WO2014160098A2

WO2014160098A2 - Bordetella specific human recombinant antibodies and uses thereof

Info

Publication number: WO2014160098A2
Application number: PCT/US2014/025811
Authority: WO
Inventors: Hugh Russell; Gwen WILMES; Vincent W. Coljee
Original assignee: Excelimmune, Inc.
Priority date: 2013-03-13
Filing date: 2014-03-13
Publication date: 2014-10-02
Also published as: WO2014160098A3

Abstract

Human antibodies that bind to and neutralize one or more strains and/or species Bordetella, for example, Bordetella pertussis and Bordetella parapertussis, strains and/or species, including antibiotic resistant strains and/or species, are disclosed. Also disclosed are human recombinant polyclonal antibody compositions, and therapeutic methods for using the antibodies.

Description

Bordetella Specific Human Recombinant Antibodies and

Uses Thereof

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Patent Application serial number 61/779,234 filed March 13, 2013, the entire contents of which are hereby incorporated by reference.

FIELD OF THE INVENTION

[0002] The field of the invention is molecular biology, immunology and infectious disease. More particularly, the field is anti-Bordetella antibodies, including the species Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiaseptica, and therapeutic human recombinant antibodies.

BACKGROUND

[0003] Bordetella bacterial species, in particular Bordetella pertussis and Bordetella parapertussis, are the major causes of pertussis, a very contagious respiratory illness commonly known as whooping cough. The bacteria attach to the cilia of the respiratory epithelial cells, produce toxins that paralyze the cilia, and cause inflammation of the respiratory tract, which interferes with the clearing of pulmonary secretions. Until recently, it was thought that B. pertussis did not invade the tissues; however, recent studies have suggested that the bacteria can survive within a variety of eukaryotic cell types, including alveolar macrophages. B. pertussis and B. parapertussis are typical human pathogens. Worldwide, there are 30-50 million cases of pertussis and about 300,000 deaths per year.

[0004] Since the 1980s, there has been an increase in the number of reported cases of pertussis in the U.S., especially among 10-19 year olds and infants younger than 6 months of age. In 2010, 27,550 cases of pertussis were reported, although many more cases go unreported. This is the highest number of cases reported in over 60 years and the highest incidence rate in over 50 years. Further, more than half of infants less than 1 year of age who get pertussis must be hospitalized.

[0005] Antibiotics, including erythromycin, clarithromycin, and azithromycin, are preferred for the treatment of pertussis in persons >1 month of age, but can have severe side effects and are not always successful in treatment with rising antibiotic resistance; nor do they counteract the toxins and immunomodulators that are already released by the bacteria. Specific antibiotic resistance by Bordetella species, especially Bordetella pertussis, has been reported against erythromycin and fluoroquinolones (Ohtsuka et al, ANTIMICROBIAL AGENTS AND CHEMOTHERAPY (2009) 53:3147- 3149). Alternative treatments are therefore needed to replace those that are increasingly ineffective.

[0006] Research has shown that humans infected with Bordetella have a diverse humoral immune reaction to the organism. Current vaccines rely on pre-acellular vaccines, e.g., Daptacel® (Sanofi-Aventis), Adacel® (Sanofi-Aventis) and Boostrix® (Glaxo SmithKline), although some countries still rely on a whole cell vaccine. Cherry et al. showed that various compositions of the acellular vaccine protect against the respective antigen used in that vaccine, for example, pertussis toxin (PTx) only, PTx/filamentous hemagglutinin (FHA), PTx/FHA/pertactin (PTN) or

PTx/FHA/PTN plus Fimbriae 2/3, but not against Bordetella antigens not included in that vaccine (CLINICAL & VACCINE IMMUNOLOGY (2010) 17:741-747). As reviewed in Gouw et al, Bordetella has a variety of different means of immune-evasion and immune modulation (FEMS MICROBIOL REV (2011) 35:441-474).

[0007] For better immune protection across a population, a greater of variety of antigens in vaccines may improve the chances for an adequate immunological response against Bordetella. For example, differences in the protective abilities of the acellular vaccines and whole cell vaccines are observed with protection against Bordetella parapertussis. Bordetella parapertussis can be prevented by whole cell vaccines; however, parapertussis infection can be enhanced by acellular vaccines even when the vaccine contains some antigens that are cross reactive (Long et al, PROC. R. SOC. B (2010) 277:2017-2025 and Lavine et al, VACCINE (2011) 29: 11-16). For Bordetella parapertussis, antibodies against the O-antigen appear to be important for humoral protection (Zhang et al, PLoS ONE 4(9):e6989).

[0008] The use of immunoglobulins or hyper-immunoglobulins to treat Bordetella pertussis infections has shown variable results. Research suggests that immunoglobulins, when administered intramuscularly, are likely of little utility (Balagtas et al. J. PEDIATRICS. 79(2): 203-208).

However, more recent data suggests a possible benefit if an immunoglobulin contains enough anti- pertussis toxin activity (Bruss et al. (1999) THE PEDIATRIC INFECTIOUS DISEASE JOURNAL 18(6): 505-511 and Halperin et al. (2007) THE PEDIATRIC INFECTIOUS DISEASE JOURNAL 26(1):79-81). [0009] Thus, there is a need for improved antibodies and antibody compositions that can be used to treat and/or prevent Bordetella infections.

SUMMARY

[00010] The invention is based, in part, upon the discovery of human antibodies and polyclonal antibody compositions that bind one or more antigens, strains and/or species of Bordetella, a gram- negative bacteria. Some embodiments comprise antibody compositions comprising one or more of the human antibodies or antigen-binding fragments thereof as disclosed herein. Other

embodiments comprise an individual human antibody, antigen-binding fragment thereof (e.g., Fab fragment or single-chain variable fragments (scFvs)), or antibodies with the same complementarity determining regions ("CDRs"); i.e., monoclonal antibody compositions. Exemplary human antibodies described herein contain one or more specific Bordetella antigen-binding CDRs. In certain embodiments, the disclosed human antibody compositions are polyclonal antibody compositions comprising at least two different antibodies that individually bind one or more Bordetella strains and/or species and/or antigens. In certain embodiments, the disclosed polyclonal antibody compositions comprise at least three different antibodies or antigen-binding fragments thereof as described herein. Exemplary polyclonal antibody compositions described herein contain specific antibodies with Bordetella binding sites based on the CDRs of the individual antibodies.

[00011] Antibodies and polyclonal human antibody compositions as described herein bind proteins that are known or likely to be involved in Bordetella virulence {e.g., pertussis toxin ("PTx") or filamentous hemagglutinin ("FHA")). Alternatively, they may bind previously uncharacterized Bordetella target proteins. The disclosed antibodies and polyclonal human antibody compositions can neutralize the activity of Bordetella toxins, cell surface antigens and/or immunomodulating antigens. For example, in some embodiments, the antibodies and polyclonal human antibody compositions can bind and neutralize PTx or FHA.

[00012] In an exemplary embodiment, the disclosed human polyclonal antibody compositions are broad-spectrum therapeutic antibodies with neutralizing activity against multiple Bordetella proteins that are present on one or more Bordetella strains and/or species. In certain embodiments, the disclosed recombinant polyclonal antibody compositions mimic the natural human immune response. It is contemplated herein that the disclosed antibodies and antibody compositions provide protection against Bordetella virulence factors. Thus, the individual antibodies and polyclonal human antibody compositions can be used as therapeutic agents to treat Bordetella infections including antibiotic-resistant Bordetella strains and/or species.

[00013] In various embodiments, the present invention provides an antibody or antigen-binding fragment thereof that binds Pertussis toxin comprising at least one heavy chain complementarity determining region (HCDR1, HCDR2 and/or HCDR3) at least 80% identical to an amino acid sequence selected from the group consisting of: a HCDR1 as set forth in SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 11, SEQ ID NO: 16, SEQ ID NO: 21, SEQ ID NO: 26, SEQ ID NO: 31, or SEQ ID NO: 36; a HCDR2 as set forth in SEQ ID NO: 2, SEQ ID NO: 7, SEQ ID NO: 12, SEQ ID NO: 17, SEQ ID NO: 22, SEQ ID NO: 27, SEQ ID NO: 32, or SEQ ID NO: 37; and/or a HCDR3 as set forth in SEQ ID NO: 3, SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 18, SEQ ID NO: 23, SEQ ID NO: 28, SEQ ID NO: 33, or SEQ ID NO: 38.

[00014] In some embodiments, there is provided an antibody or antigen-binding fragment thereof that binds Pertussis toxin comprising at least one light chain complementarity determining region (LCDR1, LCDR2 and/or LCDR3) selected from the group consisting of: a LCDR1 at least 80% identical to an amino acid sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 9, SEQ ID NO: 14, SEQ ID NO: 19, SEQ ID NO: 24, SEQ ID NO: 29, SEQ ID NO: 34, or SEQ ID NO: 39; a LCDR2 selected from the group consisting of AAS, GAS, DAS, GTS, DAS and conservatively substituted variants thereof; and/or a LCDR3 at least 80% identical to an amino acid sequence as set forth in SEQ ID NO: 5, SEQ ID NO: 10, SEQ ID NO: 15, SEQ ID NO: 20, SEQ ID NO: 25, SEQ ID NO: 30, SEQ ID NO: 35, or SEQ ID NO: 40.

[00015] In some embodiments, antibodies or antigen-binding fragments thereof are selected from the group consisting of human antibody, an antigen-binding fragment of a human antibody, a humanized antibody, an antigen-binding fragment of a humanized antibody, a chimeric antibody and an antigen-binding fragment of a chimeric antibody. In some embodiments, the antibody or antigen-binding fragment is an antigen-binding fragment selected from the group consisting of an Fab fragment, an Fab' fragment, and F(ab')₂ fragment and an Fv fragment.

[00016] In various embodiments, antibodies or antigen-binding fragments thereof competitively inhibit binding of mouse monoclonal antibody 1B7. In some embodiments, the antibody or antigen-binding fragment has an epitope specificity substantially similar to mouse monoclonal antibody 1B7.

[00017] In some embodiments of the invention, antibodies or antigen binding fragments competitively inhibit binding of mouse monoclonal antibody 11E6. In some embodiments, the antibody or antigen-binding fragment has an epitope specificity substantially similar to mouse monoclonal antibody 11E6.

[00018] In various embodiments, antibodies or antigen-binding fragments thereof bind Pertussis toxin with an equilibrium dissociation constant (KQ) of about 2-100 nM. In some embodiments, the antibody or antigen-binding fragment thereof neutralizes Pertussis toxin. In some

embodiments, the antibody or antigen binding fragment thereof binds to the catalytic SI subunit of Pertussis toxin. In some embodiments, the antibody or antigen binding fragment thereof binds to the S2 and S3 subunits of Pertussis toxin.

[00019] Various embodiments of the invention comprise antibodies and antigen-binding fragments thereof with an HCDRl that is at least 80% identical to SEQ ID NO: 1, the HCDR2 is at least 80% identical to SEQ ID NO: 2, and the HCDR3 is at least 80% identical to SEQ ID NO: 3. In some embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 6, the HCDR2 is at least 80% identical to SEQ ID NO: 7, and the HCDR3 is at least 80% identical to SEQ ID NO: 8. In some embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 11, the HCDR2 is at least 80% identical to SEQ ID NO: 12, and the HCDR3 is at least 80% identical to SEQ ID NO: 13. In certain embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 16, the HCDR2 is at least 80% identical to SEQ ID NO: 17, and the HCDR3 is at least 80% identical to SEQ ID NO: 18. In some embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 21, the HCDR2 is at least 80% identical to SEQ ID NO: 22, and the HCDR3 is at least 80% identical to SEQ ID NO: 23. In certain embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 26, the HCDR2 is at least 80% identical to SEQ ID NO: 27, and the HCDR3 is at least 80% identical to SEQ ID NO: 28. In some embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 31, the HCDR2 is at least 80% identical to SEQ ID NO: 32, and the HCDR3 is at least 80% identical to SEQ ID NO: 33. In some embodiments, the HCDRl is at least 80% identical to SEQ ID NO: 36, the HCDR2 is at least 80% identical to SEQ ID NO: 37, and the HCDR3 is at least 80% identical to SEQ ID NO: 38. [00020] Various embodiments of the invention comprise antibodies and antigen-binding fragments thereof with a LCDRl that is at least 80% identical to SEQ ID NO: 4, the LCDR2 is AAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 5. In some embodiments, the LCDRl is at least 80% identical to SEQ ID NO: 9, the LCDR2 is GAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 10. In some embodiments, the LCDRl is at least 80% identical to SEQ ID NO: 14, the LCDR2 is DAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 15. In some embodiments, the LCDRl is at least 80% identical to SEQ ID NO: 19, the LCDR2 is GTS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 20. In some embodiments, the LCDRl is at least 80% identical to SEQ ID NO: 24, the LCDR2 is DAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 25. In some

embodiments, the LCDRl is at least 80% identical to SEQ ID NO: 29, the LCDR2 is AAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 30. In some embodiments, the antibody or antigen-binding fragment thereof of claim 2, wherein the LCDRl is at least 80% identical to SEQ ID NO: 34, the HCDR2 is DAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 35. In some embodiments, the LCDRl is at least 80% identical to SEQ ID NO: 39, the LCDR2 is AAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 40.

[00021] In various embodiments of the invention, there is provided an antibody or antigen- binding fragment thereof that binds Pertussis toxin, which comprises a heavy chain variable region encoded by a nucleic acid sequence at least 80% identical to SEQ ID NO: 54 and a light chain variable region encoded by a nucleic acid sequence at least 80%> identical to SEQ ID NO: 56. In some embodiments of the invention, there is provided an antibody or antigen-binding fragment thereof that binds Pertussis toxin, which comprises a heavy chain variable region encoded by a nucleic acid sequence at least 80% identical to SEQ ID NO: 62 and a light chain variable region encoded by a nucleic acid sequence at least 80%> identical to SEQ ID NO: 64.

[00022] In some embodiments of the invention, there is provided antibodies and antigen-binding fragments thereof that bind filamentous hemagglutinin comprising at least one heavy chain complementarity determining region (HCDR1, HCDR2 and/or HCDR3) at least 80% identical to an amino acid sequence selected from the group consisting of: a HCDR1 as set forth in SEQ ID NO: 41; a HCDR2 as set forth in SEQ ID NO: 42; and/or a HCDR3 as set forth in SEQ ID NO: 43. In some embodiments, the antibody or antigen-binding fragment comprises at least one light chain complementarity determining region selected from the group consisting of: a LCDRl at least 80% identical to QSVRTN (SEQ ID NO: 44); a LCDR2 of DGF or conservatively modified variants thereof; or a LDCR3 at least 80% identical to CQQYRTWPRVTF (SEQ ID NO: 45). In some embodiments, the HCDR1 is at least 80% identical to GFTFSSFD (SEQ ID NO: 41); the HCDR2 is at least 80% identical to IRHHGTNH (SEQ ID NO: 42); the HCDR3 is at least 80% identical to CAKDLGFGELYW (SEQ ID NO: 43); the LCDRl is at least 80% identical to QSVRTN (SEQ ID NO: 44); the LCDR2 is DGF or a conservatively modified variant thereof; and the LCDR2 is at least 80% identical to CQQYRTWPRVTF (SEQ ID NO: 45).

[00023] In various aspects, the antibodies and antigen-binding fragments thereof that bind filamentous hemagglutinin are selected from the group consisting of human antibody, an antigen- binding fragment of a human antibody, a humanized antibody, an antigen-binding fragment of a humanized antibody, a chimeric antibody and an antigen-binding fragment of a chimeric antibody. In some embodiments, the antibodies and antigen-binding fragments thereof are an antigen-binding fragment selected from the group consisting of an Fab fragment, an Fab' fragment, and F(ab')2 fragment and an Fv fragment.

[00024] In some embodiments, the antibodies and antigen-binding fragments bind filamentous hemagglutinin with an equilibrium dissociation constant (KD) of about 2-100 nM. In some embodiments, the antibody or antigen binding fragment neutralizes filamentous hemagglutinin.

[00025] In another aspect of the invention, there is provided a human recombinant polyclonal antibody composition that binds to one or more Bordetella strains and/or species comprising at least three different human antibodies that individually bind one or more Bordetella strains and/or species. In certain embodiments, the human recombinant polyclonal antibody composition comprises at least 5 different human antibodies that individually bind one or more Bordetella strains and/or species. In certain embodiments, the human recombinant polyclonal antibody composition comprises at least 10 different human antibodies that individually bind one or more Bordetella strains and/or species. [00026] In some embodiments, the polyclonal antibody composition comprises at least one of the antibodies identified in Table 1. In some embodiments, the polyclonal antibody composition comprises at least 3 or more of the antibodies identified in Table 1.

[00027] In some embodiments, at least one of the antibodies in the polyclonal antibody composition binds to at least one of the Bordetella strains and/or species selected from the group consisting of: ATCC Strain BAA-589 pertussis, ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica. In some embodiments, at least one of the antibodies binds at least 5 strains and/or species, at least 6 strains and/or species, or at least 7 strains and/or species selected from the group consisting of: ATCC Strain BAA-589 pertussis, ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00028] In various aspects of the invention, at least one of the antibodies in the polyclonal antibody composition binds a cell surface antigen on one or more Bordetella strains and/or species and one of the antibodies binds a toxin produced by one or more Bordetella strains and/or species. In some of the embodiments, one of the antibodies binds a cell surface antigen on one or more Bordetella strains and/or species and one of the antibodies binds an immunomodulator produced by one or more Bordetella strains and/or species. In some embodiments, one of the antibodies binds a toxin produced by one or more Bordetella strains and/or species and one of the antibodies binds an immunomodulator produced by one or more Bordetella strains and/or species. IN certain embodiments, one of the antibodies binds a cell surface antigen on one or more Bordetella strains and/or species, one of the antibodies binds a toxin produced by one or more Bordetella strains and/or species, and one of the antibodies binds an immunomodulator produced by one or more Bordetella strains and/or species.

[00029] In various aspects of the invention, there is provided a polyclonal antibody composition wherein at least one of the antibodies in the composition is selected from the group consisting of: (a) an antibody that competes with 42.11.D4 comprising the heavy chain and light chain CDR1, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species; (b) an antibody that competes with 42.11.G2 comprising the heavy chain and light chain CDR1, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more

Bordetella strains and/or species; (c) an antibody that competes with 42.12.A12 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species; (d) an antibody that competes with 42.12.A9 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species; and (e) an antibody that competes with 42.18. E12 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species, wherein the one or more Bordetella strains and/or species are selected from the group consisting of ATCC Strain BAA-589 pertussis, ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00030] In another aspect of the invention, there is provide an expression vector comprising a recombinant nucleic acid that encodes one or more of the heavy chain complementarity

determining regions (HCDR1, HCDR2 and/or HCDR3) as described herein. In some

embodiments of the invention, there is provided an expression vector comprising a recombinant nucleic acid that encodes one or more of the light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) as described herein.

[00031] In another aspect of the invention, there is provided host cells comprising expression vectors and nucleic acids as described herein. In yet another aspect of the invention, there is provided a method of producing a polypeptide comprising an immunoglobulin heavy chain or light chain complementarity determining region, the method comprising: growing the host cell under conditions so that the host cell expresses the polypeptide comprising the immunoglobulin heavy chain or light chain complementarity determining region; and purifying the polypeptide comprising the immunoglobulin heavy or light chain complementarity determining region.

[00032] In another aspect of the invention, there is provided a method of reducing or killing a Bordetella strain by administering an effective amount of the antibody, antigen-binding fragment thereof or polyclonal antibody composition as described herein to reduce or kill the Bordetella strain. In some embodiments, there is provided a method for treating or preventing a Bordetella infection in a mammal by administering an effective amount of the antibody, antigen-binding fragment thereof or polyclonal antibody composition as described herein to a mammal in need thereof. In some embodiments, the mammal is a human. [00033] In various aspects of the invention, there is provided methods for treating or preventing a polymicrobial infection including a Bordetella infection in a mammal, the method comprising administering an effective amount of the antibody, antigen-binding fragment thereof or polyclonal antibody composition as described herein to a mammal in need thereof.

[00034] These and other aspects and advantages of the invention will become apparent upon consideration of the following figures, detailed description, and claims. As used herein,

"including" means without limitation, and examples cited are non-limiting.

DEFINITIONS

[00035] In order for the present invention to be more readily understood, certain terms are first defined. Additional definitions for the following terms and other terms are set forth throughout the specification.

[00036] Affinity: As is known in the art, "affinity" is a measure of the tightness with which a particular ligand binds to (e.g., associates non-covalently with) and/or the rate or frequency with which it dissociates from, its partner. As is known in the art, any of a variety of technologies can be utilized to determine affinity. In many embodiments, affinity represents a measure of specific binding. In some embodiments, pertussis antigen binding affinity is determined by competition ELISA using the method of Friquet et al., "Measurements of True Affinity Constant in Solution of Antigen- Antibody Complexes by Enzyme -Linked Immunosorbent Assay," J. IMMUNO METHODS 305 (1985).

[00037] The ability of an antibody to bind a specific epitope can be described by the equilibrium dissociation constant (K_D). The equilibrium dissociation constant (K_D) as defined herein is the ratio of the dissociation rate (K-off) and the association rate (K-on) of a human recombinant antibody or antigen-binding fragment thereof to a Bordetella antigen. It is described by the following formula: K_D =K-off/K-on. In some embodiments, antibodies and antibody compositions disclosed herein bind a pertussis toxin protein with an equilibrium dissociation constant (K_D) of about 100 nM, about 90 nM, about 80 nM, about 70 nM, about 60 nM, about 50 nM, about 40 nM, about 30 nM, about 20 nM, about 10 nM, about 9 nM, about 8 nM, about 7 nM, about 6 nM, about 5 nM, about 4 nM, about 3 nM, about 2 nM or less, and/or between 2-10 nM. [00038] Antibody: As used herein, the term "antibody" refers to a polypeptide consisting of one or more polypeptides substantially encoded by immunoglobulin genes or fragments of

immunoglobulin genes. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as myriad immunoglobulin variable region genes. Light chains are typically classified as either kappa or lambda. Heavy chains are typically classified as gamma, mu, alpha, delta, or epsilon, which in turn define the

immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. A typical immunoglobulin (antibody) structural unit is known to comprise a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms "variable light chain" (VL) and "variable heavy chain" (VH) refer to these light and heavy chains respectively. An antibody can be specific for a particular antigen. The antibody or its antigen can be either an analyte or a binding partner. Antibodies exist as intact immunoglobulins or as a number of well- characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)'2, a dimer of Fab which itself is a light chain joined to VH-CH1 by a disulfide bond. The F(ab)'2 may be reduced under mild conditions to break the disulfide linkage in the hinge region thereby converting the (Fab')2 dimer into an Fab' monomer. The Fab' monomer is essentially an Fab with part of the hinge region (see, FUNDAMENTAL IMMUNOLOGY, W. E. Paul, ed., Raven Press, N.Y. (1993), for a more detailed description of other antibody fragments). While various antibody fragments are defined in terms of the digestion of an intact antibody, one of ordinary skill in the art will appreciate that such Fab' fragments may be synthesized de novo either chemically or by utilizing recombinant DNA methodology. Thus, the term "antibody," as used herein also includes antibody fragments either produced by the modification of whole antibodies or synthesized de novo using recombinant DNA methodologies. In some embodiments, antibodies are single chain antibodies, such as single chain Fv (scFv) antibodies in which a variable heavy and a variable light chain are joined together (directly or through a peptide linker) to form a continuous polypeptide. A single chain Fv ("scFv") polypeptide is a covalently linked VH::VL heterodimer which may be expressed from a nucleic acid including VH- and VL-encoding sequences either joined directly or joined by a peptide-encoding linker. (See, e.g., Huston, et al. (1988) P OC. NAT. ACAD. SCI. USA, 85:5879- 5883, the entire contents of which are herein incorporated by reference.) A number of structures exist for converting the naturally aggregated, but chemically separated light and heavy polypeptide chains from an antibody V region into an scFv molecule which will fold into a three dimensional structure substantially similar to the structure of an antigen-binding site. See, e.g. U.S. Pat. Nos. 5,091,513 and 5,132,405 and 4,956,778.

[00039] CDR-grafted: As used herein, the phrase "CDR-grafted" antibody refers to an antibody or an antigen-binding fragment thereof that comprises a CDR that is not naturally associated with the framework regions of the antibody or antigen-binding fragment. In some embodiments, the CDR is from an antibody from a first species and the framework regions and constant regions (if present) are from an antibody from a different species. In some embodiments the grafted CDR and framework regions are from the same species but one or more CDRs from an individual monoclonal antibody may be grafted to the framework region of a different monoclonal antibody.

[00040] Chimeric antibody: As used herein, the term "chimeric antibody" refers to an antibody or antigen-binding fragment thereof comprising a variable region (or CDR thereof) from a first antibody and a constant region from a second antibody. In some embodiments, the variable region is from a first species and a constant region is from a different species. In other embodiments, the variable and constant regions are from the same species but are derived from different monoclonal antibodies.

[00041] Complementarity Determining Regions (CDRs): As used herein, CDRs are defined as regions within an antibody that are directly involved in antigen binding. Proceeding from the amino-terminus, these regions are designated HCDR1, HCDR2 and HCDR3 for the heavy chain, and LCDR1, LCDR2, and LCDR3 for the light chain, respectively. The CDRs are held in place by more conserved framework regions (FRs). Proceeding from the amino-terminus, these regions are designated FR HI, FR H2, FR H3, and FR H4 for the heavy chain and FR LI, FR L2, FR L3, and FR L4, for the light chain, respectively. For humanized antibodies, one or more of the CDRs are derived from a donor antibody (also referred to herein as a donor CDR), whereas the FRs are of human origin. The locations of CDR and FR regions and a numbering system may be defined by, e.g., Kabat et al. (Kabat et al, SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST, Fifth Edition, U.S. Department of Health and Human Services, U.S. Government Printing Office (1991)), incorporated by reference herein. Likewise, the positions occupied by individual residues within the heavy or the light chains of the antibodies herein may be defined by the Kabat numbering system. Therefore, the locations of residues within the human heavy and light chains required for binding may be defined by the position of the residue according to the Kabat numbering system as is well known in the art. Thus, in some embodiments, the human antibodies and antigen-binding fragments disclosed herein include substitutions, insertions and deletions within the CDRs at positions defined by the Kabat numbering system.

[00042] In some embodiments, variable regions and CDRs therein are as defined by IMGT. The IMGT unique numbering has been defined to compare the variable domains whatever the antigen receptor, the chain type, or the species (information available at

http://www.imgt.org/IMGTindex/antibodies.html). In the IMGT unique numbering system, the conserved amino acids always have the same position, for instance cysteine 23 (lst-CYS), tryptophan 41 (CONSERVED-TRP), hydrophobic amino acid 89, cysteine 104 (2nd-CYS), phenylalanine or tryptophan 118 (J-PHE or J-TRP). The IMGT unique numbering provides a standardized delimitation of the framework regions (FRl-IMGT: positions 1 to 26, FR2-IMGT: 39 to 55, FR3-IMGT: 66 to 104 and FR4-IMGT: 118 to 128) and of the complementarity determining regions: CDR1-IMGT: 27 to 38, CDR2-IMGT: 56 to 65 and CDR3-IMGT: 105 to 117. Gaps represent unoccupied positions.

[00043] Conservatively Substituted Variant: With respect to nucleic acid sequences, the phrase "conservatively substituted variant" as used herein refers to those nucleic acids that encode identical or essentially identical amino acid sequences. Because of the degeneracy of the genetic code, a number of nucleic acid sequences will encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence. Considering amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively substituted variant" where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants include polymorphic variants, sequence identity variants and affinity optimized variants as described herein. In some embodiments, the following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M).

[00044] Human Antibody: As used herein, the phrase "human antibody" refers to an antibody or an antigen-binding fragment thereof in which at least one of the CDRs of the variable regions and/or the constant regions (if present) have amino acid sequences that are encoded by nucleotide sequences derived from human (Homo sapiens) germline immunoglobulin genes. A "human antibody" can include sequences that are not encoded in the germline (e.g., due to N nucleotides, P nucleotides, and mutations that can occur as part of the processes that produce high-affinity antibodies such as, somatic mutation, affinity maturation, clonal selection) that occur as a result of biological processes in a suitable in vivo expression system (e.g., a human, a human-antibody transgenic animal). Antibodies, antigen-binding fragments of antibodies and portions or regions of human antibodies can be produced, for example, by expression of a nucleic acid of non-human origin (e.g., a synthetic nucleic acid) that has the requisite nucleotide sequence.

[00045] Identical: The terms "identical" or percent "identity," as used herein in reference to nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., 60% identity, optionally 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identity over a specified region, e.g., the entirety of the HCDR3 of the antibodies disclosed herein), when compared and aligned for maximum correspondence over a given segment or designated region, as measured using sequence comparison algorithms (e.g. BLAST) or by manual alignment and visual inspection. [00046] Percentage of sequence identity: As referred to herein, the "percentage of sequence identity" is determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window (e.g., a particular CDR) may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.

[00047] Treatment: As used herein, the term "treatment" (also "treat" or "treating") refers to any administration of a therapeutic protein (e.g., administration of an antibody or antigen binding fragment thereof that binds Pertussis toxin) that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity of and/or reduces incidence of one or more symptoms or features of a particular disease, disorder, and/or condition (e.g., whooping cough). Such treatment may be of a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition. Alternatively or additionally, such treatment may be of a subject who exhibits one or more established signs of the relevant disease, disorder and/or condition.

DESCRIPTION OF THE DRAWINGS

[00048] The drawings are for illustration purposes only not for limitation.

[00049] Figure 1 (prior art) is a schematic representation of a typical naturally-occurring antibody. Naturally occurring antibodies are multimeric proteins that contain four polypeptide chains. Two of the polypeptide chains are called heavy chains (H chains), and two of the polypeptide chains are called light chains (L chains). The immunoglobulin heavy and light chains are connected by an interchain disulfide bond. The immunoglobulin heavy chains are connected by interchain disulfide bonds. A light chain consists of one variable region (VL) and one constant region (CL). The heavy chain consists of one variable region (VH) and at least three constant regions (CHI, CH2 and CH3). The variable regions determine the specificity of the antibody. Each variable region comprises three hypervariable regions also known as complementarity determining regions (CDRs) flanked by four relatively conserved framework regions (FRs). The three CDRs, referred to as CDR1, CDR2, and CDR3, contribute to the antibody binding specificity.

[00050] Figure 2 is a series of panels showing data from a representative plate (plate 42.18) of isolated fully human antibodies. Fig. 2A is a representative FACS analysis showing the population of CD38+/CD19+ plasma cells targeted for cloning; Fig. 2B shows the amplified variable heavy and light antibody genes from individual plasma cells in a 96-well plate; Fig. 2C and 2E shows representative data from an antibody expression ELISA showing that the majority of amplified antibody cognate pairs are cloned and are able to express antibody; Figs. 2D and F show data from screening ELISA assays in which a total of 28 wells were scored as a positive against Bordetella FHA in Fig. 2D and wells A3 and E12 were scored as positive against PTx in Fig. 2F.

[00051] Figure 3 is an image of the results of an agglutination assay, which uses the

agglutination properties FHA possesses against red blood cells (RBCs) to test antibody

neutralization in a 96-well plate format. The results demonstrate that antibody 55.22.E7 works as an efficient FHA neutralizing antibody and is capable of neutralizing ^g/mL of FHA when used at a concentration of 1.5 μg/mL.

DETAILED DESCRIPTION

[00052] The human anti-Bordetella antibodies and antibody compositions disclosed herein are based, in part, on the antigen binding sites of certain human antibodies that bind and neutralize the activity of one or more Bordetella antigens, strains and/or species. The terms "individual anti- Bordetella antibody" and "anti-Bordetella recombinant antibody" describe an antibody or antigen- binding fragment thereof with at least one heavy or light chain CDR that binds a Bordetella antigen. In some embodiments, at least one of the heavy and/or light chain CDR amino acid sequences is at least 80% identical to the exemplary CDRs disclosed herein. In some

embodiments, individual anti-Bordetella antibodies described herein can bind to and neutralize antigens contributing to the pathogenic mechanisms of Bordetella, for example PTx or FHA.

[00053] The terms "anti-Bordetella polyclonal antibody" and "anti-Bordetella recombinant polyclonal antibody" describe a composition of recombinantly produced diverse antibody molecules, where the individual members of the polyclonal composition are capable of binding to at least one epitope on Bordetella or an Bordetella secreted protein {e.g., a toxin or immunomodulator) or a cell surface antigen and where the polyclonal composition as a whole is capable of neutralizing Bordetella. In some embodiments, the polyclonal anti-Bordetella antibody compositions described herein can bind to and neutralize one or more antigens contributing to the pathogenic mechanisms of Bordetella, for example PTx and FHA. In exemplary embodiments, an anti-Bordetella polyclonal antibody neutralizes Bordetella and/or one or more antibiotic-resistant Bordetella strains and/or species. It is contemplated that the disclosed anti-Bordetella polyclonal antibodies are essentially free from immunoglobulin molecules that do not bind to Bordetella or variant strains and/or species thereof (e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99% or 100% of the antibodies contained in the polyclonal composition bind to one or more strains and/or species of Bordetella).

[00054] Without wishing to be bound by any particular theory, it is contemplated that the diversity of antibodies included in an anti-Bordetella recombinant polyclonal antibody

composition provide a surprising benefit over certain monoclonal and biclonal (e.g. , a mixture of two monoclonal antibodies) antibodies because lower dosages of the polyclonal antibody composition may be administered to prevent or treat Bordetella infection. For example, it is contemplated herein that the synergistic action of the individual component antibodies in the polyclonal composition allow the polyclonal composition to be effective at lower doses than is possible with conventional monoclonal antibody therapy. In addition, because a polyclonal antibody composition binds to multiple different proteins, the composition as a whole can use lower amounts of each individual antibody to prevent or treat Bordetella infections. Further, unlike monoclonal or biclonal antibody compositions, polyclonal antibody compositions do not present the same concerns regarding the generation of drug resistance to a single or small number of agents (e.g., development of resistant Bordetella strains and/or species due to the monovalent nature of a monoclonal antibody's mode of action).

[00055] The human antibodies and polyclonal antibody compositions provided herein are capable of binding Bordetella antigens, e.g., Pertussis toxin protein (PTx) or FHA, which are in part responsible for the pathogenesis and clinical features of Bordetella infection. See, e.g., Locht, C, "Molecular Aspects of Bordetella Pertussis pathogenesis", 1999, INTERNATL. MICROBIOL, 2: 137- 144, incorporated by reference herein.

[00056] Pertussis toxin is a 105 kDa protein toxin, composed of an enzymatically active A subunit, and a B subunit primarily responsible for binding to the cell-surface. It is structurally similar to cholera and shiga toxins, conforming to the A-B5 class of toxins. The toxin is an ADP ribosylase with specificity for Gi/o proteins. The B domain of the pertussis toxin is composed of four unique proteins: a dimer of S2 and S4 subunits, a dimer of the S3 and S4 subunits, and S5. Together they form an asymmetric pentamer about a central pore. The active subunit, SI, sits atop the pore with its C-terminus penetrating halfway through the pore. The B subunit has been shown to bind with low affinity to N-linked sialoglycoproteins, including fetuin, haptoglobin, and transferrin. The S2 and S3 subunits each contain two binding clefts; an amino-terminal fold which resembles a family of mammalian calcium-dependent lectins, and a carboxy-terminal oligomer fold found in a number of proteins that bind carbohydrates. While no single universal receptor has been identified, pertussis toxin is capable of binding to all cell lines tested. Binding of the B region alone can result in cellular changes, including mitosis in lymphocytes and glucose oxidation in adipocytes, probably as a result of aggregating membrane proteins.

[00057] Cellular entry of the toxin is thought to be via endocytosis and retrograde, most likely to the endoplasmic reticulum. In this organelle, ATP and a reducing environment are present, enabling the S 1 subunit to dissociate and unfold, exposing its active site and a phospholipid binding domain which is thought to facilitate direct translocation into the cellular cytosol. Here, the S 1 subunit catalyzes the transfer of an ADP-ribose from NAD+ to cysteine-351 , near the N- termini of alpha subunits of inhibitory and olfactory G proteins and transducin. This modification blocks the G protein inhibition of adenylate cyclase, and the G proteins lose their signal transducing ability. This modification has a number of effects in vivo, including stimulation of insulin secretion, histamine sensitization and lymphocytosis, and in vitro modifies CHO cellular morphology, inducing growth in small clumps.

[00058] Filamentous hemagglutinin ("FHA") is a B. pertussis adhesion protein found at the surface of the organism. It is in part responsible for attachment to the cilia of the respiratory epithelial cells. It is produced as a 370 kDa precursor encoded by the fhaB gene. The FhaB precursor undergoes N-terminal and C-terminal proteolytic processing, resulting in the 220 kDa mature FHA. The structural model of mature FHA, based on high-resolution electron microscopy and circular dichroism measurements, as well as secondary structure predictions, is a filamentous monomeric molecule approximately 50 nm long and folded like a hairpin. It has a rod- like shape with a diameter of 4 nm, and contains at one extremity a globular structure formed by the N- terminal and C-terminal ends of the molecule. It has been suggested that 19-residue repeated motifs form an elongated amphiphilic β-sheet structure, joining the N-terminal and C-terminal moieties of the molecule by their hydrophobic interfaces to form the rod-like structure. During infection, in both humans and animal models, FHA induces a strong antibody response, both systemically and mucosally, and vaccination with purified FHA confers protection against respiratory challenge in mice.

[00059] In some embodiments, the human antibodies and polyclonal antibody compositions can neutralize (or inhibitory or antagonizing) a pertussis toxin protein (i.e. binding so as to partially or completely inhibit one or more biological activities of a pertussis toxin protein). Among the biological activities of a pertussis toxin protein that a neutralizing antibody may inhibit or block is the ability of a pertussis toxin protein to bind cellular receptors. The receptor binding region of a pertussis toxin protein consists of four polypeptide subunits referred to as subunit S2, subunit S3, subunit S4 and subunit S5, respectively. Examples of cellular receptors that are bound by the subunits S2, S3, S4, and S5 of a pertussis toxin protein are members of the N-linked

sialo glycoprotein family such as fetuin, haptoblobin, and transferrin. Another important activity of a pertussis toxin protein that may be inhibited by a neutralizing antibody is the enzymatic activity of the pertussis toxin protein as ADP ribosylase towards G proteins. The subunit conferring to the enzymatic activity as ADP-ribosylase in a pertussis toxin protein is subunit SI . In some embodiments, the pertussis toxin protein is a pertussis holotoxin. A pertussis holotoxin as referred to herein as a pertussis toxin protein that includes all five pertussis toxin protein subunits.

[00060] In some embodiments, antibodies or antigen-binding fragments thereof as disclosed herein can neutralize 1 μg/mL of PTx when used at a concentration (titer) of at least 0.1 μg/mL, for example at least 0.2 μg/mL, at least 0.3 μg/mL, at least 0.4 μg/mL, at least 0.5 μg/mL, at least 0.6 μg/mL, at least 0.7 μg/mL, at least 0.8 μg/mL, at least 0.9 μg/mL, at least 1.0 μg/mL, at least 1.5 μg/mL, at least 2.0 μg/mL, at least 2.5 μg/mL, at least 3.0 μg/mL or higher, inclusive of all ranges therein. In some embodiments, antibodies or antigen-binding fragments thereof as disclosed herein can neutralize ^g/mL of FHA when used at a concentration (titer) of at least 0.1 μg/mL, for example at least 0.2 μg/mL, at least 0.3 μg/mL, at least 0.4 μg/mL, at least 0.5 μg/mL, at least 0.6 μg/mL, at least 0.7 μg/mL, at least 0.8 μg/mL, at least 0.9 μg/mL, at least 1.0 μg/mL, at least 1.5 μg/mL, at least 2.0 μg/mL, at least 2.5 μg/mL, at least 3.0 μg/mL or higher, inclusive of all ranges therein. In some embodiments, polyclonal antibody compositions as disclosed herein can neutralize 1 μg/mL of PTx when used at a concentration (titer) of at least 0.1 μg/mL, for example at least 0.2 μ§/ηιΙ,, at least 0.3 μg/mL, at least 0.4 μ§/ηιΙ,, at least 0.5 μ§/ηιΙ,, at least 0.6 μ§/ηιΙ,, at least 0.7 μ§/ηιΙ,, at least 0.8 μ§/ηιΙ,, at least 0.9 μg/mL, at least 1.0 μ§/ηιΙ,, at least 1.5 μ§/ηιΙ,, at least 2.0 μ§/ηιΙ,, at least 2.5 μ§/ηιΙ,, at least 3.0 μg/mL or higher, inclusive of all ranges therein. In some embodiments, polyclonal antibody compositions as disclosed herein can neutralize 1 μg/mL of FHA when used at a concentration (titer) of at least 0.1 μg/mL, for example at least 0.2 μg/mL, at least 0.3 μg/mL, at least 0.4 μg/mL, at least 0.5 μg/mL, at least 0.6 μg/mL, at least 0.7 μg/mL, at least 0.8 μg/mL, at least 0.9 μg/mL, at least 1.0 μg/mL, at least 1.5 μg/mL, at least 2.0 μg/mL, at least 2.5 μg/mL, at least 3.0 μg/mL or higher, inclusive of all ranges therein.

[00061] In view of the neutralizing activity of the disclosed antibodies and polyclonal antibody compositions, they are useful for modulating the growth and/or colonization of one or more Bordetella strains and/or species including antibiotic-resistant Bordetella strains; reducing or killing one or more strains and/or species of Bordetella including antibiotic-resistant Bordetella strains and/or species; and/or treating or preventing a Bordetella infection including infection with an antibiotic-resistant strain in a mammal. In some embodiments, the human recombinant antibodies described herein can bind Bordetella antigens, e.g., PTx or FHA. An anti-Bordetella polyclonal antibody may bind to Bordetella antigens in a multivalent manner, which may result in synergistic neutralization, improved phagocytosis of infected cells by macrophages, improved antibody-dependent cellular cytotoxicity (ADCC) against infected cells, and/or increased complement activity. It is contemplated herein that Bordetella is a multifaceted pathogen that may be neutralized using a multifaceted antibody approach that targets various antigens thereby enhancing the capacity of the immune system (e.g., opsonization) to eliminate these bacteria.

[00062] In some embodiments, the antigen specificity of the individual recombinant human antibodies disclosed herein is located in the variable regions (e.g., VH and VL regions), in particular, in the CDR1, CDR2, and CDR3 regions of the immunoglobulin heavy and/or light chains. In some embodiments, the individual antibodies contain (a) an immunoglobulin heavy chain variable region comprising the structure HCDR1-HCDR2-HCDR3 and (b) an

immunoglobulin light chain variable region comprising the structure LCDR1-LCDR2-LCDR3, wherein the heavy chain variable region and the light chain variable region together define a single binding site for binding to one or more Bordetella strains and/or species by binding to an antigen of Bordetella or a Bordetella antigenic epitope. In some embodiments, the human recombinant antibodies comprise at least one heavy or light chain CDR that is at least 80% identical to the respective HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR2 amino acid sequences disclosed herein.

[00063] An individual antibody molecule may be characterized by its variable region sequences (i.e., V_H and V_L region), or by one or more heavy or light chain CDR1, CDR2, and CDR3 regions of immunoglobulin heavy and/or light chains. When the individual antibodies are defined by their heavy and light chain CDR regions, it is contemplated that the CDR regions are interposed between human immunoglobulin framework regions (FRs). Thus, in one aspect of the invention, there is provided individual heavy and light chains, or antigen-binding fragments or CDRs thereof, of recombinant human antibodies that bind Bordetella antigens. The heavy chains or light chains (and antigen-binding fragments thereof) of the invention can bind Bordetella antigens individually and/or when paired with a complementary light or heavy chain, respectively. Where necessary, complementary chains can be identified by methods known to those of skill in the art (e.g., phage display).

[00064] Antibodies of the invention include recombinant human antibodies, chimeric antibodies comprising one or more CDRs of the variable regions as disclosed herein, and humanized antibodies into which one or more CDRs as disclosed herein have been grafted. In some embodiments, the antibodies are an intact tetrameric antibody. Alternatively, the antibody can be an antigen-binding fragment of an antibody. Antibody fragments include Fab, Fab', F(ab')₂, and Fv fragments, and single chain antibodies (e.g., scFv).

[00065] In some embodiments, only one or even part of one of the CDRs disclosed herein are required for binding; i.e., there is a subset of CDRs or CDR residues required for binding. Thus, embodiments of the invention include antibodies into which have been grafted or engineered a subset of CDRs or CDR residues disclosed herein. In some embodiments, antibodies and antigen- binding fragments of the invention comprise a portion of a variable region of an antibody, said portion comprising at least one, two, preferably three CDRs selected from CDR1, CDR2, and CDR3. The antigen-binding portion can comprise a portion of an immunoglobulin heavy chain or an immunoglobulin light chain.

[00066] In some embodiments, the antibodies and antigen-binding fragments of the invention comprise one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) at least 80% identical to the heavy chain CDR1, CDR2 and CDR3 amino acid sequences disclosed herein. In some embodiments one or more amino acid residues of the heavy chain CDRs disclosed herein are conservatively substituted. For example, one amino acid residue in the heavy chain CDR1, one or two amino acid residues in the heavy chain CDR2, and/or one, two or three amino acid residues in the heavy chain CDR3 can be conservatively substituted. The antibody can further comprise one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having amino acid sequences at least 80% identical to the light chain CDR1, CDR2 and CDR3 disclosed herein. In some embodiments, one or more amino acid residues of the light chain CDRs disclosed herein are conservatively substituted. For example, one or two amino acid residues in the light chain CDR1, one amino acid residue in the light chain CDR2 and/or one or two amino acid residues in the light chain CDR3 can be conservatively substituted.

[00067] In another aspect of the invention, the antibodies and antigen-binding fragments of the invention comprise one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) at least 80% identical to the light chain CDR1, CDR2 and CDR3 amino acid sequences disclosed herein. In some embodiments one or more amino acid residues of the light chains CDRs disclosed herein are conservatively substituted. For example, one amino acid residue in the light chain CDR1, one or two amino acid residues in the light chain CDR2, and/or one, two or three amino acid residues in the light chain CDR3 can be conservatively substituted. Light chain antibodies of the invention can further comprise one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having amino acid sequences at least 80%> identical to the heavy chain CDR1, CDR2 and CDR3 sequences disclosed herein. In some embodiments one or more amino acid residues of the heavy chain CDRs disclosed herein are conservatively substituted. For example, one or two amino acid residues in the heavy chain CDR1, one amino acid residue in the heavy chain CDR2 and/or one or two amino acid residues in the heavy chain CDR3 can be conservatively substituted.

[00068] The antibodies provided herein include comprise at least one human CDR or a functional fragment thereof. A functional fragment of a CDR is a portion of a complete CDR amino acid sequence that is capable of binding to an antigen. Thus, a functional fragment of a CDR typically includes the amino acid residues required for CDR binding to the antigen. In particular embodiments, the antigen is a Bordetella antigen such as PTx or FHA. Thus, a functional fragment of an anti-Bordetella antigen antibody typically includes the amino acid residues required for CDR binding to the Bordetella antigen.

[00069] Thus, in some embodiments, a recombinant antibody or antigen-binding fragment thereof that binds a Bordetella antigen comprises a heavy chain and a light chain that includes combined one human CDR (or functional fragment thereof) as defined below or a sequence at least 80% identical thereto. In some embodiments, the heavy chain and the light chain include a combined 6 CDRs wherein at least one of the 6 CDRs is a human CDR as defined below or a sequence at least 80%> identical thereto. As described herein, antibodies within the scope of the invention may include human, humanized, mouse, sheep, rabbit, etc., and antibodies into which at least one of the human CDRs (or functional fragment thereof) as defined below, or a sequence at least 80% identical thereto, has been grafted. For example, antibodies within the scope of the invention may include a human HCDR3 derived from a donor antibody as described below and HCDR1, HCDR2, LCDR1, LCDR2, and LCDR3 derived from the acceptor antibody (e.g., a non- Bordetella human antibody or a mouse anti-Bordetella antibody).

[00070] In some embodiments, a recombinant antibody or antigen-binding fragment thereof that binds a Bordetella antigen comprises a heavy chain and a light chain that includes combined two human CDRs (or functional fragments thereof) as defined below or sequences at least 80% identical thereto. Thus, in some embodiments, the heavy chain and the light chain include a combined 6 CDRs wherein at least two of the 6 CDRs are human CDRs as defined below or a sequence at least 80% identical thereto. The human CDRs may be located on a heavy and light chain, respectively, or both may located on the heavy chain or both on the light chain. As described herein, antibodies within the scope of the invention may include human, humanized, mouse, sheep, rabbit, etc., and antibodies into which at least two of the human CDRs (or functional fragment thereof) as defined below, or sequences at least 80%> identical thereto, have been grafted. For example, antibodies within the scope of the invention may include a human HCDR3 and LCDR3 as described below derived from donor antibodies and HCDR1, HCDR2, LCDR1 and LCDR2 derived from the acceptor antibody (e.g., a non-Bordetella human antibody or a mouse anti-Bordetella antibody).

[00071] In some embodiments, a recombinant antibody or antigen-binding fragment thereof that binds a Bordetella antigen comprises a heavy chain and a light chain that includes combined three human CDRs (or functional fragments thereof) as defined below or sequences at least 80% identical thereto. Thus, in some embodiments, the heavy chain and the light chain include a combined 6 CDRs wherein at least three of the 6 CDRs are human CDRs as defined below or a sequence at least 80% identical thereto. The human CDRs may be located on the heavy and light chains in any combination, e.g., all three on the heavy chain, two on the heavy chain and one on the light chain, etc. As described herein, antibodies within the scope of the invention may include human, humanized, mouse, sheep, rabbit, etc., and antibodies into which at least three of the human CDRs (or functional fragments thereof) as defined below, or sequences at least 80%> identical thereto, have been grafted. For example, antibodies within the scope of the invention may include a human HCDR3, LCDR3 and HCDR2 as described below derived from donor antibodies and HCDR1, LCDR1 and LCDR2 derived from the acceptor antibody (e.g., a non- Bordetella human antibody or a mouse anti-Bordetella antibody).

[00072] In some embodiments, a recombinant antibody or antigen-binding fragment thereof that binds a Bordetella antigen comprises a heavy chain and a light chain that includes combined three human CDRs (or functional fragments thereof) as defined below or sequences at least 80% identical thereto. Thus, in some embodiments, the heavy chain and the light chain include a combined 6 CDRs wherein at least three of the 6 CDRs are human CDRs as defined below or a sequence at least 80% identical thereto. The human CDRs may be located on the heavy and light chains in any combination, e.g., all three on the heavy chain, two on the heavy chain and one on the light chain, etc. As described herein, antibodies within the scope of the invention may include human, humanized, mouse, sheep, rabbit, etc., and antibodies into which at least three of the human CDRs (or functional fragments thereof) as defined below, or sequences at least 80%> identical thereto, have been grafted. For example, antibodies within the scope of the invention may include a human HCDR3, LCDR3 and HCDR2 as described below derived from donor antibodies and HCDR1, LCDR1 and LCDR2 derived from the acceptor antibody (e.g., a non- Bordetella human antibody or a mouse anti-Bordetella antibody).

[00073] In some embodiments, a recombinant antibody or antigen-binding fragment thereof that binds a Bordetella antigen comprises a heavy chain and a light chain that includes combined four human CDRs (or functional fragments thereof) as defined below or sequences at least 80% identical thereto. Thus, in some embodiments, the heavy chain and the light chain include a combined 6 CDRs wherein at least four of the 6 CDRs are human CDRs as defined below or a sequence at least 80% identical thereto. The human CDRs may be located on the heavy and light chains in any combination, e.g., three on the heavy chain and one on the light chain, etc. As described herein, antibodies within the scope of the invention may include human, humanized, mouse, sheep, rabbit, etc., and antibodies into which at least four of the human CDRs (or functional fragments thereof) as defined below, or sequences at least 80% identical thereto, have been grafted. For example, antibodies within the scope of the invention may include a human HCDR3, LCDR3, HCDR2 and HCDR1 as described below derived from donor antibodies and LCDR1 and LCDR2 derived from the acceptor antibody (e.g., a non-Bordetella human antibody or a mouse anti-Bordetella antibody).

[00074] Those of skill in the art will appreciate that the heavy chain CDR3 is sufficient for most antibody specificities (Xu and Davis, Immunity, 2000, 13: 27-45, incorporated by reference in its entirety) and that any anti-Bordetella antibody library may be created using CDRH3 as the major source of diversification (see, e.g., Hoogenboom et al, J. Mol. Biol, 1992, 227: 381; Lee et al, J. Mol. Biol, 2004, 340: 1073). Thus, antibodies of the invention may be defined solely by the percentage of sequence identity to the HCDR3 sequences and conservatively substituted variants thereof disclosed herein.

[00075] In some embodiments, the antibodies or antigen-binding fragments thereof can bind the same or similar epitope as mouse monoclonal antibodies 1B7 or 11E6, or humanized versions thereof. 1B7 and 11E6 are highly neutralizing to PTx and have been isolated and extensively characterized resulting in the identification of four major neutralizing epitopes on the toxin. For example, 1B7 binds an epitope located primarily on the SI subunit, distal to the receptor binding site contained in subunits S2 and S3. It also appears to bind across the S1-S4 interface, in effect "stapling" the two subunits together. Antibodies recognizing these epitopes either block the binding of the B-oligomer to the cell surface receptors or block catalysis by preventing the dissociation of the two subunits. See, e.g., U.S. pre-grant publication 2012/0244144, incorporated herein by reference in its entirety.

[00076] Antibodies and antigen-binding fragments that bind the same or similar epitope as 1B7 or 11E6 can be identified using any suitable method, such as a competitive binding assay. For example, an antibody can be tested for the ability to competitively inhibit binding of 1B7 or 11E6 to a Bordetella antigen expressed on the surface of a cell. Competitive inhibition of binding of 1B7 or 11E6 in this type of assay indicates that the test antibody binds the same or similar epitope as 1B7 or 11E6. In particular embodiments, the antibody or antigen-binding fragment thereof can have the epitopic specificity of mouse or humanized 1B7 or 11E6. The fine epitopic specificity of an antibody can be determined using any suitable method, such as peptide competition or mutational analysis. For example, a series of PTx variants comprising amino acid replacements can be prepared and an antibody can be tested for the ability to bind each variant. Inhibited or abrogated binding to a variant comprising a particular amino acid substitution is indicative that the substituted amino acid is part of the epitope that the antibody binds.

[00077] The respective CDRs (IMGT numbering) of human recombinant antibodies according to embodiments of the invention are listed below in Table 1 :

Table 1

1 QSVSNY 14

Light 2 DAS

3 CQQRTNWLTF 15

1 GDSIGSYY 16

Heavy 2 IFYTGTI 17

3 CAREGEFFDILTGDYRGLDYW 18

1 QSVSSTY 19

Light 2 GTS

3 CQQYGTSPWTF 20

1 GFTFSNYA 21

Heavy 2 TSGSGGST 22

3 CAKDREVLRFFLLPYYFDSW 23

1 QSVSSDY 24

Light 2 DAS

3 CQQYGNSPLTF 25

1 GYTFSSYG 26

Heavy 2 ISAYNGDT 27

3 CARDTSFAPSGYGFDIS 28

1 QGIRHE 29

Light 2 AAS

3 CLQVNSYPRTF 30

1 GASVSSGTYY 31

Heavy 2 SYYTGSS 32

3 CARDRRKVRGVWAGYSGMDVW 33 1 QEITNY 34

Light 2 DAS

3 CQQYDDFPWGGVTF 35

1 GGTFGSYG 36

Heavy 2 IIPMFGTA 37

3 CARENSIVGANWFDPW 38

55.17.D8

1 QSISTY 39

Light 2 AAS

3 CQQSYSSPYTF 40

1 GFTFSSFD 41

Heavy 2 IRHHGTNH 42

3 CAKDLGFGELYW 43

55.22.E7

1 QSVRTN 44

Light 2 DGF

3 CQQYRTWPRVTF 45

[00078] An antibody, or antigen-binding fragment thereof, may also be conjugated to an effector agent such as a small molecule toxin, a drug, or a radionuclide using standard in vitro conjugation chemistries. If the effector agent is a polypeptide, the antibody can be chemically conjugated to the effector or joined to the effector as a fusion protein. Construction of fusion proteins is within ordinary skill in the art.

[00079] Individual antibodies and polyclonal antibody compositions may bind a protein associated with Bordetella virulence, such as, but not limited to various vaccine antigens including pertussis toxin (PTx), filamentous hemagglutinin (FHA), pertactin (PTN), fimbriae {e.g., fimbriae 2 and 3 (also known as Fims 2/3) (Cherry et al, (2010) CLINICAL AND VACCINE

IMMUNOLOGY, 17:741-747); Bap-5 (GenBank accession no. AF081494) or BapC (GenBank accession no. AJ277634), which has been identified as a member of the B. pertussis autotransporter family and, like BrkA, is a Bvg-regulated serum resistance factor and virulence determinant (Noofeli et al, (2011) Microbial Pathogenesis 51 :169-177); Bpl l52 (also known as IRP1-3), which shows protective immunity when used as a vaccine component (Alvarez Hayes et al, (2011) VACCINE 29(47)8731-8739; Marr et al, (2011) PLoS ONE 6(6):e20585); O-antigen, which shields B. parapertussis from binding vaccine-induced antibodies, interferes with opsonophagocytosis of B. parapertussis mediated by aP and wP -induced antibodies and blocks antibody-mediated clearance in vivo (Zhang et al, 2009 PLoS ONE 4(9):e6989; Zhang et al, (2009) INFECTION AND IMMUNITY 77:5050-5058); adenylate cyclase toxin (CyaA), which assists infection by potently suppressing the host immune response (Rossi et al, (2011) J. EXP. MED. 208: 1317-1330); glutamine-binding periplasmic protein; leu/ile/val-binding protein; one putative exported protein (BP0250); iron-superoxide dismutase (Fe-SOD) (Tefon et al, (2011) VACCINE 29:3583-3595; ATP dependent protease; 30S ribosomal protein SI; RNAP subunit; S- adenosylmethionine synthetase; EF-Tu; glutamyl t-RNA amidotransferase subunit A; ketol acid reductoisomerase; serine protease; HsplO and Hsp70 (Altindis et al, (2009) VACCINE 27:542- 548); part of the Type-Ill secretion proteins; DNT; TCT; TcfA; LPS; putative 2-hydroexyacid dehydrogenase; Sbp; amino acid-binding periplasmic protein; putative exported solute binding protein; and hypothetical proteins BP0250, BP2818 and BP3575 (Zhu et al, (2010) PLoS ONE 5(l l):el3915).

[00080] In certain embodiments, an anti-Bordetella polyclonal antibody composition binds to at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least 15, at least 20, at least 25 or more Bordetella proteins. In an exemplary embodiment, an anti-Bordetella polyclonal antibody binds to at least three Bordetella proteins. In certain embodiments, the three proteins may include a cell surface antigen, a toxin and/or an immunomodulator produced by Bordetella. In some embodiments, an anti-Bordetella polyclonal antibody binds at least a cell surface antigen on one or more strains and/or species of Bordetella and a toxin produced by one or more strains and/or species of Bordetella. In another embodiment, an anti-Bordetella polyclonal antibody binds at least a cell surface antigen on one or more strains and/or species of Bordetella and an immunomodulator produced by one or more strains and/or species of Bordetella. In yet another embodiment, an anti-Bordetella polyclonal antibody binds at least a toxin produced by one or more strains and/or species of Bordetella and an immunomodulator produced by one or more strains and/or species of Bordetella. In certain embodiments, the at least 2, 3, 4, 5, 10, 15, 20, 25 or more proteins are on more than one strain of Bordetella. It is contemplated herein that the broad spectrum nature of the disclosed anti- Bordetella polyclonal antibody compositions may be attributed to the inclusion of individual antibodies that bind a protein present on more than one strain of Bordetella {e.g., conserved proteins). In addition, the broad spectrum efficacy of the disclosed polyclonal antibodies may be attributed to the inclusion of certain antibodies that bind antigens associated with only one or two Bordetella strains and/or species.

[00081] An anti-Bordetella polyclonal antibody composition may also be composed of individual antibodies raised by the immune response of a donor (e.g., a human), which has been vaccinated or infected with Bordetella. Further, if antibodies to a particular antigen are known to be relevant and/or effective in the protection, neutralization and/or elimination of Bordetella infection, such antibodies may be raised by immunization of a donor with that particular antigen.

[00082] A human recombinant polyclonal anti-Bordetella antibody composition disclosed herein may comprise at least three, at least 4, at least 5, at least 7, at least 10, at least 15, at least 20, at least 25 or more antibodies, heavy and light chain combinations thereof, and various CDR combinations thereof as disclosed herein. In some embodiments, the recombinant polyclonal antibody compositions disclosed herein may comprise about 3 to about 30 antibodies, about 3 to about 25 antibodies, about 3 to about 20 antibodies, about 3 to about 15 antibodies, about 3 to about 10 antibodies, about 3 to about 5 antibodies, about 5 to about 30 antibodies, about 5 to about 25 antibodies, about 5 to about 20 antibodies, about 5 to about 15 antibodies, about 5 to about 10 antibodies, about 8 to about 30 antibodies, about 8 to about 25 antibodies, about 8 to about 20 antibodies, about 8 to about 15 antibodies, about 8 to about 10 antibodies, about 10 to about 30 antibodies, about 10 to about 25 antibodies, about 10 to about 20 antibodies, about 10 to about 15 antibodies, about 15 to about 25 antibodies, about 15 to about 20 antibodies, about 20 to about 25 antibodies, and about 25 to about 30 antibodies.

[00083] An anti-Bordetella antibody and polyclonal antibody composition may comprise immunoglobulin heavy and light chain variable regions or, heavy and light chain CDR regions, from two, three, four, five, or combinations thereof, of the following antibodies as disclosed herein: 42.11.D4, 42.12.G2, 42.12.A12, 42.12.A9, 42.18.E12, 55.12.A8, 55.15.H5, 55.17.D8 and 55.22.E7, wherein each of the disclosed antibodies comprises immunoglobulin heavy and light chain CDR1, CDR2, CDR3 sequences as set forth in Table 1.

[00084] Exemplary recombinant polyclonal antibody compositions including at least two antibodies disclosed herein are shown in Table 2. All other possible combinations of antibodies and the CDRs therein are within the scope of the invention.

[00085] Table 2

[00086] Yet another aspect of the invention is antibodies that compete with the antibodies disclosed herein for binding to one or more Bordetella epitopes, antigens, strains and/or species. Thus, in some embodiments, the disclosed recombinant human polyclonal antibody compositions include one or more antibodies that compete with one of the disclosed antibodies for binding to one or more Bordetella epitopes, antigens, strains and/or species, for example, under the conditions described in Example 2. Exemplary Bordetella strains and/or species for determining whether an antibody competes with binding to a disclosed antibody include ATCC Strain BAA-589 (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00087] In some embodiments, the disclosed polyclonal antibody compositions bind at least 3 different Bordetella strains and/or species or the antigens they express in their exoproteome. For example, in exemplary embodiments, the disclosed polyclonal antibody compositions bind at least 3, at least 4 or at least 5 different Bordetella strains and/or species, e.g., ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00088] In one embodiment, an antibody provided herein competes with 42.11.D4 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 47 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 49 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 42.11.D4, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00089] In one embodiment, an antibody provided herein competes with 42.11.G2 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 51 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 53 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 42.11.G2, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00090] In one embodiment, an antibody provided herein competes with 42.12.A12 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 55 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 57 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 42.12.A12, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00091] In one embodiment, an antibody provided herein competes with 42.12.A9 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 59 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 61 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 42.12.A9, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00092] In one embodiment, an antibody provided herein competes with 42.18.E12 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 63 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 65 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 42.18.E12, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica. [00093] In another embodiment, an antibody provided herein competes with 55.12.A8 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 67 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 69 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 55.12.A8, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00094] In one embodiment, an antibody provided herein competes with 55.15.H5 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 71 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 73 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 55.15.H5, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00095] In one embodiment, an antibody provided herein competes with 55.17.D8 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 75 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 77 for binding to one or more Bordetella epitopes, antigens, strains and/or species. An exemplary epitope includes the SI subunit of PTx, distal to the receptor binding site contained in subunits S2 and S3. Exemplary antigens include PTx. Exemplary Bordetella strains and/or species for a competitive binding assay with 55.17.D8, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

[00096] In one embodiment, an antibody provided herein competes with 55.22.E7 comprising CDR1, CDR2 and CDR3 of heavy chain amino acid sequence SEQ ID NO: 79 and CDR1, CDR2, CDR3 of light chain amino acid sequence SEQ ID NO: 81 for binding to one or more Bordetella epitopes, antigens, strains and/or species. Exemplary antigens include FHA. Exemplary Bordetella strains and/or species for a competitive binding assay with 55.22.E7, include ATCC Strain BAA-589 pertussis (also known as Tomaha I pertussis), ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica

[00097] Variations in the amino acid sequence of the individual antibodies and antigen-binding fragments thereof may include an amino acid addition, deletion, insertion, substitution etc., one or more modification in the backbone or side-chain of one or more amino acid, or an addition of a group or another molecule to one or more amino acids (side-chains or backbone).

[00098] Variant antibody or antigen binding fragments within the scope of the invention may have substantial sequence similarity and/or sequence identity in their amino acid sequence in comparison with that the original antibody or antigen binding fragment amino acid sequence. The degree of similarity between two sequences is based upon the percentage of identities (identical amino acids) and of conservative substitution. When determining percent identity of a framework region in comparison with another framework region (e.g., of an antibody variant), the CDRs amino acid sequence should preferably not be taken into account. Likewise, when determining percent identity of a CDR in comparison with another CDR (e.g., the HCDR3 of 42.11.D4 versus the HCDR3 of a variant of 42.11.D4), the framework amino acid sequence should preferably not be taken into account. The degree of similarity and identity between variable chains may be determined by means well known to those of skill in the art, including the BLAST algorithm.

[00099] In some embodiments, percent identity will therefore be indicative of amino acids which are identical in comparison with the original peptide and which may occupy the same or similar position. In some embodiments, percent similarity will be indicative of amino acids which are identical and those which are replaced with conservative amino acid substitution in comparison with the original peptide at the same or similar position.

[000100] Variants of the antibodies of the present invention comprise those which may have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%o, 97%), 98%o, 99%) or 100% sequence identity with an original sequence or a portion of an original sequence. Exemplary embodiments of variant antibodies within the scope of the invention are those having at least 81% sequence identity to a sequence described herein and 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence similarity with an original sequence or a portion of an original sequence. Other exemplary embodiments of variants are those having at least 82% sequence identity to a sequence described herein and 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%), 96%o, 97%), 98%o, 99% or 100% sequence similarity with an original sequence or a portion of an original sequence. Further exemplary embodiments of variants are those having at least 85% sequence identity to a sequence described herein and 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence similarity with an original sequence or a portion of an original sequence. Other exemplary embodiments of variants are those having at least 90%> sequence identity to a sequence described herein and 90%>, 91 >, 92%, 93%>, 94%), 95%), 96%o, 97%), 98%>, 99% or 100% sequence similarity with an original sequence or a portion of an original sequence. Additional exemplary embodiments of variants are those having at least 95% sequence identity to a sequence described herein and 95%, 96%, 97%, 98%, 99% or 100% sequence similarity with an original sequence or a portion of an original sequence. Yet additional exemplary embodiments of variants are those having at least 97% sequence identity to a sequence described herein and 97%, 98%, 99% or 100% sequence similarity with an original sequence or a portion of an original sequence.

[000101] Another type of variable region modification in some embodiments is mutation of amino acid residues within the VH and/or VL CDR1, CDR2 and/or CDR3 regions to thereby improve one or more binding properties (e.g., affinity) of the antibody of interest, known as "affinity maturation." Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce the mutation(s), and the effect on antibody binding, or other functional property of interest, can be evaluated in in vitro or in vivo assays as described herein. Conservative modifications can be introduced. The mutations may be amino acid substitutions, additions or deletions. Moreover, typically no more than one, two, three, four or five residues within a CDR region are altered. Methods of affinity maturation, including alanine scanning (see, e.g., international application WO1995/023813), are known to those of skill in the art. (see, e.g., U.S. Patent No. 7,117,096 and U.S. pre-grant publication 2012/0316071). Production of Antibodies

[000102] The antibodies that are disclosed herein can be made by a variety of methods familiar to those skilled in the art, including recombinant DNA methods, chemical synthesis, etc.

[000103] In some embodiments, antibodies and antigen-binding fragments thereof are produced recombinantly. In order to express the antibodies, nucleotide sequences able to encode any one of a light and heavy immunoglobulin chains described herein may be inserted into an expression vector, e.g., a vector that contains the elements for transcriptional and translational control of the inserted coding sequence in a particular host. These elements may include regulatory sequences, such as enhancers, constitutive and inducible promoters, and 5' and 3' untranslated regions.

Methods that are well known to those skilled in the art may be used to construct such expression vectors. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination.

[000104] Thus, embodiments of the invention include the use of nucleic acids that encode the heavy chains and/or heavy chain CDRs of the antibodies and antigen-binding portions described herein. For example, in some embodiments, the nucleic acid can encode a heavy chain or antigen- binding portion thereof that comprises at least one, two or three CDRs having the amino acid sequences of the human heavy chain CDRs of the anti-Bordetella antigen antibodies described herein, wherein, optionally, one, two, three or more amino acids in each CDR can be

conservatively substituted as described above.

[000105] In some embodiments, the nucleic acid encodes an antibody heavy chain or antigen- binding portion thereof that comprises three CDRs that have the amino acid sequences of the three CDRs of the heavy chains of human antibodies disclosed herein (e.g., 42.11.D4, 42.11.G2, 42.12.A12, 42.12.A9, 42.18.E12, 55.12.A8, 55.15.H5, 55.17.D8 or 55.22.E7). Nucleic acids within the scope of the invention can include any combination of the heavy chain CDRs of any of the antibodies disclosed herein. For example, a nucleic acid construct may comprise the heavy chain CDR1 of 42.11.D4, the CDR2 of 42.12.A9 and the CDR3 of 42.12.12. The antibody heavy chains and portions thereof can further comprise any suitable framework regions and/or constant region.

[000106] Thus, embodiments of the invention include the use of nucleic acids that encode the light chains and/or light chain CDRs of the antibodies and antigen-binding portions described herein. For example, in some embodiments, the nucleic acid can encode a light chain or antigen- binding portion thereof that comprises at least one, two or three CDRs having the amino acid sequences of the human light chain CDRs of the anti-Bordetella antigen antibodies described herein, wherein, optionally, one, two, three or more amino acids in each CDR can be

conservatively substituted as described above. In some embodiments, the nucleic acid encodes an antibody light chain or antigen-binding portion thereof that comprises three CDRs that have the amino acid sequences of the three CDRs of the light chains of human antibodies disclosed herein (e.g., 42.11.D4, 42.11.G2, 42.12.A12, 42.12.A9, 42.18.E12, 55.12.A8, 55.15.H5, 55.17.D8 or 55.22.E7). Nucleic acids within the scope of the invention can include any combination of the light chain CDRs of any of the antibodies disclosed herein. For example, a nucleic acid construct may comprise the light chain CDR1 of 42.11.D4, the CDR2 of 42.12.A9 and the CDR3 of 42.12.12. The antibody heavy chains and portions thereof can further comprise any suitable framework regions and/or constant region.

[000107] Expression constructs or expression vectors suitable for the expression of an antibody or antigen-binding fragment that binds a Bordetella antigen are also provided. For example, a nucleic acid encoding all or part of a desired antibody can be inserted into a nucleic acid vector, such as a plasmid or virus, for expression. The vector can be capable of replication in a suitable biological system (e.g., a replicon). A variety of suitable vectors are known in the art, including vectors which are maintained in single copy or multiple copies, or which become integrated into the host cell chromosome. Expression vectors and methods for producing recombinant antibodies for use in some embodiments of the invention are described in U.S. pre-grant publication

2012/0302464, incorporated by reference herein.

[000108] A variety of expression vector/host cell systems known to those of skill in the art may be utilized to express a polypeptide able to encode any one of a light and heavy immunoglobulin chain or CDR described herein. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with baculovirus vectors; plant cell systems transformed with viral or bacterial expression vectors; or animal cell systems. For long-term production of recombinant proteins in mammalian systems, stable expression in cell lines may be effected. For example, nucleotide sequences able to encode any one of a light and heavy immunoglobulin chains described herein may be transformed into cell lines using expression vectors that may contain viral origins of replication and/or endogenous expression elements and a selectable or visible marker gene on the same or on a separate vector. The invention is not to be limited by the vector or host cell employed. In certain embodiments of the present invention, the nucleotide sequences able to encode any one of a light and heavy immunoglobulin chains described herein may each be inserted into a separate expression vector and each chain expressed separately. In another embodiment, both the light and heavy chains able to encode any one of a light and heavy immunoglobulin chains described herein may be inserted into a single expression vector and expressed simultaneously. Nucleic acid sequences may be inserted into expression vector by methods known to those of skill in the art, include ligation and homologous recombination.

[000109] Embodiments of the invention further include expression from a vector comprising nucleotide sequences able to encode any one of a light and/or heavy immunoglobulin chains or CDRs as described herein using an in vitro transcription system or a coupled in vitro

transcription/translation system.

[000110] Suitable expression vectors can contain a number of components, for example, an origin of replication, a selectable marker gene, one or more expression control elements, such as a transcription control element (e.g., promoter, enhancer, terminator) and/or one or more translation signals, a signal sequence or leader sequence, and the like. The vector or other source, if present, can provide expression control elements and a signal or leader sequence. For example, the transcriptional and/or translational control sequences of a cloned nucleic acid encoding an antibody chain can be used to direct expression.

[000111] Suitable expression vectors for expression in mammalian cells include, for example, pCDM8, pCDNAl.l/amp, pcDNA3.1, pRc/RSV, pEF-1 (Invitrogen Life Technologies, Carlsbad, CA), pCMV-SCRIPT, pFB, pSG5, pXTl (Stratagene, La Jolla, CA), pCDEF3 (Goldman, L.A., et al, Biotechniques, 21 : 1013-1015 (1996)), pSVSPORT (GIBCO division of Invitrogen Life Technologies, Carlsbad, CA), pEF-Bos (Mizushima, S., et al, Nucleic Acids Res., 18:5322 (1990)) and the like. Expression vectors which are suitable for use in various expression hosts, such as prokaryotic cells (E. coli), insect cells (Drosophila Schnieder S2 cells, Sf9) and yeast (P.

methanolica, P. pastoris, S. cerevisiae) are also available. [000112] In another aspect, the invention relates to recombinant host cells and a method of preparing an antibody or antigen-binding fragment, antibody chain (e.g., heavy chain, light chain) or antigen-binding portion of an antibody chain, or polyclonal antibody composition of the invention. The antibody or antigen-binding fragment can be obtained, for example, by the expression of one or more recombinant nucleic acids, encoding an antibody, antigen-binding fragment of an antibody, antibody chain or antigen-binding portion of an antibody chain that binds a Bordetella antigen, in a suitable host cell, or using other suitable methods. For example, the expression constructs described herein can be introduced into a suitable host cell, and the resulting cell can be maintained (e.g., in culture) under conditions suitable for expression of the constructs. Suitable host cells can be prokaryotic, including bacterial cells such as E. coli, B. subtilis and/or other suitable bacteria; eukaryotic cells, such as fungal or yeast cells (e.g., Pichia pastoris, Aspergillus sp., Saccharomyces cerevisiae, Schizosaccharomyces pombe, Neurospora crassa), or other lower eukaryotic cells, and cells of higher eukaryotes such as those from insects (e.g., Drosophila Schnieder S2 cells, Sf9 insect cells (WO 94/26087 (O'Connor)), mammals (e.g., COS cells, such as COS-1 (ATCC Accession No. CRL-1650) and COS-7 (ATCC Accession No. CRL- 1651), CHO (e.g., ATCC Accession No. CRL-9096), 293 (ATCC Accession No. CRL-1573), HeLa (ATCC Accession No. CCL-2), CV1 (ATCC Accession No. CCL-70), NSO cells, SP2/0, HuT 78 cells and the like.

[000113] Thus, embodiments of the invention include a recombinant host cell which comprises a (one or more) recombinant nucleic acid or expression construct comprising a nucleic acid encoding an antibody, antigen-binding fragment of an antibody (e.g., a human, humanized, chimeric antibody or antigen-binding fragment of any of the foregoing), antibody chain (e.g., heavy chain, light chain), antigen-binding portion of an antibody chain that binds a Bordetella antigen.

Embodiments of the invention further include methods of producing an antibody, antigen-binding fragment of an antibody (e.g., a human, humanized, chimeric antibody or antigen-binding fragment of any of the foregoing), antibody chain (e.g., heavy chain, light chain), antigen-binding portion of an antibody chain that binds a Bordetella antigen, comprising maintaining a recombinant host cell of the invention under conditions appropriate for expression of an antibody, antigen-binding fragment of an antibody, antibody chain or antigen-binding fragment of an antibody chain.

Additional embodiments include steps of isolating or recovering the antibody, antigen-binding fragment of an antibody, antibody chain or antigen-binding fragment of an antibody chain. [000114] For example, a nucleic acid molecule (i.e., one or more nucleic acid molecules) encoding the heavy and/or light chains of a human antibody that binds a Bordetella antigen, or an expression construct (i.e., one or more constructs) comprising such nucleic acid molecule(s), can be introduced into a suitable host cell to create a recombinant host cell using any method appropriate to the host cell selected (e.g., transformation, transfection, electroporation, infection), such that the nucleic acid molecule(s) are operably linked to one or more expression control elements (e.g., in a vector, in a construct created by processes in the cell, or integrated into the host cell genome). The resulting recombinant host cell can be maintained under conditions suitable for expression (e.g., in the presence of an inducer, in a suitable non-human animal, in suitable culture media supplemented with appropriate salts, growth factors, antibiotics, nutritional supplements, etc.), whereby the encoded polypeptide(s) are produced.

[000115] In general, host cells that contain nucleotide sequences able to encode any one of a light and heavy immunoglobulin chains or CDRs described herein and/or that express a polypeptide encoded by the nucleotide sequences able to encode any one of a light and heavy immunoglobulin chains described herein, or a portion thereof, may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA/DNA or DNA/RNA hybridizations, PCR amplification, and protein bioassay or

immunoassay techniques that include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or amino acid sequences. Immunological methods for detecting and measuring the expression of polypeptides using either specific polyclonal or monoclonal antibodies are known in the art. Examples of such techniques include enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and fluorescence activated cell sorting (FACS).

[000116] Typically, antibodies expressed in mammalian cells are designed to be secreted into the culture medium, or expressed on the surface of the cell. The antibody or antibody fragments can be produced, for example, as intact antibody molecules or as individual VH and VL fragments, Fab fragments, single domains, or as single chains (scFv) (Huston et al, PNAS, 1988, 85: 5879, incorporated by reference in its entirety). The polypeptide produced by a cell may be secreted, retained on the cell surface (e.g., via a cell surface anchoring protein, or may be retained intracellularly depending on the sequence and/or the vector used. In an exemplary embodiment, expression vectors containing nucleotide sequences able to encode any one of a light and heavy immunoglobulin chains described herein may be designed to contain signal sequences that direct secretion of the polypeptide through a prokaryotic or eukaryotic cell membrane. Alternatively, antibodies can be expressed and screened by anchored periplasmic expression (APEx 2-hybrid surface display), as described, for example, in Jeong et al, PNAS, 2007, 104: 8247 or by other anchoring methods as described, for example, in Mazor et al, NATURE BIOTECHNOLOGY, 2007, 25: 563. Additional embodiments of the invention utilize yeast cell surface display (see, e.g. U.S. Patent No. 6,699,658 and references therein).

[000117] In certain embodiments, an anti-Bordetella antibody and polyclonal antibody composition may be produced from a single manufacturing cell line or a mixture of cell lines producing individual monoclonal antibodies. In other embodiments, DNA molecules encoding light chain variable regions and heavy chain variable regions can be chemically synthesized using the sequence information provided herein. Synthetic DNA molecules can be ligated to other appropriate nucleotide sequences, including, e.g., constant region coding sequences, and expression control sequences, to produce conventional gene expression constructs encoding the desired antibodies. Production of defined gene constructs is within routine skill in the art.

Alternatively, the sequences provided herein can be cloned out of hybridomas or B-cells by conventional hybridization techniques or polymerase chain reaction (PCR) techniques, using synthetic nucleic acid probes whose sequences are based on sequence information provided herein, or prior art sequence information regarding genes encoding the heavy and light chains of human antibodies in hybridoma cells.

[000118] Specific expression and purification conditions will vary depending upon the expression system employed. For example, if a gene is to be expressed in E. coli, it is first cloned into an expression vector by positioning the engineered gene downstream from a suitable bacterial promoter, e.g., Trp or Tac, and a prokaryotic signal sequence. The expressed secreted protein accumulates in retractile or inclusion bodies, and can be harvested after disruption of the cells by French press or sonication. The refractile bodies then are solubilized, and the proteins refolded and cleaved by methods known in the art.

[000119] If the engineered gene is to be expressed in eukaryotic host cells, e.g., CHO cells, it is first inserted into an expression vector containing a suitable eukaryotic promoter, a secretion signal, IgG enhancers, and various introns. This expression vector optionally contains sequences encoding all or part of a constant region, enabling an entire, or a part of, a heavy or light chain to be expressed. The gene construct can be introduced into eukaryotic host cells using conventional techniques. The host cells express VL or VH fragments, VL-VH heterodimers, VH-VL or VL-VH single chain polypeptides, complete heavy or light immunoglobulin chains, or portions thereof, each of which may be attached to a moiety having another function (e.g., cytotoxicity). In some embodiments, a host cell is transfected with a single vector expressing a polypeptide expressing an entire, or part of, a heavy chain (e.g., a heavy chain variable region) or a light chain (e.g., a light chain variable region). In other embodiments, a host cell is transfected with a single vector encoding (a) a polypeptide comprising a heavy chain variable region and a polypeptide comprising a light chain variable region, or (b) an entire immunoglobulin heavy chain and an entire

immunoglobulin light chain. In still other embodiments, a host cell is co-transfected with more than one expression vector (e.g., one expression vector expressing a polypeptide comprising an entire, or part of, a heavy chain or heavy chain variable region, and another expression vector expressing a polypeptide comprising an entire, or part of, a light chain or light chain variable region).

[000120] The expression vector may also include constant regions for the heavy and/or light chain. It is contemplated that the choice of the constant region may vary for the individual antibodies included in the polyclonal composition. For example, it may be desirous to have IgGl constant regions for certain antibodies and IgG2 constant regions for other antibodies depending on the desired effector function to clear or destroy antigen (e.g., ADCC, phagocytosis, increased complement activity (e.g., via the classic and/or alternative complement pathways), binding to mass cells and/or basinophiles). Heavy chain constant regions may be selected from the isotypes IgGl, IgG2, IgG3, IgG4, IgAl, IgA2, IgM, IgD, and IgE. Light chain constant regions may be either kappa or lambda.

[000121] A human antibody that binds to one or more Bordetella strains and/or species, or an antigen-binding fragment of the antibody, can be produced by growing a host cell transfected with: (a) an expression vector that encodes a complete or partial immunoglobulin heavy chain, and a separate expression vector that encodes a complete or partial immunoglobulin light chain; or (b) a single expression vector that encodes both chains, under conditions that permit expression of both chains. The intact antibody (or antigen-binding fragment) can be harvested and purified using techniques well known in the art, e.g., Protein A, Protein G, affinity tags such as glutathione- S- transferase (GST) and histidine tags. It is within ordinary skill in the art to express the heavy chain and the light chain from a single expression vector or from two separate expression vectors.

Use of Antibodies

[000122] Anti-Bordetella antibodies, antigen-binding fragments thereof and polyclonal antibody compositions as described herein can be used to treat one or more strains and/or species of Bordetella and/or prevent infection of one or more strains and/or species of Bordetella, including antibiotic resistant strains and/or species of Bordetella, e.g., macrolide-resistant strains and/or species, and erythromycin-resistant strains and/or species. The disclosed antibodies may also be used to treat antibiotic-sensitive strains and/or species. Bordetella infected host cells (e.g., mammalian host cells, e.g., human host cells) are exposed to a therapeutically effective amount of the antibody so as to inhibit infectivity of Bordetella by inhibiting growth and/or colonization, and/or by inducing phagocytosis, and/or killing the Bordetella. In some embodiments, the antibodies inhibit infectivity of Bordetella by at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%.

[000123] In certain embodiments, individual human anti-Bordetella antibodies may be used to treat one or more strains and/or species of Bordetella and/or prevent infection of one or more strains and/or species of Bordetella.

[000124] As used herein, "treat", "treating" and "treatment" mean the treatment of a disease in a mammal, e.g., in a human. This includes: (a) inhibiting the disease or infection, i.e., arresting its development or progression; (b) relieving the disease or infection, i.e., causing regression of the disease state or infection; and/or (c) curing the disease or infection.

[000125] Exemplary diseases that can be treated or prevented using the disclosed antibodies and antigen-binding fragments thereof include, but are not limited to, invasive or toxigenic diseases associated with pathogenic Bordetella strains and/or species. Invasive diseases include pneumonia. It is contemplated herein that certain disorders associated with polymicrobial infections including a Bordetella infection may be treated or prevented using the disclosed antibodies. Exemplary disorders associated with polymicrobial infections (including Bordetella infections) include cystic fibrosis (e.g., infections with Bordetella and Pseudomonas), upper and lower respiratory tract infections, meningitis and pneumonia. [000126] Generally, a therapeutically effective amount of active component is in the range of 0.1 mg/kg to 100 mg/kg, e.g., 1 mg/kg to 100 mg/kg, 1 mg/kg to 10 mg/kg. In an exemplary embodiment, a human recombinant polyclonal antibody may be administered at 1 mg/kg. The amount administered will depend on variables such as the type and extent of disease or indication to be treated, the overall health of the patient, the in vivo potency of the antibody, the

pharmaceutical formulation, and the route of administration. The initial dosage can be increased beyond the upper level in order to rapidly achieve the desired blood-level or tissue-level.

Alternatively, the initial dosage can be smaller than the optimum, and the dosage may be progressively increased during the course of treatment. Human dosage can be optimized, e.g. , in a conventional Phase I dose escalation study designed to run from 0.5 mg/kg to 20 mg/kg. Dosing frequency can vary, depending on factors such as route of administration, dosage amount and the disease being treated. Exemplary dosing frequencies are once per day, once every 2 days, once every three days, once every four days, once every five days, once every six days, once per week, once every two weeks, once every month, once every six months, and once a year. In some embodiments, dosing is once every two weeks. A preferred route of administration is parenteral, e.g., intravenous or subcutaneous. Formulation of antibody-based drugs is within ordinary skill in the art. In some embodiments, the antibody is lyophilized and reconstituted in buffered saline at the time of administration.

[000127] For therapeutic use, one or more disclosed antibodies, or antigen-binding fragments thereof, can be combined with a pharmaceutically acceptable carrier. As used herein,

"pharmaceutically acceptable carrier" means buffers, carriers, and excipients suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. The carrier(s) should be "acceptable" in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient. Pharmaceutically acceptable carriers include buffers, solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is known in the art.

[000128] Pharmaceutical compositions containing one or more of the disclosed antibodies can be presented in a dosage unit form and can be prepared by any suitable method. A pharmaceutical composition should be formulated to be compatible with its intended route of administration. Examples of routes of administration are intravenous (IV), intradermal, inhalation, transdermal, topical, transmucosal, and rectal administration. An exemplary route of administration for monoclonal antibodies is IV infusion. Useful formulations can be prepared by methods well known in the pharmaceutical art. For example, see Remington's Pharmaceutical Sciences, 18th ed. (Mack Publishing Company, 1990). Formulation components suitable for parenteral

administration include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetates, citrates or phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose.

[000129] For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). The carrier should be stable under the conditions of manufacture and storage, and should be preserved against microorganisms. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol), and suitable mixtures thereof.

[000130] Pharmaceutical formulations preferably are sterile. Sterilization can be accomplished, for example, by filtration through sterile filtration membranes. Where the composition is lyophilized, filter sterilization can be conducted prior to or following lyophilization and reconstitution.

EXAMPLES

[000131] The following Examples are merely illustrative and are not intended to limit the scope or content of the invention in any way.

Example 1: Sequence Analysis of Αη -Bordetella Antibodies

[000132] This example describes the sequence analysis of the anti-Bordetella antibodies disclosed herein.

[000133] Individual anti-Bordetella antibodies were isolated from human donor individuals vaccinated with either the acellular vaccines Boostrix and Adacel. Antibody genes were amplified by multiplex PCR. In order to capture all families of variable chains representing the various kappa, lambda and heavy variable families naturally present, several primers were utilized in order to capture every possible gene family. The PCR primers each comprised unique nucleotide sequences; however, some similarities were shared and primers with degenerate bases were used.

[000134] During the PCR reaction, it was possible that a primer unintended for a particular gene family could become incorporated into the 5 ' or 3 ' end of the amplified sequence, which would not affect the CDRs. Regardless of whether a primer was incorporated into the final PCR product cloned, antibody families were identified based on the framework sequences that exist between the CDRs in each of heavy, lambda and kappa chains. To ensure the most natural human antibody, "misprimed" sequences (i.e. those with a primer incorporated for a gene family for which it was not intended) were corrected. As the framework sequences identify the gene families for each of the antibody families, a PCR reaction was utilized to change the incorrectly incorporated primer. The reactions utilizing a PCR reaction with two primers to amplify the gene as is common in the art. Those two primers would be specific to the correct gene family. Alternatively, corrected sequences could be synthesized de novo.

[000135] Individual antibodies (e.g., VH and VL regions) were sequenced by Sanger dideoxy- sequencing and analyzed using IMGT/V -Quest software (Montpellier, France) to identify and confirm variable region sequences.

[000136] The nucleic acid sequences encoding and the protein sequences defining heavy and light chain variable regions of the anti-Bordetella antibodies are shown below (amino terminal signal peptide sequences are not shown). CDR sequences (IMGT definition) are indicated by bold font and underlining in the nucleic acid and amino acid sequences. [000137] Nucleic Acid Sequence Encoding the 42.1 1.D4 Heavy Chain Variable Region (SEQ ID NO: 46)

CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGT GCAGGCTCTGGATTCATCTTCAGTTCGTATGCTATGAACTGGGTCCGCCAGACTCCAGTCAAGGGG CTGGAGTGGGTGGCAGTTATATCATATGACGGAAGTAATAAATACTACGCAGACTCCGTGAAGGGC CGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGGGAGGT GAGGACACGGCTGTCTATTTTTGTGCGCGAGGCCGCGTACATAGCAGTAGCTGGCCCACCCCCTCA GACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGTG

[000138] Protein Sequence Defining the 42.1 1.D4 Heavy Chain Variable Region (SEQ ID NO: 47)

QVQLVESGGGVVQPGRSLRLSCAGSGFIFSSYAMNWVRQTPVKGLEWVAVISYDGSNKYYADSVKG RFTI SRDNSKNTLYLQMNSLGGEDTAVYFCARGRVHSSSWPTPSDLWGQGTLVTVSS

[000139] Nucleic Acid Sequence Encoding the 42.1 1.D4 Kappa Chain Variable Region (SEQ ID

NO: 48)

GACATCCAGTTGACCCAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACT TGCTGGGCCAGTCAGGACATTAGCAGTTATTTAGCCTGGTATCAACAAAGACCAGGGGAAGCCCCT GAGCTCCTCATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCGGTGGA TCTGGGACAGACTTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTCGCAACTTATTACTGT CAACAGCTTAAGAGTTACCCTCCCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAAC

[000140] Protein Sequence Defining the 42.1 1.D4 Kappa Chain Variable Region (SEQ ID NO: 49)

DIQLTQSPSFLSASVGDRVTITCWASQDISSYLAWYQQRPGEAPELLIYAASTLQSGVPSRFSGGG SG DF L I SSLQPEDFA YYCQQLKSYPPTFGGG KVEIK

[000141] Nucleic Acid Sequence Encoding the 42.12.G2 Heavy Chain Variable Region (SEQ ID NO: 50)

CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCTCTGAGAGTCTCCTGT GAAGCCTCTGGACTCAACTTCAGTAACCATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGG CTGGAATGGTTGGCCATTATCTCATATGATGGAACTAATAAGTTCTATGCAGACTCCGTGAAGGGC CGGTTCACCATCTCCAGAGACAATTCCAGGAACACGGTGTTTCTGGAAATGAACAGCCTGAGAGCT GAAGACACGGCTGTGTATTACTGTGCGAAAGATGTAGTGGTGGTAGTACCCCGCCCCTCACGGGAC TCCTACTACTCCGCTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCGAGTG [000142] Protein Sequence Defining the 42.12.G2 Heavy Chain Variable Region (SEQ ID NO: 51)

QVQLVESGGGVVQPGRSLRVSCEASGLNFSNHGMHWVRQAPGKGLEWLAIISYDGTNKFYADSVKG RFTI SRDNSRNTVFLEMNS LRAE DTAVYYCAKDVWWPRPSRDSYYSAMDVWGQGT TVTVS S

[000143] Nucleic Acid Sequence Encoding the 42.12.G2 Kappa Chain Variable Region (SEQ ID NO: 52)

GAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTTTCCAGGGGAAAGAGCCACCCTCTCC TGCAGGGCCAGTCAGAGTGTTAGCAGCAACTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCT CCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGT GGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTAC TGTCAGCTGTTTGGTAGCCCGTTCGGCCAAGGGACCAAGGTGGAAATCAAAC

[000144] Protein Sequence Defining the 42.12.G2 Kappa Chain Variable Region (SEQ ID NO: 53)

EIVLTQSPGTLSLFPGERATLSCRASQSVSSNYLAWYQQKPGQAPRLLIYGASSRATGI PDRFSGS GSGTDFTLTI SRLEPEDFAVYYCQLFGSPFGQGTKVEIK

[000145] Nucleic Acid Sequence Encoding the 42.12.A12 Heavy Chain Variable Region (SEQ

ID NO: 54)

CAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTCAAGCCTGGAGGGTCCCTGAGACTCTCCTGT GCAGCCTCTGGATTCACCTTCACTGACTACTACATGACTTGGATCCGCCAGGCTCCAGGGAAGGGG CTGGAGTGGGTTTCATGCGTTGGTAGTAGTGGTACTTATACAAACTACGCAGACTCTGTGAAGGGC CGATTCACCATCTCCAGAGACAACGCCAAGAACTCACTGTATCTGCAAATGAACAGCCTGAGAACC GAGGACACGGCTGTATAT TACTGTGCGAGAGTGACTGGTTCAGGGAATTTTTATGGCTCTTACCAC TACTACTACATGGACGTCTGGGGCAAAGGGACCACGGTCACCGTCTCGAGTG

[000146] Protein Sequence Defining the 42.12.A12 Heavy Chain Variable Region (SEQ ID NO: 55)

QVQLLESGGGLVKPGGSLRLSCAASGFTFTDYYMTWIRQAPGKGLEWVSCVGSSGTYTNYADSVKG RFTI SRDNAKNSLYLQMNSLRTEDTAVYYCARVTGSGNFYGSYHYYYMDVWGKGTTVTVSS

[000147] Nucleic Acid Sequence Encoding the 42.12.A12 Kappa Chain Variable Region (SEQ

ID NO: 56) GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGAAAAAAGCCACCCTCTCC TGCAGGGCCAGTCAGAGTGTTAGCAACTACTTAGCCTGGTACCAACAGAAACCTGGCCAGTCTCCC AGGCTCCTCATCTTTGATGCCTCCAAGAGGGCCGCTGGCATCCCAGCCAGGTTCAGTGGCAGTGGG TCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGT CAGCAGCGTACCAACTGGCTCACTTTCGGCGGGGGGACCAAGGTGGAGATCAAAC

[000148] Protein Sequence Defining the 42.12.A12 Kappa Chain Variable Region (SEP ID NO: 57)

EIVLTQSPATLSLSPGKKATLSCRASQSVSNYLAWYQQKPGQSPRLLIFDASKRAAGIPARFSGSG SG DFTL I SSLEPEDFAVYYCQQRT WLTFGGGTKVEIK

[000149] Nucleic Acid Sequence Encoding the 42.12.A9 Heavy Chain Variable Region (SEQ ID

NO: 58)

CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACGCTGTCCCTCACCTGC ACTGTCTCTGGTGACTCCATCGGTAGTTACTACTGGAGCTGGATCCGGCAGACCCCAGCGAAGGGA CTGGAGTGGATTGGGAATATCTTTTACACGGGGACTATCAACTATAACCCCTCCCTCAAGAGTCGA GTCACCATTTCAGTCGACACGTTCAAGAACCAGTTCTCCCTGACGCTGACCTCTGTGACCGCTGCG GACACGGCCGTCTATTATTGTGCGAGAGAAGGGGAGTTTTTCGATATTTTGACTGGAGATTATAGG GGCTTAGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGTG

[000150] Protein Sequence Defining the 42.12.A9 Heavy Chain Variable Region (SEQ ID NO: 59)

QVQLQESGPGLVKPSETLSLTCTVSGDSIGSYYWSWIRQTPAKGLEWIGNIFYTGTINYNPSLKSR VTI SVDTFKNQFSLTLTSVTAADTAVYYCAREGEFFDILTGDYRGLDYWGQGTLVTVSS

[000151] Nucleic Acid Encoding the 42.12.A9 Kappa Chain Variable Region (SEP ID NO: 60)

GAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCC TGCAGGGCCAGTCAGAGTGTTAGCAGCACCTACTTAGCCTGGTTCCAGCAGAGGCCTGGCCAGGCT CCCAGGCTCCTCATCTATGGTACATCCAGCCGGGCCACTGGCATCCCAGACAGATTCAGTGGCGGT GGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTTC TGTCAGCAGTATGGTACCTCACCGTGGACGTTCGGCCAGGGGACCAAGGTGGAAATCAAAC

[000152] Protein Sequence Defining the 42.12.A9 Kappa Chain Variable Region (SEQ ID NP: 61)

EIVLTQSPGTLSLSPGERATLSCRASQSVSSTYLAWFQQRPGQAPRLLIYGTSSRATGIPDRFSGG GSGTDFTLTISRLEPEDFAVYFCQQYGTSPWTFGQGTKVEIK [000153] Nucleic Acid Sequence Encoding the 42.18.E12 Heavy Chain Variable Region (SEQ ID NO: 62)

GAGGTGCAGCTGGTGGAGTCTGGGGGAGACTTGGTTCAGCCGGGGGGGTCCCTGAGACTCTCCTGT GAAGCCTCTGGATTCACCTTTAGCAACTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGG CTGGAGTGGGTCTCAGGTACTAGTGGGAGTGGAGGCAGCACATACTACGCAGACTCCGTGAAGGGC CGGTTCACCCTCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCC GAGGACACGGCCGTATATTACTGTGCGAAGGACCGAGAGGTATTACGATTTTTTCTTCTGCCGTAC TACTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGTG

[000154] Protein Sequence Defining the 42.18.E12 Heavy Chain Variable Region (SEQ ID NO: 63)

EVQLVESGGDLVQPGGSLRLSCEASGFTFSNYAMSWVRQAPGKGLEWVSGTSGSGGSTYYADSVKG RFTLSRDNSKNTLYLQMNSLRAEDTAVYYCAKDREVLRFFLLPYYFDSWGQGTLVTVSS

[000155] Nucleic Acid Sequence Encoding the 42.18.E12 Kappa Chain Variable Region (SEQ

ID NO: 64)

GAAATTGTGTTGACGCAGTCTCCAGGCTCCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCC TGCAGGGCCAGTCAGAGTGTTAGCAGCGACTACGTGGCCTGGTATCAGCAGAAACCTGGCCAGGCT CCCCGGCTCCTCATCTTTGATGCATCCACCAGGGCCACCGGCATCCCGGACAGGTTCAGTGGCAGT GGGTCTGGGACAGACTTTACTCTCACCATCGGCAGACTGGAGCCTGAAGATTTTGCCGTCTATTAC TGTCAGCAATATGGTAACTCACCCCTCACTTTCGGGGGAGGGGCCAAGGTGGAGATCAAAC

[000156] Protein Sequence Defining the 42.18.E12 Kappa Chain Variable Region (SEP ID NO: 65)

EIVLTQSPGSLSLSPGERATLSCRASQSVSSDYVAWYQQKPGQAPRLLIFDASTRATGIPDRFSGS GSG DFTL IGRLEPEDFAVYYCQQYG SPLTFGGGAKVEIK

[000157] Nucleic Acid Sequence Encoding the 55.12.A8 Heavy Chain Variable Region (SEQ ID NO: 66)

CAGGTGCAGCTGGTGGAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGC AAGACTTCTGGGTACACATTTTCAAGTTATGGTATCACCTGGGTGCGACAGGCCCCTGGACAAGGG CTTGAGTGGATGGGATGGATCAGCGCCTACAATGGTGACACAAATTATGCACAGAATGTCCAGGGC AGAGTCACCATGACCACAGATACATCCACGAGCACAGCCTACATGGAACTGAGGAGCCTCAGATTT GACGACACGGCCGTTTATTACTGTGCGAGAGATACGAGTTTCGCTCCCAGTGGTTATGGCTTTGAT ATCTCGGGCCAGGGGACCACGGTCACCGTCTCGAGTG [000158] Protein Sequence Defining the 55.12.A8 Heavy Chain Variable Region (SEP ID NO: 67)

QVQLVESGAEVKKPGASVKVSCKTSGYTFSSYGITWVRQAPGQGLEWMGWISAYNGDTNYAQNVQG RVTMTTDTSTSTAYMELRSLRFDDTAVYYCARDTSFAPSGYGFDISGQGTTVTVSS

[000159] Nucleic Acid Sequence Encoding the 55.12.A8 Kappa Chain Variable Region (SEQ ID NO: 68)

GACATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACT TGCCGGGCAAGTCAGGGCATTAGACATGAGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCT CAGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGA TCTGGGACAGATTTCACTCTCACAATCAATAGCCTGCAGCCTGAAGATTCTGCAACTTATTACTGT CTACAGGTTAATAGTTATCCCCGGACGTTCGGCCAAGGGACCAGGGTGGACATCAAAC

[000160] Protein Sequence Defining the 55.12.A8 Kappa Chain Variable Region (SEQ ID NO: 69)

DIQLTQSPSSLSASVGDRVTITCRASQGIRHELAWYQQKPGKAPQRLIYAASSLQSGVPSRFSGSG SGTDFTLTINSLQPEDSATYYCLQV SYPRTFGQGTRVDIK

[000161] Nucleic Acid Sequence Encoding the 55.15.H5 Heavy Chain Variable Region (SEQ ID NO: 70)

CAGGTGCAGCTGGTGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGC ACAGTCTCTGGTGCCTCCGTCAGCAGTGGCACTTACTACTGGAGCTGGATCCGGCAGCCCCCAGGG AAGGGACTGGAGTGGATTGGGTATAGTTATTACACTGGAAGTAGCGACTCCAACGCCTCCCTCAAG AGTCGAGTCACCTTATCAATAGACACGTCCCGGAACCTGTTGTCCCTGAAGCTGACCTCTGTGACC GCTGCGGACACGGCCGTTTACTACTGTGCGAGAGATAGGCGGAAGGTTCGGGGAGTTTGGGCCGGG TACTCCGGTATGGACGTCTGGGGCCCAGGGACCACGGTCACCGTCTCGAGTG

[000162] Protein Sequence Defining the 55.15.H5 Heavy Chain Variable Region (SEP ID NO: 71)

QVQLVESGPGLVKPSETLSLTCTVSGASVSSGTYYWSWIRQPPGKGLEWIGYSYYTGSSDSNASLK SRVTLS ID SRNLLSLKL SVTAADTAVYYCARDRRKVRGVWAGYSGMDVWGPG VTVSS

[000163] Nucleic Acid Sequence Encoding the 55.15.H5 Kappa Chain Variable Region (SEQ ID

NP: 72) GAAATTGTAATGACGCAGTCTCCATCCTCCCTGTCTGCATCTGTTGGAGACAGAGTCACCATCACT TGCCAGGCGAGTCAGGAGATTACCAACTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCT AAACTCCTGATCTATGATGCTTCCCATTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGA TCTGGGACAGATTTTTCTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGT CAACAGTATGATGATTTCCCTTGGGGTGGGGTTACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA

C

[000164] Protein Sequence Defining the 55.15.H5 Kappa Chain Variable Region (SEQ ID NO: 73)

EIVMTQSPSSLSASVGDRVTITCQASQEITNYLNWYQQKPGKAPKLLIYDASHLETGVPSRFSGSG SGTDFSFTISSLQPEDIATYYCQQYDDFPWGGVTFGGGTKVEIK

[000165] Nucleic Acid Sequence Encoding the 55.17.D8 Heavy Chain Variable Region (SEQ ID NO: 74)

CAGATCACCTTGAGGGAGTCTGGGGCTGAGGTGACGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGC AAGGCTTCTGGAGGCACCTTCGGCAGCTATGGTATCAGATGGGTGCGACAGGCCCCTGGACAAGGG CTTGAGTGGATGGGAGGGATCATCCCTATGTTTGGAACAGCAAATTACGCTCAGAGGTTCCAGGGC AGAGTCACTATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGTAGCCTGAAATCT GAGGACACGGCCGTGTATTATTGTGCGAGAGAGAATTCCATAGTGGGGGCTAATTGGTTCGACCCC TGGGGCCAGGGAACCCTGGTCACCGTCTCGAGTG

[000166] Protein Sequence Defining the 55.17.D8 Heavy Chain Variable Region (SEP ID NO: 75)

QITLRESGAEVTKPGSSVKVSCKASGGTFGSYGIRWVRQAPGQGLEWMGGIIPMFGTANYAQRFQG RVT I TADKSTSTAYMELS SLKSEDTAVYYCARENSIVGA WFDPWGQGTLVTVS S

[000167] Nucleic Acid Sequence Encoding the 55.17.D8 Kappa Chain Variable Region (SEQ ID NO: 76)

GAAATTGTGATGACGCAGTCTCCATCCTCCCTGTCTGCATCTGTCGGAGACAGAGTCACCTTCACT TGCCGGGCAAGTCAGAGCATTAGCACCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCT ACCCTCCTGATCTATGCTGCATCCACTTTGCGAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGA TCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGTAGATTTTGCAACTTACTTCTGT CAACAGAGTTACAGTTCCCCGTACACTTTTGGCCAGGGGACCACGTTGGAGATCAAAC

[000168] Protein Sequence Defining the 55.17.D8 Kappa Chain Variable Region (SEQ ID NO: 77) EIVMTQSPSSLSASVGDRVTFTCRASQSISTYLNWYQQKPGKAPTLLIYAASTLRSGVPSRFSGSG SG DF L I S SLQPVDFA YFCQQSYSSPYTFGQG LE IK

[000169] Nucleic Acid Sequence Encoding the 55.22.E7 Heavy Chain Variable Region (SEQ ID NO: 78)

GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAACCTGGGGGCTCCCTGAGACTCTCCTGT GCAGCGTCTGGATTCACCTTCAGTAGCTTTGACATGCACTGGGTCCGCCAGGCTCCAGGCAAAGGG CTGCAGTGGGTGGCAT T TATCCGGCATCATGGAACAAATCATTCCTATGTGGACTCCGTGAAGGGC CGCTTTACCATCTCCAGAGACAATTCCAGGAACACCCTGTATCTACAGATGAACAGCCTGAGACCT GAGGACACGGCTCTCTAT TACTGTGCGAAAGACCTAGGCTTCGGGGAGTTGTACTGGGGCCAGGGC ACCCTGGTCACCGTCTCGAGTG

[000170] Protein Sequence Defining the 55.22.E7 Heavy Chain Variable Region (SEQ ID NO: 79)

EVQLVESGGGVVQPGGSLRLSCAASGFTFSSFDMHWVRQAPGKGLQWVAFIRHHGTNHSYVDSVKG RFTISRDNSRNTLYLQMNSLRPEDTALYYCAKDLGFGELYWGQGTLVTVSS

[000171] Nucleic Acid Sequence Encoding the 55.22.E7 Kappa Chain Variable Region (SEQ ID NO: 80)

GACATCGTGATGACCCAGTCTCCAGCCACCCTGTCTGTGTCTCCAGGGGACAGGGCCACCCTCTCA TGCAGGGCCAGTCAGAGTGTCAGGACCAACT TAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCC AGGCTCCTCATTCACGATGGATTCACCAGGTCCGCTGGTATCCCAGCCAGGTTCAGTGGCAGTGGG TCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGTCTGAAGATTTTGCCGTTTATTACTGT CAGCAATATCGTACTTGGCCTCGGGTCACTTTCGGCCCTGGGACCAAGGTGGATATCAAAC

[000172] Protein Sequence Defining the 55.22.E7 Kappa Chain Variable Region (SEQ ID NO: 81)

DIVMTQSPATLSVSPGDRATLSCRASQSVRTNLAWYQQKPGQAPRLLIHDGFTRSAGIPARFSGSG SGTEFTLTISSLQSEDFAVYYCQQYRTWPRVTFGPGTKVDIK

[000173] Table 1 summarizes the heavy chain and light chain CDR sequences (IMGT definition) of the disclosed Bordetella antibodies. To create complete heavy and/or light chain antibody sequences, each variable sequence above can be combined with a constant region. Human constant regions for heavy chain, kappa chain, and lambda chain are known in the art. For example, a complete heavy chain comprises a heavy variable sequence followed by a human heavy chain constant sequence such as IgGl, IgG2, IgG3, IgG4, IgAl, IgA2, IgM, IgD, or IgE. A complete kappa chain comprises a kappa variable sequence followed by the human kappa light chain constant sequence, and a complete lambda chain comprises a lambda variable sequence followed by the human lambda light chain constant sequence. Exemplary human heavy chain, kappa chain, and lambda chains are shown below.

[000174] Nucleic Acid Sequence Encoding the genomic Human IgGl Heavy Chain Constant

Region (SEP ID NO: 82)

CCTCCACCAAGGGCCCATCGGTCT TCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGG GCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACT TCCCCGAACCGGTGACGGTGTCGT GGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCT TCCCGGCTGTCCTACAGTCCTCAG GACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCT TGGGCACCCAGACCT ACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGT TGGTGAGA GGCCAGCACAGGGAGGGAGGGTGTCTGCTGGAAGCAGGCTCAGCGCTCCTGCCTGGACGC ATCCCGGCTATGCAGCCCCAGTCCAGGGCAGCAAGGCAGGCCCCGTCTGCCTCT TCACCC GGAGCCTCTGCCCGCCCCACTCATGCTCAGGGAGAGGGTCT TCTGGCT T T T TCCCAGGCT CTGGGCAGGCACAGGCTAGGTGCCCCTAACCCAGGCCCTGCACACAAAGGGGCAGGTGCT GGGCTCAGACCTGCCAAGAGCCATATCCGGGAGGACCCTGCCCCTGACCTAAGCCCACCC CAAAGGCCAAACTCTCCACTCCCTCAGCTCGGACACCT TCTCTCCTCCCAGAT TCCAGTA ACTCCCAATCT TCTCTCTGCAGAGCCCAAATCT TGTGACAAAACTCACACATGCCCACCG TGCCCAGGTAAGCCAGCCCAGGCCTCGCCCTCCAGCTCAAGGCGGGACAGGTGCCCTAGA GTAGCCTGCATCCAGGGACAGGCCCCAGCCGGGTGCTGACACGTCCACCTCCATCTCT TC CTCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCT TCCCCCCAAAACCCAAGGA CACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGA AGACCCTGAGGTCAAGT TCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC AAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCT GCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCC AGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGTGGGACCCGTGGGGTGCGAGGGCC ACATGGACAGAGGCCGGCTCGGCCCACCCTCTGCCCTGAGAGTGACCGCTGTACCAACCT CTGTCCTACAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGA GCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCT TCTATCCCAGCGACAT CGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCT TCT TCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTG GCAGCAGGGGAACGTCT TCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACAC GCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA

[000175] Nucleic Acid Sequence Encoding the Human IgGl Heavy Chain Constant Region cDNA (SEP ID NO: 83)

CCTCCACCAAGGGCCCATCGGTCT TCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAG CGGCCCTGGGCTGCCTGGTCAAGGACTACT TCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCG CCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCAC CGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA CCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTG AGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGG AGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATG GCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCA AAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCA AGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG AGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCT TCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCT CCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA

[000176] Protein Sequence Defining the Human IgGl Heavy Chain Constant Region (SEQ ID NO: 84)

STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEK I SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[000177] Nucleic Acid Sequence Encoding the genomic Human IgG2 Heavy Chain Constant gion (SEQ ID NO: 85)

CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGA GCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGT GGAACTCAGGCGCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAG GACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCT ACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGGTGAGA GGCCAGCTCAGGGAGGGAGGGTGTCTGCTGGAAGCCAGGCTCAGCCCTCCTGCCTGGACG CACCCCGGCTGTGCAGCCCCAGCCCAGGGCAGCAAGGCAGGCCCCATCTGTCTCCTCACC CGGAGGCCTCTGCCCGCCCCACTCATGCTCAGGGAGAGGGTCTTCTGGCTTTTTCCACCA GGCTCCAGGCAGGCACAGGCTGGGTGCCCCTACCCCAGGCCCTTCACACACAGGGGCAGG TGCTTGGCTCAGACCTGCCAAAAGCCATATCCGGGAGGACCCTGCCCCTGACCTAAGCCG ACCCCAAAGGCCAAACTGTCCACTCCCTCAGCTCGGACACCTTCTCTCCTCCCAGATCCG AGTAACTCCCAATCTTCTCTCTGCAGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCC AGGTAAGCCAGCCCAGGCCTCGCCCTCCAGCTCAAGGCGGGACAGGTGCCCTAGAGTAGC CTGCATCCAGGGACAGGCCCCAGCTGGGTGCTGACACGTCCACCTCCATCTCTTCCTCAG CACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCA TGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCG AGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCAC GGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGG ACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCA TCGAGAAAACCATCTCCAAAACCAAAGGTGGGACCCGCGGGGTATGAGGGCCACATGGAC AGAGGCCGGCTCGGCCCACCCTCTGCCCTGGGAGTGACCGCTGTGCCAACCTCTGTCCCT

ACAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACC AAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTG GAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGAC TCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAG GGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAG AGCCTCTCCCTGTCTCCGGGTAAA

[000178] Nucleic Acid Sequence Encoding the Human IgG2 Heavy Chain Constant Region cDNA (SEQ ID NO: 86)

CCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAG CCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCG CTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCA GCGTGGTGACCGTGCCCTCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGC CCAGCAACACCAAGGTGGACAAGACAGTTGAGCGCAAATGTTGTGTCGAGTGCCCACCGTGCCCAG CACCACCTGTGGCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCT CCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCA ACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCACGGGAGGAGCAGTTCAACA GCACGTTCCGTGTGGTCAGCGTCCTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTACA AGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGC AGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCA GCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC AGCCGGAGAACAACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCTACA GCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATG AGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA

[000179] Protein Sequence Defining the Human IgG2 Heavy Chain Constant Region (SEQ ID NO: 87)

STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYK CKVSNKGLPAPIEK I SKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[000180] Nucleic Acid Sequence Encoding the Human Kappa Light Chain Constant Region (SEQ ID NO: 88)

GAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTG CCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATA ACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACA GCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAG TCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT [000181] Protein Sequence Defining the Human Kappa Light Chain Constant Region (SEQ ID

NO: 89)

RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTY SLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[000182] Nucleic Acid Sequence Encoding the Human Lambda Light Chain Constant Region (SEQ ID NO: 90)

GTCAGCCCAAGGCCAACCCCACTGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTCCAAGCCAACA AGGCCACACTAGTGTGTCTGATCAGTGACTTCTACCCGGGAGCTGTGACAGTGGCCTGGAAGGCAG ATGGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCAAACCCTCCAAACAGAGCAACAACAAGTACG CGGCCAGCAGCTACCTGAGCCTGACGCCCGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGG TCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA

[000183] Protein Sequence Defining the Human Lambda Light Chain Constant Region (SEQ ID NO: 91)

GQPKANPTVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADGSPVKAGVE KPSKQSNNKY AASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[000184] The variable region sequences described herein can be ligated to each of a number of other constant region sequences known to those skilled in the art to produce active full length immunoglobulin heavy and light chains.

Example 2: Identification of Human Bordetella Antibodies

[000185] Blood from qualified subjects was processed through a Ficoll gradient to isolate PBMCs. Antibody-secreting plasma cells, identified by cell surface markers, were single-cell sorted into 96-well plates and subjected to RT-PCR to amplify antibody genes (Fig. 2A). As shown in Fig. 2A, the upper two panels show plasma cells with a clear CD38high/CD19+ population (R3 gate). The lower two panels show the lambda positive and kappa positive plasma cell populations (R4). Cells from the lambda and kappa positive populations were sorted into individual wells of 96-well PCR plates and heavy and light chains were recovered from single cells by RT-PCR with a cognate pair recovery rate of 95% (Fig. 2B). The variation in size between individual amplicons in the heavy chain gel suggests that a single heavy chain is being amplified in each well as suggested by the sequence data from cloned antibodies. Amplified products (Fig. 2B) were cloned and expressed in mammalian cells (CHO-K1 cells) in 96-well format for screening.

[000186] Expressed antibodies were screened by ELISA to identify Bordetella-specific binders (Fig. 2C-F). Wells in which an antibody was detected (values greater than two times background) are shaded, with color indicating relative expression levels with respect to background.

Background is calculated as the average value of negative control wells, left empty during sorting. Fig. 2D and 2F are tables showing data from plate 42.18 from a screening ELISA against

Bordetella FHA and PTx antigens. Wells displaying 3x background (average value of purple wells) were scored as positive.

Example 3: Identification of FHA Neutralizing Antibodies

[000187] The process set forth in Example 2 identified several anti-filamentous hemagglutinin (FHA) antibodies. An assay was developed test if these antibodies were capable of neutralizing FHA. The assays used the agglutination properties FHA possesses against red blood cells (RBCs) to test antibody neutralization in a 96-well plate format. Of the eight anti-FHA antibodies tested, 55.22.E7 proved to be an efficient FHA neutralizing antibody. It was capable of neutralizing 1 μg/mL of FHA when used at a concentration of 0.3-1.5 μg/mL.

[000188] The 55.22.E7 antibody was serially diluted to give a range of concentrations from 10 μg/mL to 0.05 μg/mL in either PBS or RPMI 1640 medium and then allowed to incubate with FHA (1 μg/mL) in U-bottomed 96-well plates for 30 minutes. After incubating, red blood cells were added to the wells and the mix was allowed to incubate at room temperature for 1 hour. After an hour the plates were observed for pelleting or agglutination of RBCs at the bottom of the wells to indicate if any neutralization occurred. Results are shown in Figure 3

[000189] In conclusion, the 55.22.E7 antibody is an effective neutralizing antibody against FHA, with a titer of O^g/mL remaining effective at neutralizing ^g/mL of FHA. INCORPORATION BY REFERENCE

[000190] The entire disclosure of each of the patent documents and scientific articles referred to herein is incorporated by reference for all purposes.

EQUIVALENTS

[000191] The invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting on the invention described herein. Scope of the invention is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and the range of equivalency of the claims are intended to be embraced therein.

Claims

WHAT IS CLAIMED IS:

1. An antibody or antigen-binding fragment thereof that binds Pertussis toxin comprising at least one heavy chain complementarity determining region (HCDRl, HCDR2 and/or HCDR3) at least 80% identical to an amino acid sequence selected from the group consisting of: a. a HCDRl as set forth in SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 11, SEQ ID NO: 16, SEQ ID NO: 21, SEQ ID NO: 26, SEQ ID NO: 31, or SEQ ID NO: 36; b. a HCDR2 as set forth in SEQ ID NO: 2, SEQ ID NO: 7, SEQ ID NO: 12, SEQ ID NO: 17, SEQ ID NO: 22, SEQ ID NO: 27, SEQ ID NO: 32, or SEQ ID NO: 37; and/or c. a HCDR3 as set forth in SEQ ID NO: 3, SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 18, SEQ ID NO: 23, SEQ ID NO: 28, SEQ ID NO: 33, or SEQ ID NO: 38.

2. An antibody or antigen-binding fragment thereof that binds Pertussis toxin comprising at least one light chain complementarity determining region (LCDRl, LCDR2 and/or LCDR3) selected from the group consisting of: a. a LCDRl at least 80% identical to an amino acid sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 9, SEQ ID NO: 14, SEQ ID NO: 19, SEQ ID NO: 24, SEQ ID NO: 29, SEQ ID NO: 34, or SEQ ID NO: 39; b. a LCDR2 selected from the group consisting of AAS, GAS, DAS, GTS, DAS and conservatively substituted variants thereof; and/or c. a LCDR3 at least 80% identical to an amino acid sequence as set forth in SEQ ID NO: 5, SEQ ID NO: 10, SEQ ID NO: 15, SEQ ID NO: 20, SEQ ID NO: 25, SEQ ID NO: 30, SEQ ID NO: 35, or SEQ ID NO: 40.

3. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein said antibody or antigen-binding fragment thereof is selected from the group consisting of human antibody, an antigen-binding fragment of a human antibody, a humanized antibody, an antigen-binding fragment of a humanized antibody, a chimeric antibody and an antigen-binding fragment of a chimeric antibody.

4. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen-binding fragment is an antigen-binding fragment selected from the group consisting of an Fab fragment, an Fab' fragment, and F(ab')₂ fragment and an Fv fragment.

5. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen binding fragment competitively inhibits binding of mouse monoclonal antibody 1B7.

6. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen-binding fragment has an epitope specificity substantially similar to mouse monoclonal antibody 1B7.

7. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen binding fragment competitively inhibits binding of mouse monoclonal antibody 11E6.

8. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen-binding fragment has an epitope specificity substantially similar to mouse monoclonal antibody 11E6.

9. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen binding fragment binds Pertussis toxin with an equilibrium dissociation constant (K_D) of about 2-100 nM.

10. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen-binding fragment neutralizes Pertussis toxin.

11. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen binding fragment thereof binds to the catalytic SI subunit of Pertussis toxin.

12. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein the antibody or antigen binding fragment thereof binds to the S2 and S3 subunits of Pertussis toxin.

13. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 1, the HCDR2 is at least 80% identical to SEQ ID NO: 2, and the HCDR3 is at least 80% identical to SEQ ID NO: 3.

14. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 6, the HCDR2 is at least 80% identical to SEQ ID NO: 7, and the HCDR3 is at least 80% identical to SEQ ID NO: 8.

15. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 11, the HCDR2 is at least 80% identical to SEQ ID NO: 12, and the HCDR3 is at least 80% identical to SEQ ID NO: 13.

16. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 16, the HCDR2 is at least 80% identical to SEQ ID NO: 17, and the HCDR3 is at least 80% identical to SEQ ID NO: 18.

17. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 21, the HCDR2 is at least 80% identical to SEQ ID NO: 22, and the HCDR3 is at least 80% identical to SEQ ID NO: 23.

18. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 26, the HCDR2 is at least 80% identical to SEQ ID NO: 27, and the HCDR3 is at least 80% identical to SEQ ID NO: 28.

19. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 31, the HCDR2 is at least 80% identical to SEQ ID NO: 32, and the HCDR3 is at least 80% identical to SEQ ID NO: 33.

20. The antibody or antigen-binding fragment thereof of claim 1, wherein the HCDR1 is at least 80% identical to SEQ ID NO: 36, the HCDR2 is at least 80% identical to SEQ ID NO: 37, and the HCDR3 is at least 80% identical to SEQ ID NO: 38.

21. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80%) identical to SEQ ID NO: 4, the LCDR2 is AAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 5.

22. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80% identical to SEQ ID NO: 9, the LCDR2 is GAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 10.

23. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80% identical to SEQ ID NO: 14, the LCDR2 is DAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 15.

24. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80% identical to SEQ ID NO: 19, the LCDR2 is GTS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 20.

25. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80% identical to SEQ ID NO: 24, the LCDR2 is DAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 25.

26. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80% identical to SEQ ID NO: 29, the LCDR2 is AAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 30.

27. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80% identical to SEQ ID NO: 34, the HCDR2 is DAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 35.

28. The antibody or antigen-binding fragment thereof of claim 2, wherein the LCDR1 is at least 80%) identical to SEQ ID NO: 39, the LCDR2 is AAS or a conservatively substituted variant thereof, and the LCDR3 is at least 80% identical to SEQ ID NO: 40.

29. An antibody or antigen-binding fragment thereof that binds Pertussis toxin, wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable region encoded by a nucleic acid sequence at least 80% identical to SEQ ID NO: 54 and a light chain variable region encoded by a nucleic acid sequence at least 80%> identical to SEQ ID NO: 56.

30. An antibody or antigen-binding fragment thereof that binds Pertussis toxin, wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable region encoded by a nucleic acid sequence at least 80% identical to SEQ ID NO: 62 and a light chain variable region encoded by a nucleic acid sequence at least 80%> identical to SEQ ID NO: 64.

31. An antibody or antigen-binding fragment thereof that binds filamentous hemagglutinin comprising at least one heavy chain complementarity determining region (HCDRl, HCDR2 and/or HCDR3) at least 80% identical to an amino acid sequence selected from the group consisting of: a. a HCDRl as set forth in SEQ ID NO: 41; b. a HCDR2 as set forth in SEQ ID NO: 42; and/or c. a HCDR3 as set forth in SEQ ID NO: 43.

32. The antibody or antigen-binding fragment of claim 31 , wherein the antibody or antigen- binding fragment comprises at least one light chain complementarity determining region selected from the group consisting of: a. a LCDR1 at least 80% identical to QSVRTN (SEQ ID NO: 44); b. a LCDR2 of DGF or conservatively modified variants thereof; or c. a LDCR3 at least 80% identical to CQQYRTWPRVTF (SEQ ID NO: 45).

33. The antibody or antigen-binding fragment of claim 32, wherein: a. the HCDR1 is at least 80% identical to GFTFSSFD (SEQ ID NO: 41); b. the HCDR2 is at least 80% identical to IRHHGTNH (SEQ ID NO: 42); c. the HCDR3 is at least 80% identical to CAKDLGFGELYW (SEQ ID NO: 43); d. the LCDR1 is at least 80% identical to QSVRTN (SEQ ID NO: 44); e. the LCDR2 is DGF or a conservatively modified variant thereof; and f. the LCDR2 is at least 80% identical to CQQYRTWPRVTF (SEQ ID NO : 45).

34. The antibody or antigen-binding fragment thereof of claim 31 , wherein said antibody or antigen-binding fragment thereof is selected from the group consisting of human antibody, an antigen-binding fragment of a human antibody, a humanized antibody, an antigen-binding fragment of a humanized antibody, a chimeric antibody and an antigen-binding fragment of a chimeric antibody.

35. The antibody or antigen-binding fragment thereof of claim 31 , wherein the antibody or antigen-binding fragment is an antigen-binding fragment selected from the group consisting of an Fab fragment, an Fab' fragment, and F(ab')₂ fragment and an Fv fragment.

36. The antibody or antigen-binding fragment of claim 31 , wherein the antibody or antigen binding fragment binds filamentous hemagglutinin with an equilibrium dissociation constant (K_D) of about 2-100 nM.

37. The antibody or antigen-binding fragment of claim 31 , wherein the antibody or antigen binding fragment neutralizes filamentous hemagglutinin.

38. A human recombinant polyclonal antibody composition that binds to one or more Bordetella strains and/or species comprising at least three different human antibodies that individually bind one or more Bordetella strains and/or species.

39. The polyclonal antibody composition of claim 38, comprising at least 5 different human antibodies that individually bind one or more Bordetella strains and/or species.

40. The polyclonal antibody composition of claim 38, comprising at least 10 different human antibodies that individually bind one or more Bordetella strains and/or species.

41. The polyclonal antibody composition of claim 38, comprising at least one of the antibodies identified in Table 1.

42. The polyclonal antibody composition of claim 38, comprising at least 3 or of the antibodies identified in Table 1.

43. The polyclonal antibody composition of claim 38, wherein at least one of the antibodies binds to at least one of the Bordetella strains and/or species selected from the group consisting of: ATCC Strain BAA-589 pertussis, ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

44. The polyclonal antibody composition of claim 38, wherein at least one of the antibodies binds at least 5 strains and/or species, at least 6 strains and/or species, or at least 7 strains and/or species selected from the group consisting of: ATCC Strain BAA-589 pertussis, ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

45. The polyclonal antibody composition of claim 38, wherein one of the antibodies binds a cell surface antigen on one or more Bordetella strains and/or species and one of the antibodies binds a toxin produced by one or more Bordetella strains and/or species.

46. The polyclonal antibody composition of claim 38, wherein one of the antibodies binds a cell surface antigen on one or more Bordetella strains and/or species and one of the antibodies binds an immunomodulator produced by one or more Bordetella strains and/or species.

47. The polyclonal antibody composition of claim 38, wherein one of the antibodies binds a toxin produced by one or more Bordetella strains and/or species and one of the antibodies binds an immunomodulator produced by one or more Bordetella strains and/or species.

48. The polyclonal antibody composition of claim 38, wherein one of the antibodies binds a cell surface antigen on one or more Bordetella strains and/or species, one of the antibodies binds a toxin produced by one or more Bordetella strains and/or species, and one of the antibodies binds an immunomodulator produced by one or more Bordetella strains and/or species.

49. The polyclonal antibody composition of claim 38, wherein at least one of the antibodies is selected from the group consisting of:

(a) an antibody that competes with 42.11.D4 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species;

(b) an antibody that competes with 42.11.G2 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species;

(c) an antibody that competes with 42.12.A12 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species;

(d) an antibody that competes with 42.12.A9 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species;

(e) an antibody that competes with 42.18.E12 comprising the heavy chain and light chain CDRl, CDR2, and CDR3 sequences set forth in Table 1 for binding to one or more Bordetella strains and/or species; wherein the one or more Bordetella strains and/or species are selected from the group consisting of ATCC Strain BAA-589 pertussis, ATCC Strain 10380 pertussis, BAA-1335 pertussis, BAA-587 parapertussis, ATCC Strain 15989 parapertussis, ATCC Strain 10580 bronchiseptica, and ATCC Strain 31437 bronchiseptica.

50. An expression vector comprising a recombinant nucleic acid that encodes one or more of the heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) of claim 1.

51. An expression vector comprising a recombinant nucleic acid that encodes one or more of the light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) of claim 2.

52. A host cell comprising the expression vector of claim 50 or 51.

53. A method of producing a polypeptide comprising an immunoglobulin heavy chain or light chain complementarity determining region, the method comprising:

(a) growing the host cell of claim 52 under conditions so that the host cell expresses the polypeptide comprising the immunoglobulin heavy chain or light chain complementarity determining region; and

(b) purifying the polypeptide comprising the immunoglobulin heavy or light chain complementarity determining region.

54. A method of reducing or killing a Bordetella strain, the method comprising administering an effective amount of the antibody, antigen-binding fragment thereof or polyclonal antibody composition of any one of claims 1-49 to reduce or kill the Bordetella strain.

55. A method for treating or preventing a Bordetella infection in a mammal, the method comprising administering an effective amount of the antibody, antigen-binding fragment thereof or polyclonal antibody composition of any one of claims 1-49 to a mammal in need thereof.

56. The method of claim 55 wherein the mammal is a human.

57. A method for treating or preventing a polymicrobial infection including a Bordetella infection in a mammal, the method comprising administering an effective amount of the antibody, antigen-binding fragment thereof or polyclonal antibody composition of any one of claims 1-49 to a mammal in need thereof.