloning of their V H and V L regions, 134 V H and V L successfully paired Kp-reactive mAb sequences were recovered. To verify the binding properties of the isolated mAbs, they were expressed in small scale and tested by ELISA against the epidemic-associated ST147 NDM-1 Kp strain (B). Figure 1

[0543] 1.2. Selection of functional ST147 NDM-1 Kp-specific mAbs by high-throughput luminescent serum bactericidal assay (L-SBA)

[0544] Further, since it is generally accepted that pathogens can elicit both functional and non-functional antibodies, mAbs capable of inducing complement-dependent ST147 NDM-1 Kp killing were screened.

[0545] To this end, a high-throughput luminescent serum bactericidal assay (L-SBA) was developed that utilizes luminescence readout of ATP generation as an indicator of Kp viability. Four dilutions of each mAb were tested, with a reduction in bacterial viability of more than 30% being considered a positive result. Out of the 134 Kp-binding mAbs screened, 25 candidates with bactericidal activity against ST147 NDM-1 were selected (Table 3, C). Figure 1 ​​C) and were subjected to larger scale expression. After excluding 5 mAbs that failed to be reconfirmed for their activity, we continued to evaluate the binding and functional properties of 20 bactericidal mAbs targeting ST147 NDM-1 .

[0546] 1.3. Klebsiella library analysis revealed that bactericidal mAbs have different degrees of cross-reactivity and target ST147 NDM-1 capsular and O antigens

[0547] To characterize the specificity and cross-reactivity of the isolated bactericidal mAbs, their binding to a panel of bacterial strains was analyzed by flow cytometry. Pathogenic Kp belonging to different STs at a distant genetic distance, as well as non-pathogenic Klebsiella species and several commensal strains were included (Table 2). The mAbs that bound to the bacterial surface showed different levels of intensity, from almost zero to very high, indicating differential expression of the target antigen Figure 2 A). In addition, mAbs showed a high degree of specificity for pathogenic Kp strains in the strict sense, as they did not react with other Kp phylogenetic groups or commensal bacteria Figure 8 . Based on the binding profile, mAbs were grouped into two main clusters Figure 2 A).

[0548] The first cluster contained mAbs with limited binding properties, recognizing only epidemic ST147 carrying bla NDM-1 or bla NDM-9 genes (doi:10.2807 / 1560-7917.ES.2020.25.48.2001779). The binding pattern of the second group of mAbs was more extensive, recognizing up to seven pathogenic Kp strains at a distant genetic distance.

[0549] Comparative whole-genome sequencing analysis of the Kp library allowed the prediction of the surface antigen targeted by the mAbs. We deduced that mAbs of cluster 1 could be directed against the ST147 NDM-1 capsular (K antigen), as these mAbs recognized strains carrying genes of the capsular type 64 biosynthetic locus (KL64) (https: / / pubmed.ncbi.nlm.nih.gov / 487363 / ). In contrast, mAbs of cluster 2 could recognize the ST147 NDM-1 O antigen, as they bound to the surface of strains carrying genes of the O2a locus encoding O antigen biosynthesis genes (Table S4). To test these hypotheses, we evaluated the binding of mAbs to purified Kp capsules and several types of O antigens in ELISA Figure 9 and Figure 10 ). We observed that the KL64 capsule was only recognized by mAbs of cluster 1 Figure 2B). Immunoblotting of total sugar extracts from Kp further confirmed that mAbs of cluster 1 recognized only high molecular weight (MW) molecular species in Kp strains carrying KL64 Figure 2 C).

[0550] In contrast, ELISA against purified O antigens revealed a heterogeneous picture among mAbs of cluster 2. Three mAbs recognized purified O2a O antigens with different degrees of cross-reactivity with O2afg and O1 O antigens Figure 2 B). Immunoblotting analysis demonstrated that these mAbs displayed a stepwise binding pattern, consistent with the typical O antigen immunoblotting profile Figure 2 D). The additional five mAbs from cluster 2 did not recognize purified O antigens in ELISA, but they displayed a similar stepwise binding profile in the intermediate MW range with total sugar extracts from Kp strains Figure 2 E). Binding was restricted to strains expressing O2a O antigens. Moreover, no binding was observed with ST147 NDM-9 , which is closely related at the genomic level to ST147 NDM-1 . Comparative genetic analysis showed that, among all the genes encoding surface Kp antigens, ST147 NDM-9 exhibits a single point deletion in the wbbO gene, which encodes a glycosyltransferase essential for O2a biosynthesis (REF). The mutation results in a frameshift and premature termination, hampering the production of a complete O antigen. HPLC and SDS-PAGE analysis of total sugar extracts from ST147 NDM-9 confirmed that ST147 NDM-9 is a natural O antigen-deficient mutant Figure 11 , which explains its inability to bind mAbs of cluster 2.

[0551] Therefore, in summary, our analysis allowed us to classify the isolated ST147 NDM-1 targeting mAbs into KL64-specific mAbs and O2a O antigen-specific mAbs, some of which also had cross-reactivity with O1 -type O antigens. High-resolution and high-content analysis of mAb binding to the ST147 NDM-1 surface revealed that the pattern of mAbs targeting the capsule and O antigens did not overlap. Anti-capsule mAb 08O09 displayed higher intensity binding and distributed over a larger area compared to anti-O antigen mAbs 05D08 and 05N02 Figure 3 B, Figure 3 C, Figure 12 ). Moreover, the 08O09 signal was located at the outermost layer of the bacteria Figure 3D). mAb clonal composition analysis revealed a clonal usage bias for anti-capsular mAbs, while 3 clonal pairs were detected among O antigen-specific mAbs (Figure 13). Thus, we gathered information on the cross-binding properties, specificity, and sequence features of 20 bactericidal mAbs targeting ST147 NDM-1 .

[0552] 1.4. Functional analysis revealed that the isolated mAbs have very strong bactericidal activity, but only anti-capsular mAbs have polyfunctionality

[0553] To better assess the protective properties of anti-capsular and anti-O antigen bactericidal antibodies, we used several in vitro assays related to antibody-mediated control of bacteria in vivo. Bactericidal mAbs were expressed as human IgGl with an E430G mutation in the Fc domain, which enhances the naturally occurring hexamerization process between Fc domains, thereby enhancing antibody effector functions. We also improved the processivity of L-SBA by designing a fluorescent SBA (F-SBA) in 384-well format and using resazurin staining for fluorescent readout of bacterial viability. F-SBA analysis of mAbs against the complement-sensitive pathogenic Kp strains (ST147 NDM-1 , ST147 NDM-9 , and ST307 NDM-5 ) revealed that most antibodies were functional in the picomolar concentration range, with the most potent mAbs exhibiting IC 50 values of 0.5-6 ng / mL ( Figure 4 A; Figure 14 ). The ability to kill different Kp strains correlated with the cross-binding properties of mAbs, with anti-KL64 mAbs being effective against ST147 NDM-1 and ST147 NDM-9 carrying KL64, and anti-O antigen mAbs being effective against ST147 NDM-1 and ST307 NDM-5 carrying O2 O antigen ( Figure 4 A). F-SBA analysis allowed us to rank mAbs according to the cumulative bactericidal potency in each cluster ( Figure 15 ). Additional refinement of the ranking was also based on the maximum capacity of mAbs to reduce bacterial viability ( Figure 15 ).

[0554] Subsequently, the antibodies exhibiting the highest bactericidal efficiency were evaluated in a macrophage-mediated opsonophagocytosis assay, as phagocytes play a crucial role in Kp clearance during infection (Broug-Holub et al., 1997; Cheung et al., 2000). Prior to incubation with differentiated THP1 cells, recombinant ST147 NDM-1 overexpressing superfolded (sf)mCherry were opsonized with increasing doses of mAbs. The fluorescence of internalized Kp was then measured as a readout of the mAbs’ ability to promote opsonophagocytosis. We found that anti-KL64 mAbs increased Kp uptake by THP-1 cells at doses greater than 0.6 mg / mL, but 10H18, which exhibited only a very slight effect Figure 4 B). However, no Kp uptake was detected for anti-O-antigen mAbs Figure 4 C). This indicates, on the one hand, that anti-capsular mAbs (but not anti-O-antigen mAbs) are able to promote bacterial phagocytosis. On the other hand, this shows that the opsonizing effect of anti-capsular mAbs is able to overcome the inhibition of opsonophagocytosis by Kp surface virulence factors (Alvarez et al., 2000; Cortes et al., 2002; Regueiro et al., 2006; March et al., 2013).

[0555] Finally, to analyze the impact of Kp culture over time in the presence of bactericidal mAbs, we performed time-lapse imaging of ST147 NDM-1 -sfmCherry incubated with increasing doses of mAbs. We found that Cluster 1 mAbs induced bacterial chain growth at 10 and 100 mg / mL Figure 4 D). Mechanistically, this phenotype is associated with the accumulation of concatenated replicating bacteria, without physical division of sister cells. In contrast, Cluster 2 antibodies did not induce chain growth at any of the tested doses Figure 4 D). Bacterial concatenation induced by dimeric IgA has previously been proposed as a protective mechanism that limits pathogen growth and promotes its clearance from the gut (Moor et al., 2017; Bansept et al., 2019). Interestingly, we observed this phenomenon in the presence of monomeric IgG.

[0556] In summary, the multi-layered analysis indicates that the isolated mAbs have a very strong bactericidal activity, whose function can go beyond opsonophagocytosis and can involve other effector mechanisms, such as chain Kp growth. However, surprisingly, only anti-capsular mAbs have such multifunctionality.

[0557] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24 functional mAbs.

[0558] Table 3. Sequence analysis of the 24

[0559]

[0560]

[0561] 1.5. mAbs offer multifunctional protection to the body against explosive ST147 attacks. NDM-1 Bloodstream infection is necessary

[0562] To answer the fundamental question of whether picomolar bactericidal activity alone is sufficient to make therapeutic mAbs effective in vivo, or whether they need to be multifunctional, we evaluated the protective properties of the most potent mAb candidates (anti-capsular O8O09, anti-O2 O antigen-specific O5D08, and anti-O1 / O2 O antigen-specific O5N02) in prophylactic (PRO) and therapeutic (THR) studies in an immunocompetent mouse sepsis model. A fulminant bloodstream infection model was chosen to reflect the clinical history of donors from whom mAbs were isolated, and the model was set to achieve a 90-100% mortality rate within 24 hours of bacterial challenge. Figure 16 A) This leaves a narrow time window for the therapeutic effect of mAbs.

[0563] To evaluate ST147 NDM-1 The in vivo PRO activity was measured in three groups of mice (n=10 per group) 24 hours before bacterial inoculation, with doses of 1, 5, 10, and 20 mg / kg mAb. Figure 5 A). Compared with the placebo control group, 5 mg / kg and higher doses of 08O09 significantly prolonged survival (P<0.05), with a survival rate of 30%. Figure 5 B). Consistent with this observation, the CFU count in the spleen was reduced compared to the control group. Figure 16 B). Conversely, 05D08 and 05N02 mAb did not significantly prolong animal survival at any dosage. Figure 5 CD). A prophylactic test at 5 mg / kg 08O09 was repeated in a total of 30 animals, and the results showed consistency and reproducibility, with mice surviving for at least 35 hours and 50% surviving for up to 96 hours. Figure 5 E). The plasma concentration of 08O09 associated with optimal in vivo efficacy was 74.3 (±21.7) μg / mL. Figure 17 ).

[0564] Taking these results into account, we evaluated the therapeutic (THR) efficacy of intravenous administration of 08O09 at doses of 1, 5, 10, and 20 mg / kg one hour after bacterial challenge. Figure 5 F). In this case, doses of 5 mg / kg and above can achieve moderate protection (F). Figure 5G), which correlates with lower CFU counts in the spleen Figure 16 C) Finally, we evaluated the protective properties of 08009 in a combined prophylactic / therapeutic (PRO+THR) regimen, in which the first dose of 08009 was administered intraperitoneally 24 hours before bacterial inoculation and the second dose was administered intravenously one hour after bacterial challenge Figure 5 H) Interestingly, the advantage of using a combined treatment was only observed at the 1 mg / kg dose, with a 30% survival rate at 96 hours, while the effect of the single treatment regimens was not optimal Figure 5 I and 16D). However, when 08009 was administered at 5 mg / kg, the effect of the prophylactic monotherapy was clearly superior to the combined treatment or the therapeutic monotherapy Figure 5 I and 16E), indicating that while a suboptimal mAb dose can be compensated by a combined prophylactic treatment regimen, the optimal mAb dose can perform well as a single monotherapy in a prophylactic regimen. Altogether, these experiments demonstrate for the first time the potential use of anti-KL64 specific multifunctional mAbs as therapeutic agents for the treatment of pan-resistant ST147 NDM-1

[0565] 1.6 Discussion

[0566] We here present a panel of 25 potent mAbs that exhibit different properties. The mAbs of cluster 1, and in particular 08009, have been shown to promote the opsonic phagocytosis of Kp and to have a protective effect in in vivo models. Although this mAb targets a specific target, i.e. CPS type 64, another practical situation is that the Tuscany outbreak is still ongoing and the diversity of the isolates is not significant (data not shown). Moreover, to the best of our knowledge, this is the first report of an anti-K64 mAb.

[0567] From a therapeutic point of view, AMR Kp infections represent a huge clinical challenge. K. pneumoniae has developed virulence factors (e.g. capsule and LPS) that prevent its uptake by phagocytes (Alvarez et al., 2000; Cortes et al., 2002; Regueiro et al., 2006; March et al., 2013). Our aim was to explore whether the mAbs of cluster 1 and cluster 2 could impair the protective effect of the capsule and LPS and promote macrophage-dependent uptake. We found that anti-capsule mAbs were able to block the protective effect of the capsule itself in preventing phagocytosis and macrophage-mediated killing.

[0568] ​Computational models suggest that chain-like bacterial growth triggered by the host immune response involves only rapidly replicating bacteria. Thus, through this mechanism, infected organisms would concentrate bacteria, thus facilitating their clearance and protecting their microbiota. It should also be noted that the strain used in the in vivo challenge is a pan-resistant strain, which is not only spreading in Italy but also worldwide, highlighting the potential of 0809 in the fight against MDR infections.

[0569] 2. Materials and methods

[0570] 2.1. Single cell sorting of memory B cells from convalescent donors

[0571] Peripheral blood mononuclear cells (PBMCs) were isolated from heparin-treated whole blood using density gradient centrifugation (Ficoll-Paque PREMIUM, GE Healthcare). After isolation, PBMCs were stained with 100 pL final volume (1 : 1000 dilution) of Live / Dead FSV780 (BD Horizon) at room temperature (RT). After 20 minutes of incubation, cells were washed with IX Phosphate Buffered Saline (PBS) and saturated for non-specific binding with 100 pL of 20% normal rabbit serum (Life Technologies). After 30 minutes of incubation at 4°C, cells were washed with PBS IX, centrifuged at 1200 rpm for 8 minutes, and stained with CD19 APC (BD cat#561742), IgM PeCF594 (BD cat#562539), CD27 APC R700 (BD cat#565116), IgD PE (BD cat#562024), CD3 PE-Cy7 (BD cat#560910), CD14 PE-Cy7 (BD cat#560919), and CD56 PECy7 (BD cat#560916) in staining buffer (1% fetal bovine serum in PBS IX) for 30 minutes at 4°C. After a further wash, cells were resuspended in sorting buffer (PBS / EDTA 2.5 mM). Stained memory B cells (MBCs) were single cell sorted into 384-well plates containing 3T3-CD40L feeder cells using a FACS Aria III Fusion BD (BD Biosciences) and incubated with IL-2 and IL-21 for 14 days (Andreano et al., 2021).

[0572] ELISA screening of mAbs binding to K. pneumoniae

[0573] MBC supernatant generated by single cell sorting was used as a source of IgGl and IgA monoclonal antibodies (mAbs) to detect their binding to whole bacteria in a high-throughput enzyme-linked immunosorbent assay (ELISA). ST147 NDM-1 c.i.1 and ST147 NDM- 1 The pooled liquid of c.i.2 (Table S2) was grown in LB medium, pelleted at the exponential phase (OD 600 = 0.5), resuspended in PB and inoculated in 384-well plastic plates (Greiner, ref. 781101). The inoculated bacteria were fixed with 0.5% paraformaldehyde for 30 min at room temperature (RT). Then the plates were blocked in PBS plus 1% fetal bovine serum for 1 h at RT. After incubation, the bacteria were incubated with MBC supernatant diluted in sample buffer (PBS-BSA 1% - Tween 20 0.05%) for 1 h. Next, anti-human IgG and anti-human IgA secondary antibodies conjugated with alkaline phosphatase (Southern Biotech) were added and incubated for 45 min. To detect bacteria-mAb binding, pNPP (para-nitrophenyl phosphate; Sigma-Aldrich) was used as soluble substrate and the final reaction was quantitatively analyzed using a Varioskan Lux reader (Thermo Fisher Scientific) at a wavelength of 405 nm. After each incubation step, the plates were washed three times with 100 μΐ per well of wash buffer (phosphate-buffered saline and 0.05% Tween-20). Using sample buffer as a blank, we considered as positive results the wells with an OD 405 value at least 2-fold higher than the blank. The cells corresponding to the positive results were lysed in 25 μΐ of buffer containing RNAsi Out 0.2 U / μΐ, ultrapure BSA 1 mg / mL and H2O DPC (Thermo Fisher Scientific) and stored at -80 °C for the following steps.

[0574] 2.2. Single cell RT-PCR and nested PCR for amplification of V H and V L

[0575] cDNA was synthesized from 5 pL of MBC lysate. Reverse transcription (RT) reactions were performed in a total volume of 25 pL per well containing 1 pL random hexamer primers (50 ng / mL), 1 pL dNTPs (10 mM), 2 pL of 0.1 M DTT, 40 U / pL of RNase OUT, MgCl2(25 mM), 5 pL of 5X buffer, 0.25 pL of Superscript IV Reverse Transcriptase (Invitrogen) and nuclease-free water (DEPC). The RT-PCR reaction conditions were 42 °C / 10 min, 25 °C / 10 min, 50 °C / 60 min and 94 °C / 5 min. After cDNA synthesis, two additional rounds of PCR were performed to obtain the variable regions of the heavy chain (VH H ) and light chain (VL L ). Briefly, in the first round of PCR (PCR I), a total volume of 25 pL per well was added containing 4 pL of cDNA, 10 pM of VH or 10 pM of VL / VK primer mix (Table 4), 0.5 pL of dNTPs (10 mM), 1.5 pL of MgCl2(25 mM), 5 pL of 5X Kapa Long Range Buffer and 0.125 pL of Kapa Long Range Polymerase (Sigma) and amplification was performed under the following conditions: 95 °C / 3’, 5 cycles of (95 °C / 30”, 57 °C / 30”, 72 °C / 30”) and 30 cycles of (95 °C / 30”, 60 °C / 30”, 72 °C / 30”) and 72 °C / 2’. Nested PCR (PCR II) was performed using the same cycling conditions and primers listed in Table 5, using 3 pL of unpurified PCR I product as template. PCR II products were purified using Millipore MultiScreen PCR 96-well plates according to the manufacturer’s instructions. Samples were eluted with 30 pL of 50 °C pre-warmed nuclease-free water and quantified using a NanoDrop One (Thermo Fisher Scientific).

[0576] Table 4. List of primer mixtures used for PCR I.

[0577]

[0578]

[0579] Table 5. List of primer mixtures used for PCR II.

[0580]

[0581]

[0582] 2.3. V Hand V L Cloning and expression of TAP-transfected recombinant antibodies

[0583] Amplified antibody sequences were ligated into human IgGl expression vectors and used to generate transcriptionally active PCR (TAP) products. Briefly, IgGl, IgK or IgA expression vectors were digested with Age I, Sail and Xho I restriction enzymes, respectively. Gibson Assembly (New England BioLabs) was used to ligate 25 ng of linearized plasmid with 75 ng of purified V H and V L in a final volume of 5 μΐ. The ligation product was diluted 10-fold with DEPC water and used as a template for a transcriptionally active PCR (TAP) reaction, which allows for direct in vitro expression using linear DNA fragments. Functional promoter (human CMV) and terminator sequences (SV40) were directly ligated to the PCR II product by amplification using specific primers (Table 6).

[0584] Table 6. List of primer mixtures used for PCR II.

[0585] Name Sequence SEQ ID CMV_TAP_FW TTAGGCACCCCAGGCTTTAC SEQ ID NO: 501 polyA_TAP_Rev AGATGGTTCTTTCCGCCTCA SEQ ID NO: 502

[0586] TAP-PCR was performed in a total volume of 25 μΐ containing 0.25 μΐ Q5 polymerase (NEB), 5 μΐ GC enhancer (NEB), 5 μΐ 5X buffer, 0.5 μΐ dNTPs (10 mM), 0.125 μΐ forward / reverse primers and 3 μΐ ligation product, using the following conditions: one step 98°C / 2', 35 cycles of 98°C / 10", 61°C / 20", 72°C / 1' and extension 72°C / 5'. After purification and quantification, TAP products were transfected into Expi293F cells for small-scale production of recombinant mAbs. Briefly, cells were transfected with both TAP amplification products in a final volume of 1 ml in 96-deep well plates (Eppendorf) and, after 7 days of expression, mAb-containing supernatants were collected by centrifugation.

[0587] 2.4. Quantification of TAP-produced mAbs by ELISA

[0588] To quantify the mAb concentration in each supernatant, ELISA plates were coated overnight at 4°C with 2 pg / ml goat anti-human IgG (Southern Biotech). Plates were washed 3 times with PBS supplemented with 0.05% Tween20 and blocked for 1 hour at 37°C in PBS supplemented with 1% BSA. Samples were then washed and incubated with TAP-produced mAbs diluted in sample buffer (PBS IX-BSA 1%-Tween200.05%) for 1 hour at 37°C. After washing again, a 1-hour incubation at 37°C with AP-conjugated anti-goat IgG secondary antibody (Southern Biotech) was performed and absorbance was read by addition of PNPP. Concentrations were assessed by linear regression analysis established by fitting the OD 405 values to a standard curve generated by titration of an unlabelled antibody to human IgG (Southern Biotech).

[0589] 2.5. Large-scale expression and purification of mAbs

[0590] Expi293F cells (Thermo Fisher) were transiently transfected with plasmids carrying each antibody heavy and light chain (1 :2 weight / weight ratio). Cells were grown for six days at 37°C, 8% CO2 and 125 rpm shaking, with the addition of an optimized mix of Enhancer 1 and 2 (Thermo Fisher) the day after transfection. mAbs were harvested twice by cell pelleting at RT at 1,100 x g for 10 min on the third and sixth day, and supernatants were then pooled and clarified by centrifugation (4°C, 3,000 x g, 15 min) followed by 0.45 mm filtration. mAbs were purified at RT by affinity chromatography on an AKTA go purification system (Cytiva) using a HiTrap Protein G HP column (Cytiva) binding to Fc domains. Specifically, the column was equilibrated in 0.02 M sodium phosphate buffer, pH 7, at a flow rate of 1 mL / min, which was also used for the following steps. After injection, the column was rinsed with 10 column volumes (CV) and then mAbs were eluted with 10 CV of 0.1 M glycine-HCl, pH 2.7. The mAb pool was dialyzed overnight at 4°C in PBS, pH 7.4, using Slide-A-Lyzer G2 Dialysis Cassettes 3.5K (Thermo Scientific). Concentration was determined by measuring absorbance at 280 nm on a Nanodrop for each purified antibody. All purified antibodies were aliquoted and stored at -80°C.

[0591] 2.6. Serum bactericidal assay (SBA)

[0592] Two days prior to the assay, Kp stock glycerol solution was streaked onto LB agar plates and incubated overnight at 37°C (ON). The next day, single colonies were inoculated into 4 mL LB agar and incubated at 37°C (ON). The culture was then amplified to 10 mL LB agar in 125 mL Erlenmeyer flasks until the OD600 reached 0.05, and then incubated at 37°C with shaking until the exponential growth phase (OD600 ≥ 0.05). 600 (0.4-0.6). The bacteria were then diluted with PBS at a ratio of 1:10.

[0593] The appropriate concentration of rabbit complement (BRC) used in this assay was determined by complement sensitivity testing. In short, 2 x 10⁻⁶... 5 Bac-Titer-Globe bacteria resuspended in PBS were inoculated into 96-well round-bottom plates, starting at 50% and screened for BRC concentration using 11 consecutive 2-fold serial dilutions. After incubation at 37°C for 2 hours, the bacteria were centrifuged to precipitate. The supernatant was discarded, and the bacteria were resuspended in 30 μl of PBS and transferred to white Optiplate plates (Perkin Elmer). 30 μl of BacTiter-Globe was added to each well. TM 1X (Promega) was used, and luminescence values ​​were measured using a Varioskan Lux microplate reader (Thermo Fisher Scientific) with an exposure time of 500 ms. Data were plotted and analyzed using a GraphPad Prism. 12.5% ​​BRC was established as a universal BRC concentration, allowing for complement-based antibody killing without significant toxicity.

[0594] For luminescent SBA (L-SBA) produced using mAb expressed with TAP, in the presence of 12.5% ​​BRC, 2 x 10⁻⁶ cells were used. 6 ST147 bacteria / mL NDM-1 Inoculate into 96-well U-shaped round-bottom plates and add four serially diluted TAP expression mAbs (1:10, 1:50, 1:250, 1:1250), for a total volume of 100 μl per well. After incubation at 37°C for 2 hours, centrifuge at 4000 x g to pellet the bacteria. Resuspend the bacterial pellet in 30 μl of PBS per well and transfer to white Optiplate (Perkin Elmer). Add 30 μl of BacTiter-Glo to each well. TM 1X (Promega) was used, and luminescence was measured using a Varioskan Lux microplate reader (ThermoFisher Scientific) with an exposure time of 500 ms. In each experiment, the obtained luminescence values ​​were used to calculate the median for each dilution factor. The difference between the luminescence signal of each mAb and the median was measured, plotted as a percentage, and 30% was used as the cutoff value.

[0595] For fluorescent SBA (F-SBA), 2x10 6 6 bacteria per ml PBS were inoculated in a 384-well black clear bottom plate (ViewPlate -384F TC, PerkinElmer) in a final volume of 50 μΙ per well. Purified recombinant mAbs were added in a 3-fold serial dilution series. After 2 hours of incubation at 37°C, 40 μΙ LB and 10 μΙ 0.025% resazurin (Sigma-Aldrich) were added per well. After 2 hours of incubation at 37°C, fluorescence (λ Ex 560 nm and λ Em 590 nm) was measured using a Varioskan Lux microplate reader (Thermo Fisher Scientific). Exposure time: 250 ms. Width: 12 nm). GraphPad Prism was used to plot and analyze data and to deduce IC 50 values.

[0596] 2.7. Flow cytometry analysis of mAb binding to the bacterial surface

[0597] Binding of mAbs to the bacterial strain panel (Table S2) was performed on bacteria in exponential growth phase (OD 600 0.4-0.6) in LB. Bacteria were pelleted and resuspended in an equal volume of PBS-BSA 1%. 100 μΙ_ of bacteria were inoculated into each well of a round-bottom 96-well plate, centrifuged at 4000 x g for 5 min, and then incubated with 5 μg / ml purified antibody for 1 h at RT. The plates were centrifuged, the pellets were washed three times with PBS-BSA 1%, and then incubated with a 1:2000 dilution of Alexa Fluor® 488-conjugated α-human IgG secondary antibody for 30 min at RT in the dark. After another centrifugation and washing, the bacteria were fixed with 0.5% paraformaldehyde for 30 min at RT, washed again, resuspended in a PBS solution containing 1% BSA until an OD 600 of 0.05 was reached, and read by fluorescence. Samples were acquired using a BD FACSCanto II (BD Biosciences, USA). Data were analyzed using FlowJo software v10 (BD Biosciences, USA).

[0598] 2.8. Antibody sequence analysis

[0599] Immunoglobulin genes were identified using a custom python script that employed NCBI IgBlast and IMGT nomenclature (time frame 2020-2022). IGHV gene somatic hypermutation counts were from the start of FWR1 to the end of FWR3. Insertions or deletions were counted as one single mutation.

[0600] 2.9Genomic analysis of Kp isolates.

[0601] ST147 strain was sequenced using short and long read technologies. High-throughput sequencing was performed on a MiSeq platform (Illumina, San Diego, CA, USA) in paired-end mode with read length of 150 bp. Paired-end short reads were quality checked using fastp v0.20.1 (REF) and poor quality reads were filtered out. Long read libraries were prepared using the lllumina®M lllumina®MinION®multi-sequencing technology and sequenced according to the manufacturer’s guidelines using a flow cell R9.4.1 (Oxford Nanopore). Long reads were mapped to the corresponding short reads using Filtlong v0.2.0 (https: / / github.com / rrwick / Filtlong) to control the quality of long reads with a minimum quality of Q8 and length of 2,000 bp. Long reads were inputted into the hybrid assembly software Unicycler v0.4.8 (REF) along with the corresponding clean reads and run in the conservative mode. ST and capsule typing prediction was performed using Kleborate v2.0.0 (REF) and Kaptive (REF).

[0602] 2.10Western blot analysis of mAb binding to Kp lysates

[0603] For sample preparation, bacteria from glycerol stocks were grown on LB agar plates overnight. A single colony of Kp was picked from the plate and grown in LB at 37°C statically overnight. The next day, bacteria were grown in LB at 37°C, 150 rpm starting from OD 0.05 until reaching exponential phase (OD 0.4-0.6). Glycocalyx extracts for immunoblotting were prepared using a lipopolysaccharide (LPS) extraction kit (Sigma). Total bacterial lysates were prepared by centrifuging the inoculum at 2500 g for 10 min at 4°C and then filtering with a Nanosep 0.2 pm column at 14000 g. A fraction of the sample was treated with proteinase K at 60°C for 1 hour to remove proteins. SDS-PAGE samples were prepared by adding loading buffer and 1:10 reducing agent at a 1:4 ratio and incubating at 95°C for 5 min. iBlot TMGel Transfer Device and Stacks (Termofisher) to transfer SDS-PAGE gels to PVDF membranes. Membranes were blocked in TBS lx / 0.1% Tween-20 / 5% milk for 1 hour at room temperature. Purified mAbs were 1 pg / mL in TBS lx / 0.1% Tween-20 / 5% milk. After overnight incubation at 4°C, membranes were washed 3 times with TBS lx / 0.1% Tween. Incubation with secondary antibody (goat anti-human Fab) diluted 1 :75,000 in TBS lx / 0.1% Tween / 5% milk was performed for 1 hour at room temperature. Membranes were washed 3 times in TBS lx / 0.1% Tween and developed with chemiluminescent reading.

[0604] 2.11 mAb binding was monitored by high resolution and high content confocal microscopy

[0605] Single colonies of stable over-folded (sf) mCherry expressing bacteria were picked from LB agar plates and grown ON in LB containing 150 pg / mL hygromycin. Overnight cultures were diluted in antibiotic-free LB and grown until OD reached 0.025 or 0.05. Then, 50 mL of bacteria were transferred to 96-well Phenoplate (Perkin Elmer, 6055300) and incubated for 2 hours at 37°C in static conditions without CO2. Subsequently, supernatant was discarded and adherent bacteria were fixed in 4% paraformaldehyde (PFA) / PBS (Thermo Fisher) or Cytofix (BD) for 15 minutes at RT.

[0606] For staining experiments with single anti-Kp mAbs, selected antibodies were diluted in PBS / 1% BSA (bovine serum albumin) solution at a concentration of 0.5 pg / mL and incubated for 30 minutes at RT. Then, samples were washed with PBS and a mixture of goat anti-human Alexa 488 conjugated secondary antibody (1 :2000 dilution) and DAPI (1 :2000 dilution) in PBS / BSA 1% solution was added and incubated for 30 minutes at RT. Finally, samples were washed with PBS and 50 pL of 1% low melting (LM) agarose was added in each well. Samples were stored at 4°C and imaged within the following 24 hours.

[0607] For simultaneous staining experiments with 08O09, 05N02 and 05D08 mAbs, Zip Alexa Fluor TMRapid Antibody Labeling Kits (Thermo Fisher) were used to conjugate antibodies with Alexa 488, Alexa 555 and Alexa 647 fluorophores, respectively, following the manufacturer’s instructions. After fixation of samples with PFA, they were first blocked with PBS / 1% BSA for 30 min at RT, then incubated with a mixture of fluorophore-conjugated mAbs (0.5 mg / mL concentration for each mAb in PBS / 1% BSA) and DAPI (1 :2000). After 30 min incubation at RT, samples were washed with PBS and 50 μL of 1% LM agarose was added for imaging preparation. The next day, samples were imaged.

[0608] 2.12 Opsonophagocytic assay

[0609] THP1 cells (ATCC) were cultured in RPMI 1640 containing GlutaMax (Thermo Fisher) and supplemented with 10 mM Hepes (Thermo Fisher), 1 mM sodium pyruvate (Thermo Fisher) and 10% fetal bovine serum (FBS) (Thermo Fisher). Three days before the assay, 50,000 cells / well were seeded in 96-well black shielded optiplate (PerkinElmer) in the presence of 20 ng / mL phorbol-12-myristate-13-acetate (PMA) to promote monocyte differentiation into macrophages. The next day, PMA was washed away and cells were incubated for two more days in fresh culture medium.

[0610] Macrophages were infected with Kp expressing ST147 NDM1 -sfmCherry. Bacteria were ON-cultured and re-started the next morning to reach OD 6000.5. After centrifugation, bacteria were resuspended in phagocytosis medium (RPMI 1640 + GlutaMax, 10 mM Hepes, 1 mM sodium pyruvate) and the initial culture volume was diluted 1 :2. To perform bacterial opsonization, mAb dilutions were prepared in 25 mL of phagocytosis medium and 25 mL of bacteria were incubated at 37°C with shaking (500 rpm) for 30 minutes. This mixture was then added to differentiated macrophages and centrifuged for 3 minutes at room temperature to synchronize infection. After 1 hour of incubation at 37°C in the presence of 5% CO2, samples were treated with 150 mg / mL streptomycin for an additional hour to kill non-phagocytosed bacteria. Finally, samples were incubated with PBS / 0.1% X100-Triton for 5 minutes at room temperature to permeabilize the cells and allow the release of endocytosed bacteria into the supernatant. Bacterial fluorescence was read using a Varioskan Lux microplate reader (Thermo Fisher) as a readout of bacteria taken up.

[0611] 2.13 Live bacteria imaging

[0612] ST147 NDM1 were grown as previously described and cultures were re-started until OD 600 0.5 was reached. Assays were performed in a final volume of 50 mL containing a 1 : 100 dilution of exponential Kp dilution, 12.5% BRC and increasing doses of anti-Kp mAb (in PBS). Samples were prepared in a 96-well Phenoplate (Perkin Elmer, 6055300) and centrifuged briefly at 1000 rpm to bring the bacteria closer to the well bottom. Acquisition was started immediately after centrifugation and was performed at 37°C, with an image acquisition every 2 minutes for 2 hours.

[0613] 2.14 Microscopy and image analysis

[0614] Imaging was performed using an Opera Phenix platform (Perkin Elmer) and a 63X N.A. 1.15 water objective was used to acquire all samples.

[0615] For mAb binding experiments performed under fixation conditions, 20 fields of view (FOV) per well were selected. For each FOV, 5 z-axis stacks were acquired with a z-axis step size of 0.5 pm between z-axis stacks. Imaging was performed in confocal mode, exciting the samples with lasers at 425 nm, 488 nm and 561 nm and collecting the emitted light with bandpass filters at 435-480 nm, 500-550 nm and 570-630 nm, respectively.

[0616] For bacterial live imaging experiments, 3 FOVs per well were acquired and imaged with three Z-axis stacks with 0.5 pm z-axis step spacing between stacks. Imaging was performed in widefield mode, exciting the sample with a 561 nm laser and collecting light with a 570-630 nm bandpass emission filter.

[0617] Images were analyzed using Harmony (v4.9) provided by Perkin Elmer, in combination with a custom image analysis pipeline described in the supplementary material. Individual bacteria were detected using the DAPI channel and filtered based on their morphology according to data available in the literature. To measure mAb intensity levels, the average intensity of the A488 signal within the ROI drawn around each bacterium was measured. To measure mAb occupancy area, A488 spots were detected and their morphological features were measured. A488 spots with an area smaller than 0.5 mm 2 were excluded from the analysis as they could not represent the mAb signal distribution around the bacteria.

[0618] ST147 NDM-1 Purification of the capsule

[0619] To extract the capsule, ST147 NDM-1Kp was grown in LB in presence of meropenem at 37°C overnight. 200 pL of liquid culture was inoculated on Worfel-Ferguson agar plates enriched with carbohydrates to increase the production of Kp capsule. Worfel-Ferguson agar plates contain 0.2% yeast extract, 0.2% sodium chloride, 0.1% potassium sulfate, 0.025% magnesium sulfate, 2% sucrose and 1.5% agar. After overnight incubation at 37°C, bacteria were mechanically collected from the plates and resuspended in LB. Samples were centrifuged at 4000 rpm for 8 minutes, the pellet was collected and resuspended in H2O. After incubation at 99°C for 6 hours, samples were centrifuged at 4000 rpm for 8 minutes. The supernatant was collected and filtered with a 0.22 pM filter. To precipitate the DNA, 1% CTAB, 5 mM Na2S04 and 0.24 mM NaCl were added to the solution. The sample was vortexed and incubated at 100°C at 550 rpm for 4 hours, then centrifuged at 14000 rpm for 15 minutes. The supernatant was collected, diluted 1 : 10 with 5 mM Na2S04 and incubated at 37°C for 1 hour. After centrifugation at 4000 x g for 15 minutes, the pellet was resuspended in 4 mL of 1 M CaCl2. To remove impurities, 25% of EtOH was added to the sample and left on an inclined plane at room temperature for 30 minutes. The sample was centrifuged at 4000 x g for 15 minutes and the supernatant was collected. To precipitate the capsule, 80% of EtOH was added to the collected supernatant and left on an inclined plane at room temperature for 1 hour. After incubation, it was centrifuged at 4000 x g for 15 minutes. The pellet obtained was resuspended in 1 mL of 1 M NaCl and purified with an Amicon 10K filter, washed at least 10 times in MilliQ H2O to remove protein contaminants and other impurities. The sugar content was evaluated by phenol-H2S04 assay, the protein content was estimated by microBCA (Thermo Fisher) and the presence of DNA was measured using the Qubit kit (add more kit information). Protein and DNA contaminants were <1% compared to the sugar concentration, the mass to estimate the molecular weight distribution was done with SEC-HPLC using a tandem of Tosoh TSKgel PWXL guard-G3000 PWXL columns (4.0 cm x 6.0 mm; cod. 808033 and 30 cm x 7.8 mm; cod. 808021, respectively), 0.1 M NaCl, 0.1 M NaH2P04, pH 7.2 as running buffer.

[0620] 2.16 Purification of Kp strain O antigen

[0621] Extraction of O antigen was based on a published protocol (https: / / pubmed.ncbi.nlm.nih.gov / 32959249 / ) with slight modifications. 300 ml of LB supplemented with 0.4% glucose was inoculated with Kp, which was allowed to reach log phase growth, after which it was pelleted by centrifugation at 4,000 rpm for 15 minutes. The SN was discarded and the pellet was resuspended in 15 ml of 3% acetic acid, the solution was incubated at 90°C with 500 rpm shaking for at least 4 hours, after which the SN was collected and its pH was neutralized with 100 mM NaOH. The sample was desalted using a Disposable PD 10 (Cytiva). 100 mM citrate buffer pH 2.7 was added under constant stirring to a final concentration of 20 mM citrate, incubated at RT for 30 minutes, then centrifuged at 12,000 rpm, 15°C for 30 minutes, the resulting SN was subjected to cation exchange purification with a Sartobind MA75 (Sartorius). After neutralization, a second desalting step was performed, then the sample was subjected to anion exchange with HiTrap Q FF (Cytiva) from GE Healthcare Life Sciences. The column was previously equilibrated with 5 CV of buffer A (10 mM Tris-HCl pH 8), the sample was diluted to 100 mL in buffer A final concentration and loaded on the column with the sample pump. The column was washed with 5 CV of buffer A and eluted with 10 CV of buffer B (10 mM TRIS pH8 + 200 mM NaCl). The flow rate was 5 mL / min for all steps. O antigen elution was in the flow-through, concentrated with Amicon 10K. Total sugar quantification was measured with phenol-sulfuric acid assay (https: / / pubs.acs.org / doi / 10.1021 / ac60111a017), the mass for the estimation of the molecular weight distribution was done with SEC-HPLC using a tandem of Tosoh TSK gel PWXL guard-G3000PWXL columns (4.0 cm x 6.0 mm; cod. 808033 and 30 cm x 7.8 mm; cod. 808021, respectively) with 0.1 M NaCl, 0.1 M NaH2P04, pH 7.2 as running buffer. Detection was done with UV and refractive index detectors. Protein impurities were measured with microBCA (Thermo Fisher).

[0622] 2.17 Purification of Kp O antigen and ELISA of the capsule

[0623] For ELISA, LPS as well as O2a and O2afg antigens (these antigens have been characterized by the team of Chris Whitfield and provided) were used, in addition to in-house purified material, capsule and O antigen to coat high binding 384 well plates (Greiner ref. 781061) and incubated overnight at 4°C. The next day, plates were blocked with PBS plus 1% BSA for 1 hour at 37°C. After blocking, plates were incubated with primary antibody (in-house production) at a final concentration of 10 pg / ml in the presence of PBS, 1% BSA and 0.05% Tween-20 for 2 hours at RT. Next, anti-human IgG secondary antibody conjugated to alkaline phosphatase (Southern Biotech) was added and incubated for 1 hour at 37°C. To detect antigen-mAb binding, pNPP (para-nitrophenyl phosphate; Sigma-Aldrich) was used as soluble substrate and the final reaction was quantitatively analyzed at 405 nm wavelength using a Varioskan Lux reader (Thermo Fisher Scientific). After each incubation step, plates were washed three times with 100 pl / well of wash buffer (PBS plus 0.05% Tween-20). As blank, sample buffer and an irrelevant mAb were used and we considered as positive result the wells with an OD 405 value 3 times higher than the blank. Plasma mix at 1 : 100 dilution was used as positive control.

[0624] 2.18 Immunocompetent ST147 NDM-1 Bacteremia model

[0625] This study was performed in the laboratories of Dr. David P. Nicolau and Dr. Kamilia Abdelraouf at the Center for Anti-infective Research and Development Hartford Hospital. Based on the average body weight of the mouse population, 08009, 05N02 and 05D08 were reconstituted in PBS to the desired concentrations to provide 1, 5, 10 and 20 mg / kg doses. For the prophylaxis (PRO) studies, a single dose of test mAb was administered at the indicated dose by the intraperitoneal (IP) route 24 hours prior to bacterial inoculation. For the therapy (THR) studies, a single dose of test mAb was administered at the indicated dose by the intravenous (IV) route 1 hour after bacterial inoculation. For the prophylaxis plus therapy (PRO+THR) studies, one dose of mAb was administered IP 24 hours prior to bacterial inoculation and a second dose of the same mAb was administered IV 1 hour after bacterial inoculation. All dosing solutions were kept on ice and the syringes containing the dosing solutions were refrigerated until use. PBS solution at pH 7.4 was used as vehicle for dosing control animals throughout the study.

[0626] Specific pathogen-free, female ICR mice weighing 20-22 grams were obtained from Charles River Laboratories, Inc. (Wilmington, MA). The animals were allowed to acclimate to the environment for at least 48 hours prior to the start of the experiment and had free access to food and water. The experimental protocol was reviewed and approved by the Institutional Animal Care and Use Committee at Hartford Hospital. Mice received a single dose of 5 mg / kg uranyl nitrate three days prior to inoculation to induce a controlled degree of renal impairment. A total of 322 mice were used in the experiments described in this report. NDM-1 were previously frozen in defibrinated bovine milk (BD Biosciences, Sparks, MD) at -80°C. Just prior to inoculation of the mice, the organisms were transferred twice onto trypticase soy agar plates containing 5% sheep blood (TSA II TM ; Becton, Dickinson & Co.; Sparks, MD) and incubated at 37°C for approximately 24 hours. After 18-24 hours of secondary transfer, inocula were prepared at approximately the target number of colony forming units (CFU) / ml in 5% porcine gastric mucin for inoculation. Final inoculum concentrations were determined by serial dilution and plating techniques. Sepsis was induced by IP injection of 0.5 ml of inoculum.

[0627] SEQUENCE LISTING IN THE SPECIFICATION

[0628] Antibody sequence identified herein as SBJ03-F18

[0629] Amino acid sequence of SBJ03-F18

[0630] >SEQ ID NO: 1 CDR1 of the heavy chain variable domain of SBJ03-F18

[0631] GFTFSSYS

[0632] >SEQ ID NO: 2 CDR2 of the heavy chain variable domain of SBJ03-F18

[0633] ISGNSNYI

[0634] >SEQ ID NO: 3 CDR3 of the heavy chain variable domain of SBJ03-F18

[0635] ARGTIIGAAGYDC

[0636] >SEQ ID NO: 4 CDR1 of the light chain variable domain of SBJ03-F18

[0637] QDISTS

[0638] >SEQ ID NO: 5 CDR2 of the light chain variable domain of SBJ03-F18, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0639] AAS (Ala-Ala-Ser)

[0640] >SEQ ID NO: 6 CDR3 of the light chain variable domain of SBJ03-F18

[0641] QQLKSYPLA

[0642] >SEQ ID NO: 7 Heavy chain variable domain of SBJ03-F18

[0643] EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISGNSNYIYYADSVKGRFTISRDNAKNSLYLQTSSLRAEDTALYFCARGTIIGAAGYDCWGQGTLV

[0644] TVSS

[0645] >SEQ ID NO: 8 Light chain variable domain of SBJ03-F18

[0646] AIQMTQSPSFLSASVGDRVTLTCRASQDISTSLAWYHQKPGKAPERLVYAASTLQSGVPSRFSGSGSGTGFTLTISSLQPEDFGTYYCQQLKSYPLAFGGGTKVEIK

[0647] >SEQ ID NO: 9 SBJ03-F18 heavy chain

[0648] MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISGNSNYIYYADSVKGRFTISRDNAKNSLYLQTSSLRAEDTALYFCARGTIIGAAGYDCWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG

[0649] >SEQ ID NO: 10 SBJ03-F18 light chain

[0650] MGWSCIILFLVATATGVHSAIQMTQSPSFLSASVGDRVTLTCRASQDISTSLAWYHQKPGKAPERLVYAASTLQSGVPSRFSGSGSGTGFTLTISSLQPEDFGTYYCQQLKSYPLAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0651] Nucleic acid sequence of SBJ03-F18:

[0652] >SEQ ID NO: 11 SBJ03-F18_Heavy chain leader sequence

[0653] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0654] >SEQ ID NO: 12 SBJ03-F18_Heavy chain variable domain

[0655] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTTCCTATAGCATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTGGCAATAGTAATTACATATATTACGCCGACTCAGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCACTCTATCTGCAAACGAGCAGCCTGAGAGCCGAGGACACGGCTCTCTATTTCTGTGCGAGGGGGACAATTATTGGAGCTGCGGGATATGACTGCTGGGGACAGGGAACCCTTGTCACTGTCTCCTCAG

[0656] >SEQ ID NO: 13 SBJ03-F18_Heavy chain constant domain

[0657] CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0658] >SEQ ID NO: 14 SBJ03-F18_Heavy chain complete

[0659]

[0660] >SEQ ID NO: 15 SBJ03-F18_light kappa chain leader sequence

[0661] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0662] >SEQ ID NO: 16 SBJ03-F18_light kappa chain variable domain

[0663] GCCATCCAGATGACCCAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACAGAGTCACACTCACTTGCCGGGCCAGTCAGGACATTAGCACTTCTTTAGCCTGGTATCATCAGAAACCAGGGAAAGCCCCTGAACGCCTGGTCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGGTCTGGGACAGGATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGGAACTTATTACTGTCAACAACTTAAAAGTTACCCTCTCGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAC

[0664] >SEQ ID NO: 17 SBJ03-F18_light kappa chain constant domain

[0665] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0666] >SEQ ID NO: 18 SBJ03-F18_light kappa chain complete

[0667] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGCCATCCAGATGACCCAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACAGAGTCACACTCACTTGCCGGGCCAGTCAGGACATTAGCACTTCTTTAGCCTGGTATCATCAGAAACCAGGGAAAGCCCCTGAACGCCTGGTCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGGTCTGGGACAGGATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGGAACTTATTACTGTCAACAACTTAAAAGTTACCCTCTCGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0668] Antibody sequence identified herein as SBJ03-L02

[0669] Amino acid sequence of SBJ03-L02:

[0670] >SEQ ID NO: 19 CDR1 of SBJ03-L02 heavy chain variable domain

[0671] GFTFSNYG

[0672] >SEQ ID NO: 20 CDR2 of SBJ03-L02 heavy chain variable domain

[0673] ISYDGRNR

[0674] >SEQ ID NO:21 CDR3 of SBJ03-L02 heavy chain variable domain

[0675] AKKILDNGTFQGSY

[0676] >SEQ ID NO:22 CDR1 of SBJ03-L02 light chain variable domain

[0677] RGFGNY

[0678] >SEQ ID NO:23 CDR2 of SBJ03-L02 light chain variable domain, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0679] GAS (Gly-Ala-Ser)

[0680] >SEQ ID NO:24 CDR3 of SBJ03-L02 light chain variable domain

[0681] QKYDNDPWA

[0682] >SEQ ID NO:25 SBJ03-L02 heavy chain variable domain

[0683] EVQLVESGGGVVQPGRSLRLSCTASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGRNRFYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKKILDNGTFQGSYWGQGTLLTVSS

[0684] >SEQ ID NO:26 SBJ03-L02 light chain variable domain

[0685] AIQMTQSPSSLSASVGDRVTITCRASRGFGNYLAWYQQMPGKVPKLLIYGASTLQSGVPSRFSGSGSGTDFSLTISSLQPEDVATYYCQKYDNDPWAFGQGTKVEIK

[0686] >SEQ ID NO:27 SBJ03-L02 heavy chain

[0687] MGWSCIILFLVATATGVHSEVQLVESGGGVVQPGRSLRLSCTASGFTFSNYGMHWVRQAPGKGLEWVAVISYDGRNRFYAESVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKKILDNGTFQGSYWGQGTLLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG

[0688] >SEQ ID NO:28 SBJ03-L02 light chain

[0689] MGWSCIILFLVATATGVHSAIQMTQSPSSLSASVGDRVTITCRASRGFGNYLAWYQQMPGKVPKLLIYGASTLQSGVPSRFSGSGSGTDFSLTISSLQPEDVATYYCQKYDNDPWAFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0690] Nucleic acid sequence of SBJ03-L02:

[0691] >SEQ ID NO:29 SBJ03-L02 heavy chain leader sequence

[0692] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0693] >SEQ ID NO:30 SBJ03-L02_Heavy chain variable domain

[0694] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACTTTCAGTAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGGAATAGATTCTATGCAGAGTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCTAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTCTACTACTGTGCGAAAAAGATACTCGACAACGGTACTTTTCAGGGGAGCTATTGGGGCCAGGGAACCCTACTCACCGTCTCCTCAG

[0695] >SEQ ID NO:31 SBJ03-L02_Heavy chain constant domain

[0696] CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0697] >SEQ ID NO: 32 SBJ03-L02_Heavy chain complete

[0698]

[0699] >SEQ ID NO:33 SBJ03-L02_light kappa chain leader sequence

[0700] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0701] >SEQ ID NO:34 SBJ03-L02_light kappa chain variable domain

[0702] GCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCGAGTCGGGGCTTCGGTAATTATTTAGCCTGGTATCAGCAGATGCCAGGGAAAGTTCCTAAGCTCCTGATCTATGGTGCATCCACTTTGCAATCAGGGGTCCCATCTCGCTTCAGTGGCAGTGGATCTGGGACAGATTTCAGTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAAGTATGACAATGACCCTTGGGCGTTCGGCCAGGGGACCAAGGTGGAAATCAAAC

[0703] >SEQ ID NO:35 SBJ03-L02_light kappa chain constant domain

[0704] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0705] >SEQ ID NO:36 SBJ03-L02_light kappa chain complete

[0706] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCGAGTCGGGGCTTCGGTAATTATTTAGCCTGGTATCAGCAGATGCCAGGGAAAGTTCCTAAGCTCCTGATCTATGGTGCATCCACTTTGCAATCAGGGGTCCCATCTCGCTTCAGTGGCAGTGGATCTGGGACAGATTTCAGTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAAGTATGACAATGACCCTTGGGCGTTCGGCCAGGGGACCAAGGTGGAAATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0707] Antibody sequence identified herein as SBJ05-B17

[0708] Amino acid sequence of SBJ05-B17:

[0709] >SEQ ID NO:37 CDR1 of SBJ05-B17 heavy chain variable domain

[0710] GFPFSSRW

[0711] >SEQ ID NO:38 CDR2 of SBJ05-B17 heavy chain variable domain

[0712] IDTNGRTT

[0713] >SEQ ID NO:39 CDR3 of the heavy chain variable domain of SBJ05-B17

[0714] ARDLPNFDSSDAGWGHGVDV

[0715] >SEQ ID NO:40 CDR1 of the light chain variable domain of SBJ05-B17

[0716] QTISSH

[0717] >SEQ ID NO:41 CDR2 of the light chain variable domain of SBJ05-B17, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0718] AAS (Ala-Ala-Ser)

[0719] >SEQ ID NO:42 CDR3 of the light chain variable domain of SBJ05-B17 LQTYTSLPT

[0720] >SEQ ID NO:43 Heavy chain variable domain of SBJ05-B17

[0721] EVQLVESGGGLVQPGGSLRLSCEASGFPFSSRWIHWVRQGPGKGLVWLSRIDTNGRTTNYADSVNGRFTISKDNGKSTVYLQMNSLRAEDTAVYYCARDLPNFDSSDAGWGHGVDVWGQGTTVIVSS

[0722] >SEQ ID NO:44 Light chain variable domain of SBJ05-B17

[0723] AIQMTQSPSSLSASVGDRVTITCRARQTISSHLSWYQQKPGKPPKSLIYAASHLQSGVPSRFSGSGSGTDFTLTISRLQPEDSATYYCLQTYTSLPTFGPGTKVEIK

[0724] >SEQ ID NO:45 Heavy chain of SBJ05-B17

[0725] MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGGSLRLSCEASGFPFSSRWIHWVRQGPGKGLVWLSRIDTNGRTTNYADSVNGRFTISKDNGKSTVYLQMNSLRAEDTAVYYCARDLPNFDSSDAGWGHGVDVWGQGTTVIVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0726] >SEQ ID NO:46 SBJ05-B17 light chain

[0727] MGWSCIILFLVATATGVHSAIQMTQSPSSLSASVGDRVTITCRARQTISSHLSWYQQKPGKPPKSLIYAASHLQSGVPSRFSGSGSGTDFTLTISRLQPEDSATYYCLQTYTSLPTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0728] Nucleic acid sequence of SBJ05-B17:

[0729] >SEQ ID NO:47 SBJ05-B17 heavy chain leader sequence

[0730] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0731] >SEQ ID NO:48 SBJ05-B17_Heavy chain variable domain

[0732] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTTCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGAAGCCTCTGGGTTCCCCTTCAGTAGTCGCTGGATTCACTGGGTCCGCCAGGGTCCCGGGAAGGGGCTGGTGTGGCTCTCACGTATTGACACTAATGGGAGGACAACAAACTACGCGGACTCCGTAAATGGCCGATTCACCATCTCCAAAGACAACGGCAAGAGCACGGTGTATCTGCAAATGAATAGCCTGAGAGCCGAGGACACGGCTGTGTATTATTGTGCAAGAGATTTGCCCAATTTTGACTCCTCCGATGCAGGATGGGGCCACGGTGTGGACGTCTGGGGCCAAGGGACCACGGTCATCGTCTCCTCA

[0733] >SEQ ID NO:49 SBJ05-B17_Heavy chain constant domain

[0734] GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0735] >SEQ ID NO: 50 SBJ05-B17_Heavy chain complete

[0736]

[0737] >SEQ ID NO: 51 SBJ05-B17_light kappa chain leader sequence

[0738] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0739] >SEQ ID NO: 52 SBJ05-B17_light kappa chain variable domain

[0740] GCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCCGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGGCAGACCATTAGTAGTCACTTAAGTTGGTATCAGCAGAAACCAGGGAAGCCGCCTAAGTCCCTGATCTATGCTGCATCCCACTTGCAAAGTGGGGTCCCATCACGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCCGTCTGCAACCTGAAGATTCTGCTACTTATTATTGTCTACAGACTTATACTTCCCTTCCCACATTCGGCCCTGGGACCAAAGTGGAAATTAAAC

[0741] >SEQ ID NO: 53 SBJ05-B17_light kappa chain constant domain

[0742] GAACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0743] >SEQ ID NO:54 SBJ05-B17_light kappa chain complete

[0744] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCCGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGGCAGACCATTAGTAGTCACTTAAGTTGGTATCAGCAGAAACCAGGGAAGCCGCCTAAGTCCCTGATCTATGCTGCATCCCACTTGCAAAGTGGGGTCCCATCACGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCCGTCTGCAACCTGAAGATTCTGCTACTTATTATTGTCTACAGACTTATACTTCCCTTCCCACATTCGGCCCTGGGACCAAAGTGGAAATTAAACGAACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0745] Antibody sequence identified herein as SBJ05-C11

[0746] Amino acid sequence of SBJ05-C11

[0747] >SEQ ID NO:55 CDR1 of SBJ05-C11 heavy chain variable domain

[0748] GYIFSNNG

[0749] >SEQ ID NO:56 CDR2 of SBJ05-C11 heavy chain variable domain

[0750] ISPYTGHT

[0751] >SEQ ID NO: 57 CDR3 of the heavy chain variable domain of SBJ05-C11

[0752] ATDYQDGVRALAH

[0753] >SEQ ID NO: 58 CDR1 of the light chain variable domain of SBJ05-C11

[0754] NSNIGNNH

[0755] >SEQ ID NO: 59 CDR2 of the light chain variable domain of SBJ05-C11, which sequence is not included in the sequence listing because it is less than 4 amino acids

[0756] DDY (Asp-Asp-Tyr)

[0757] >SEQ ID NO: 60 CDR3 of the light chain variable domain of SBJ05-C11

[0758] GTWDTSLNARV

[0759] >SEQ ID NO: 61 Heavy chain variable domain of SBJ05-C11

[0760] QVQLVQSGADVENPGASVKVSCRSSGYIFSNNGITWVRQVPGQGLEWMGWISPYTGHTNYAQTLQGRVAMTTDTSTSIFYMELRSLRSDDTAVYYCATDYQDGVRALAHWGQGTLVTVSS

[0761] >SEQ ID NO: 62 Light chain variable domain of SBJ05-C11

[0762] QAVVTQEPSMSAAPGQKVTISCSGINSNIGNNHVSWYQQLPGTAPKLLIYDDYKRPSGIPDRFSGSKSGTSATLGITGLQTGDEGDYYCGTWDTSLNARVFGGGTKLTVL

[0763] >SEQ ID NO: 63 Heavy chain of SBJ05-C11

[0764] MGWSCIILFLVATATGVHSQVQLVQSGADVENPGASVKVSCRSSGYIFSNNGITWVRQVPGQGLEWMGWISPYTGHTNYAQTLQGRVAMTTDTSTSIFYMELRSLRSDDTAVYYCATDYQDGVRALAHWGQGTLVTVSSASPTSPKSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0765] >SEQ ID NO:64 SBJ05-C11 light chain

[0766] MGWSCIILFLVATATGSWAQAVVTQEPSMSAAPGQKVTISCSGINSNIGNNHVSWYQQLPGTAPKLLIYDDYKRPSGIPDRFSGSKSGTSATLGITGLQTGDEGDYYCGTWDTSLNARVFGGGTKLTVLSQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS*

[0767] Nucleic acid sequence of SBJ05-C11:

[0768] >SEQ ID NO:65 SBJ05-C11 heavy chain leader sequence

[0769] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0770] >SEQ ID NO:66 SBJ05-C11_Heavy chain variable domain

[0771] CAGGTGCAGCTGGTGCAGTCTGGGGCTGACGTGGAGAACCCTGGGGCCTCAGTGAAAGTCTCCTGCAGGTCTTCTGGTTATATTTTTAGCAACAATGGCATCACCTGGGTGCGACAGGTCCCTGGACAAGGCCTTGAGTGGATGGGGTGGATCAGCCCTTACACTGGTCACACAAACTATGCACAGACGCTCCAGGGCAGAGTCGCCATGACCACAGACACATCCACGAGTATATTCTACATGGAGCTGAGGAGCCTGAGGTCTGACGACACGGCCGTCTATTACTGCGCGACCGATTACCAGGATGGTGTCCGAGCATTGGCCCACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG

[0772] >SEQ ID NO:67 SBJ05-C11_Heavy chain constant domain

[0773] CATCCCCGACCAGCCCCAAGTCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0774] >SEQ ID NO: 68 SBJ05-C11_Heavy chain complete

[0775]

[0776] >SEQ ID NO: 69 SBJ05-C11_light kappa chain leader sequence

[0777] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTTCCTGGGCC

[0778] >SEQ ID NO: 70 SBJ05-C11_light kappa chain variable domain

[0779] CAGGCTGTGGTGACTCAGGAGCCCTCAATGTCTGCGGCCCCAGGACAGAAGGTCACCATCTCCTGCTCTGGAATCAACTCCAACATTGGAAATAATCATGTTTCCTGGTATCAGCAGCTCCCAGGAACAGCCCCCAAACTCCTCATTTATGACGATTATAAGCGACCCTCAGGGATTCCTGACCGATTCTCTGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAGACTGGGGACGAGGGCGATTATTATTGCGGGACATGGGATACCAGCCTGAATGCCCGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA

[0780] >SEQ ID NO: 71 SBJ05-C11_light kappa chain constant domain

[0781] AGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCGAGTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATAG

[0782] >SEQ ID NO:72 SBJ05-C11_light kappa chain complete

[0783] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTTCCTGGGCCCAGGCTGTGGTGACTCAGGAGCCCTCAATGTCTGCGGCCCCAGGACAGAAGGTCACCATCTCCTGCTCTGGAATCAACTCCAACATTGGAAATAATCATGTTTCCTGGTATCAGCAGCTCCCAGGAACAGCCCCCAAACTCCTCATTTATGACGATTATAAGCGACCCTCAGGGATTCCTGACCGATTCTCTGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAGACTGGGGACGAGGGCGATTATTATTGCGGGACATGGGATACCAGCCTGAATGCCCGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAAGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCGAGTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATAG

[0784] Antibody sequence identified herein as SBJ05-D08

[0785] Amino acid sequence of SBJ05-D08

[0786] >SEQ ID NO:73 CDR1 of SBJ05-D08 heavy chain variable domain

[0787] GFTFTKTW

[0788] >SEQ ID NO:74 CDR2 of SBJ05-D08 heavy chain variable domain

[0789] IKSKIDGETT

[0790] >SEQ ID NO:75 CDR3 of SBJ05-D08 heavy chain variable domain

[0791] TSRVLTTNDY

[0792] >SEQ ID NO:76 CDR1 of SBJ05-D08 light chain variable domain

[0793] SSDVGGYDY

[0794] >SEQ ID NO:77 CDR2 of SBJ05-D08 light chain variable domain, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0795] DVS (Asp-Val-Ser)

[0796] >SEQ ID NO:78 CDR3 of SBJ05-D08 light chain variable domain

[0797] CSFTTSGTFV

[0798] >SEQ ID NO:79 SBJ05-D08 heavy chain variable domain

[0799] EVQLVESGGGLVKPGGSLRLSCAASGFTFTKTWMNWVRQAPGKGLEWLGRIKSKIDGETTDYAAPVKGRFTISRDDSKNTVYLQMNSLGTEDTALYYCTSRVLTTNDYWGQGTLVTVSS

[0800] >SEQ ID NO:80 SBJ05-D08 light chain variable domain

[0801] QSALTQPASVSGSPGQSIAISCTGTSSDVGGYDYVSWYQQHPGKVPKHMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQPEDEADYYCCSFTTSGTFVFGTGTKVTVL

[0802] >SEQ ID NO:81 SBJ05-D08 heavy chain

[0803] MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGSLRLSCAASGFTFTKTWMNWVRQAPGKGLEWLGRIKSKIDGETTDYAAPVKGRFTISRDDSKNTVYLQMNSLGTEDTALYYCTSRVLTTNDYWGQGTLVTVSSASPTSPKSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0804] >SEQ ID NO:82 SBJ05-D08 light chain

[0805] MGWSCIILFLVATATGSWAQSALTQPASVSGSPGQSIAISCTGTSSDVGGYDYVSWYQQHPGKVPKHMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQPEDEADYYCCSFTTSGTFVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS*

[0806] Nucleic acid sequence of SBJ05-D08:

[0807] >SEQ ID NO:83 SBJ05-D08 heavy chain leader sequence

[0808] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0809] >SEQ ID NO: 84 SBJ05-D08_Heavy chain variable domain

[0810] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTAAAGCCTGGGGGGTCCCTTAGACTCTCCTGTGCGGCCTCTGGATTCACTTTCACTAAGACCTGGATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGCTTGGTCGTATTAAAAGCAAAATTGATGGTGAGACAACAGACTACGCTGCACCCGTGAAAGGCAGATTCACCATCTCAAGAGATGATTCAAAAAACACGGTGTATCTGCAAATGAACAGCCTGGGAACCGAGGACACAGCCCTATATTACTGTACCTCTCGAGTCCTGACTACGAATGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG

[0811] >SEQ ID NO: 85 SBJ05-D08_Heavy chain constant domain

[0812] CATCCCCGACCAGCCCCAAGTCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0813] >SEQ ID NO: 86 SBJ05-D08_Heavy chain complete

[0814]

[0815] >SEQ ID NO: 87 SBJ05-D08_light kappa chain leader sequence

[0816] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTTCCTGGGCC

[0817] >SEQ ID NO: 88 SBJ05-D08_light kappa chain variable domain

[0818] CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCGCCATCTCCTGCACTGGAACGAGCAGTGACGTTGGTGGTTATGACTATGTCTCCTGGTACCAACAACACCCCGGCAAAGTCCCCAAACACATGATTTATGATGTCAGTAATCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGACTCCAGCCTGAGGACGAGGCTGATTATTACTGCTGCTCATTTACAACCAGCGGAACTTTTGTCTTCGGAACTGGGACCAAGGTCACCGTCCT

[0819] >SEQ ID NO: 89 SBJ05-D08_light kappa chain constant domain

[0820] TGGTCAGCCCAAGGCCAACCCCACTGTCACTCTGTTCCCACCCTCGAGTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATAG

[0821] >SEQ ID NO: 90 SBJ05-D08_light kappa chain complete

[0822] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTTCCTGGGCCCAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCGCCATCTCCTGCACTGGAACGAGCAGTGACGTTGGTGGTTATGACTATGTCTCCTGGTACCAACAACACCCCGGCAAAGTCCCCAAACACATGATTTATGATGTCAGTAATCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGACTCCAGCCTGAGGACGAGGCTGATTATTACTGCTGCTCATTTACAACCAGCGGAACTTTTGTCTTCGGAACTGGGACCAAGGTCACCGTCCTTGGTCAGCCCAAGGCCAACCCCACTGTCACTCTGTTCCCACCCTCGAGTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATAG

[0823] Antibody sequence identified herein as SBJ05-D14

[0824] Amino acid sequence of SBJ05-D14:

[0825] >SEQ ID NO: 91 CDR1 of SBJ05-D14 heavy chain variable domain

[0826] GFTFSNTW

[0827] >SEQ ID NO:92 CDR2 of SBJ05-D14 heavy chain variable domain

[0828] IKRKVDGETT

[0829] >SEQ ID NO:93 CDR3 of SBJ05-D14 heavy chain variable domain

[0830] TSRVLTTNDH

[0831] >SEQ ID NO:94 CDR1 of SBJ05-D14 light chain variable domain

[0832] SSDVGRYDY

[0833] >SEQ ID NO:95 CDR2 of SBJ05-D14 light chain variable domain, which sequence is not included in the sequence listing because it is less than 4 amino acids

[0834] DVS (Asp-Val-Ser)

[0835] >SEQ ID NO:96 CDR3 of SBJ05-D14 light chain variable domain

[0836] CSFTGGEIFV

[0837] >SEQ ID NO:97 SBJ05-D14 heavy chain variable domain

[0838] EVQLVESGGGLVKPGGSLRLSCAASGFTFSNTWMNWVRQAPGKGLEYLGRIKRKVDGETTHYAAPAKGRFTISRDDSKNTLYLQLSSLGTEDTALYYCTSRVLTTNDHWGQGTLVTV

[0839] >SEQ ID NO:98 SBJ05-D14 light chain variable domain

[0840] QSALTQPASVSGSPGQSITISCTGTSSDVGRYDYVSWYQQHPGKVPKLLIYDVSNRPSGGSNRFSGSKSGNTASLTISGLQPEDEADYYCCSFTGGEIFVFGTGTKVTVL

[0841] >SEQ ID NO:99 SBJ05-D14 heavy chain

[0842] MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGSLRLSCAASGFTFSNTWMNWVRQAPGKGLEYLGRIKRKVDGETTHYAAPAKGRFTISRDDSKNTLYLQLSSLGTEDTALYYCTSRVLTTNDHWGQGTLVTVHSSVRSTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0843] >SEQ ID NO: 100 SBJ05-D14 light chain

[0844] MGWSCIILFLVATATGSWAQSALTQPASVSGSPGQSITISCTGTSSDVGRYDYVSWYQQHPGKVPKLLIYDVSNRPSGGSNRFSGSKSGNTASLTISGLQPEDEADYYCCSFTGGEIFVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS*

[0845] Nucleic acid sequence of SBJ05-D14:

[0846] >SEQ ID NO: 101 SBJ05-D14 heavy chain leader sequence

[0847] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0848] >SEQ ID NO: 102 SBJ05-D14_Heavy chain variable domain

[0849] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTAAAGCCTGGGGGGTCCCTTAGACTCTCCTGTGCGGCCTCTGGATTCACTTTCAGTAACACCTGGATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACCTTGGTCGTATTAAAAGGAAAGTTGATGGTGAGACAACACACTACGCTGCACCCGCGAAAGGCAGATTCACCATCTCAAGAGATGATTCCAAAAACACTCTGTATCTGCAACTGAGCAGCCTGGGAACCGAGGACACAGCCCTATATTATTGCACCTCTCGAGTCCTGACTACCAATGACCACTGGGGCCAGGGAACCCTGGTCACCGT

[0850] >SEQ ID NO: 103 SBJ05-D14_Heavy chain constant domain

[0851]

[0852] >SEQ ID NO: 104 SBJ05-D14_Heavy chain complete

[0853]

[0854] >SEQ ID NO: 105 SBJ05-D14_light kappa chain leader sequence

[0855] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTTCCTGGGCC

[0856] >SEQ ID NO: 106 SBJ05-D14_light kappa chain variable domain

[0857] CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGGTCGTTATGACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGTCCCCAAATTGTTGATTTATGATGTCAGTAATCGGCCCTCAGGGGGATCCAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGACTCCAGCCTGAGGACGAGGCCGATTATTACTGCTGCTCATTTACTGGCGGCGAAATTTTTGTCTTCGGAACTGGGACCAAGGTCACTGTCCT

[0858] >SEQ ID NO: 107 SBJ05-D14_light kappa chain constant domain

[0859] TGGTCAGCCCAAGGCCAACCCCACTGTCACTCTGTTCCCACCCTCGAGTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATAG

[0860] >SEQ ID NO: 108 SBJ05-D14_light kappa chain complete

[0861] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTTCCTGGGCCCAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGGTCGTTATGACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGTCCCCAAATTGTTGATTTATGATGTCAGTAATCGGCCCTCAGGGGGATCCAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGACTCCAGCCTGAGGACGAGGCCGATTATTACTGCTGCTCATTTACTGGCGGCGAAATTTTTGTCTTCGGAACTGGGACCAAGGTCACTGTCCTTGGTCAGCCCAAGGCCAACCCCACTGTCACTCTGTTCCCACCCTCGAGTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATAG

[0862] Antibody sequence identified herein as SBJ05-K07

[0863] Amino acid sequence of SBJ05-K07:

[0864] >SEQ ID NO: 127 CDR1 of SBJ05-K07 heavy chain variable domain

[0865] EFTFSSYA

[0866] >SEQ ID NO: 128 CDR2 of SBJ05-K07 heavy chain variable domain

[0867] ISTGGDRT

[0868] >SEQ ID NO: 129 CDR3 of SBJ05-K07 heavy chain variable domain

[0869] AKSLESGSIPTRVRALDY

[0870] >SEQ ID NO: 130 CDR1 of SBJ05-K07 light chain variable domain

[0871] QGITNF

[0872] >SEQ ID NO: 131 CDR2 of SBJ05-K07 light chain variable domain, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0873] AAS (Ala-Ala-Ser)

[0874] >SEQ ID NO: 132 CDR3 of SBJ05-K07 light chain variable domain

[0875] QKYNSAPWT

[0876] >SEQ ID NO: 133 SBJ05-K07 heavy chain variable domain

[0877] EVQLVESGGDLVQPGGSLRLSCAASEFTFSSYAMSWVRQAPGKGLEWVAVISTGGDRT YYADSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVYYCAKSLESGSIPTRVRALDYWG QGTLVTVSS

[0878] >SEQ ID NO: 134 SBJ05-K07 light chain variable domain

[0879] AIQMTQSPSSLSASVGDRVTITCRASQGITNFLAWYQQKPGKVPKLLIYAASTLQSGVPS RFSGSGSGTDFTLTISSLQPEDVATYYCQKYNSAPWTFGQGTKVEIK

[0880] >SEQ ID NO: 135 SBJ05-K07 heavy chain

[0881] MGWSCIILFLVATATGVHSEVQLVESGGDLVQPGGSLRLSCAASEFTFSSYAMSWVRQAPGKGLEWVAVISTGGDRTYYADSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVYYCAKSLESGSIPTRVRALDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG

[0882] >SEQ ID NO: 136 SBJ05-K07 light chain

[0883] MGWSCIILFLVATATGVHSAIQMTQSPSSLSASVGDRVTITCRASQGITNFLAWYQQKPGKVPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQKYNSAPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0884] Nucleic acid sequence of SBJ05-K07:

[0885] >SEQ ID NO: 137 SBJ05-K07 heavy chain leader sequence

[0886] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0887] >SEQ ID NO: 138 SBJ05-K07_Heavy chain variable domain

[0888] GAGGTGCAGCTGGTGGAGTCTGGGGGAGACTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGAATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGCAGTTATTAGTACTGGTGGTGATAGGACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGTTGTATCTGCAAATGAACAGCCTGAGAGTCGAGGACACGGCCGTATATTACTGTGCGAAATCCCTCGAAAGTGGTTCGATACCGACTCGGGTCCGCGCTTTGGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG

[0889] >SEQ ID NO: 139 SBJ05-K07_Heavy chain constant domain

[0890] CTTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0891] >SEQ ID NO: 140 SBJ05-K07_Heavy chain complete

[0892]

[0893] >SEQ ID NO: 141 SBJ05-K07_light kappa chain leader sequence ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0894] >SEQ ID NO: 142 SBJ05-K07_light kappa chain variable domain

[0895] GCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGAGTCACCATCACTTGCCGGGCGAGTCAGGGCATTACCAATTTTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATGCTGCATCCACTTTGCAATCAGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAAGTATAACAGTGCCCCGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAAC

[0896] >SEQ ID NO: 143 SBJ05-K07_light kappa chain constant domain

[0897] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0898] >SEQ ID NO: 144 SBJ05-K07_light kappa chain complete

[0899] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGAGTCACCATCACTTGCCGGGCGAGTCAGGGCATTACCAATTTTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATGCTGCATCCACTTTGCAATCAGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAAGTATAACAGTGCCCCGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0900] Antibody sequence identified herein as SBJ05-M13

[0901] Amino acid sequence of SBJ05-M13

[0902] >SEQ ID NO: 145 CDR1 of the heavy chain variable domain of SBJ05-M13

[0903] GFSFSDYA

[0904] >SEQ ID NO: 146 CDR2 of the heavy chain variable domain of SBJ05-M13

[0905] IDNNGANT

[0906] >SEQ ID NO: 147 CDR3 of SBJ05-M13 heavy chain variable domain

[0907] VRGTTT

[0908] >SEQ ID NO: 148 CDR1 of SBJ05-M13 light chain variable domain

[0909] QSLVHSDGNTY

[0910] >SEQ ID NO: 149 CDR2 of SBJ05-M13 light chain variable domain, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0911] EIS (Glu-Ile-Ser)

[0912] >SEQ ID NO: 150 CDR3 of SBJ05-M13 light chain variable domain

[0913] LQATHFPHGT

[0914] >SEQ ID NO: 151 SBJ05-M13 heavy chain variable domain

[0915] QVQLVQSGGGLVQPGGSLRLSCSASGFSFSDYAVHWVRQPPGKGLEYLSAIDNNGANTFYVDSVKGRFTISRDNSKNMLYLQMSGLRVDDTAVYYCVRGTTTWGQGTLVTVSS >SEQ ID NO: 152 SBJ05-M13 light chain variable domain

[0916] DIVMTQSPLSSSVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPGQPPRLLISEISKRFSGVPDRFSGSGAGTDFTLKISRVEAEDVGVYYCLQATHFPHGTFGQGTKLEIK

[0917] >SEQ ID NO: 153 SBJ05-M13 heavy chain

[0918] MGWSCIILFLVATATGVHSQVQLVQSGGGLVQPGGSLRLSCSASGFSFSDYAVHWVRQPPGKGLEYLSAIDNNGANTFYVDSVKGRFTISRDNSKNMLYLQMSGLRVDDTAVYYCVRGTTTWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0919] >SEQ ID NO: 154 SBJ05-M13 light chain

[0920] MGWSCIILFLVATATGVHSDIVMTQSPLSSSVTLGQPASISCRSSQSLVHSDGNTYLSWLQQRPGQPPRLLISEISKRFSGVPDRFSGSGAGTDFTLKISRVEAEDVGVYYCLQATHFPHGTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0921] Nucleic acid sequence of SBJ05-M13:

[0922] >SEQ ID NO: 155 SBJ05-M13 heavy chain leader sequence

[0923] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0924] >SEQ ID NO: 156 SBJ05-M13 heavy chain variable domain

[0925] CAGGTGCAGCTGGTGCAGTCTGGGGGCGGCTTGGTCCAGCCGGGGGGGTCCCTGAGACTCTCCTGTTCAGCCTCTGGATTCAGCTTCAGTGACTATGCTGTGCACTGGGTCCGCCAGCCTCCAGGGAAGGGACTGGAATACCTTTCAGCTATTGATAATAATGGGGCTAACACATTCTACGTAGACTCCGTGAAGGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACATGTTGTATCTTCAGATGAGTGGGCTGAGAGTTGACGACACCGCTGTGTATTACTGTGTGAGGGGGACTACCACCTGGGGCCAGGGAACCCTAGTCACCGTCTCCTCAG

[0926] >SEQ ID NO: 157 SBJ05-M13 heavy chain constant domain

[0927] CTTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0928] >SEQ ID NO: 158 SBJ05-M13 heavy chain complete

[0929]

[0930] >SEQ ID NO: 159 SBJ05-M13_light kappa chain leader sequence

[0931] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0932] >SEQ ID NO: 160 SBJ05-M13_light kappa chain variable domain

[0933] GATATTGTGATGACTCAGTCTCCACTCTCCTCATCTGTCACCCTTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTACACAGTGATGGAAACACTTACTTGAGTTGGCTTCAGCAGAGGCCAGGCCAGCCACCAAGACTCCTAATTTCTGAGATTTCTAAGCGGTTCTCTGGGGTCCCAGACAGATTCAGTGGCAGTGGGGCAGGGACAGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAAGATGTCGGGGTTTATTATTGCTTGCAAGCTACACATTTTCCTCATGGGACTTTTGGCCAGGGGACCAAGCTGGAGATCAAAC

[0934] >SEQ ID NO: 161 SBJ05-M13_light kappa chain constant domain

[0935] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0936] >SEQ ID NO: 162 SBJ05-M13_light kappa chain complete

[0937] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGATATTGTGATGACTCAGTCTCCACTCTCCTCATCTGTCACCCTTGGACAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAAAGCCTCGTACACAGTGATGGAAACACTTACTTGAGTTGGCTTCAGCAGAGGCCAGGCCAGCCACCAAGACTCCTAATTTCTGAGATTTCTAAGCGGTTCTCTGGGGTCCCAGACAGATTCAGTGGCAGTGGGGCAGGGACAGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAAGATGTCGGGGTTTATTATTGCTTGCAAGCTACACATTTTCCTCATGGGACTTTTGGCCAGGGGACCAAGCTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0938] Antibody sequence identified herein as SBJ05-N02

[0939] Amino acid sequence of SBJ05-N02

[0940] >SEQ ID NO: 163 CDR1 of SBJ05-N02 heavy chain variable domain

[0941] GFTFSNHW

[0942] >SEQ ID NO: 164 CDR2 of SBJ05-N02 heavy chain variable domain

[0943] INPDGTYT

[0944] >SEQ ID NO: 165 CDR3 of SBJ05-N02 heavy chain variable domain

[0945] ARDLPRSDPPGVGWGSGMDV

[0946] >SEQ ID NO: 166 CDR1 of SBJ05-N02 light chain variable domain

[0947] QTISTY

[0948] >SEQ ID NO: 167 CDR2 of SBJ05-N02 light chain variable domain, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0949] ASS (Ala-Ser-Ser)

[0950] >SEQ ID NO: 168 CDR3 of SBJ05-N02 light chain variable domain

[0951] QQSYSTPPT

[0952] >SEQ ID NO: 169 SBJ05-N02 heavy chain variable domain

[0953] EVQLVESGGGLVQPGGSLRLSCAASGFTFSNHWMHWVRQGPGKGLVWVSRINPDGTYTSYADSVSGRFTIARDNAKNTLYLHMNSLRDEDTAVYYCARDLPRSDPPGVGWGSGMDVWGQGTRVTVSS

[0954] >SEQ ID NO: 170 SBJ05-N02 light chain variable domain

[0955] AIQMTQSPSSLSASAGDRVTITCRASQTISTYLSWYQQKPGKAPKMLIYASSSLQSGVPSRFSGSGSGTDFTLTISRLQPEDFATYYCQQSYSTPPTFGPGTKVEIK

[0956] >SEQ ID NO: 171 SBJ05-N02 heavy chain

[0957] MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGGSLRLSCAASGFTFSNHWMHWVRQGPGKGLVWVSRINPDGTYTSYADSVSGRFTIARDNAKNTLYLHMNSLRDEDTAVYYCARDLPRSDPPGVGWGSGMDVWGQGTRVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0958] >SEQ ID NO: 172 SBJ05-N02 light chain

[0959] MGWSCIILFLVATATGVHSAIQMTQSPSSLSASAGDRVTITCRASQTISTYLSWYQQKPGKAPKMLIYASSSLQSGVPSRFSGSGSGTDFTLTISRLQPEDFATYYCQQSYSTPPTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0960] Nucleic acid sequence of SBJ05-N02:

[0961] >SEQ ID NO: 173 SBJ05-N02 heavy chain leader sequence

[0962] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0963] >SEQ ID NO: 174 SBJ05-N02_Heavy chain variable domain

[0964] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTTCAGCCGGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAACCACTGGATGCACTGGGTCCGCCAAGGTCCAGGGAAGGGACTGGTGTGGGTCTCACGAATTAATCCTGATGGGACTTACACAAGTTACGCGGACTCCGTGAGTGGCCGATTCACCATCGCCAGAGACAACGCCAAGAACACCCTGTACCTGCACATGAACAGTCTGAGAGACGAGGACACGGCTGTATATTACTGTGCAAGAGACTTGCCCAGATCTGACCCCCCCGGTGTGGGATGGGGCTCCGGTATGGACGTCTGGGGCCAAGGGACCAGGGTCACCGTCTCCTCA

[0965] >SEQ ID NO: 175 SBJ05-N02_Heavy chain constant domain

[0966] GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[0967] >SEQ ID NO: 176 SBJ05-N02_Heavy chain complete

[0968]

[0969] >SEQ ID NO: 177 SBJ05-N02_light kappa chain leader sequence

[0970] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[0971] >SEQ ID NO: 178 SBJ05-N02_light kappa chain variable domain

[0972] GCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGCAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGACCATTAGCACCTATTTAAGTTGGTATCAGCAAAAACCAGGGAAAGCCCCTAAGATGTTGATCTATGCTTCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCCGTCTGCAACCTGAAGACTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCCACTTTCGGCCCTGGGACCAAAGTGGAGATCAAA

[0973] >SEQ ID NO: 179 SBJ05-N02_light kappa chain constant domain

[0974] AGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0975] >SEQ ID NO: 180 SBJ05-N02_light kappa chain complete

[0976] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGCAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGACCATTAGCACCTATTTAAGTTGGTATCAGCAAAAACCAGGGAAAGCCCCTAAGATGTTGATCTATGCTTCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCCGTCTGCAACCTGAAGACTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCCACTTTCGGCCCTGGGACCAAAGTGGAGATCAAAAGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0977] Antibody sequence identified herein as SBJ08-D18

[0978] Amino acid sequence of SBJ08-D18

[0979] >SEQ ID NO: 181 CDR1 of heavy chain variable domain of SBJ08-D18

[0980] GFTFSNYV

[0981] >SEQ ID NO: 182 CDR2 of heavy chain variable domain of SBJ08-D18

[0982] ISTTGYTT

[0983] >SEQ ID NO: 183 CDR3 of the heavy chain variable domain of SBJ08-D18

[0984] GTLGTTARDFDY

[0985] >SEQ ID NO: 184 CDR1 of the light chain variable domain of SBJ08-D18

[0986] QDIRND

[0987] >SEQ ID NO: 185 CDR2 of the light chain variable domain of SBJ08-D18, this sequence is not included in the sequence listing because it is less than 4 amino acids

[0988] AAS (Ala-Ala-Ser)

[0989] >SEQ ID NO: 186 CDR3 of the light chain variable domain of SBJ08-D18

[0990] LQHNSHPFA

[0991] >SEQ ID NO: 187 Heavy chain variable domain of SBJ08-D18

[0992] EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYVMSWVRQAPGKGLEWVSAISTTGYTTPYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTALYFCGTLGTTARDFDYWGQGTLVTVSS

[0993] >SEQ ID NO: 188 Light chain variable domain of SBJ08-D18

[0994] AIQMTQSPSSLSASVGDRVTITCRASQDIRNDLGWYQQKPGKAPKRLIYAASNLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSHPFAFGPGTKVDIK

[0995] >SEQ ID NO: 189 Heavy chain of SBJ08-D18

[0996] MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGGSLRLSCAASGFTFSNYVMSWVRQAPGKGLEWVSAISTTGYTTPYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTALYFCGTLGTTARDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[0997] >SEQ ID NO: 190 SBJ08-D18 light chain

[0998] MGWSCIILFLVATATGVHSAIQMTQSPSSLSASVGDRVTITCRASQDIRNDLGWYQQKPGKAPKRLIYAASNLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSHPFAFGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[0999] Nucleic acid sequence of SBJ08-D18:

[1000] >SEQ ID NO: 191 SBJ08-D18 heavy chain leader sequence

[1001] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[1002] >SEQ ID NO: 192 SBJ08-D18_Heavy chain variable domain

[1003] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGTTTCACCTTTAGTAACTATGTCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCAATTAGTACTACTGGTTATACCACACCCTACGCAAACTCTGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCCTTTATTTCTGTGGGACCCTGGGGACAACAGCACGCGATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG

[1004] >SEQ ID NO: 193 SBJ08-D18_Heavy chain constant domain

[1005] CTTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[1006] >SEQ ID NO: 194 SBJ08-D18_Heavy chain complete

[1007]

[1008] >SEQ ID NO: 195 SBJ08-D18_light kappa chain leader sequence

[1009] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[1010] >SEQ ID NO: 196 SBJ08-D18_light kappa chain variable domain

[1011] GCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTCGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGACATTAGAAATGACTTAGGCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCGCCTGATCTATGCTGCATCCAATTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAACATAATAGTCACCCATTCGCTTTCGGCCCTGGGACCAAAGTGGATATCAAAC

[1012] >SEQ ID NO: 197 SBJ08-D18_light kappa chain constant domain

[1013] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[1014] >SEQ ID NO: 198 SBJ08-D18_light kappa chain complete ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTCGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGACATTAGAAATGACTTAGGCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCGCCTGATCTATGCTGCATCCAATTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAACATAATAGTCACCCATTCGCTTTCGGCCCTGGGACCAAAGTGGATATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[1015] Antibody sequence identified herein as SBJ08-F04

[1016] Amino acid sequence of SBJ08-F04

[1017] >SEQ ID NO: 199 CDR1 of SBJ08-F04 heavy chain variable domain

[1018] GFTFSSYG

[1019] >SEQ ID NO: 200 CDR2 of SBJ08-F04 heavy chain variable domain

[1020] IWYDGSNK

[1021] >SEQ ID NO: 201 CDR3 of the heavy chain variable domain of SBJ08-F04

[1022] ASEYYYGSSGHLPIDC

[1023] >SEQ ID NO: 202 CDR1 of the light chain variable domain of SBJ08-F04

[1024] QSVSSY

[1025] >SEQ ID NO: 203 CDR2 of the light chain variable domain of SBJ08-F04, this sequence is not included in the sequence listing because it is less than 4 amino acids

[1026] DAS (Asp-Ala-Ser)

[1027] >SEQ ID NO: 204 CDR3 of the light chain variable domain of SBJ08-F04

[1028] QQRSNWPRT

[1029] >SEQ ID NO: 205 Heavy chain variable domain of SBJ08-F04

[1030] EVQLVESGGGVVQPGRSLRLSCVASGFTFSSYGMHWVRQAPGKGLEWVAIIWYDGSNK YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASEYYYGSSGHLPIDCWGQGTLVTVSS

[1031] >SEQ ID NO: 206 Light chain variable domain of SBJ08-F04

[1032] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLVIYDASNRATGIPARF SGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKVEIK

[1033] >SEQ ID NO: 207 Heavy chain of SBJ08-F04

[1034] MGWSCIILFLVATATGVHSEVQLVESGGGVVQPGRSLRLSCVASGFTFSSYGMHWVRQAPGKGLEWVAIIWYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASEYYYGSSGHLPIDCWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[1035] >SEQ ID NO:208 SBJ08-F04 light chain

[1036] MGWSCIILFLVATATGVHSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLVIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*

[1037] Nucleic acid sequence of SBJ08-F04:

[1038] >SEQ ID NO:209 SBJ08-F04 heavy chain leader sequence

[1039] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[1040] >SEQ ID NO: 210 SBJ08-F04_Heavy chain variable domain

[1041] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGCGTCTGGATTCACCTTCAGTAGTTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAATTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTCTATTATTGTGCGAGCGAGTATTACTATGGTAGTAGTGGTCATTTACCCATTGACTGTTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAG

[1042] >SEQ ID NO: 211 SBJ08-F04_Heavy chain constant domain

[1043] CTTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[1044] >SEQ ID NO: 212 SBJ08-F04_Heavy chain complete

[1045]

[1046] >SEQ ID NO: 213 SBJ08-F04_light kappa chain leader sequence

[1047] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[1048] >SEQ ID NO: 214 SBJ08-F04_light kappa chain variable domain

[1049] GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGTTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCGTCATCTATGATGCATCCAACAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAAC

[1050] >SEQ ID NO: 215 SBJ08-F04_light kappa chain constant domain

[1051] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[1052] >SEQ ID NO:216 SBJ08-F04 light kappa chain complete

[1053] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCCGAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGTTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCGTCATCTATGATGCATCCAACAGGGCCACTGGCATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[1054] Antibody sequence identified herein as SBJ08-H10

[1055] Amino acid sequence of SBJ08-H10:

[1056] >SEQ ID NO:217 CDR1 of SBJ08-H10 heavy chain variable domain

[1057] GFTFSSHA

[1058] >SEQ ID NO:218 CDR2 of SBJ08-H10 heavy chain variable domain

[1059] ISGTGDRT

[1060] >SEQ ID NO: 219 CDR3 of SBJ08-H10 heavy chain variable domain

[1061] ARAPIGTFYYDN

[1062] >SEQ ID NO: 220 CDR1 of SBJ08-H10 light chain variable domain

[1063] QSVGTRN

[1064] >SEQ ID NO: 221 CDR2 of SBJ08-H10 light chain variable domain, this sequence is not included in the sequence listing because it is less than 4 amino acids

[1065] GAS (Gly-Ala-Ser)

[1066] >SEQ ID NO: 222 CDR3 of SBJ08-H10 light chain variable domain

[1067] QQYGISPRT

[1068] >SEQ ID NO: 223 SBJ08-H10 heavy chain variable domain

[1069] EVQLVESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAISGTGDRTDDTGSVRGRFTISRDNSKNTVYLQMFFLTVEDTALYYCARAPIGTFYYDNWGQGTLVTVSS

[1070] >SEQ ID NO: 224 SBJ08-H10 light chain variable domain

[1071] ETTLTQSPGTLSLSPGERATLSCRASQSVGTRNLAWYQQKPGQSPRLLIYGASSRATGIPDRFSGGGSGTDFTLTISRLEPEDFAVYYCQQYGISPRTFGQGTKVEIK

[1072] >SEQ ID NO: 225 SBJ08-H10 heavy chain

[1073] HGMVMYHPFSSTTATGVHSEVQLVESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAISGTGDRTDDTGSVRGRFTISRDNSKNTVYLQMFFLTVEDTALYYCARAPIGTFYYDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHGALHNHYTQKSLSLFPG*

[1074] >SEQ ID NO: 226 SBJ08-H10 light chain

[1075] MGWSCIILFLVATATGVHSETTLTQSPGTLSLSPGERATLSCRASQSVGTRNLAWYQQKPGQSPRLLIYGASSRATGIPDRFSGGGSGTDFTLTISRLEPEDFAVYYCQQYGISPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1076] Nucleic acid sequence of SBJ08-H10:

[1077] >SEQ ID NO: 227 SBJ08-H10 heavy chain leader sequence

[1078] CATGGGATGGTCATGTATCATCCTTTTTCTAGTACAACTGCAACCGGTGTACATTCC

[1079] >SEQ ID NO: 228 SBJ08-H10_Heavy chain variable domain

[1080] GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTAGTAGTCATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCCATAAGTGGCACTGGTGATAGAACAGACGACACAGGCTCCGTGAGGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGGTGTATCTGCAAATGTTTTTTTTGACAGTCGAGGACACGGCCCTATATTACTGTGCGAGAGCCCCTATCGGAACTTTCTACTATGACAACTGGGGCCAGGGAACCCTGGTCACCGTCTCATCAG

[1081] >SEQ ID NO: 229 SBJ08-H10_Heavy chain constant domain

[1082] CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGGCGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTTCCCCGGGTAA

[1083] >SEQ ID NO: 230 SBJ08-H10_Heavy chain complete

[1084]

[1085] >SEQ ID NO: 231 SBJ08-H10_light kappa chain leader sequence

[1086] ATGGGATGGTCATGTATCATCCTTTTTCTAGTAGCAACTGCAACCGGTGTACATTCC

[1087] >SEQ ID NO: 232 SBJ08-H10_light kappa chain variable domain

[1088] GAAACGACACTCACGCAGTCTCCAGGCACCCTGTCTCTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTGGCACCAGGAACTTAGCCTGGTACCAGCAGAAACCTGGCCAGTCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCGGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCAGTATGGTATCTCACCTCGCACGTTCGGCCAAGGGACCAAGGTGGAAATCAAAC

[1089] >SEQ ID NO: 233 SBJ08-H10_light kappa chain constant domain

[1090] GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCA...

Claims

1. A human monoclonal antibody or an antigen-binding portion thereof that specifically binds to a surface antigen of Klebsiella pneumoniae.

2. The human monoclonal antibody or an antigen-binding portion thereof of claim 1 that specifically binds to a surface antigen of at least one drug resistant or multi-drug resistant Klebsiella pneumoniae strain.

3. The human monoclonal antibody or an antigen-binding portion thereof of claim 2 that specifically binds to a surface antigen of at least one New Delhi metallo-beta-lactamase (NDM)-producing Klebsiella pneumoniae strain.

4. The human monoclonal antibody or an antigen-binding portion thereof of claim 2 or 3, wherein the at least one Klebsiella pneumoniae strain is a Klebsiella pneumoniae capsular type K64 strain.

5. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 2 to 4, wherein the at least one Klebsiella pneumoniae strain is selected from the group consisting of Klebsiella pneumoniae sequence type 147 (ST147), Klebsiella pneumoniae sequence type 258 (ST258), Klebsiella pneumoniae sequence type 493 (ST493), Klebsiella pneumoniae sequence type 307 (ST307), and Klebsiella pneumoniae sequence type 13 (ST13).

6. The human monoclonal antibody or an antigen-binding portion thereof of claim 5, wherein the at least one Klebsiella pneumoniae strain is Klebsiella pneumoniae sequence type 147 (ST147).

7. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 2 to 6, wherein the at least one Klebsiella pneumoniae strain is selected from a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain or a NDM-9 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

8. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 2 to 7 that specifically binds to and is bactericidal against the capsular antigen of at least one of the following strains: a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain and a NDM-9 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

9. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 2 to 8 that specifically binds to the capsular antigen of a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain and is bactericidal against the strain.

10. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 1 to 8, wherein the surface antigen is selected from a capsular antigen or an O antigen.

11. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 1 to 10, wherein the surface antigen is a capsular antigen.

12. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 1 to 11, wherein the surface antigen is a capsular polysaccharide.

13. The human monoclonal antibody or an antigen-binding portion thereof of any one of claims 1 to 12, wherein the surface antigen is a K64 type capsular polysaccharide.

14. The human monoclonal antibody or antigen-binding portion thereof according to any one of claims 1 to 13, wherein the antibody or antigen-binding portion thereof shows a 50% inhibitory concentration (IC50) of less than 100 ng / ml, preferably less than 10 ng / ml, more preferably less than 5 ng / ml, when detected by an in vitro fluorescent serum bactericidal assay (F-SBA).

15. The human monoclonal antibody or antigen-binding portion thereof according to claim 14, wherein the antibody or antigen-binding portion is detected in an in vitro F-SBA assay against the ST147 NDM-1 strain, the ST147 NDM-9 strain and / or the ST307 NDM-5 strain of Klebsiella pneumoniae.

16. The human monoclonal antibody or antigen-binding portion thereof according to claim 14, wherein the antibody or antigen-binding portion is detected in an in vitro F-SBA assay against the ST147 NDM-1 strain and / or the ST147 NDM-9 strain of Klebsiella pneumoniae.

17. The human monoclonal antibody or antigen-binding portion thereof according to any one of claims 1 to 16, comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH and VL comprise the following complementarity determining regions (CDRs): - a VH CDR1 of SEQ ID NO: 325, - a VH CDR2 of SEQ ID NO: 326, - a VH CDR3 of SEQ ID NO: 327, - a VL CDR1 of SEQ ID NO: 328, - a VL CDR2 of the sequence DAS (Asp-Ala-Ser) and - a VL CDR3 of SEQ ID NO: 330; or - a VH CDR1 of SEQ ID NO: 181, - a VH CDR2 of SEQ ID NO: 182, - a VH CDR3 of SEQ ID NO: 183, - a VL CDR1 of SEQ ID NO: 184, - a VL CDR2 of the sequence AAS (Ala-Ala-Ser) and - a VL CDR3 of SEQ ID NO: 186; or - a VH CDR1 of SEQ ID NO: 1, - a VH CDR2 of SEQ ID NO: 2, - a VH CDR3 of SEQ ID NO: 3, - a VL CDR1 of SEQ ID NO: 4, - a VL CDR2 of the sequence AAS (Ala-Ala-Ser) and - a VL CDR3 of SEQ ID NO: 6; or - a VH CDR1 of SEQ ID NO: 19, - a VH CDR2 of SEQ ID NO: 20, - a VH CDR3 of SEQ ID NO: 21, - a VL CDR1 of SEQ ID NO: 22, - a VL CDR2 of the sequence AAS (Ala-Ala-Ser) and - a VL CDR3 of SEQ ID NO:

24. -VL CDR2 with sequence GAS (Gly-Ala-Ser) and -VL CDR3 with SEQ ID NO:24; or -VH CDR1 with SEQ ID NO:37 -VH CDR2 with SEQ ID NO:38 -VH CDR3 with SEQ ID NO:39 -VL CDR1 with SEQ ID NO:40 -VL CDR2 with sequence AAS (Ala-Ala-Ser) and - VL CDR3 with SEQ ID NO:42; or -VH CDR1 with SEQ ID NO:55 -VH CDR2 with SEQ ID NO:56 -VH CDR3 with SEQ ID NO:57 -VL CDR1 with SEQ ID NO:58 -VL CDR2 with the sequence DDY(Asp-Asp-Tyr) and -VL CDR3 with SEQ ID NO:60; or -VH CDR1 with SEQ ID NO:73, -VH CDR2 with SEQ ID NO:74 -VH CDR3 with SEQ ID NO:75 -VL CDR1 with SEQ ID NO:76 -VL CDR2 with sequence DVS (Asp-Val-Ser) and - VL CDR3 with SEQ ID NO:78; or -VH CDR1 with SEQ ID NO:91, -VH CDR2 with SEQ ID NO:92, -VH CDR3 with SEQ ID NO:93 -VL CDR1 with SEQ ID NO:94 -VL CDR2 with sequence DVS (Asp-Val-Ser) and - VL CDR3 with SEQ ID NO:96; or -VH CDR1 with SEQ ID NO:127, -VH CDR2 with SEQ ID NO:128, -VH CDR3 with SEQ ID NO:129 -VL CDR1 with SEQ ID NO:130, -VL CDR2 with sequence AAS (Ala-Ala-Ser) and - VL CDR3 with SEQ ID NO:132; or -VH CDR1 with SEQ ID NO:145, -VH CDR2 with SEQ ID NO:146, -VH CDR3 with SEQ ID NO:147 -VL CDR1 with SEQ ID NO:148 -VL CDR2 with sequence EIS (Glu-Ile-Ser) and - VL CDR3 with SEQ ID NO:150; or - a VH CDR1 having SEQ ID NO: 163, - a VH CDR2 having SEQ ID NO: 164, - a VH CDR3 having SEQ ID NO: 165, - a VL CDR1 having SEQ ID NO: 166, - a VL CDR2 having the sequence ASS (Ala-Ser-Ser) and - a VL CDR3 having SEQ ID NO: 168; or - a VH CDR1 having SEQ ID NO: 199, - a VH CDR2 having SEQ ID NO: 200, - a VH CDR3 having SEQ ID NO: 201, - a VL CDR1 having SEQ ID NO: 202, - a VL CDR2 having the sequence DAS (Asp-Ala-Ser) and - a VL CDR3 having SEQ ID NO: 204; or - a VH CDR1 having SEQ ID NO: 217, - a VH CDR2 having SEQ ID NO: 218, - a VH CDR3 having SEQ ID NO: 219, - a VL CDR1 having SEQ ID NO: 220, - a VL CDR2 having the sequence GAS (Gly-Ala-Ser) and - a VL CDR3 having SEQ ID NO: 222; or - a VH CDR1 having SEQ ID NO: 235, - a VH CDR2 having SEQ ID NO: 236, - a VH CDR3 having SEQ ID NO: 237, - a VL CDR1 having SEQ ID NO: 238, - a VL CDR2 having the sequence QIS (Gln-Ile-Ser) and - a VL CDR3 having SEQ ID NO: 240; or - a VH CDR1 having SEQ ID NO: 253, - a VH CDR2 having SEQ ID NO: 254, - a VH CDR3 having SEQ ID NO: 255, - a VL CDR1 having SEQ ID NO: 256, - a VL CDR2 having the sequence DAS (Asp-Ala-Ser) and - a VL CDR3 having SEQ ID NO: 258; or - a VH CDR1 having SEQ ID NO: 271, - a VH CDR2 having SEQ ID NO: 272, - a VH CDR3 having SEQ ID NO: 273, - a VL CDR1 having SEQ ID NO: 274, - a VL CDR2 having the sequence AAS (Ala-Ala-Ser) and - a VL CDR3 having SEQ ID NO: 276; or - a VH CDR1 having SEQ ID NO: 289, - a VH CDR2 having SEQ ID NO: 290, - a VH CDR3 having the sequence of SEQ ID NO: 291, - a VL CDR1 having the sequence of SEQ ID NO: 292, - a VL CDR2 having the sequence KVS (Lys-Val-Ser) and - a VL CDR3 having the sequence of SEQ ID NO: 294; or - a VH CDR1 having the sequence of SEQ ID NO: 307, - a VH CDR2 having the sequence of SEQ ID NO: 308, - a VH CDR3 having the sequence of SEQ ID NO: 309, - a VL CDR1 having the sequence of SEQ ID NO: 310, - a VL CDR2 having the sequence GVS (Gly-Val-Ser) and - a VL CDR3 having the sequence of SEQ ID NO: 312; or - a VH CDR1 having the sequence of SEQ ID NO: 343, - a VH CDR2 having the sequence of SEQ ID NO: 344, - a VH CDR3 having the sequence of SEQ ID NO: 345, - a VL CDR1 having the sequence of SEQ ID NO: 346, - a VL CDR2 having the sequence EVS (Glu-Val-Ser) and - a VL CDR3 having the sequence of SEQ ID NO: 348; or - a VH CDR1 having the sequence of SEQ ID NO: 361, - a VH CDR2 having the sequence of SEQ ID NO: 362, - a VH CDR3 having the sequence of SEQ ID NO: 363, - a VL CDR1 having the sequence of SEQ ID NO: 364, - a VL CDR2 having the sequence GVS (Gly-Val-Ser) and - a VL CDR3 having the sequence of SEQ ID NO: 366; or - a VH CDR1 having the sequence of SEQ ID NO: 379, - a VH CDR2 having the sequence of SEQ ID NO: 380, - a VH CDR3 having the sequence of SEQ ID NO: 381, - a VL CDR1 having the sequence of SEQ ID NO: 382, - a VL CDR2 having the sequence EVS (Glu-Val-Ser) and - a VL CDR3 having the sequence of SEQ ID NO: 383; or - a VH CDR1 having the sequence of SEQ ID NO: 397, - a VH CDR2 having the sequence of SEQ ID NO: 398, - a VH CDR3 having the sequence of SEQ ID NO: 399, - a VL CDR1 having the sequence of SEQ ID NO: 400, - a VL CDR2 having the sequence KVS (Lys-Val-Ser) and - a VL CDR3 having the sequence of SEQ ID NO: 402; or - a VH CDR1 having the sequence of SEQ ID NO: 415, - a VH CDR2 having the sequence of SEQ ID NO: 416, - a VH CDR3 having the sequence of SEQ ID NO: 417, - a VL CDR1 having the sequence of SEQ ID NO: 418, ​ ​ ​ ​ ​ ​ -VL CDR2 with sequence GAS (Gly-Ala-Ser) and - VL CDR3 with SEQ ID NO:420; or -VH CDR1 with SEQ ID NO:433, -VH CDR2 with SEQ ID NO:434, -VH CDR3 with SEQ ID NO:435 -VL CDR1 with SEQ ID NO:436 -VL CDR2 with sequence RVS (Arg-Val-Ser) and - VL CDR3 with SEQ ID NO:

438.

18. The human monoclonal antibody or its antigen-binding moiety according to any one of claims 1 to 17, comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), Wherein the VH has SEQ ID NO:331 and the VL has SEQ ID NO:332; or Wherein the VH has SEQ ID NO:187 and the VL has SEQ ID NO:188; or Wherein the VH has SEQ ID NO:7 and the VL has SEQ ID NO:8; or Wherein the VH has SEQ ID NO:25 and the VL has SEQ ID NO:26; or Wherein the VH has SEQ ID NO:43 and the VL has SEQ ID NO:44; or Wherein the VH has SEQ ID NO:61 and the VL has SEQ ID NO:62; or Wherein the VH has SEQ ID NO:79 and the VL has SEQ ID NO:80; or Wherein the VH has SEQ ID NO:97 and the VL has SEQ ID NO:98; or Wherein the VH has SEQ ID NO:133 and the VL has SEQ ID NO:134; or Wherein the VH has SEQ ID NO:151 and the VL has SEQ ID NO:152; or Wherein the VH has SEQ ID NO:169 and the VL has SEQ ID NO:170; or Wherein the VH has SEQ ID NO:205 and the VL has SEQ ID NO:206; or Wherein the VH has SEQ ID NO:223 and the VL has SEQ ID NO:224; or Wherein the VH has SEQ ID NO:241 and the VL has SEQ ID NO:242; or Wherein the VH has SEQ ID NO:259 and the VL has SEQ ID NO:260; or Wherein the VH has SEQ ID NO:277 and the VL has SEQ ID NO:278; or wherein the VH has SEQ ID NO: 295 and the VL has SEQ ID NO: 296; or wherein the VH has SEQ ID NO: 313 and the VL has SEQ ID NO: 314; or wherein the VH has SEQ ID NO: 349 and the VL has SEQ ID NO: 350; or wherein the VH has SEQ ID NO: 367 and the VL has SEQ ID NO: 368; or wherein the VH has SEQ ID NO: 385 and the VL has SEQ ID NO: 386; or wherein the VH has SEQ ID NO: 403 and the VL has SEQ ID NO: 404; or wherein the VH has SEQ ID NO: 421 and the VL has SEQ ID NO: 422; or wherein the VH has SEQ ID NO: 439 and the VL has SEQ ID NO:

440.

19. The human monoclonal antibody or an antigen binding portion thereof according to any one of claims 1 to 18, wherein the VL and the VH are at least 85%, preferably at least 95%, more preferably at least 99% identical to the amino acid sequences of the following VH and VL: the VH having SEQ ID NO: 331 and the VL having SEQ ID NO: 332; or the VH having SEQ ID NO: 187 and the VL having SEQ ID NO: 188; or the VH having SEQ ID NO: 7 and the VL having SEQ ID NO: 8; or the VH having SEQ ID NO: 25 and the VL having SEQ ID NO: 26; or the VH having SEQ ID NO: 43 and the VL having SEQ ID NO: 44; or the VH having SEQ ID NO: 61 and the VL having SEQ ID NO: 62; or the VH having SEQ ID NO: 79 and the VL having SEQ ID NO: 80; or the VH having SEQ ID NO: 97 and the VL having SEQ ID NO: 98; or the VH having SEQ ID NO: 133 and the VL having SEQ ID NO: 134; or the VH having SEQ ID NO: 151 and the VL having SEQ ID NO: 152; or the VH having SEQ ID NO: 169 and the VL having SEQ ID NO: 170; or the VH having SEQ ID NO: 205 and the VL having SEQ ID NO: 206; or the VH having SEQ ID NO: 223 and the VL having SEQ ID NO: 224; or the VH having SEQ ID NO: 241 and the VL having SEQ ID NO: 242; or the VH having SEQ ID NO: 259 and the VL having SEQ ID NO: 260; or the VH having SEQ ID NO: 277 and the VL having SEQ ID NO: 278; or the VH having SEQ ID NO: 295 and the VL having SEQ ID NO: 296; or the VH having SEQ ID NO: 313 and the VL having SEQ ID NO: 314; or the VH having SEQ ID NO: 331 and the VL having SEQ ID NO: 332; or the VH having SEQ ID NO: 349 and the VL having SEQ ID NO: 350; or the VH having SEQ ID NO: 367 and the VL having SEQ ID NO: 368; or the VH having SEQ ID NO: 385 and the VL having SEQ ID NO: 386; or the VH having SEQ ID NO: 403 and the VL having SEQ ID NO: 404; or the VH having SEQ ID NO: 421 and the VL having SEQ ID NO: 422; or the VH having SEQ ID NO: 439 and the VL having SEQ ID NO:

440. the VHhas SEQ ID NO: 277 and the VLhas SEQ ID NO: 278; or the VHhas SEQ ID NO: 295 and the VLhas SEQ ID NO: 296; or the VHhas SEQ ID NO: 313 and the VLhas SEQ ID NO: 314; or the VHhas SEQ ID NO: 349 and the VLhas SEQ ID NO: 350; or the VHhas SEQ ID NO: 367 and the VLhas SEQ ID NO: 368; or the VHhas SEQ ID NO: 385 and the VLhas SEQ ID NO: 386; or the VHhas SEQ ID NO: 403 and the VLhas SEQ ID NO: 404; or the VHhas SEQ ID NO: 421 and the VLhas SEQ ID NO: 422; or the VHhas SEQ ID NO: 439 and the VLhas SEQ ID NO:

440.

20. The human monoclonal antibody of any one of claims 1 to 19, wherein the heavy chain of the antibody has SEQ ID NO: 333 and the light chain of the antibody has SEQ ID NO: 334, or the heavy chain of the antibody has SEQ ID NO: 189 and the light chain of the antibody has SEQ ID NO: 190, or the heavy chain of the antibody has SEQ ID NO: 9 and the light chain of the antibody has SEQ ID NO: 10; or the heavy chain of the antibody has SEQ ID NO: 27 and the light chain of the antibody has SEQ ID NO: 28, or the heavy chain of the antibody has SEQ ID NO: 45 and the light chain of the antibody has SEQ ID NO: 46, or the heavy chain of the antibody has SEQ ID NO: 63 and the light chain of the antibody has SEQ ID NO: 64, the heavy chain of the antibody has SEQ ID NO: 81 and the light chain of the antibody has SEQ ID NO: 82, the heavy chain of the antibody has SEQ ID NO: 99 and the light chain of the antibody has SEQ ID NO: 100, or the heavy chain of the antibody has SEQ ID NO: 135 and the light chain of the antibody has SEQ ID NO: 136, or the heavy chain of the antibody has SEQ ID NO: 153 and the light chain of the antibody has SEQ ID NO: 154, or the heavy chain of the antibody has SEQ ID NO: 171 and the light chain of the antibody has SEQ ID NO: 172, or the heavy chain of the antibody has SEQ ID NO: 207 and the light chain of the antibody has SEQ ID NO: 208, or the heavy chain of the antibody has SEQ ID NO: 225 and the light chain of the antibody has SEQ ID NO: 226, or the heavy chain of the antibody has SEQ ID NO: 243 and the light chain of the antibody has SEQ ID NO: 244, or the heavy chain of the antibody has SEQ ID NO: 261 and the light chain of the antibody has SEQ ID NO: 262, or the heavy chain of the antibody has SEQ ID NO: 279 and the light chain of the antibody has SEQ ID NO: 280, or the heavy chain of the antibody has SEQ ID NO: 297 and the light chain of the antibody has SEQ ID NO: 298, or the heavy chain of the antibody has SEQ ID NO: 315 and the light chain of the antibody has SEQ ID NO: 316, or the heavy chain of the antibody has SEQ ID NO: 333 and the light chain of the antibody has SEQ ID NO: 334, or the heavy chain of the antibody has SEQ ID NO: 351 and the light chain of the antibody has SEQ ID NO: 352, or the heavy chain of the antibody has SEQ ID NO: 370 and the light chain of the antibody has SEQ ID NO: 371, or the heavy chain of the antibody has SEQ ID NO: 389 and the light chain of the antibody has SEQ ID NO: 390, or the heavy chain of the antibody has SEQ ID NO: 407 and the light chain of the antibody has SEQ ID NO: 408, or the heavy chain of the antibody has SEQ ID NO: 425 and the light chain of the antibody has SEQ ID NO: 426, or the heavy chain of the antibody has SEQ ID NO: 443 and the light chain of the antibody has SEQ ID NO:

444. the heavy chain of said antibody has the sequence of SEQ ID NO: 225 and the light chain of said antibody has the sequence of SEQ ID NO: 226, or the heavy chain of said antibody has the sequence of SEQ ID NO: 243 and the light chain of said antibody has the sequence of SEQ ID NO: 244, or the heavy chain of said antibody has the sequence of SEQ ID NO: 261 and the light chain of said antibody has the sequence of SEQ ID NO: 262, or the heavy chain of said antibody has the sequence of SEQ ID NO: 279 and the light chain of said antibody has the sequence of SEQ ID NO: 280, or the heavy chain of said antibody has the sequence of SEQ ID NO: 297 and the light chain of said antibody has the sequence of SEQ ID NO: 298, or the heavy chain of said antibody has the sequence of SEQ ID NO: 315 and the light chain of said antibody has the sequence of SEQ ID NO: 316, or the heavy chain of said antibody has the sequence of SEQ ID NO: 351 and the light chain of said antibody has the sequence of SEQ ID NO: 352, or the heavy chain of said antibody has the sequence of SEQ ID NO: 369 and the light chain of said antibody has the sequence of SEQ ID NO: 370, or the heavy chain of said antibody has the sequence of SEQ ID NO: 387 and the light chain of said antibody has the sequence of SEQ ID NO: 388, or the heavy chain of said antibody has the sequence of SEQ ID NO: 405 and the light chain of said antibody has the sequence of SEQ ID NO: 406, or the heavy chain of said antibody has the sequence of SEQ ID NO: 423 and the light chain of said antibody has the sequence of SEQ ID NO: 424, or the heavy chain of said antibody has the sequence of SEQ ID NO: 441 and the light chain of said antibody has the sequence of SEQ ID NO:

442.

21. An immunospecific polypeptide comprising at least a variable domain, in particular at least a heavy chain variable domain (VH) and a light chain variable domain (VL) as defined in any one of claims 17 to 20, or any one of the CDRs as defined in claim 17, in particular said immunospecific polypeptide is a multispecific antibody, a bispecific antibody, a trispecific antibody, a monoclonal antibody, a scFV, a diabody, a triabody, a tetrabody, a minibody, a linear antibody, a chelating recombinant antibody, a triabody, a diabody, an intrabody, a nanobody, a binding domain immunoglobulin fusion protein, a fusion antibody, an immunoadhesin, or an antigen binding fragment thereof.

22. The human monoclonal antibody or antigen binding portion thereof according to any one of claims 1 to 20 or the immunospecific polypeptide according to claim 21 for use in the prophylactic or therapeutic treatment of a Klebsiella pneumoniae infection or a condition or disease caused by such an infection, in particular for use in the prophylaxis and / or treatment of a drug resistant or multi-drug resistant Klebsiella pneumoniae strain infection.

23. The human monoclonal antibody or antigen binding portion thereof for use or the immunospecific polypeptide for use according to claim 22, wherein the drug resistant or multi-drug resistant strain is a Klebsiella pneumoniae capsular type K64 strain.

24. The human monoclonal antibody or antigen-binding portion thereof according to any one of claims 1 to 20 or the antibody or antigen-binding portion thereof according to claim 21 for use in the prophylactic or therapeutic treatment of an infection with NDM-producing Klebsiella pneumoniae strains, in particular NDM-1, NDM-5 and / or NDM-9 positive strains, or a condition or disease caused by such an infection.

25. The human monoclonal antibody or antigen-binding portion thereof for use according to claim 24 or the immunospecific polypeptide, wherein the strain is at least one of the following strains: a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain and a NDM-9 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

26. The human monoclonal antibody or antigen-binding portion thereof for use according to claim 24, wherein the strain is a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

27. A pharmaceutical composition comprising one or more human monoclonal antibodies or antigen-binding portions thereof according to any one of claims 1 to 20 and / or one or more immunospecific polypeptides according to claim 21, and a pharmaceutically acceptable carrier.

28. The composition according to claim 27 for use in the prophylaxis and / or treatment of Klebsiella pneumoniae infection.

29. The composition for use according to claim 28, wherein the infection is an infection with a drug-resistant or multidrug-resistant Klebsiella pneumoniae capsular type K64 strain.

30. The composition for use according to claim 29, wherein the strain is a NDM-1 positive Klebsiella pneumoniae sequence type 147 (ST147) strain.

31. An in vitro method for diagnosing a Klebsiella pneumoniae infection in a subject, comprising the following steps: i) contacting an antibody or antigen-binding portion thereof according to any one of claims 1 to 20 or an immunospecific polypeptide according to claim 21 with a biological sample of the subject; ii) detecting the binding of the antibody or antigen-binding portion thereof to a Klebsiella pneumoniae surface antigen.

32. The in vitro method according to claim 31, wherein step ii) comprises detecting the binding of the antibody or antigen-binding portion thereof to a Klebsiella pneumoniae capsular antigen, preferably to a K64 type capsular antigen.

33. A diagnostic kit comprising an antibody or antigen-binding portion thereof according to any one of claims 1 to 20 and / or an immunospecific polypeptide according to claim 21 as a specific reagent, in particular intended to be used in a method for detecting or quantifying anti-Klebsiella pneumoniae antibodies, in particular anti-NDM-producing Klebsiella pneumoniae antibodies, in a patient biological sample.

34. Use of an antibody or antigen-binding portion thereof according to any one of claims 1 to 20 or an immunospecific polypeptide according to claim 21 for the design of a vaccine against Klebsiella pneumoniae, in particular against NDM-producing Klebsiella pneumoniae strains.

35. The use of claim 34, wherein the vaccine is a vaccine against a K. pneumoniae capsular type K64 strain.

36. The use of claim 34 or 35, wherein the vaccine is a vaccine against a NDM-1 positive K. pneumoniae sequence type 147 (ST147) strain.

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