Antibody compositions for disrupting biofilms

Abstract

To provide antibody compositions for disrupting biofilms.SOLUTION: Disclosed herein are methods and compositions for disrupting biofilms in vitro and in vivo. Also disclosed is an antibody comprising a specific heavy chain (HC) immunoglobulin variable domain sequence and / or a specific light chain (LC) immunoglobulin variable domain sequence. Specifically disclosed is an antibody comprising: (i) a heavy chain (HC) immunoglobulin variable domain sequence comprising a sequence from amino acid (aa) 25 to aa 144 of SEQ ID NO:13 and or an equivalent thereof and (ii) a light chain (LC) immunoglobulin variable domain sequence comprising a sequence from aa 21 to aa 132 of SEQ ID NO: 14 or an equivalent thereof, or a fragment of the antibody.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 63 / 033,109, filed June 1, 2020, and U.S. Provisional Application No. 62 / 871,457, filed July 8, 2019, the contents of each of which are hereby incorporated by reference in their entirety.

[0002] Statement of government support This invention was made with government support under Grant No. R01 DC011818 awarded by the National Institute on Deafness and Other Communication Disorders (NIDCD) of the National Institutes of Health (NIH). The government has certain rights in this invention.

[0003] Field of Disclosure The present disclosure relates generally to methods and compositions for reducing and / or eradicating bacterial biofilms. [Background technology]

[0004] background Proteins of the DNABII family are naturally found on the outside of bacterial cells and contribute to the formation of biofilms. At least one protein from the DNABII family is found in all known eubacteria. These proteins induce strong innate and adaptive immune responses, but host subjects are unable to naturally produce immune protective antibodies against members of the family as a result of infection. A major problem with bacterial biofilms is the inability of the host immune system and / or antibiotics and other antimicrobial agents to access the bacteria protected within the biofilm.

[0005] Biofilms exist in industrial environments as well. For example, they are responsible for a wide range of oil processing problems, from production sites to gas station storage tanks. Here, sulfate-reducing biofilm bacteria produce hydrogen sulfide (sour oil). In processing pipelines, biofilm activity produces slime that clogs filters and orifices. Biofilms and biofilm microorganisms also cause corrosion in pipelines and oil processing equipment. These problems can be manifested throughout oil or gas production facilities, to the point where fouling and corrosive biofilm microorganisms can even be found on the surfaces of end-product storage tanks.

[0006] Biofilms are involved in a wide range of water treatment processes, both domestic and industrial. They grow on treatment equipment surfaces and can disrupt equipment performance, such as reducing heat transfer and clogging filters and membranes. Biofilm growth on cooling tower fill can impart enough weight to cause the fill to collapse. Biofilms can corrode even highly specialized stainless steel. Biofilms in water treatment, for example, biofilm contamination in paper processing or even the attachment of single cells to silicon chips, can degrade the value of the end product. Biofilms growing in drinking water distribution systems can contain potentially pathogenic microorganisms, corrosive microorganisms, or bacteria that degrade the aesthetic properties of the water. In the home, biofilms are found on any surface that supports microbial growth, such as in drains, food preparation surfaces, toilets, and swimming pools and spas.

[0007] Therefore, there is a need to penetrate the protective barrier of biofilms to treat or kill associated bacterial infections and remove them from surfaces and water systems. Summary of the Invention [Means for solving the problem]

[0008] overview In bacterial cells, DNABII proteins are DNA-binding proteins that necessarily bend the DNA substrate upon binding. Similarly, DNA that is already in a bent conformation is a preferred substrate because it eliminates the energy required for bending.

[0009] The DNABII family is a member of a class of proteins called nucleoid-associated proteins (NAPs), bacterial proteins that partially form the intracellular bacterial nucleoid (Browning et al. (2010) Curr. Opin. Microbiol. 13:773-780). In addition, this family The RH family is ubiquitous and expressed by virtually all eubacteria. All family members characterized to date function as either homodimers or heterodimers of subunits. This family is divided into two types: HU (histone-like protein) and IHF (integration host factor). While B. cenocepacia is capable of expressing both of these proteins (J2315 genetic strains: BCAL3530, hupA; BCAL1585, hupB; BCAL1487, ihfA, and BCAL2949, ihfb), many other bacteria also express both HU and IHF. The main difference between these family members is that HU binds DNA in a sequence-independent manner, while IHF binds to a consensus sequence (WATCAANNNNTTR (where W is A or T and R is a purine, SEQ ID NO: 28) that is conserved across genera (Swinger et al. (2004) Curr. Opin. Struct. Biol. 14:28-35). All DNABII proteins bind to DNA and bend it significantly (e.g., E. coli IHF can bend DNA into a substantial U-turn (Rice et al. (1996) Cell 87: 1295-1306)). In addition, all family members bind to pre-bent DNA. They prefer curved or curved DNA structures (e.g., Holliday junctions, the cruciform structures central to DNA recombination). Indeed, DNABII proteins function as cofactors that facilitate all intracellular DNA functions, including gene expression, recombination, repair, and replication (Swinger et al. (2004) Curr. Opin. Struct. Biol. 14:28-35).

[0010] Proteins of the DNABII family are found on the outside of bacterial cells in biofilms, and applicants have shown that these proteins indeed bind to extracellular DNA at critical branch junctions.

[0011] Thus, provided herein is an antibody or antigen-binding fragment thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of, the sequence of amino acids (aa) 25 to aa 144 of SEQ ID NO: 13, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of, a sequence selected from the group of aa 21 to aa 132 of SEQ ID NO: 14, or an equivalent thereof. Further provided herein is an antibody or antigen-binding fragment thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of a sequence selected from the group of aa25 to aa144 of SEQ ID NO: 13, 24, or 26, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of a sequence selected from the group of aa21 to aa132 of SEQ ID NO: 14 or 25, aa21 to aa126 of SEQ ID NO: 27, or an equivalent of each thereof. In certain embodiments, the antibody or fragment thereof is a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI and / or the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail region of a DNABII peptide comprising the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI or tail chimeric peptide IhfA3-IhfB2 NTHI Combine with.

[0012] Also provided herein are antibodies or antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of a sequence selected from the group of aa25 to aa144 of SEQ ID NO: 13 or 24, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of a sequence selected from the group of aa21 to aa132 of SEQ ID NO: 14 or 25, or an equivalent of each thereof. In certain embodiments, the antibody or fragment thereof is a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI In one embodiment, the antibody or fragment thereof binds to the chimeric peptide IhfA5-mIhfB4. NTHI Combine with.

[0013] Still further provided are antibodies or antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) immunoglobulin variable domain sequence selected from the group consisting of aa25 to aa144 of SEQ ID NO: 1-6, 24, or 26, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence selected from the group consisting of aa21 to aa132 of SEQ ID NO: 7-9, or 25, aa21 to aa126 of SEQ ID NO: 10-12, or 27, or an equivalent of each thereof. In certain embodiments, the antibody or fragment thereof is a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHIand / or the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail region of a DNABII peptide comprising the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI or tail chimeric peptide IhfA3-IhfB2 NTHI Combine with.

[0014] Also provided are antibodies or antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of a sequence selected from the group consisting of aa 25 to aa 473 of SEQ ID NO: 1-6, 13, 24, or 26, or an equivalent of each thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of a sequence selected from the group consisting of aa 21 to aa 239 of SEQ ID NO: 7-9, 14, or 25, aa 21 to aa 233 of SEQ ID NO: 10-12, or 27, or an equivalent of each thereof. In certain embodiments, the antibody or fragment thereof binds to a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI and / or the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail region of a DNABII peptide comprising the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI or tail chimeric peptide IhfA3-IhfB2 NTHI Combine with.

[0015] Additionally, antibodies or fragments thereof are provided that comprise, consist essentially of, or even consist of any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 1-6, 13, 24, or 26, and / or any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 7-12, 14, 25, or 27. In certain embodiments, the antibodies or fragments thereof are directed against a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI and / or the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail region of a DNABII peptide comprising the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI or tail chimeric peptide IhfA3-IhfB2 NTHI Combine with.

[0016] Non-limiting examples of antigen-binding fragments are selected from Fab, F(ab')2, Fab', scFv or Fv.

[0017] In another embodiment, a Fab fragment of an antibody, wherein the antibody is a Fab fragment of a DNABII peptide, including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4 NTHI Also provided herein are Fab fragments that specifically bind to the antibody disclosed herein. In one embodiment, the fragment is an antigen-binding fragment selected from the group of Fab, F(ab')2, Fab', scFv, or Fv. In a further embodiment, the fragment specifically binds to the tip region of the DNABII peptide.

[0018] Further provided is a formulation comprising the Fab fragment.

[0019] In another embodiment, the antibody or fragment thereof is modified, non-limiting examples of which include PEGylation, PEG mimetics, polysialylation, HESylation, or glycosylation.

[0020] The antibodies and antigen-binding fragments may be detectably labeled or may include a detectable label and / or a purification marker.

[0021] Also provided herein is a polypeptide comprising, consisting essentially of, or consisting of the complementarity determining region (CDR) of the antibody or antigen-binding fragment or region disclosed above. The CDR can be selected from the group of CDR1, CDR2, or CDR3. The CDR can be detectably labeled or contain a detectable label and / or a purification marker.

[0022] Further provided are isolated polypeptides comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-14 or 24-27; amino acids (aa) 25 to 144 of SEQ ID NOs: 1-6, 13, 24, or 26; aa21 to aa132 of SEQ ID NOs: 7-9, 14, or 25; or aa21 to aa126 of SEQ ID NOs: 10-12 or 27; or equivalents of each. Also provided are isolated polypeptides comprising, alternatively consisting essentially of, or alternatively consisting of one or more of the amino acid sequences of the antibodies or fragments thereof disclosed herein. The polypeptides may be detectably labeled and / or include a detectable label and / or purification marker. In further embodiments, the polypeptides further include a signal peptide.

[0023] Further provided are isolated polynucleotides encoding an antibody or antigen-binding fragment thereof, or the CDR of the antigen-binding fragment or antibody, and a polypeptide of the present disclosure, or equivalents thereof, optionally operably linked to a promoter and / or enhancer element. The polynucleotides may be detectably labeled and / or include a detectable label and / or purification marker. The polynucleotides may further include a polynucleotide sequence of a signal peptide located upstream of the immunoglobulin variable domain of the antibody. The polynucleotides may be contained within vectors and / or host cells. Thus, further provided are vectors comprising, alternatively consisting essentially of, or even consisting of one or more of the isolated polynucleotides disclosed herein. Such vectors may be plasmids or viral vectors, optionally selected from the group consisting of retroviral vectors, lentiviral vectors, adenoviral vectors, and adeno-associated viral vectors. Also provided are host cells comprising one or more of the polynucleotides and / or vectors disclosed herein.

[0024] Further provided is a composition comprising, alternatively consisting of, or even consisting of a carrier and one or more of an antibody or fragment thereof, a CDR, a polypeptide, an isolated polynucleotide, a vector, or a host cell. In one embodiment, the carrier is a pharmaceutically acceptable carrier. Additionally or alternatively, the carrier is a solid phase carrier or solid support. In another embodiment, the carrier is suitable for use in an industrial environment. Also provided is a non-physiological surface coated with the antibody or fragment thereof disclosed herein. In one embodiment, the surface is in an industrial environment. In certain embodiments, the fragment is an antigen-binding fragment.

[0025] Also provided is a method for producing an antibody, fragment, CDR, or polypeptide, comprising, alternatively consisting of, or further consisting of culturing a host cell containing a polynucleotide encoding the antibody, antigen-binding fragment, polypeptide, or CDR under conditions for expression of the polynucleotide, and optionally isolating the antibody, fragment, CDR, and / or polypeptide from the cell and / or culture. In one embodiment, the host cell is a mammalian cell.

[0026] In one aspect, a method is provided for inhibiting or competing with the binding of a DNABII polypeptide or protein to microbial DNA. The method comprises, consists essentially of, or even consists of contacting a DNABII polypeptide or protein with one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein. In one embodiment, the antibody, fragment thereof, polypeptide, or CDR is a DNABII peptide with a tip region (including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI In a further embodiment, the antibody, fragment thereof, polypeptide or CDR binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI In still further embodiments, the contacting is in vivo or in vitro.

[0027] In another aspect, a method for disrupting a biofilm is provided. The method comprises, consists essentially of, or further consists of contacting the biofilm with one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein. In one embodiment, the antibody, fragment, polypeptide, or CDR is a peptide that binds to the tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHIIn a further embodiment, the antibody, fragment thereof, polypeptide or CDR binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI In still further embodiments, the contacting is in vivo or in vitro.

[0028]

[0013] A method for preventing biofilm formation or disrupting biofilms on a surface, comprising, or alternatively consisting of, or still further consisting of treating a surface prone to or containing biofilm with, for example, one or more of the antibodies, antigen-binding fragments, polypeptides or CDRs described herein, wherein the antibody, fragment thereof, polypeptide or CDR binds to the tip region of a DNABII peptide (including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI In a further embodiment, the antibody, fragment thereof, polypeptide or CDR binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI In still further embodiments, the treating is in vivo or in vitro.

[0029] Also provided is a method for detecting biofilm on a surface, comprising, or alternatively consisting of, or still further consisting of contacting a surface (in one embodiment, susceptible to or containing biofilm) with one or more of the antibodies, antigen-binding fragments, polypeptides or CDRs described herein, wherein the antibody, fragment thereof, polypeptide or CDR binds to the tail or tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHIIn a further embodiment, the antibody, fragment thereof, polypeptide or CDR binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI In one embodiment, the contacting is in vivo or in vitro.

[0030] In one aspect, a method for detecting a biofilm-producing microbial infection in a subject is provided. The method comprises, or alternatively consists of, or further consists of contacting one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein with a biological sample suspected of containing a biofilm and isolated from the subject, and detecting binding of the antibody, fragment thereof, polypeptide, or CDR to any biofilm in the sample. The amount, e.g., an effective amount for a subject, can be determined by the treating physician or veterinarian. In one embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the tail or tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, or the like). NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In a further embodiment, the contacting is in vivo or in vitro.

[0031] In another aspect, a method for screening a subject for having a biofilm is provided, comprising, alternatively consisting of, or even further consisting of contacting one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein with a biological sample containing a biofilm and isolated from the subject, and detecting binding of the antibody, fragment thereof, polypeptide, or CDR to any biofilm in the sample. In one embodiment, the antibody, fragment thereof, polypeptide, or CDR is selected from the group consisting of a tail or tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, and the like). NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In a further embodiment, the subject in whom binding is detected is selected for administration of one or more of the antibodies, fragments thereof, polypeptides or CDRs disclosed herein, and / or one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides or CDRs, wherein the antibody, fragment, polypeptide or CDR binds to the tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4 NTHI (including). The amount, e.g., an effective amount for a subject, can be determined by a treating physician or veterinarian. In yet a further embodiment, the contacting is in vivo or in vitro.

[0032] Provided herein are methods for detecting a biofilm in a subject by administering to the subject one or more of the antibodies or fragments thereof disclosed herein and detecting any binding of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein to the biofilm. In one aspect, the antibody, fragment, polypeptide, or CDR binds to the tail or tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In another embodiment, the method further comprises detecting binding of the antibody, fragment thereof, polypeptide, or CDR to the biofilm.

[0033] In one aspect, a method for preventing or disrupting biofilms in a subject comprises, or alternatively consists of, or still further consists of administering to the subject one or more of the antibodies, fragments thereof, polypeptides or CDRs disclosed herein, and / or one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides or CDRs, wherein such comprises a tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI

[0023] Methods are provided for detecting a biofilm by contacting one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein with a sample suspected of containing a biofilm and detecting binding of the biofilm to the antibody, fragments thereof, polypeptides, or CDRs, wherein the antibody, fragments thereof, polypeptides, or CDRs bind to a tip or tail region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI (including

[0034] In another aspect, a method for inhibiting, preventing, or treating a biofilm-producing microbial infection in a subject comprises, or alternatively consists of, or still further consists of administering to the subject one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein, and / or one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides, or CDRs, wherein such comprises a tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI

[0023] Methods are provided for detecting a biofilm by contacting one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein with a sample suspected of containing a biofilm and detecting binding of the biofilm to the antibody, fragments thereof, polypeptides, or CDRs, wherein the antibody, fragments thereof, polypeptides, or CDRs bind to a tip or tail region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI (including

[0035] In yet another aspect, a method for preventing or treating a condition characterized by biofilm formation in a subject by administering to the subject one or more of the antibodies, fragments thereof, polypeptides or CDRs disclosed herein, and / or one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides or CDRs, wherein such comprises a tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI(including, for example, an effective amount for a subject) are provided. The amount, e.g., an effective amount for a subject, can be determined by the treating physician or veterinarian. Non-limiting examples of conditions include chronic non-healing wounds, lung infections caused by Burkholderia species, venous ulcers, diabetic foot ulcers, ear infections, sinus infections, urinary tract infections, gastrointestinal tract diseases, hospital-acquired pneumonia, ventilator-associated pneumonia, surgical implant-associated infections, lung infections, airway infections, cystic fibrosis, chronic obstructive pulmonary disease, catheter-associated infections, indwelling device-associated infections, implantable prosthesis-associated infections, osteomyelitis, cellulitis, abscesses, and periodontal disease. In one embodiment, the method further comprises detecting a biofilm by contacting one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein with a sample suspected of containing a biofilm and detecting binding of the biofilm to the antibody, fragment, polypeptide, or CDR, wherein the antibody, fragment, polypeptide, or CDR binds to a tip region or tail region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI (including

[0036] In certain embodiments of the methods disclosed herein, administration of one or more of the antibodies, fragments thereof, polypeptides, or CDRs reduces one or more of pro-inflammatory cytokines in a subject. Non-limiting examples of pro-inflammatory cytokines include IL-1β, IL6, IL8, IL12p70, IL17A, interferon (IFN), and tumor necrosis factor (TNF). Additionally or alternatively, administration of one or more of the antibodies, fragments thereof, polypeptides, or CDRs increases one or more of the anti-inflammatory cytokines in a subject. In one embodiment, anti-inflammatory cytokines include, but are not limited to, IL10, IL13, IL-1ra, IL-4, IL-11, and transforming growth factor-β (TGF-β).

[0037] Also provided is a method for providing passive immunity in a subject, comprising, alternatively consisting essentially of, or alternatively consisting of, one or more of the antibodies, fragments thereof, polypeptides or CDRs disclosed herein, and / or one or more of the polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides or CDRs, wherein the antibody, fragment, polypeptide or CDR is a polypeptide that encodes a DNABII peptide tip region (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI The amount, e.g., an effective amount for a subject, can be determined by the treating physician or veterinarian.

[0038] In one embodiment, therapeutic methods can be combined with diagnostic methods to detect and / or monitor biofilm formation and disruption using the antibodies, fragments thereof, polypeptides or CDRs disclosed herein.

[0039] In certain embodiments, the biofilm is derived from (i.e., produced by) Gram-negative or Gram-positive biofilm-producing bacteria. In certain embodiments, the biofilm comprises a DNABII protein. In further embodiments, the biofilm comprises a histone-like protein or an integration host factor (IHF)-binding protein from E. coli strain U93(HU). In certain embodiments, the DNABII peptide is an IHF peptide. Additionally or alternatively, the DNABII peptide is an HU peptide. In certain embodiments, the tip region of the DNABII peptide is an IHFA tip region and / or an IHFB tip region. In further embodiments, the tip region of the DNABII peptide is an IHFA tip region conjugated directly or indirectly (e.g., via a linker) to an IHFB tip region. In yet further embodiments, the tip region of the DNABII peptide is IhfA5-mIhfB4 NTHI In a specific embodiment, the tail region of the DNABII peptide is the tail region of IHFA and / or the tail region of IHFB. In a further embodiment, the tail region of the DNABII peptide is the tail region of IHFA conjugated directly or indirectly (e.g., via a linker) to the tail region of IHFB. In yet a further embodiment, the tail region of the DNABII peptide is IhfA3-IhfB2 NTHI It is a tail chimeric peptide.

[0040] Still further provided is a method for preparing an interfering nucleic acid, the method comprising, or further comprising, preparing a nucleic acid of about 10-20 nucleotides that specifically binds to a specific binding partner for an antibody or fragment thereof disclosed herein, and, optionally, isolating the interfering nucleic acid prepared by the method.

[0041] In another aspect, provided herein is a non-physiological surface coated with one or more of the antibodies, fragments thereof, polypeptides or CDRs disclosed herein, optionally in one aspect in an industrial setting.

[0042] Also provided is a method for obtaining an antiserum effective in disrupting a biofilm, comprising immunizing a subject with a small molecule, collecting antiserum from the subject, and, optionally, isolating polyclonal antisera or monoclonal antibodies from the subject.

[0043] Kits are also provided that include one or more of the antibodies, fragments thereof, polypeptides or CDRs, polynucleotides, vectors, host cells, or compositions disclosed herein, and, optionally, instructions for use. [Brief explanation of the drawings]

[0044] [Figure 1] FIG. 1 provides a diagram showing how a DNABII protein (eg, IHF) binds to double-stranded DNA as well as the two arms of the DNABII protein.

[0045] [Figure 2A-2B]Figures 2A-2D show the results of direct binding ELISAs performed to evaluate the binding of humanized monoclonal antibody variants to their corresponding peptides. Chip or tail chimeric peptides were coated onto 96-well plates at 2 μg / mL, and an eight-point dilution series of antibody was added (starting concentration 50 μg / mL, 1:3 dilution). Anti-human IgG Fc HRP-conjugated antibody (1:7000, Jackson ImmunoResearch Laboratories, 109-035-098) was used as the secondary detection antibody. Absorbance was read at 450 nm using a microplate reader according to standard direct ELISA protocols. EC50 values ​​were calculated, and the humanized variants were determined to have comparable binding. The humanized chip-chimeric peptide antibodies are shown in Figures 2A and 2B. The humanized tail-chimeric peptide antibodies are shown in Figures 2C and 2D. [Figures 2C-2D] Figures 2A-2D show the results of direct binding ELISAs performed to evaluate the binding of humanized monoclonal antibody variants to their corresponding peptides. Chip or tail chimeric peptides were coated onto 96-well plates at 2 μg / mL, and an eight-point dilution series of antibody was added (starting concentration 50 μg / mL, 1:3 dilution). Anti-human IgG Fc HRP-conjugated antibody (1:7000, Jackson ImmunoResearch Laboratories, 109-035-098) was used as the secondary detection antibody. Absorbance was read at 450 nm using a microplate reader according to standard direct ELISA protocols. EC50 values ​​were calculated, and the humanized variants were determined to have comparable binding. The humanized chip-chimeric peptide antibodies are shown in Figures 2A and 2B. The humanized tail-chimeric peptide antibodies are shown in Figures 2C and 2D.

[0046] [Figure 3] FIG. 3 is an in vitro model for disruption of established biofilms in 8-well chamber slides using humanized tip or tail chimeric peptide monoclonal antibodies of the present disclosure.

[0047] [Figure 4A] Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp. [Figure 4B] Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp. [Figure 4C] Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp. [Figure 4D] Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp. [Figure 4E] Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp. [Figure 4F]Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp. [Figure 4G] Figures 4A-4G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the CHIP chimeric peptide IhfA5-mIhfB4NTH I (tabled in Figure 4A, and detailed alignments shown in Figures 4B-4G). Amino acid alignments were performed by NCBI blastp.

[0048] [Figure 5A] Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp. [Figure 5B] Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp. [Figure 5C] Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp. [Figure 5D]Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp. [Figure 5E] Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp. [Figure 5F] Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp. [Figure 5G] Figures 5A-5G provide a comparison between the amino acid identities of the heavy and light chains of a humanized monoclonal antibody designed to target the tail chimeric peptide IhfA3-IhfB2NTHI (tabled in Figure 5A, with detailed alignments shown in Figures 5B-5G). Amino acid alignments were performed using NCBI blastp.

[0049] [Figure 6]Figure 6 provides comparative biofilm disruption data using a humanized monoclonal antibody designed to target the IhfA5-mIhfB4 NTHI chip chimeric peptide. Biofilm disruption: NTHI 86-028NP colonies were harvested from overnight cultures on chocolate agar and suspended in brain heart perfusion broth (sBHI) supplemented with 2 μg β-NAD and heme / ml medium. The absorbance at 490 nm was then adjusted to 0.65, and the culture was diluted 1:6 in sBHI and then incubated statically for 3 hours at 37°C and 5% CO2. The culture was then diluted 1:2500 in fresh sBHI, and 200 μl of the suspension was aliquoted into each well of an 8-well chamber slide. The slide was then incubated statically for 3 hours at 37°C and 5% CO2. After 16 hours, 200 μl of fresh sBHI was added to each well, and the slide was incubated for an additional 8 hours. At this point, the medium was aspirated from each well, and 5 μg of monoclonal antibody was added per well. The biofilms were incubated for an additional 16 h. The biofilms were then washed, stained with FM1-43FX bacterial cell membrane stain (Invitrogen), and fixed overnight at 4°C with 16% paraformaldehyde, 2.5% glutaraldehyde, and 4.0% acetic acid in 0.1 M phosphate buffer (pH 7.4). The fixative was aspirated, and 200 μl of Hank's buffered saline was added to each well. The biofilms were then observed using a Zeiss 800 Meta laser scanning confocal microscope. Images were edited using Zeiss Zen Black software, and the biofilm biomass was calculated using COMSTAT 2.1 software. The Kα (1 / M) of the IhfA5-mIhfB4NTHI chip chimeric peptide ranged from approximately 4E+05 to approximately 2E+08.

[0050] [Figure 7]Figure 7 provides comparative biofilm disruption data using a humanized monoclonal antibody designed to target the IhfA3-IhfB2 NTHI tail chimeric peptide. Biofilm disruption: NTHI 86-028NP colonies were harvested from overnight cultures on chocolate agar and suspended in brain heart perfusion broth (sBHI) supplemented with 2 μg β-NAD and heme / ml medium. The absorbance at 490 nm was then adjusted to 0.65, and the culture was diluted 1:6 in sBHI and then incubated statically for 3 hours at 37°C and 5% CO2. The culture was then diluted 1:2500 in fresh sBHI, and 200 μl of the suspension was aliquoted into each well of an 8-well chamber slide. The slide was then incubated statically for 3 hours at 37°C and 5% CO2. After 16 hours, 200 μl of fresh sBHI was added to each well, and the slide was incubated for an additional 8 hours. At this point, the medium was aspirated from each well, and 5 μg of monoclonal antibody was added per well. The biofilms were incubated for an additional 16 h. Next, the biofilms were washed, stained with FM1-43FX bacterial cell membrane stain (Invitrogen), and fixed overnight at 4°C with 16% paraformaldehyde, 2.5% glutaraldehyde, and 4.0% acetic acid in 0.1 M phosphate buffer (pH 7.4). The fixative was aspirated, and 200 μl of Hank's buffered saline was added to each well. The biofilms were then observed using a Zeiss 800 Meta laser scanning confocal microscope. Images were edited using Zeiss Zen Black software, and the biofilm biomass was calculated using COMSTAT 2.1 software. The Kα (1 / M) of the IhfA3-IhfB2NTHI tail chimeric peptide ranged from approximately 8E+06 to approximately 2E+09.

[0051] [Figure 8A-8B]Figures 8A-8B show that Fab fragments derived from a mouse monoclonal antibody against β-tip disrupted biofilms formed by five human pathogens in vitro. Figure 8A shows representative images of bacterial biofilms (pseudocolored white) revealing significant biofilm disruption by intact IgG or Fab against the β-tip domain of IHF at 170 nM. Orthogonal projections show top-down views to depict the spatial distribution of biofilms in the xy plane, while side views indicate biofilm height in the z plane (arrowheads). Scale bar, 20 μm. Figure 8B shows the biomass within each image, as quantified by COMSTAT2 software. Each assay was repeated three times on different days, and the mean ± SEM is shown. *P ≤ 0.05, **P ≤ 0.01 compared to intact IgG or Fab, respectively (one-way ANOVA with multiple comparisons).

[0052] [Figures 9A-9D]Figures 9A-9D show that murine monoclonal antibody β-tip Fab mediated eradication of biofilm-resident NTHI and dissolution of mucosal biofilm from the middle ear. Figure 9A shows the study timeline and treatments administered. A dose of 342 nM Fab was delivered to each middle ear. Figure 9B shows the relative amount of NTHI adhered to the middle ear mucosa within the mucosal biofilm in the treated cohort. Figure 9C shows the relative mucosal biofilm scores determined by six reviewers blinded to treatment in the treated cohort. Figures 9B and 9C show six to eight middle ears per cohort, individual ear values, and cohort means. The cohorts in each panel are, from left to right, isotype control Fab, β-tail Fab, and β-tip Fab. *P≦0.05, **P≦0.01, ***P≦0.001 (one-way ANOVA with multiple comparisons). Figure 9D shows representative images of chinchilla middle ears from each cohort, with the average mucosal biofilm score of the ear indicated in the box in the lower right corner. TM, tympanic membrane; S, bony septum; MEM, middle ear mucosa; B, biofilm (circled). In one embodiment, delivery of both mouse monoclonal antibody β-tip Fabs significantly reduced bacterial burden and eradicated mucosal biofilm in the middle ear, whereas β-tail Fabs not only failed to do so but were associated with significant inflammation.

[0053] [Figures 10A-10B]Figures 10A-10B show the pro-inflammatory cytokines predominant in middle ear effusions from animals treated with 342 nM murine monoclonal antibody-derived β-tail Fab. Figure 10A shows the relative abundance of pro-inflammatory cytokines in chinchilla middle ear effusions after NTHI exposure and Fab fragment treatment, as determined by cytometric bead array. *P≦0.05, **P≦0.01 compared to β-tip Fab (one-way ANOVA with multiple comparisons), ‡P≦0.05 compared to isotype control Fab (unpaired t-test). Figure 10B shows the relative mean concentrations of the anti-inflammatory cytokine IL-10 in the three Fab fragment-treated cohorts. *P≦0.05 compared to β-tail Fab, ++P≦0.01 compared to β-tail Fab or isotype control Fab (one-way ANOVA with multiple comparisons). Five to seven middle ear effusions were tested per cohort. Mean cytokine concentrations ± SD are shown. For either panel (FIG. 10A or 10B), cytokine results are shown from left to right for a cohort of isotype control Fab, β-tail Fab, and β-tip Fab. In one embodiment, delivery of β-tip Fab resulted in significant reductions in the concentrations of six pro-inflammatory cytokines and significantly higher concentrations of the anti-inflammatory cytokine IL-10, supporting relative inflammation or lack thereof, as shown in FIG. 9D.

[0054] [Figures 11A-11D]Figures 11A-11D show that tip-chimeric Fab derived from polyclonal rabbit IgG mediated clearance of biofilm-resident NTHI, eradication of established mucosal biofilms, and resolution of experimental disease. Figure 11A shows the study timeline. One dose of 342 nM Fab was delivered to each middle ear. Figure 11B shows the relative amount of NTHI resident within the mucosal biofilm and attached to the middle ear mucosa 1 or 7 days after the end of antibody treatment. Figure 11C shows the relative amount of remaining mucosal biofilm as determined by six reviewers blinded to the delivered treatment. Figures 11B and 11C show six middle ears per cohort, with values ​​for individual ears and the mean for each cohort. Cohorts in each plot are, from left to right, untreated serum Fab, tail-chimeric Fab, and tip-chimeric Fab. *P≦0.05, **P≦0.01, ***P≦0.001 (one-way ANOVA with multiple comparisons). Figure 11D shows representative images of middle ear mucosal biofilms; the average mucosal biofilm score is indicated in the box in the lower right corner. TM, tympanic membrane; S, bony septum; MEM, middle ear mucosa; B, biofilm (circled). Rabbit polyclonal antibody Fab against the tip chimeric peptide mediated rapid and sustained clearance of biofilm-resident NTHI, eradication of established mucosal biofilms, and recovery of experimental OM. Rabbit polyclonal antibody Fab against the tail chimeric peptide did not induce clearance of biofilm-resident NTHI or disease recovery; instead, it was associated with significant inflammation (Figure 11D), similar to their mouse monoclonal antibody Fab against the β-tail (see Figure 9D).

[0055] [Figures 12A-12B]Figures 12A-12B show that humanized Tip-chimeric peptide monoclonal antibodies disrupted bacterial biofilms in vitro. Figure 12A shows representative images of bacterial biofilms (pseudocolored white) after 16 hours of incubation with HuTipMab or HuTailMab. The orthogonal projection shows a top-down view to depict the spatial distribution of the biofilm in the xy plane, while the side view indicates the biofilm height in the z plane (arrowhead). Scale bar, 20 μm. As used herein, the term "HuTipMab" refers to monoclonal antibodies (MAbs or mAbs) including anti-TipHC1 having the amino acid sequence of SEQ ID NO: 1 and anti-TipLC1 having the amino acid sequence of SEQ ID NO: 7. Figure 12B shows the percentage of biofilm biomass remaining after exposure to HuTipMab compared to HuTailMab, as determined by COMSTAT2 analysis. Mean values ​​± SD are shown. Experiments were performed three times on separate days. ***P≦0.001 (unpaired t-test). HuTipMab significantly disrupted biofilms formed by various respiratory tract pathogens in vitro. As used herein, the term "HuTailMab" refers to a monoclonal antibody (Mab or mAb) comprising an anti-tail HC1 having the amino acid sequence of SEQ ID NO: 4 and an anti-tail LC1 having the amino acid sequence of SEQ ID NO: 10.

[0056] [Figures 13A-13D]Figures 13A-13D show that a humanized monoclonal antibody (HuTipMab) against the tip chimeric peptide lysed pre-existing NTHI biofilms present in the middle ear during experimental NTHI-induced OM. Figure 13A shows the study timeline and treatments. Figure 13B shows the relative amount of NTHI in the middle ear. Figure 13C shows the relative amount of NTHI mucosal biofilm remaining in the middle ear after treatment with the human monoclonal antibody. Figures 13B and 13C show six middle ears per cohort, individual ear values, and cohort means. The cohorts in each plot are, from left to right, saline, humanized monoclonal antibody against the tail chimeric peptide, and humanized monoclonal antibody against the tip chimeric peptide. ****P≦0.0001 (one-way ANOVA with multiple comparisons). Figure 13D provides representative images of middle ears from each cohort; the mean mucosal biofilm score for the ears is indicated in the box in the lower right corner. S, bony septum; MEM, middle ear mucosa; B, biofilm, circled area. HuTipMab eradicated NTHI by lysis of the mucosal biofilm, with rapid and sustained results. Note also that once humanized, the monoclonal antibody against the tail chimeric peptide was no longer associated with significant inflammatory changes, as consistently observed for either a mouse monoclonal antibody against the β-tail (see Figure 9D) or a rabbit polyclonal antibody Fab against the tail chimeric peptide (see Figure 11D).

[0057] [Figures 14A-14B]Figures 14A-14B show that pro-inflammatory cytokines were significantly less abundant in middle ear effusions from animals treated with HuTipMab. Relative abundance of pro- and anti-inflammatory cytokines in chinchilla middle ear effusions after NTHI challenge and treatment with humanized monoclonal antibodies at (Figure 14A) day 6 after NTHI challenge (day 1 after end of treatment) and (Figure 14B) day 13 after NTHI challenge (day 7 after end of treatment), as determined by cytometric bead assay. *P≦0.05 compared to saline, **P≦0.01 compared to saline, ***P≦0.001 compared to saline, ****P≦0.0001 compared to saline, +P≦0.05 compared to HuTailMab, ++P≦0.01 compared to HuTailMab, ++P≦0.001 compared to HuTailMab, ++++P≦0.0001 compared to HuTailMab (one-way ANOVA with multiple comparisons). Three to five middle ear effusions per cohort were tested. Mean cytokine concentrations ± SD are shown. Pro-inflammatory cytokine levels in animals treated with HuTailMab were now equivalent to those in animals treated with saline, providing evidence of no association with treatment with this now-humanized monoclonal antibody compared to the murine monoclonal antibody.

[0058] [Figures 15A-15D]Figures 15A-15D show that immunization with the tip chimeric peptide prevents ascending experimental OM in a virus-bacteria co-infection model. Figure 15A shows the study timeline and delivered vaccine formulation. Figure 15B shows the relative amount of NTHI in nasopharyngeal tears at day 1 after bacterial challenge to ensure comparable engraftment in all cohorts. Eight lavage fluids per cohort; NS, not significant; one-way ANOVA. Figure 15C shows the number of animals in each cohort with signs of experimental OM, i.e., inflammation and / or middle ear effusion, as visualized by blinded otoendoscopy. Eight animals per cohort; ****P≦0.001 compared with dmLT or tail chimeric peptide; Mantel-Cox test. Figure 15D shows representative images of tympanic membranes from each cohort on day 11, which was the day of highest disease incidence in the cohort immunized with the tip chimeric peptide. TM, tympanic membrane. Immunization with the CHIP chimeric peptide prevented ascending polymicrobial OM and promoted rapid resolution of the limited disease observed. DETAILED DESCRIPTION OF THE INVENTION

[0059] Detailed Description Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All nucleotide sequences provided herein are shown in the 5' to 3' direction. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, certain non-limiting exemplary methods, devices, and materials are described herein. All technical and patent publications cited herein are incorporated by reference in their entirety. Nothing herein should be construed as an admission that the present disclosure is not entitled to antedate such disclosure by virtue of prior invention.

[0060] The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of tissue culture, immunology, molecular biology, microbiology, cell biology, and recombinant DNA, which are within the skill of the art. See, e.g., Sambrook and Russell eds., (2001) Molecular Cloning: A Laboratory Manual, 3 rd edition; Series Ausubel et al. eds. (2007) Current Protocols in Molecular Biology; Series Methods in Enzymology (Academic Press, Inc., NY); MacPherson et al. (1991) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual;Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5 th edition; Gait ed. (1984) Oligonucleotide Synthesis; U.S. Patent No. 4,683,195; Hames and Higgins eds. (1984) Nucleic Acid Hybridization;Anderson (1999) Nucleic Acid Hybridization; Hames and Higgins eds. (1984) Transcription and Translation; Immobilized Cells and Enzymes (IRL Press (1986)); Perbal (1984) A Practical Guide to Molecular Cloning; Miller and Calos eds, (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); and Herzenberg et al. eds. (1996) Weir's Handbook of Experimental Immunology.

[0061] All numerical designations, including ranges, e.g., pH, temperature, time, concentration, and molecular weight, are approximations which are varied (+) or (-) by increments of 1.0 or 0.1, as appropriate, or by a variation of + / - 15%, or alternatively 10%, or alternatively 5%, or alternatively 2%. It is to be understood, although not always expressly indicated, that all numerical designations are preceded by the term "about." It is also to be understood, although not always expressly indicated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0062] As used in this specification and claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a polypeptide" includes multiple polypeptides, including mixtures thereof.

[0063] As used herein, the term "comprising" is intended to mean that the compositions and methods include the recited elements, but do not exclude others. "Consisting essentially of," when used to define compositions and methods, means excluding any other elements that are essential to the combination for its intended use. Thus, a composition consisting essentially of the elements defined herein does not exclude trace contaminants from isolation and purification methods, and pharmaceutically acceptable carriers, e.g., phosphate-buffered saline, preservatives, and the like. "Consisting of" means excluding more than trace elements of other ingredients and substantial method steps for administering the compositions disclosed herein. Embodiments defined by each of these transition terms are within the scope of this disclosure.

[0064] The term "biofilm" refers to an organized microbial community in which microorganisms adhere to structural surfaces, which may be organic or inorganic, along with macromolecules such as DNA that are secreted, released, and / or made available to the extracellular environment by bacterial lysis. Biofilms are highly resistant to microbiotics and antimicrobial agents. They survive on gingival tissue, teeth, and restorations, causing dental caries and periodontal disease, also known as periodontal plaque disease. They also cause chronic middle ear infections. Biofilms can also form on the surfaces of dental implants, stents, catheter systems, and contact lenses. They grow on pacemakers, heart valve replacements, artificial joints, and other surgical implants. The Centers for Disease Control estimates that more than 65% of nosocomial (hospital-acquired) infections are caused by biofilms. They cause chronic vaginal infections and life-threatening systemic infections in people with compromised immune systems. Biofilms are also involved in many diseases. For example, cystic fibrosis patients often have Pseudomonas infections, which result in antibiotic-resistant biofilms. In one embodiment, the biofilm comprises a DNABII polypeptide or protein. In a further embodiment, the biofilm comprises IHF and / or HU. In yet a further embodiment, the biofilm comprises IHFA and / or IHFB.

[0065] The term "disrupt" refers to the reduction of the formation of the DNA / protein matrix that is a component of a microbial biofilm. In certain embodiments, disrupting a biofilm refers to dispersing the biofilm, releasing the microorganisms from the DNA / protein matrix of the biofilm, and, if necessary, allowing the microorganisms to be killed by the host's immune effectors and / or antibiotics.

[0066] "DNABII polypeptide or protein" refers to a DNA-binding protein or polypeptide that consists of a DNA-binding domain and therefore has specific or general affinity for microbial DNA. In one embodiment, they bind to the minor groove of DNA. Non-limiting examples of DNABII proteins are integration host factor (IHF) protein and histone-like protein from E. coli strain U93 (HU). Other DNA-binding proteins that may be associated with biofilms include DPS (Genbank Accession No. CAA49169), H-NS (Genbank Accession No. CAA47740), Hfq (Genbank Accession No. ACE63256), CbpA (Genbank Accession No. BAA03950), and CbpB (Genbank Accession No. NP_418813).

[0067] "Integration host factor" or "IHF" proteins are bacterial proteins used by bacteriophages to integrate their DNA into host bacteria. They also bind to extracellular microbial DNA. The genes encoding IHF protein subunits in E. coli are the himA (GenBank accession number POA6X7.1) and himD (POA6Y1.1) genes. Homologs of these genes have been found in other organisms. In certain embodiments, the term "IHF" refers to one or both of the two IHF subunits: integration host factor subunit alpha (IHFA or IhfA) and integration host factor subunit beta (IHFB or IhfB).

[0068] "HU" or "histone-like protein from E. coli strain U93" refers to a class of heterodimeric proteins typically associated with E. coli. HU proteins are known to bind to DNA junctions. Related proteins have been isolated from other microorganisms. The complete amino acid sequence of E. coli HU was reported by Laine et al. (1980) Eur. J. Biochem 103(3)447-481. Antibodies against the HU protein are commercially available from Abeam. The genes encoding the HU protein subunits in E. coli are hupA and hupB, which correspond to SEQ ID NOs: 29 and 30, respectively. Homologs of these genes have been found in other organisms, and peptides corresponding to these genes from other organisms can be found in Table 10 of WO 2011 / 123396.

[0069] The term "surface antigen" or "surface protein" refers to a protein or peptide on the surface of a cell, e.g., a bacterial cell. Examples of outer membrane proteins are OMP P5 (GenBank Accession No. YP_004139079.1), OMP P2 (GenBank Accession No. ZZX87199.1), and OMP P26 (GenBank Accession No. YP_665091.1), while examples of surface antigens are rsPilA or recombinant soluble PilA (GenBank Accession No. EFU96734.1) and type IV Pilin (GenBank Accession No. Yp_003864351.1).

[0070] The term "Haemophilus influenzae" refers to a pathogenic bacterium that can cause many different infections, such as ear infections, eye infections, and sinusitis. Many different strains of Haemophilus influenzae have been isolated and contain IhfA, ihfB, and hupA genes or proteins. Some non-limiting examples of different strains of Haemophilus influenzae include Rd KW20, 86-028NP, R2866, PittGG, PittEE, R2846, and 2019.

[0071] By "microbial DNA" is intended single-stranded or double-stranded DNA from microorganisms incorporated into a biofilm.

[0072] By "inhibiting, preventing or disrupting" a biofilm is intended a prophylactic or therapeutic reduction in biofilm structure.

[0073] By "bent polynucleotide" is intended a double-stranded polynucleotide containing a small loop in one strand that does not pair with the other strand. In some embodiments, the loop is 1 to about 20 bases in length, or alternatively 2 to about 15 bases in length, or alternatively about 3 to about 12 bases in length, or alternatively about 4 to about 10 bases in length, or alternatively has about 4, 5, 6, 7, 8, 9, or 10 bases.

[0074] "A polypeptide that competes with DNABII binding, e.g., IHF in binding to DNA" refers to a protein or peptide that competes with DNABII (e.g., IHF) in binding to bent or distorted DNA structures, but does not form a biofilm with DNA. Examples include, but are not limited to, fragments of IHF containing one or more DNA-binding domains of IHF, or biological equivalents thereof.

[0075] A "subject" of diagnosis or treatment is a cell or an animal, e.g., a mammal or a human. Non-human animals for diagnosis or treatment include non-human animals or animal models subject to infection, such as primates, murines, e.g., rats, mice, chinchillas, canines, e.g., dogs, lagomorphs, e.g., rabbits, livestock, sport animals, and pets. The terms "subject," "host," "individual," and "patient," when used interchangeably herein, refer to an animal, typically a mammal. Non-limiting examples of mammals include humans, non-human primates (e.g., apes, gibbons, chimpanzees, orangutans, monkeys, macaques, etc.), domestic animals (e.g., dogs and cats), livestock animals (e.g., horses, cows, goats, sheep, pigs), and laboratory animals (e.g., mice, rats, rabbits, guinea pigs). In some embodiments, the mammal is a human. The mammal may be of any age or at any stage of development (e.g., an adult, adolescent, child, juvenile, or in utero mammal). The mammal may be male or female. In some embodiments, the subject is a human.

[0076] The terms "protein," "peptide," and "polypeptide" are used interchangeably and in their broadest sense to refer to a compound of two or more subunit amino acids, amino acid analogs, or peptidomimetics. The subunits may be linked by peptide bonds. In alternative embodiments, the subunits may be linked by other bonds, such as esters, ethers, etc. A protein or peptide must contain at least two amino acids, and no limit is placed on the maximum number of amino acids that may make up a protein's or peptide's sequence. As used herein, the term "amino acid" refers to any natural and / or unnatural or synthetic amino acids, amino acid analogs, and peptidomimetics, including glycine and both D and L optical isomers.

[0077] By "C-terminal polypeptide" is intended at least 10, or alternatively at least 15, or alternatively at least 20, or at least 25 C-terminal amino acids, or alternatively half of the polypeptide. In another embodiment, for a polypeptide containing 90 amino acids, the C-terminal polypeptide would include amino acids 46 to 90. In one embodiment, the term contemplates the carboxy-terminal to C-terminal 20 amino acids.

[0078] By "chip fragment" of a DNABII polypeptide is intended a DNABII polypeptide that forms the two arms of the protein, using IHF alpha and IHF beta as examples. (See Figure 1). Non-limiting examples of such include IhfA, A chip fragment: NFELRDKSSRPGRNPKTGDVV, SEQ ID NO: 31, and IhfB, B chip fragment: SLHHRQPRLGRNPKTGDSVNL, SEQ ID NO: 32.

[0079] By "tail fragment" of a DNABII polypeptide is intended the region of the protein that is exposed to the bulk medium and is not occluded by DNA or other polypeptides.

[0080] An immunodominant antigen is intended to be a region of a protein that is recognized by an antibody and binds to it with high affinity.

[0081] An immunoprotective antigen refers to a region of a protein that is recognized by an antibody, binds to it with high affinity, and interferes with the function of the protein; antibodies generated against an immunoprotective antigen are characterized by an enhanced or optimal effect on the target indication, in this case, an improved ability to clear biofilm, as a result of interfering with the function of the protein.

[0082] The terms "polynucleotide" and "oligonucleotide" are used interchangeably and refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or their analogs. Polynucleotides can have any three-dimensional structure and can perform any function, known or unknown. The following are non-limiting examples of polynucleotides: genes or gene fragments (e.g., probes, primers, ESTs, or SAGE tags), exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, RNAi, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. Polynucleotides can contain modified nucleotides, such as methylated nucleotides and nucleotide analogs. If present, modifications to the nucleotide structure can be imparted before or after assembly of the polynucleotide. The sequence of nucleotides can be interrupted by non-nucleotide components. Polynucleotides may be further modified after polymerization, for example, by conjugation with a labeling component. The term also refers to both double-stranded and single-stranded molecules. Unless otherwise specified or required, any embodiment disclosed herein that is a polynucleotide encompasses both the double-stranded form and each of the two complementary single-stranded forms that are known or predicted to constitute the double-stranded form.

[0083] A polynucleotide consists of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); thymine (T); and, if the polynucleotide is RNA, uracil (U) instead of thymine. Thus, the term "polynucleotide sequence" is an alphabetic representation of a polynucleotide molecule. This alphabetic representation can be input into a database on a computer with a central processing unit and used in bioinformatics applications, such as functional genomics and homology searching.

[0084] The terms "isolated" or "recombinant," as used herein with respect to nucleic acids, e.g., DNA or RNA, refer to molecules separated from other DNAs or RNAs and polypeptides, respectively, present in the natural source of the polymer. The term "isolated or recombinant nucleic acid" is intended to include nucleic acid fragments not naturally occurring as fragments, not found in the natural state. The term "isolated" is also used herein to refer to polynucleotides, polypeptides, and proteins isolated from other cellular proteins and is intended to encompass both purified and recombinant polypeptides. In other embodiments, the term "isolated or recombinant" refers to being separated from cellular components and other cells, tissues, polynucleotides, peptides, polypeptides, proteins, antibodies, or fragment(s) thereof, with which it is otherwise normally associated in nature. For example, an isolated cell is one that is separated from tissues or cells of a dissimilar phenotype or genotype. An isolated polynucleotide is separated from the 3' and 5' flanking nucleotides with which it is normally associated in its native or natural environment, e.g., on a chromosome. As will be apparent to one of skill in the art, a non-naturally occurring polynucleotide, peptide, polypeptide, protein, antibody or fragment(s) thereof does not require "isolation" to distinguish it from its naturally occurring counterpart.

[0085] Where the present disclosure relates to a polypeptide, protein, polynucleotide, or antibody, it should be assumed that equivalents or biological equivalents of such are intended within the scope of the present disclosure, without explicit recitation and unless otherwise intended. As used herein, the term "biological equivalent thereof" when referring to a reference protein, antibody, fragment, polypeptide, or nucleic acid is intended to be synonymous with "equivalent thereof," and is intended to be synonymous with "equivalent thereof," and to have minimal homology while still maintaining the desired structure or functionality. Unless otherwise recited herein, any polynucleotide, polypeptide, or protein listed herein is also intended to include its equivalent. In one aspect, an equivalent polynucleotide is a polynucleotide that hybridizes under stringent conditions to the polynucleotide or complement of the polynucleotide described herein for use in the described method. In another embodiment, an equivalent antibody or antigen-binding polypeptide is intended to be an antibody or antigen-binding polypeptide that binds to a reference antibody or antigen-binding fragment with at least 70%, or alternatively at least 75%, or alternatively at least 80%, or alternatively at least 85%, or alternatively at least 90%, or alternatively at least 95% affinity or higher. In another embodiment, the equivalent competes with the antibody or antigen-binding fragment's binding to its antigen in a competitive ELISA assay. In another embodiment, an equivalent is intended to be at least about 80% homology or identity, and alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 95%, or alternatively 98% percent homology or identity, and exhibits substantially equivalent biological activity to the reference protein, polypeptide, or nucleic acid. Examples of biologically equivalent polypeptides are provided in Table 9 of WO2011 / 123396, which identifies conservative amino acid substitutions for the disclosed amino acid sequences.

[0086] A polynucleotide or polynucleotide region (or polypeptide or polypeptide region) having a certain percentage (e.g., 80%, 85%, 90%, or 95%) of "sequence identity" to another sequence means that, when aligned, that percentage of bases (or amino acids) are the same in comparing the two sequences. Alignment and percent homology or sequence identity can be determined using software programs known in the art, such as the software programs described in Current Protocols in Molecular Biology (Ausubel et al., eds. 1987) Supplement 30, section 7.7.18, Table 7.7.1. In certain embodiments, default parameters are used for the alignment. A non-limiting exemplary alignment program is BLAST, using default parameters. In particular, exemplary programs include BLASTN and BLASTP, using the following default parameters: Genetic coding = standard; Filter = none; Strand = both; Cutoff = 60; Prediction = 10; Matrix = BLOSUM62; Description = 50 sequences; Classification = high score; Database = non-redundant GenBank + EMBL + DDBJ + PDB + GenBank CDS translation + SwissProtein + SPupdate + PIR. Details of these programs can be found at the following internet address: ncbi.nlm.nih.gov / cgi-bin / BLAST. Sequence identity and percent identity were determined by incorporating them into clustalW (available at the web address: align.genome.jp (last accessed March 7, 2011)).

[0087] "Homology" or "identity" or "similarity" refers to the sequence similarity between two peptides or two nucleic acid molecules. Homology can be determined by comparing a position in each sequence that can be aligned for comparison purposes. If a position in the compared sequences is occupied by the same base or amino acid, the molecules are homologous at that position. The degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. An "unrelated" or "non-homologous" sequence shares less than 40% identity, or alternatively less than 25% identity, with one of the sequences of the present disclosure.

[0088] "Homology" or "identity" or "similarity" can also refer to two nucleic acid molecules that hybridize under stringent conditions.

[0089] "Hybridization" refers to a reaction in which one or more polynucleotides react to form a complex stabilized through hydrogen bonds between the bases of nucleotide residues. Hydrogen bonds can occur through Watson-Crick base pairing, Hoogsteen binding, or any other sequence-specific manner. The complex can include two strands forming a double-stranded structure, three or more strands forming a multi-stranded complex, a single self-hybridizing strand, or any combination thereof. A hybridization reaction can constitute a step in a more extensive process, such as the initiation of a PCR reaction or the enzymatic cleavage of a polynucleotide by a ribozyme.

[0090] Examples of stringent hybridization conditions include an incubation temperature of about 25°C to about 37°C, a hybridization buffer concentration of about 6xSSC to about 10xSSC, a formamide concentration of about 0% to about 25%, and a wash solution of about 4xSSC to about 8xSSC. Examples of moderate hybridization conditions include an incubation temperature of about 40°C to about 50°C, a buffer concentration of about 9xSSC to about 2xSSC, a formamide concentration of about 30% to about 50%, and a wash solution of about 5xSSC to about 2xSSC. Examples of high stringency conditions include an incubation temperature of about 55°C to about 68°C, a buffer concentration of about 1xSSC to about 0.1xSSC, a formamide concentration of about 55% to about 75%, and a wash solution of about 1xSSC, 0.1xSSC, or deionized water. Generally, hybridization incubation times range from 5 minutes to 24 hours, with one, two, or more wash steps, with wash incubation times of about 1, 2, or 15 minutes. SSC is a 0.15M NaCl and 15mM citrate buffer. It is understood that equivalents of SSC using other buffer systems can be used.

[0091] As used herein, "expression" refers to the process by which a polynucleotide is transcribed into mRNA and / or the process by which the transcribed mRNA is then translated into a peptide, polypeptide, or protein. If the polynucleotide is derived from genomic DNA, expression may include splicing of the mRNA in a eukaryotic cell.

[0092] The term "encoding," when applied to a polynucleotide, refers to a polynucleotide that is said to "encode" a polypeptide when, in its native state, or when manipulated by methods well known to those of skill in the art, it is capable of being transcribed and / or translated to produce mRNA and the polypeptide and / or fragment thereof. The antisense strand is the complement of such a nucleic acid, and the coding sequence can be deduced therefrom.

[0093] As used herein, the terms "treat," "treatment," and the like are used herein to mean obtaining a desired pharmacological and / or physiological effect. The effect may be preventative, in that a disorder or its signs or symptoms are completely or partially prevented, and / or therapeutic, in that a disorder and / or adverse effects resulting from the disorder are partially or completely cured. As used herein, "treating" or "treatment" of a disease in a subject may refer to (1) preventing a symptom or disease from occurring in a subject who is susceptible to the disease or who does not yet exhibit symptoms of the disease; (2) inhibiting or arresting the development of the disease; or (3) ameliorating or causing regression of the disease or symptoms of the disease. As understood in the art, "treatment" is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of the present technology, beneficial or desired results may include, but are not limited to, one or more of: alleviation or amelioration of one or more symptoms; attenuation of the severity of a condition (including a disease); stabilization (i.e., non-worsening) of a condition (including a disease); delay or slowing of a condition (including a disease); progression, improvement or alleviation of a condition (including a disease), condition and remission (whether partial or total), whether detectable or undetectable. In one embodiment, treatment excludes prevention.

[0094] By prevent is intended preventing a disorder or effect in vitro or in vivo in a system or subject susceptible to the disorder or effect, one example of which is preventing biofilm formation in a system infected with a microorganism known to produce biofilms.

[0095] A "composition" is intended to mean a combination of an active agent with another compound or composition, inert (e.g., a detectable agent or label) or active, such as an adjuvant, diluent, binder, stabilizer, buffer, salt, lipophilic solvent, preservative, adjuvant, etc., and includes a pharmaceutically acceptable carrier. Carriers also include pharmaceutical excipients and additives, proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, di-, tri-, tetra-oligosaccharides, and oligosaccharides; derivatized sugars, e.g., alditols, aldonic acids, esterified sugars, etc.; and polysaccharides or sugar polymers), which may be present alone or in combination, including 1-99.99% by weight or volume. Exemplary protein excipients include serum albumin, e.g., human serum albumin (HSA), and the like. These include recombinant human albumin (rHA), gelatin, casein, etc. Representative amino acids / antibody components that may also function in buffering capacity include alanine, arginine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, etc. Carbohydrate excipients are also contemplated within the scope of this technology, examples of which include, but are not limited to, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, etc.; disaccharides such as lactose, sucrose, trehalose, cellobiose, etc.; polysaccharides such as raffinose, melezitose, maltodextrin, dextran, starch, etc.; and alditols such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol), and myo-inositol.

[0096] A "pharmaceutical composition" is intended to include a combination of an active agent with a carrier, inert or active, that makes the composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

[0097] "Pharmaceutically acceptable carrier" refers to any diluent, excipient or carrier that can be used in the compositions disclosed herein. Pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphate, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, wax, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference textbook in this field. They may be selected with regard to the intended dosage form, ie, oral tablet, capsule, elixir, syrup, etc., and consistent with conventional pharmaceutical practice.

[0098] Compositions used in accordance with the present disclosure may be packaged in dosage unit form for ease of administration and uniformity of dosage. The term "unit dose" or "dosage" refers to a physically discrete unit suitable for use in a subject, each unit containing a predetermined amount of composition calculated to produce the desired response in association with its administration, i.e., an appropriate route and regimen. The amount to be administered, both by number of treatments and unit dose, depends on the desired outcome and / or protection. The precise amount of the composition also depends on the judgment of the practitioner and is peculiar to each individual. Factors affecting dosage include the subject's physical and clinical condition, the route of administration, the intended purpose of treatment (relief of symptoms versus cure), and the efficacy, stability, and toxicity of the particular composition. Upon formulation, solutions are administered in a manner compatible with the administration formulation and in an amount that is therapeutically or prophylactically effective. The formulations are easily administered in a variety of dosage forms, for example, the types of injectable solutions described herein.

[0099] By "bioactive agent" or active agent as disclosed herein is intended one or more of an isolated or recombinant polypeptide, an isolated or recombinant polynucleotide, a vector, an isolated host cell, or an antibody, and compositions comprising one or more of these.

[0100] "Administration" can be achieved in one dose, continuously, or intermittently throughout the course of treatment. Methods for determining the most effective means and dosage of administration are known to those skilled in the art and vary depending on the composition used for treatment, the purpose of treatment, the target cells being treated, and the subject being treated. Single or multiple administrations can be performed at dose levels and patterns selected by the treating physician. Suitable administration formulations and methods for administering the active substance are known in the art. In addition, the route of administration can be determined, and methods for determining the most effective route of administration are known to those skilled in the art and vary depending on the composition used for treatment, the purpose of treatment, the health condition or disease stage of the subject being treated, and the target cells or tissues being treated. Non-limiting examples of administration routes include oral administration, nasal administration, injection, and topical application.

[0101] The agents of the present disclosure may be administered for therapy by any suitable route of administration, it being understood that the most suitable route will vary with the condition and age of the recipient and the disease being treated.

[0102] As used herein, the term "contacting" refers to direct or indirect binding or interaction between two or more molecules. A specific example of direct interaction is binding. A specific example of indirect interaction is when one entity acts on an intermediate molecule, which then acts on a second reference entity. As used herein, contacting includes contacting in solution, in solid phase, in vitro, ex vivo, in cells, and in vivo. In vivo contacting can be referred to as administering or administration.

[0103] The term "effective amount" refers to an amount sufficient to achieve a desired effect. For therapeutic or prophylactic applications, the effective amount depends on the type and severity of the condition in question, as well as the characteristics of the individual subject, such as general health, age, sex, weight, and tolerance to the pharmaceutical composition. For immunogenic compositions, in some embodiments, the effective amount is an amount sufficient to produce a protective response against a pathogen. In other embodiments, the effective amount of an immunogenic composition is an amount sufficient to produce antibody production against an antigen. In some embodiments, the effective amount is the amount necessary to confer passive immunity to a subject in need thereof. For immunogenic compositions, in some embodiments, the effective amount depends on the intended use, the degree of immunogenicity of the particular antigenic compound, and the health / responsiveness of the subject's immune system, in addition to the factors described above. Those skilled in the art will be able to determine the appropriate amount depending on these and other factors.

[0104] In the case of intravital application, in some embodiments, the effective amount depends on the size and nature of the application in question. It also depends on the nature and sensitivity of the intravital target and the method of use. Those skilled in the art can determine the effective amount based on these and other considerations. The effective amount may include one or more administrations of the composition depending on the embodiment.

[0105] The term "contacting" refers to direct or indirect binding or interaction between two or more molecules. A specific example of direct interaction is binding. A specific example of indirect interaction is when one entity acts on an intermediate molecule, which in turn acts on a second reference entity. Contacting, as used herein, includes contacting in solution, in solid phase, in vitro, ex vivo, in cells, and in vivo. In vivo contacting can be referred to as administering or administration.

[0106] The term "conjugated moiety" refers to a moiety that can be added to an isolated chimeric polypeptide by forming a covalent bond with a residue of the chimeric polypeptide. The moiety can be directly attached to the residue of the chimeric polypeptide or can form a covalent bond with a linker that then forms a covalent bond with the residue of the chimeric polypeptide.

[0107] A "peptide conjugate" refers to the association of one or more polypeptides with another chemical or biological compound, either covalently or non-covalently. In a non-limiting example, the "conjugation" of a polypeptide with a chemical compound results in improved stability or efficacy of the polypeptide for its intended purpose. In one embodiment, the peptide is conjugated to a carrier, and the carrier is a liposome, a micelle, or a pharmaceutically acceptable polymer.

[0108] "Liposomes" are microscopic vesicles composed of concentric lipid bilayers. Structurally, liposomes range in size and shape from elongated tubes to spheres, with dimensions ranging from hundreds of angstroms to fractions of a millimeter. Vesicle-forming lipids are selected to achieve a specified degree of fluidity or rigidity in the final complex, providing the lipid composition of the outer layer. They can be neutral (cholesterol) or bipolar, and include phospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), and sphingomyelin (SM), as well as other types of bipolar lipids, including, but not limited to, dioleoylphosphatidylethanolamine (DOPE), with hydrocarbon chain lengths ranging from 14 to 22 and either saturated or containing one or more double C=C bonds. Examples of lipids that can form stable liposomes, alone or in combination with other lipid components, are phospholipids, such as hydrogenated soy phosphatidylcholine (HSPC).soy phosphatidylcholine), lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cephalin, cardiolipin, phosphatidic acid, cerebrosides, distearoylphosphatidylethanolamine (DSPE), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), palmitoyloleoylphosphatidylcholine (POPC) These are palmitoyloteoylphosphatidylcholine, palmitoyloleoylphosphatidylethanolamine (POPE), and dioleoylphosphatidylethanolamine 4-(N-maleimido-triethyl)cyclohexane-1-carboxylate (DOPE-mal). Additional non-phosphorus-containing lipids that can be incorporated into liposomes include stearylamine, dodecylamine, hexadecylamine, isopropyl myristate, triethanolamine-lauryl sulfate, alkyl-aryl sulfates, acetyl palmitate, glycerol ricinoleate, hexadecyl stereate, amphoteric acrylic polymers, polyethyloxylated fatty acid amides, and the cationic lipids listed above (DDAB, DODAC, DMRIE, DMTAP, DOGS, DOTAP (DOTMA), DOSPA, DPTAP, DSTAP, DC-Chol).Negatively charged lipids include phosphatidic acid (PA), dipalmitoylphosphatidylglycerol (DPPG), dioteoylphosphatidylglycerol (DOPG), and dicetyl phosphate, which can form vesicles. Typically, liposomes can be divided into three categories based on their overall size and the nature of their lamellar structure. The three classifications developed by the December 1977 New York Academy of Sciences conference, "Liposomes and Their Use in Biology and Medicine," are multilamellar vesicles (MLVs), small unilamellar vesicles (SUVs), and large unilamellar vesicles (LUVs). Bioactive agents can be encapsulated in such vesicles for administration according to the methods described herein.

[0109] "Micelles" are aggregates of surfactant molecules dispersed in a liquid colloid. Typical micelles in aqueous solution form aggregates with their hydrophilic "head" regions in contact with the surrounding solvent and their hydrophobic tail regions sequestered in the center of the micelle. This type of micelle is known as a normal-phase micelle (oil-in-water micelle). Reverse micelles have head groups in the center and tails pointing outward (water-in-oil micelle). Micelles can be used to bind polynucleotides, polypeptides, antibodies, or compositions described herein to facilitate efficient delivery to target cells or tissues.

[0110] The phrase "pharmaceutically acceptable polymer" refers to a group of compounds that can be conjugated to one or more polypeptides described herein. It is contemplated that conjugation of a polymer to a polypeptide can extend the half-life of the polypeptide in vivo and in vitro. Non-limiting examples include polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, cellulose derivatives, polyacrylates, polymethacrylates, sugars, polyols, and mixtures thereof. Bioactive agents can be conjugated to pharmaceutically acceptable polymers for administration according to the methods described herein.

[0111] " Gene delivery vehicle " is defined as any molecule that can carry inserted polynucleotide into host cell. Examples of gene delivery vehicles include liposomes, micelles, biocompatible polymers including natural and synthetic polymers, lipoproteins, polypeptides, polysaccharides, lipopolysaccharides, artificial virus envelopes, metal particles, and bacteria or viruses, such as baculoviruses, adenoviruses and retroviruses, bacteriophages, cosmids, plasmids, fungal vectors, and other recombinant vehicles typically used in the art, which have been described for expression in various eukaryotic and prokaryotic hosts and can be used for gene therapy and simple protein expression.

[0112] The polynucleotides disclosed herein can be delivered to cells, tissues, or subjects using gene delivery vehicles. As used herein, "gene delivery," "gene transfer," "transduction," and the like are terms that refer to the introduction of an exogenous polynucleotide (sometimes referred to as a "transgene") into a host cell, regardless of the method used for transfer. Such methods include various well-known techniques, such as vector-mediated gene transfer (e.g., by viral infection / transfection or various other protein-based or lipid-based gene delivery complexes) and techniques that facilitate the delivery of "naked" polynucleotides (e.g., electroporation, "gene gun" delivery, and various other techniques used to introduce polynucleotides). The introduced polynucleotide can be stably or transiently maintained in the host cell. Stable maintenance typically requires that the introduced polynucleotide either contain a replication origin compatible with the host cell or be integrated into a host cell replicon, such as an extrachromosomal replicon (e.g., a plasmid) or a nuclear or mitochondrial chromosome. As known in the art and described herein, several vectors are known to be capable of mediating the transfer of genes into mammalian cells.

[0113] As used herein, the term "eDNA" refers to extracellular DNA found as a component of pathogenic biofilms.

[0114] A "plasmid" is an extrachromosomal DNA molecule that is separate from chromosomal DNA and can replicate independently of it. It is often circular and double-stranded. Plasmids provide a mechanism for horizontal gene transfer within a population of microorganisms and typically provide a selective advantage under given environmental conditions. Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche, or alternatively, the proteins produced may act as toxins under similar conditions.

[0115] "Plasmids" used in genetic engineering are called "plasmid vectors." Many plasmids are commercially available for such use. The gene to be replicated is inserted into a copy of the plasmid, which also contains a gene that makes the cell resistant to a particular antibiotic and a multiple cloning site (MCS, or polylinker), a short region containing several commonly used restriction sites and allowing easy insertion of DNA fragments at this location. Another major use of plasmids is to produce large amounts of proteins. In this case, researchers grow bacteria containing a plasmid harboring the gene of interest. Just as the bacterium produces the protein that confers its antibiotic resistance, it can also be induced to produce large amounts of the protein from the inserted gene. This is an inexpensive and easy way to mass-produce a gene or the protein it encodes.

[0116] "Yeast artificial chromosome" or "YAC" refers to a vector used to clone large DNA fragments (greater than 100 kb, up to 3000 kb). It is an artificially constructed chromosome that contains telomere, centromere, and origin of replication sequences required for replication and storage in yeast cells. Once assembled using initial circular plasmids, they can be linearized using restriction enzymes, and then DNA ligase can be used to add sequences or genes of interest into the linear molecule using the cohesive ends. Yeast expression vectors, such as YACs, YIp (yeast integrating plasmid), and YEp (yeast episomal plasmid), are very useful because yeast is itself a eukaryotic cell, allowing for the production of eukaryotic protein products with post-translational modifications. However, YACs are known to be more unstable than BACs and produce chimeric effects.

[0117] A "viral vector" is defined as a recombinant virus or virus particle containing a polynucleotide to be delivered to a host cell, either in vivo, ex vivo, or in vitro. Examples of viral vectors include retroviral vectors, adenoviral vectors, adeno-associated viral vectors, and alphavirus vectors. Infectious tobacco mosaic virus (TMV)-based vectors are sometimes used to manufacture proteins and have been reported to express Griffithsin in tobacco leaves (O'Keefe et al. (2009) Proc. Nat. Acad. Sci. USA 106(15):6099-6104). Alphavirus vectors, such as Semliki Forest virus-based vectors and Sindbis virus-based vectors, have also been developed for use in gene therapy and immunotherapy. See Schlesinger & Dubensky (1999) Curr. Opin. Biotechnol. 5:434-439 and Ying et al. (1999) Nat. Med. 5(7):823-827. In embodiments where gene transfer is mediated by a retroviral vector, vector construct refers to a polynucleotide comprising the retroviral genome or portion thereof and a therapeutic gene.

[0118] As used herein, "retroviral-mediated gene transfer" or "retroviral transduction" have the same meaning and refer to the process by which a gene or nucleic acid sequence is stably transferred into a host cell by a virus that enters the cell and integrates the viral genome into the host cell genome. The virus may enter the host cell through its normal infection mechanism, or the virus may be modified to bind to a different host cell surface receptor or ligand and enter the cell. As used herein, a retroviral vector refers to a viral particle that can introduce exogenous nucleic acid into a cell through a viral or viral-like entry mechanism.

[0119] Retroviruses carry their genetic information in the form of RNA; however, when the virus infects a cell, the RNA is reverse transcribed into a DNA form that integrates into the genomic DNA of the infected cell. The integrated DNA form is called a provirus.

[0120] In embodiments in which gene transfer is mediated by a DNA viral vector, such as an adenovirus (Ad) or an adeno-associated virus (AAV), the term vector construct refers to a polynucleotide comprising the viral genome or a portion thereof and a transgene. Adenoviruses (Ad) are a relatively well-characterized, homogenous group of viruses, including over 50 serotypes. See, e.g., PCT International Application Publication No. WO95 / 27071. Ad does not require integration into the host cell genome. Recombinant Ad-derived vectors have also been constructed, particularly those with reduced potential for recombination and wild-type virus generation. See, e.g., PCT International Application Publication Nos. WO95 / 00655 and WO95 / 11984. Wild-type AAV has high infectivity and specificity for integration into the host cell genome. See Hermonat & Muzyczka (1984) Proc. Natl. Acad. Sci. USA 81:6466-6470 and Lebkowski et al. (1988) Mol. Cell. Biol. 8:3988-3996.

[0121] Vectors containing both a promoter and a cloning site to which a polynucleotide can be operably linked are well known in the art. Such vectors are capable of transcribing RNA in vitro or in vivo and are commercially available from suppliers such as Stratagene (La Jolla, Calif.) and Promega Biotech (Madison, Wis.). To optimize expression and / or in vitro transcription, it may be necessary to remove, add, or modify the 5' and / or 3' untranslated portions of the clone to eliminate extra, potentially inappropriate alternative translation initiation codons or other sequences that may interfere with or reduce expression at either the transcriptional or translational level. Alternatively, a consensus ribosome binding site can be inserted immediately 5' of the initiation codon to improve expression.

[0122] Gene delivery vehicles also include DNA / liposome complexes, micelles, and targeted viral protein-DNA complexes. Liposomes containing targeting antibodies or fragments thereof can also be used in the methods disclosed herein. In addition to delivery of polynucleotides to cells or cell populations, direct introduction of the proteins described herein into cells or cell populations can be achieved by, but is not limited to, protein transfection techniques, or culture conditions that allow for enhanced expression and / or activity of the proteins disclosed herein, among other techniques.

[0123] As used herein, the terms "antibody," "antibodies," and "immunoglobulin" include whole antibodies and any antigen-binding fragments or single chains thereof. Accordingly, the term "antibody" includes any protein- or peptide-containing molecule comprising at least a portion of an immunoglobulin molecule. The terms "antibody," "antibodies," and "immunoglobulin" also include immunoglobulins of any isotype; fragments of antibodies that retain specific binding to an antigen, including, but not limited to, Fab, Fab', F(ab)2, Fv, scFv, dsFv, Fd fragments, dAb, VH, VL, VhH, and V-NAR domains; minibodies, diabodies, triabodies, tetrabodies, and kappabodies; antibody fragments and multispecific antibody fragments formed from one or more isolated fragments. Examples of such include, but are not limited to, the complementarity determining regions (CDRs) of a heavy or light chain or its ligand-binding portion, a heavy or light chain variable region (also referred to herein as a variable domain), a heavy or light chain constant region (also referred to herein as a constant domain), a framework (FR) region, or any portion thereof, at least a portion of a binding protein, a chimeric antibody, a humanized antibody, a single-chain antibody, and a fusion protein comprising an antigen-binding portion of an antibody and a non-antibody protein. The variable regions of the heavy and light chains of an immunoglobulin molecule contain binding domains that interact with antigens. The constant region of an antibody (Ab) can mediate the binding of the immunoglobulin to host tissue. The term "anti-" when used before a protein name, e.g., anti-DNABII, anti-IHF, anti-HU, anti-OMP P5, refers to a monoclonal or polyclonal antibody that exhibits binding to and / or affinity for a specific protein. For example, "anti-IHF" refers to an antibody that binds to the IHF protein. A specific antibody has affinity for or is capable of binding to a protein other than the protein against which the antibody was raised.For example, an anti-IHF may be raised specifically against the IHF protein, but may also bind to other proteins related through sequence homology or through structural homology.

[0124] Complementarity-determining regions (CDRs) are portions of the variable regions of antibodies or T cell receptors produced by B cells and T cells, respectively, where these molecules bind to their specific antigen (also called epitope). In certain embodiments, the terms "variable region" and "variable domain" are used interchangeably to refer to polypeptides in the light or heavy chain of an antibody that vary greatly in sequence of amino acid residues from one antibody to another and determine the conformation of the binding site that confers the antibody's specificity for a particular antigen. In further embodiments, the variable region is about 90 to about 200 amino acids in length, including, but not limited to, about 100 amino acids in length, or alternatively about 110 amino acids in length, or alternatively about 120 amino acids in length, or alternatively about 130 amino acids in length, or alternatively about 140 amino acids in length, or alternatively about 150 amino acids in length, or alternatively about 160 amino acids in length, or alternatively about 170 amino acids in length, or alternatively about 180 amino acids in length, or alternatively about 190 amino acids in length. In certain embodiments, a variable region of an amino acid sequence, as used herein, refers to the first about 100 amino acids, or alternatively about 110 amino acids, or alternatively about 120 amino acids, or alternatively about 130 amino acids, or alternatively about 140 amino acids, or alternatively about 150 amino acids (including or excluding the signal peptide, if applicable) of an amino acid sequence being the variable region.

[0125] A set of CDRs constitutes a paratope, also called an antigen-binding site, which is the part of an antibody that recognizes and binds to an antigen. Three CDRs (CDR1, CDR2, and CDR3) are present in the amino acid sequence of the variable region of an antigen receptor, such as a heavy or light chain, non-contiguously arranged, optionally from the amino terminus to the carboxy terminus. As used herein, CDRn refers to a CDRn in an immunoglobulin chain or a CDRn derived from an immunoglobulin chain, where the number n is selected from 1 to 3. In one embodiment, CDRLn refers to a CDRn in a light chain or a CDRn derived from a light chain, where the number n is selected from 1 to 3, while CDRHn refers to a CDRn in a heavy chain or a CDRn derived from a heavy chain, where the number n is selected from 1 to 3. In certain embodiments, framework region (FR) refers to a portion of the variable region that is not a CDR. In certain embodiments, FRn refers to a FR in a heavy or light chain or a FR derived from a heavy or light chain, where the number n is selected from 1 to 4. In certain embodiments, the variable region comprises, consists essentially of, or even consists of the following (optionally in the order provided, and optionally from amino terminus to carboxy terminus): FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0126] The variable regions and / or CDRs of an antibody or fragment thereof can be determined by one of skill in the art using, for example, publicly available or commercially available tools. Non-limiting examples of such tools include IgBlast (accessible at www.ncbi.nlm.nih.gov / igblast / ). ), Scaligner (available from drugdesigntech at www.scaligner.com / ), IMGT rules and / or tools (see, e.g., www.imgt.org / IMGTScientificChart / Nomenclature / IMGT-FRCDRdefinition.html or accessible at www.imgt.org / ), Chothia Canonical Assignment (accessible at www.bioinf.org.uk / abs / chothia.html), Antigen Receptor Numbering And Receptor Calibration (ANARCI, accessible at opig.stats.ox.ac.uk / webapps / newsabdab / sabpred / anarci / ), Kabat numbering method / scheme (e.g., Kabat, EA, et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242 ), or the Paratome web server (accessible at www.ofranlab.org / paratome / ; see Vered Kunik, et al, Nucleic Acids Research, Volume 40, Issue W1, 1 July 2012, Pages W521-W524).

[0127] Antibodies can be polyclonal, monoclonal, multispecific (e.g., bispecific) and antibody fragments, so long as they exhibit the desired biological activity. Antibodies can be isolated from any suitable biological source, for example, murines, rats, sheep and canines.

[0128] The terms "polyclonal antibody" or "polyclonal antibody composition," as used herein, refer to a preparation of antibodies derived from different B-cell lines. They are a mixture of immunoglobulin molecules secreted against a particular antigen, each recognizing a different epitope.

[0129] As used herein, "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population. Because each monoclonal antibody is directed against a single determinant on the antigen, monoclonal antibodies are highly specific. Antibodies can be detectably labeled, for example, with a radioisotope, an enzyme that generates a detectable product, a fluorescent protein, etc. Antibodies can be further conjugated to other moieties, such as members of specific binding pairs, for example, biotin (a member of the biotin-avidin specific binding pair), etc. Antibodies can also be bound to solid supports, including, but not limited to, polystyrene plates or beads.

[0130] Monoclonal antibodies can be produced using hybridoma technology or recombinant DNA methods known in the art. Hybridomas are cells produced in a laboratory from the fusion of antibody-producing lymphocytes with non-antibody-producing cancer cells, usually myelomas or lymphomas. The hybridomas grow and produce continuous samples of specific monoclonal antibodies. Alternative techniques for producing or selecting antibodies include in vitro exposure of lymphocytes to the antigen of interest and screening antibody display libraries in cell, phage, or similar systems.

[0131] The term "human antibody," as used herein, is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies disclosed herein may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody," as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, e.g., a mouse, have been grafted onto human framework sequences. Thus, as used herein, the term "human antibody" refers to an antibody in which substantially any portion of the protein (e.g., CDRs, framework, C L , C H Domain (e.g., C H1 , C H2 , C H3 ), hinge, (VL, VH)) also refer to antibodies that are substantially non-immunogenic in humans and involve only minor sequence changes or variations. Similarly, primate (monkey, baboon, chimpanzee, etc.), rodent (mouse, rat, rabbit, guinea pig, hamster, etc.), and other mammalian designations of antibodies refer to such species-, subgenus-, genus-, subfamily-, and family-specific antibodies. Furthermore, chimeric antibodies include any combination of the above. Such changes or variations optionally retain or reduce immunogenicity in humans or other species compared to unmodified antibodies. Thus, human antibodies are distinct from chimeric or humanized antibodies. It is noted that human antibodies can be produced by non-human animals or prokaryotic or eukaryotic cells capable of expressing functionally rearranged human immunoglobulin (e.g., heavy and / or light chain) genes. Furthermore, when a human antibody is a single-chain antibody, it may contain a linker peptide not found in native human antibodies. For example, an Fv may include a linker peptide, e.g., two to about eight glycine or other amino acid residues, which connects the heavy chain variable region and the light chain variable region, and such a linker peptide may be of human origin.

[0132] As used herein, a human antibody is "derived" from a particular germline sequence if the antibody is obtained from a system that uses human immunoglobulin sequences, for example, by immunizing a transgenic mouse harboring human immunoglobulin genes or by screening a human immunoglobulin gene library. A human antibody "derived" from a human germline immunoglobulin sequence can itself be identified by comparing the amino acid sequence of the human antibody to the amino acid sequence of a human germline immunoglobulin. Typically, the selected human antibody is at least 90% identical in amino acid sequence to the amino acid sequence encoded by the human germline immunoglobulin gene and contains amino acid residues that identify the human antibody as a human antibody when compared to the germline immunoglobulin amino acid sequence of another species (e.g., a murine germline sequence). In certain cases, the human antibody may be at least 95%, or even at least 96%, 97%, 98%, or 99% identical in amino acid sequence to the amino acid sequence encoded by the germline immunoglobulin gene. Typically, a human antibody derived from a particular human germline sequence displays no more than 10 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene. In certain cases, the human antibody may display no more than 5, or even no more than 4, 3, 2, or 1 amino acid differences from the amino acid sequence encoded by the germline immunoglobulin gene.

[0133] As used herein, the term "humanized antibody" or "humanized immunoglobulin" refers to a human / non-human chimeric antibody containing minimal sequence derived from a non-human immunoglobulin. Generally, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a variable region or fragment thereof (e.g., one, two, three, four, five, or all six CDRs) of the recipient are replaced by residues from a variable region or fragment thereof (e.g., one, two, three, four, five, or all six CDRs) of a non-human species (donor antibody), such as mouse, rat, rabbit, or non-human primate, having the desired specificity, affinity, and capacity. Humanized antibodies can contain residues that are not found in the recipient antibody or in the donor antibody. Humanized antibodies can also optionally comprise non-human antibodies containing one or more amino acids substituted with correspondingly positioned amino acids from a human antibody in at least a portion of an immunoglobulin constant region (Fc), typically a framework region, constant region, or CDR, of a human immunoglobulin. Without wishing to be bound by theory, humanized antibodies result in a reduced immune response in a human host compared to a non-humanized version of the same antibody. Humanized antibodies may have conservative amino acid substitutions that have no substantial effect on antigen binding or other antibody functions. Conservative substitution classes include glycine-alanine, valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, serine-threonine, and asparagine-glutamine. Specifically, the humanized antibodies disclosed herein contain, within certain ranges, the following: EC 50 , K. on , K. off , K. A and / or K DIn one or more of the above, the humanized antibody specifically binds to the DNABII polypeptide or a fragment thereof (such as a tip chimeric peptide or a tail chimeric peptide), and inhibits or releases a specific cytokine when treating a subject. In a further embodiment, a humanized antibody that specifically binds to the tip region (such as a tip chimeric peptide) of the DNABII polypeptide destroys biofilms both in vivo and in vitro. In addition, although the humanization process is a rational design process, it may produce unexpected changes (positive or negative) in binding affinity, antigen specificity, or physical properties such as solubility or aggregation, and therefore the properties of the humanized antibody cannot be essentially predicted from the properties of the starting non-human antibody.

[0134] In one embodiment, an antibody, as used herein, may be a recombinant antibody. The term "recombinant antibody," as used herein, includes all antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies isolated from transgenic or transchromosomic animals (e.g., mice) for immunoglobulin genes or hybridomas prepared therefrom, antibodies isolated from host cells transformed to express the antibody, e.g., from transfectomas, antibodies isolated from recombinant combinatorial antibody libraries, and antibodies prepared, expressed, generated, or isolated by any other means involving splicing of immunoglobulin (Ig) gene sequences into other DNA sequences. However, in certain embodiments, such recombinant antibodies may be subjected to in vitro mutagenesis (or, if an animal transgenic for Ig sequences is used, in vivo somatic mutagenesis), such that the amino acid sequences of the VH and VL regions of the recombinant antibody are sequences that may not naturally exist within the in vivo antibody germline repertoire. Methods for producing these antibodies are described herein.

[0135] In one embodiment, an antibody, as used herein, may be a chimeric antibody. As used herein, a chimeric antibody is an antibody whose light and heavy chain genes have been constructed, typically by genetic engineering, from antibody variable and constant region genes belonging to different species.

[0136] As used herein, the term "antibody derivative" includes a full-length antibody or a fragment of an antibody in which one or more of the amino acids have been chemically modified, for example, by alkylation, pegylation, acylation, ester formation, or amide formation, to link the antibody to a second molecule, including, but not limited to, pegylated antibodies, cysteine-pegylated antibodies, and variants thereof.

[0137] As used herein, the term "label" refers to a directly or indirectly detectable compound or composition, e.g., an N-terminal histidine tag (N-His), a magnetically rotatory isotope, e.g., a nucleotide sequence, which is directly or indirectly conjugated to a composition to be detected to produce a "labeled" composition. 115 Sn, 117 Sn and 119 Sn, non-radioactive isotopes, e.g. 13 C and 15The term also contemplates a polynucleotide or protein, such as an antibody. The term also encompasses sequences conjugated to a polynucleotide that provide a signal upon expression of the inserted sequence, such as green fluorescent protein (GFP). Labels may be detectable by themselves (e.g., radioisotope labels or fluorescent labels) or, in the case of enzymatic labels, may catalyze a chemical change in a detectable substrate compound or composition. Labels may be suitable for small-scale detection or may be more suitable for high-throughput screening. Suitable labels thus include, but are not limited to, magnetically rotatory isotopes, non-radioactive isotopes, radioisotopes, fluorescent dyes, chemiluminescent compounds, dyes, and proteins, including enzymes. Labels may be simply detected, and labels may be quantified. A simply detected response generally includes a response whose presence is simply confirmed, while a quantified response generally includes a response that has a quantifiable (e.g., numerically reportable) value, such as intensity, polarization, and / or other property. In luminescent or fluorescent assays, the detectable response can be generated directly using a luminophore or fluorophore associated with the assay component actually involved in binding, or indirectly using a luminophore or fluorophore associated with another (e.g., reporter or indicator) component. Examples of luminescent labels that produce a signal include, but are not limited to, bioluminescence and chemiluminescence. The detectable luminescent response generally involves a change in, or the generation of, a luminescent signal. Suitable methods and luminophores for luminescently labeling assay components are known in the art, see, for example, Haugland, Richard P. (1996) Handbook of Fluorescent Probes and Research Chemicals (6 th Examples of luminescent probes include, but are not limited to, aequorin and luciferase.

[0138] As used herein, the term "immunoconjugate" includes an antibody or antibody derivative associated with or linked to a second agent, e.g., a cytotoxic agent, a detectable agent, a radioactive agent, a targeting agent, a human antibody, a humanized antibody, a chimeric antibody, a synthetic antibody, a semi-synthetic antibody, or a multispecific antibody.

[0139] Examples of suitable fluorescent labels include, but are not limited to, fluorescein, rhodamine, tetramethylrhodamine, eosin, erythrosine, coumarin, methyl-coumarin, pyrene, malachite green, stilbene, Lucifer Yellow, Cascade Blue™, and Texas Red. Other suitable optical dyes are described in Haugland, Richard P. (1996) Handbook of Fluorescent Probes and Research Chemicals (6 th ed.).

[0140] In another embodiment, the fluorescent label is functionalized to facilitate covalent binding to cellular components present in cells or tissues, or on the cell or tissue surface, such as cell surface markers.Suitable functional groups include, but are not limited to, isothiocyanate groups, amino groups, haloacetyl groups, maleimides, succinimidyl esters, and sulfonyl halides, all of which can be used to bind the fluorescent label to a second molecule.The choice of functional group of the fluorescent label depends on the binding site to either a linker, an agent, a marker, or a second labeling agent.

[0141] "Eukaryotic cells" include all kingdoms of life except the kingdom Monera. They can be easily distinguished by a membrane-bound nucleus. Animals, plants, fungi, and protists are eukaryotes, or organisms in which the cell is organized into a complex structure by internal membranes and a cytoskeleton. The most characteristic membrane-bound structure is the nucleus. Unless otherwise recited, the term "host" includes eukaryotic hosts, including, for example, yeast, higher plants, insects, and mammalian cells. Non-limiting examples of eukaryotic cells or hosts include primates, bovines, porcines, murines, rats, birds, reptiles, and humans.

[0142] "Prokaryotic cells" typically lack a nucleus or any other membrane-bound organelles and are divided into two domains: bacteria and archaea. In addition to chromosomal DNA, these cells may also contain genetic information in circular loops called episomes. Bacterial cells are very small, roughly the size of animal mitochondria (approximately 1-2 μm in diameter and 10 μm in length). Prokaryotic cells are characterized by three main shapes: rod-shaped, spherical, and spiral. Instead of undergoing an elaborate replication process like eukaryotes, bacterial cells divide by binary fission. Examples include, but are not limited to, Bacillus, E. coli, and Salmonella.

[0143] A "native" or "natural" antigen is one that has been isolated from a natural biological source and that is capable of binding to an antigen receptor, particularly a T cell antigen receptor (TCR), in a subject. A polypeptide, protein, or fragment containing an epitope capable of specifically binding to a target polypeptide (e.g., a polypeptide-binding protein or fragment containing an epitope) is a polypeptide, protein, or fragment containing an epitope capable of specifically binding to a target polypeptide (e.g., a polypeptide-binding protein or fragment containing an epitope) or ...

[0144] The terms "antigen" and "antigenic" refer to a molecule capable of being recognized by an antibody or otherwise acting as a member of an antibody-ligand pair. "Specific binding" or "binding" refers to the interaction of an antigen with the variable regions of immunoglobulin heavy and light chains. Antibody-antigen binding can occur in vivo or in vitro. Those skilled in the art will understand that macromolecules, including proteins, nucleic acids, fatty acids, lipids, lipopolysaccharides, and polysaccharides, have the potential to act as antigens. Those skilled in the art will further understand that nucleic acids encoding proteins with the potential to act as antibody ligands necessarily encode antigens. Those skilled in the art will further understand that antigens are not limited to full-length molecules but can also include partial molecules. The term "antigenic" is an adjective that refers to a molecule having the properties of an antigen. This term encompasses substances that are immunogenic, i.e., immunogens, and substances that induce immunological unresponsiveness or anergy, i.e., anergens.

[0145] An "altered antigen" is an antigen having a primary sequence different from that of the corresponding wild-type antigen. Altered antigens can be produced by synthetic or recombinant methods and include, but are not limited to, antigenic peptides that are differentially modified during or after translation, for example, by phosphorylation, glycosylation, cross-linking, acylation, proteolytic cleavage, or linkage to antibody molecules, membrane molecules, or other ligands. (Ferguson et al. (1988) Ann. Rev. Biochem. 57:285-320) The synthetic or altered antigens disclosed herein are intended to bind to the same TCR as the natural epitope.

[0146] An "autoantigen," also referred to herein as a native or wild-type antigen, is an antigenic peptide that elicits little or no immune response in a subject due to self-tolerance to that antigen. An example of an autoantigen is the melanoma-specific antigen gp100.

[0147] "Immune response" broadly refers to an antigen-specific response of lymphocytes to a foreign substance. The terms "immunogen" and "immunogenic" refer to a molecule capable of inducing an immune response. All immunogens are antigens, however, not all antigens are immunogenic. The immune responses disclosed herein can be humoral (via antibody activity) or cell-mediated (via T cell activation). The response can occur in vivo or in vitro. Those skilled in the art will understand that various macromolecules, including proteins, nucleic acids, fatty acids, lipids, lipopolysaccharides, and polysaccharides, have the potential to be immunogenic. Those skilled in the art will further understand that a nucleic acid encoding a molecule capable of inducing an immune response necessarily encodes an immunogen. Those skilled in the art will further understand that an immunogen is not limited to a full-length molecule but can include a partial molecule.

[0148] The term "passive immunity" refers to the transfer of immunity from one subject to another through the transfer of antibodies. Passive immunity can occur naturally when maternal antibodies are transferred to a fetus. Passive immunity can also occur artificially when an antibody composition is administered to a non-immunized subject. Antibody donors and recipients can be human or non-human subjects. Depending on the embodiment, antibodies can be polyclonal or monoclonal, generated in vitro or in vivo, and purified, partially purified, or unpurified. In some embodiments described herein, passive immunity is provided to a subject in need thereof through the administration of an antibody or antigen-binding fragment that specifically recognizes or binds to a particular antigen. In some embodiments, passive immunity is provided through the administration of an isolated or recombinant polynucleotide encoding an antibody or antigen-binding fragment that specifically recognizes or binds to a particular antigen.

[0149] In the context of the present disclosure, a "ligand" is a polypeptide. In one aspect, the term "ligand" as used herein refers to any molecule that binds to a specific site on another molecule. In other words, the ligand confers specificity to the protein in its reaction with immune effector cells or antibodies to the protein or DNA to the protein. In one aspect, it is the ligand site within the protein that directly combines with the complementary binding site on the immune effector cell.

[0150] As used herein, the terms "solid phase support" or "solid support," which are used interchangeably, are not limited to a particular type of support. Rather, numerous supports are available and known to those skilled in the art. Solid phase supports include silica gel, resins, derivatized plastic films, glass beads, cotton, plastic beads, and alumina gel. As used herein, "solid support" also includes synthetic antigen-presenting matrices, cells, and liposomes. A suitable solid phase support can be selected based on the desired end use and suitability for various protocols. For example, for peptide synthesis, a solid support may refer to a resin such as polystyrene (e.g., PAM-resin available from Bachem Inc., Peninsula Laboratories, etc.), POLYHIPE® resin (available from Aminotech, Canada), polyamide resin (available from Peninsula Laboratories), polystyrene resin grafted with polyethylene glycol (TentaGel®, Rapp Polymere, Tübingen, Germany) or polydimethylacrylamide resin (available from Milligen / Biosearch, Calif.).

[0151] Examples of solid supports include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylase, natural and modified cellulose, polyacrylamide, gabbro, and magnetite. The nature of the support can be either soluble to some extent or insoluble. The support material can have virtually any possible structural configuration, so long as the attached molecule can bind to a polynucleotide, polypeptide, or antibody. Thus, the support configuration can be spherical, as in a bead, or cylindrical, as in the inner surface of a test tube or the outer surface of a rod. Alternatively, the surface can be flat, such as a sheet, test strip, or polystyrene beads. Those skilled in the art will know many other suitable supports for binding antibodies or antigens, or can ascertain such by use of routine experimentation.

[0152] As used herein, a biological sample, or sample, can be obtained from a subject, a cell line, or a cultured cell or tissue. Exemplary samples include, but are not limited to, cell samples, tissue samples, liquid samples such as blood and other liquid samples of biological origin (including, but not limited to, ocular fluid (aqueous humor and vitreous humor), peripheral blood, serum, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humor, amniotic fluid, earwax, breast milk, bronchoalveolar lavage fluid, semen, prostatic fluid, Cowper's fluid or pre-ejaculatory fluid, female ejaculate, sweat, tears, cyst fluid, pleural and peritoneal fluid, pericardial fluid, ascites, lymph, chyme, chyle, bile, interstitial fluid, menstrual fluid, pus, sebum, vomit, vaginal secretions / washings, synovial fluid, mucosal secretions, stool water, pancreatic juice, nasal lavage fluid, bronchopulmonary aspirate, blastocyst cavity fluid, blastocyst fluid, cy ... In one embodiment, the biological sample is suspected of having a biofilm. In another embodiment, the biological sample comprises a biofilm.

[0153] As used herein, the term "signal peptide" or "signal polypeptide" refers to an amino acid sequence that is usually present at the N-terminus of a newly synthesized secreted or membrane polypeptide or protein. It acts to direct the polypeptide to a specific cellular location, for example, across the cell membrane, into the cell membrane, or into the nucleus. In some embodiments, the signal peptide is removed from the following locations: Examples of signal peptides are well known in the art; non-limiting examples are those described in U.S. Patent Nos. 8,853,381, 5,958,736, and 8,795,965.

[0154] As used herein, the term "chimera" or "chimeric peptide" refers to a recombinant polypeptide comprising, or alternatively essentially consisting of, or even consisting of, two or more fragments or domains of a DNABII polypeptide that are directly or indirectly (e.g., via a linker) conjugated to each other. In one embodiment, the domain is a conformational tip domain and / or a conformational tail domain. Additionally or alternatively, the two or more fragments or domains are derived from the same or different DNABII polypeptides. In one embodiment, the chimeric peptide comprises, or alternatively essentially consisting of, or even consisting of, the tip domain of IhfA and the tip domain of IhfB that are directly or indirectly (e.g., via a linker) conjugated to each other. In another embodiment, the chimeric peptide comprises, or alternatively essentially consisting of, or even consisting of, the tail domain of IhfA and the tail domain of IhfB that are directly or indirectly (e.g., via a linker) conjugated to each other. A "conformational tip domain" of a polypeptide refers to a polypeptide that comprises a primary amino acid sequence whose structure has an antiparallel beta ribbon with a sharp turn, typically mediated by a proline residue. The "tip" of an IHF polypeptide is shown in Figure 1 of WO2018 / 129078.

[0155] In certain embodiments, the chip-chimeric peptide IhfA5-mIhfB4 NTHI teeth, [ka] (wherein "X" is optionally an amino acid linker sequence comprising, consisting essentially of, or alternatively consisting of, 1 to 20 amino acids, where "X1" is any amino acid, or alternatively, "X1" is selected from the amino acids Q, R, K, S, or T). In a further aspect, "X1" is K or Q. In a further embodiment, the chip-chimeric peptide IhfA5-mIhfB4 NTHI teeth, [ka] (where "X" is optionally an optional amino acid linker sequence comprising, consisting essentially of, or even consisting of 1-20 amino acids). In yet a further embodiment, the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI teeth, [ka] The polypeptide sequence of the present invention may comprise, consist essentially of, or even consist of, the polypeptide sequence of

[0156] In certain embodiments, the tail chimeric peptide IhfA3-IhfB2 NTHI is FLEEIRLSLESGQDVKLSGF-X-TLSAKEIENMVKDILEFISQ (SEQ ID NO: 41) (where "X" is optionally an amino acid linker sequence comprising, consisting essentially of, or even consisting of 1-20 amino acids). In certain embodiments, the linker is selected from any one or more of SEQ ID NOs: 42-49. In one embodiment, the tail chimeric peptide IhfA3-IhfB2 NTHI comprises, consists essentially of, or alternatively consists of FLEEIRLSLESGQDVKLSGFGPSLTLSAKEIENMVKDILEFISQ (SEQ ID NO: 50).

[0157] As used herein, "EC 50 " refers to the concentration of an antibody or fragment thereof that induces a response (e.g., binding between an antibody or fragment thereof and its target) intermediate between baseline and maximum after a specified exposure time.

[0158] Several parameters are used herein to describe the binding and non-binding reactions of receptor (R, e.g., an antibody or fragment thereof) and ligand (L, e.g., the target of the antibody or fragment thereof) molecules, and are formalized as R + L ⇔ RL. This reaction is expressed as M -1 s -1 and s -1 The on-rate constant k has units of on and the off-rate constant k off At equilibrium, the forward bonding transition R+L → RL must be balanced by the backward non-bonding transition RL → R+L. That is, k on [R][L]=k off [RL], where [R], [L], and [RL] represent the concentration of unbound free receptor, the concentration of unbound free ligand, and the concentration of the receptor-ligand complex. Furthermore, the equilibrium dissociation constant, K D " is [R] × [L] / [RL] k off / k on whereas the equilibrium binding constant "K A" is [RL] / ([R] × [L]) k on / k off It can be calculated as:

[0159] As used herein, the term "cytokine" refers, without limitation, to small proteins (approximately 5-20 kDa) important in cell signaling, including, but not limited to, chemokines, interferons, interleukins (ILs), lymphokines, and tumor necrosis factors, generally non-hormonal. Cytokines are peptides that cannot cross the lipid bilayer of cells and enter the cytoplasm. Inflammatory or proinflammatory cytokines are a type of signaling molecule (cytokine) secreted by immune cells such as helper T cells (Th) and macrophages, as well as certain other cell types, that promote inflammation. These include (but are not limited to) interleukin-1 (IL-1), IL-12, and IL-18, tumor necrosis factor alpha (TNF-α), interferon gamma (IFNγ), and granulocyte-macrophage colony-stimulating factor (GM-CSF), and play an important role in mediating the innate immune response. Proinflammatory cytokines are preferentially produced by and are involved in the upregulation of inflammatory responses. The term "anti-inflammatory cytokine" includes immunoregulatory molecules that control proinflammatory cytokine responses. Cytokines act together with specific cytokine inhibitors and soluble cytokine receptors to regulate human immune responses. Major anti-inflammatory cytokines include interleukin (IL)-1 receptor antagonists, IL-4, IL-6, IL-10, IL-11, and IL-13. Specific cytokine receptors for IL-1, tumor necrosis factor-alpha, and IL-18 also function as pro-inflammatory cytokine inhibitors. Methods for measuring cytokine levels, including anti-inflammatory and pro-inflammatory cytokines, are well known in the art. For example, serum cytokine levels can be measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits.

[0160] As used herein, the term "anti-infective agent" refers to an agent capable of inhibiting the spread of or completely killing infectious organisms. This term includes, but is not limited to, antibiotics, antifungals, anthelmintics, antimalarials, antiprotozoal agents, antituberculosis agents, and antivirals. Antifungal agents are also called antimycotic agents. They kill or inactivate fungi and are used to treat fungal infections (including yeast infections). One non-limiting example, polyene antifungals, are used to treat fungal infections of the gastrointestinal tract, such as thrush, because they are not absorbed when administered orally. Other non-limiting examples include azole antifungals, which are synthetic fungistatic agents with broad-spectrum activity; echinocandins, which are lipopeptide molecules that non-competitively inhibit the (1,3) beta-d-glucan synthase enzyme and target fungal cell walls; Fulvicin U / F (i.e., griseofulvin); Grifulvin V(Pro) (i.e., griseofulvin); Lamisil(Pro) (i.e., terbinafine); Gris-PEG(Pro) (i.e., griseofulvin); Ancobon(Pro) (i.e., flucytosine); Fulvicin P / G (i.e., griseofulvin); and Terbinex(Pro) (i.e., terbinafine). Additional anti-infective agents can be found, for example, at www.drugbank.ca / categories / DBCAT000065. The terms "anti-viral" or "antiviral" refer to a class of drugs used to treat viral infections. Most antiviral drugs target specific viruses, while broad-spectrum antivirals are effective against a wide range of viruses. Unlike most antibiotics, antiviral drugs do not destroy their target pathogens, but instead inhibit their development.Some of the ways in which they can act include inhibiting viral DNA polymerase; binding to specific cell surface receptors to inhibit viral penetration or uncoating; inhibiting viral protein synthesis; or preventing viral replication by blocking late stages of viral assembly. Non-limiting examples of antiviral agents can be found, for example, at www.drugbank.ca / categories / DBCAT000066.

[0161] As used herein, the term "antiparasitic drug" refers to a class of drugs indicated for the treatment of parasitic diseases, such as diseases caused by helminths, amoebae, ectoparasites, parasitic fungi, and protozoa, among others. Non-limiting examples of antiparasitic drugs can be found, for example, at www.drugbank.ca / categories / DBCAT000522. Modes for Carrying Out This Disclosure antibody composition

[0162] The present disclosure provides an isolated antibody comprising a heavy chain (HC) variable domain sequence and a light chain (LC) variable domain sequence, wherein the heavy and light chain immunoglobulin variable domain sequences form an antigen-binding site that binds to an epitope of a DNABII protein. In certain embodiments, the antibody or fragment thereof binds to a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI and / or the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail region of a DNABII peptide comprising the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI In another embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2NTHI Combine with.

[0163] In one aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of amino acids (aa) 25 to aa 144 of SEQ ID NO: 1 to 6, 13, 24 or 26, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of aa 21 to aa 132 of SEQ ID NO: 7 to 9, 14 or 25, aa 21 to aa 126 of SEQ ID NO: 10 to 12 or 27, or an equivalent of each thereof.

[0164] In a further aspect, antibodies and antigen-binding fragments thereof are provided which comprise, consist essentially of, or alternatively consist of a heavy chain (HC) comprising, consisting essentially of, or consisting of a sequence selected from the group of aa25 to aa473 of SEQ ID NO: 1 to 6, 13, 24 or 26, or an equivalent of each thereof, and / or a light chain (LC) comprising, consisting essentially of, or consisting of a sequence selected from the group of aa21 to aa239 of SEQ ID NO: 7 to 9, 14 or 25, aa21 to aa233 of SEQ ID NO: 10 to 12 or 27, or an equivalent of each thereof.

[0165] In a still further aspect, there is provided antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) comprising, consisting essentially of, or consisting of a sequence selected from the group of SEQ ID NOs: 1 to 6, 13, 24 or 26, or an equivalent of each thereof, and / or a light chain (LC) comprising, consisting essentially of, or consisting of a sequence selected from the group of SEQ ID NOs: 7 to 12, 14, 25 or 27, or an equivalent of each thereof.

[0166] In another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of amino acids (aa) 25 to 144 of SEQ ID NO: 1 to 6, 13, 24 or 26, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of aa 21 to aa 132 of SEQ ID NO: 6 to 9, 14 or 25, aa 21 to aa 126 of SEQ ID NO: 10 to 12 or 27, or an equivalent of each thereof.

[0167] In yet another aspect, there is provided an antibody or fragment thereof comprising, consisting essentially of, or even consisting of, any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 1 to 6, 13, 24 or 26, and / or any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 7 to 12, 14, 25 or 27.

[0168] In another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of aa25 to aa144 of SEQ ID NO: 13, 24 or 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of aa21 to aa132 of SEQ ID NO: 14 or 25, aa21 to aa126 of SEQ ID NO: 27, or an equivalent thereof. In a further aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of aa25 to aa144 of SEQ ID NO: 1-6, 13, 24 or 26, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of a sequence selected from the group of aa21 to aa132 of SEQ ID NO: 7-9, 14 or 25, aa21 to aa126 of SEQ ID NO: 10-12 or 27, or an equivalent of each thereof.

[0169] Also provided are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 1, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of any one of aa21 to aa132 of SEQ ID NO: 7 to 9, 14 or 25, or aa21 to aa126 of SEQ ID NO: 10 to 12 or 27, or an equivalent thereof. In another aspect, provided herein is an antibody or fragment thereof comprising, or alternatively consisting essentially of, or consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 2, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of any one of aa21 to aa132 of SEQ ID NO: 7-9, 14 or 25, aa21 to aa126 of SEQ ID NO: 10-12 or 27, or an equivalent of each thereof. In a further aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of any one of aa21 to aa132 of SEQ ID NO: 7 to 9, 14 or 25, aa21 to aa126 of SEQ ID NO: 10 to 12 or 27, or an equivalent of each thereof.

[0170] In a still further aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 4, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of any one of aa21 to aa132 of SEQ ID NO: 7 to 9, 14 or 25, aa21 to aa126 of SEQ ID NO: 10 to 12 or 27, or an equivalent of each thereof. Also provided are antibodies and antigen-binding fragments thereof comprising, alternatively consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence of any one of aa21 to aa132 of SEQ ID NO: 7 to 9, 14 or 25, or aa21 to aa126 of SEQ ID NO: 10 to 12 or 27, or an equivalent of each thereof. Furthermore, antibodies and antigen-binding fragments thereof are provided which comprise, or alternatively consist essentially of, or consist of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consisting of, or consisting of the amino acid sequence of any one of aa21 to aa132 of SEQ ID NO: 7 to 9, 14 or 25, or aa21 to aa126 of SEQ ID NO: 10 to 12 or 27, or an equivalent thereof.

[0171] In another aspect, provided herein is an antibody or antigen-binding fragment thereof comprising, consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence from aa25 to aa144 of any one of SEQ ID NOs: 1-6, 13, 24 or 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence from aa21 to aa132 of SEQ ID NO: 7, or an equivalent thereof. Furthermore, there is provided an antibody or antigen-binding fragment thereof comprising, consisting essentially of, or additionally consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of any one of SEQ ID NOs: 1 to 6, 13, 24 or 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In another aspect, also provided is an antibody or antigen-binding fragment thereof comprising, consisting essentially of, or additionally consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence from aa25 to aa144 of any one of SEQ ID NOs: 1 to 6, 13, 24 or 26, or an equivalent of each thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence from aa21 to aa132 of SEQ ID NO: 9, or an equivalent thereof.

[0172] In a further aspect, there is also provided an antibody or antigen-binding fragment thereof comprising, or alternatively consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, or alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence from aa25 to aa144 of any one of SEQ ID NOs: 1 to 6, 13, 24 or 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, or alternatively consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 10, or an equivalent thereof. Also provided are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or additionally consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of any one of SEQ ID NOs: 1 to 6, 13, 24 or 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. Also provided are antibodies and antigen-binding fragments thereof comprising, consisting essentially of, or additionally consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, or consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of any one of SEQ ID NOs: 1 to 6, 13, 24 or 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising the amino acid sequence of aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof.

[0173] In one aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, alternatively consisting essentially of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence of aa25 to aa144 of SEQ ID NO: 1, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence of aa21 to aa132 of SEQ ID NO: 7, or an equivalent thereof. In one embodiment, provided are antibodies and antigen-binding fragments thereof comprising, alternatively essentially consisting of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence of aa25 to aa144 of SEQ ID NO: 1, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence of aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In another embodiment, provided herein are antibodies and antigen-binding fragments thereof comprising, alternatively essentially consisting of, or alternatively consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence of aa25 to aa144 of SEQ ID NO: 1, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, alternatively consisting essentially of, or alternatively consisting of, the amino acid sequence of aa21 to aa132 of SEQ ID NO: 9, or an equivalent thereof.

[0174] In another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, or alternatively consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 2, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, or alternatively consisting essentially of, or consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 7, or an equivalent thereof. In another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, or alternatively consisting essentially of, or even additionally of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In yet another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO:2, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO:9, or an equivalent thereof.

[0175] In another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or even further of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 7, or an equivalent thereof. In a further aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or even further of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In one embodiment, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, or consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising the amino acid sequence of aa21 to aa132 of SEQ ID NO: 9, or an equivalent thereof.

[0176] Also provided are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 4, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 10, or an equivalent thereof. In another aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 4, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. In a further aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, or consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 4, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, or consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof.

[0177] In one embodiment, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 10, or an equivalent thereof. In another embodiment, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. In a further aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, or consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, or consisting essentially of, the amino acid sequence of aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof.

[0178] In another embodiment, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively essentially consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 10, or an equivalent thereof. In one aspect, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively essentially consisting of, a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. In one embodiment, provided herein are antibodies and antigen-binding fragments thereof comprising, or alternatively consisting essentially of, or consisting of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof.

[0179] In one aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 24, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 25, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof is a fragment of an antibody or fragment thereof that comprises, consists essentially of, or alternatively consists of the tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI In one embodiment, the antibody or fragment thereof binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHIIn yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa 25 to aa 144 of SEQ ID NO: 1, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa 21 to aa 132 of SEQ ID NO: 7, or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 1, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 1, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 9, or an equivalent thereof.In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 2, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa132 of SEQ ID NO: 7, or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 2, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 2, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa132 of SEQ ID NO: 9, or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa132 of SEQ ID NO: 7, or an equivalent thereof.In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 8, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 3, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa132 of SEQ ID NO: 9, or an equivalent thereof.

[0180] In another aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa25 to aa144 of SEQ ID NO: 26, or an equivalent thereof, and a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa21 to aa126 of SEQ ID NO: 27, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof is coupled to a tail region of a DNABII peptide (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2 NTHIIn yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa 25 to aa 144 of SEQ ID NO: 4, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa 21 to aa 126 of SEQ ID NO: 10, or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 4, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 4, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof.In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 10, or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 5, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 10, or an equivalent thereof.In another embodiment, the antibody or fragment thereof comprises, essentially consists of, or further consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 11, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence from aa25 to aa144 of SEQ ID NO: 6, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence from aa21 to aa126 of SEQ ID NO: 12, or an equivalent thereof.

[0181] In one aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 24 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 25 or an equivalent thereof. In a further embodiment, the antibody or fragment thereof is a fragment of an antibody or fragment thereof that binds to a DNABII peptide tip region (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI In one embodiment, the antibody or fragment thereof binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHIIn yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 7 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 8 or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 1 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 9 or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 7 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or further consists of a heavy chain (HC) comprising, consisting essentially of, or further consisting of the amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or further consisting of the amino acid sequence of SEQ ID NO: 8 or an equivalent thereof.In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 2 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 9 or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 7 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 8 or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 3 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 9 or an equivalent thereof.

[0182] In another aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 26 or an equivalent thereof, and a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 27 or an equivalent thereof. In a further embodiment, the antibody or fragment thereof is fused to a tail region of a DNABII peptide (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2 NTHIIn yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 10 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 11 or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 4 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 12 or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 10 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or further consists of a heavy chain (HC) comprising, consisting essentially of, or further consisting of the amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or further consisting of the amino acid sequence of SEQ ID NO: 11 or an equivalent thereof.In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 5 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 12 or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 10 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 11 or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 6 or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 12 or an equivalent thereof.

[0183] In one aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa 25 to aa 473 of SEQ ID NO: 24, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa 21 to aa 239 of SEQ ID NO: 25, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof is a fragment of an antibody or fragment thereof that binds to a DNABII peptide tip region (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI In one embodiment, the antibody or fragment thereof binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHIIn yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or consists of a heavy chain (HC) comprising, essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 1 or an equivalent thereof, and / or a light chain (LC) comprising, essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 7 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 1 or an equivalent thereof, and / or a light chain (LC) comprising, essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 8 or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 1, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 9, or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 2, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 7, or an equivalent thereof.In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 2, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 8, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 2, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 9, or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 3, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 7, or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 3, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa239 of SEQ ID NO: 8, or an equivalent thereof.In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or further consists of a heavy chain (HC) comprising, consisting essentially of, or further consisting of the amino acid sequence from aa 25 to aa 473 of SEQ ID NO: 3, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or further consisting of the amino acid sequence from aa 21 to aa 239 of SEQ ID NO: 9, or an equivalent thereof.

[0184] In another aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa 25 to aa 473 of SEQ ID NO: 26, or an equivalent thereof, and a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of aa 21 to aa 233 of SEQ ID NO: 27, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof is fused to a tail region of a DNABII peptide (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2 NTHIIn yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, the antibody or fragment thereof comprises, essentially consists of, or alternatively consists of a heavy chain (HC) comprising, essentially consists of, or alternatively consists of the amino acid sequence of aa25 to aa473 of SEQ ID NO: 4 or an equivalent thereof, and / or a light chain (LC) comprising, essentially consists of, or alternatively consists of the amino acid sequence of aa21 to aa233 of SEQ ID NO: 10 or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, essentially consists of, or alternatively consists of a heavy chain (HC) comprising, essentially consists of, or alternatively consists of the amino acid sequence of aa25 to aa473 of SEQ ID NO: 4 or an equivalent thereof, and / or a light chain (LC) comprising, essentially consists of, or alternatively consists of the amino acid sequence of aa21 to aa233 of SEQ ID NO: 11 or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 4, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa233 of SEQ ID NO: 12, or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 5, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa233 of SEQ ID NO: 10, or an equivalent thereof.In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 5, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa233 of SEQ ID NO: 11, or an equivalent thereof. In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 5, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa233 of SEQ ID NO: 12, or an equivalent thereof. In one embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 6, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa233 of SEQ ID NO: 10, or an equivalent thereof. In another embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa25 to aa473 of SEQ ID NO: 6, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or alternatively consisting of the amino acid sequence from aa21 to aa233 of SEQ ID NO: 11, or an equivalent thereof.In yet another embodiment, the antibody or fragment thereof comprises, consists essentially of, or further consists of a heavy chain (HC) comprising, consisting essentially of, or further consisting of the amino acid sequence from aa 25 to aa 473 of SEQ ID NO: 6, or an equivalent thereof, and / or a light chain (LC) comprising, consisting essentially of, or further consisting of the amino acid sequence from aa 21 to aa 233 of SEQ ID NO: 12, or an equivalent thereof.

[0185] In one aspect, an antibody or fragment thereof is provided that comprises, consists essentially of, or even consists of any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 1-3 or 24, and / or any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 7-9 or 25. In a further embodiment, the antibody or fragment thereof is provided with a CDR that is a CDR of a DNABII peptide (including, but not limited to, the CDR of IHF or HU, the CDR of IHFA or IHFB, and / or the CDR of ... NTHI In one embodiment, the antibody or fragment thereof binds to the CHIP-chimeric peptide IhfA5-mIhfB4 NTHI In yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, an antibody or fragment thereof is provided that comprises, consists essentially of, or even consists of all three CDRs of a sequence selected from the group of SEQ ID NOs: 1-3 or 24, or equivalents of each of them, and / or all three CDRs of a sequence selected from the group of SEQ ID NOs: 7-9 or 25, or equivalents of each of them.

[0186] In another embodiment, an antibody or fragment thereof is provided that comprises, consists essentially of, or even consists of any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 4-6 or 26, and / or any one or any two or all three CDRs or their respective equivalents of a sequence selected from the group of SEQ ID NOs: 10-12 or 27. In a further embodiment, the antibody or fragment thereof is provided with a CDR comprising the tail region of a DNABII peptide (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody or fragment thereof binds to the tail chimeric peptide IhfA3-IhfB2 NTHI In yet a further embodiment, the fragment is an antigen-binding fragment. In one embodiment, an antibody or fragment thereof is provided that comprises, consists essentially of, or even consists of all three CDRs of a sequence selected from the group of SEQ ID NOs: 4-6 or 26, or equivalents of each of them, and / or all three CDRs of a sequence selected from the group of SEQ ID NOs: 10-12 or 27, or equivalents of each of them.

[0187] In certain embodiments, the antibodies or fragments thereof provided herein further comprise one or more signal peptides. In one embodiment, the signal peptide comprises, consists essentially of, or even consists of amino acids (aa) 1 to aa24 of any one of SEQ ID NOs: 1-6, 13, 24, or 26. In another embodiment, the signal peptide comprises, consists essentially of, or even consists of aa1 to aa20 of any one of SEQ ID NOs: 7-12, 14, 25, and 27. In a further embodiment, the signal peptide is located at the amino terminus of the light chain variable region. Additionally or alternatively, the same or a different signal peptide is located at the amino terminus of the heavy chain variable region.

[0188] The antibodies or fragments thereof provided herein may be monospecific or bispecific. In one embodiment, the antibodies or fragments thereof are trispecific, tetraspecific, or pentaspecific. Additionally or alternatively, the antibodies are selected from the group of IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., IgG1, IgG2, IgG3, or IgG4), or IgM antibodies. In one embodiment, the antibody further comprises a constant region selected from the group of an IgA constant region (e.g., an IgA1 constant region or an IgA2 constant region), an IgD constant region, an IgE constant region, an IgG constant region (e.g., an IgG1 constant region, an IgG2 constant region, an IgG3 constant region, or an IgG4 constant region), or an IgM constant region.

[0189] In certain embodiments, an equivalent to an amino acid sequence comprises, consists essentially of, or even consists of a polypeptide having at least about 80% (including about 80%-100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) amino acid identity to the amino acid sequence. Additionally or alternatively, an equivalent to an amino acid sequence comprises, consists essentially of, or even consists of a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions to the complement of a polynucleotide encoding the amino acid sequence. In further embodiments, an equivalent to an amino acid sequence comprises, consists essentially of, or even consists of a polypeptide that is at least 80% (including about 80%-100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical to the amino acid sequence. In certain embodiments, an equivalent to an amino acid sequence (e.g., an antibody or fragment thereof, or any one or more of SEQ ID NOs: 1-14 and 24-26 disclosed herein, or a fragment thereof, including but not limited to, aa25 to aa144 of SEQ ID NO: 13, 24 or 26, aa21 to aa132 of SEQ ID NO: 14 or 25, aa21 to aa126 of SEQ ID NO: 27, aa25 to aa473 of SEQ ID NO: 13, 24 or 26, aa21 to aa239 of SEQ ID NO: 14 or 25, aa21 to aa233 of SEQ ID NO: 27) comprises, consists essentially of, or even consists of a polypeptide comprising one or more or all CDRs of that amino acid sequence.Additionally or alternatively, the polypeptide has at least about 80% (including about 80%-100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) amino acid identity to the amino acid sequence.

[0190] In certain embodiments, an equivalent to an amino acid sequence, e.g., an antibody, a fragment thereof, a complementarity determining region (CDR) thereof, or a CDR-containing polypeptide, lacks amino acid differences relative to the amino acid sequence in the CDR. However, an equivalent to an amino acid sequence, e.g., an antibody, a fragment thereof, a CDR thereof, or a CDR-containing polypeptide, contains one or more (e.g., without limitation, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acid differences compared to the amino acid sequence in a non-CDR region, provided that the tertiary arrangement of the CDR and / or the CDR is retained. In certain embodiments, an equivalent polypeptide to an amino acid sequence, e.g., an antibody, fragment thereof, CDR thereof, or CDR-containing polypeptide, has at least about 80% amino acid identity to the amino acid sequence (including about 80-100%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%), provided that the tertiary arrangement of the CDRs and / or the CDRs is retained.

[0191] Non-limiting examples of such non-CDR regions include framework regions (FR), constant regions, Fc regions, pFc' regions, constant heavy chain (CH) domains (e.g., CH1, CH2, CH3, or CH4), constant light chain (CL) domains, or hinge regions. In one embodiment, such amino acid differences may be conservative amino acid substitutions and / or do not alter the three-dimensional sequence of the antibody, fragment thereof, its CDRs, or CDR-containing polypeptide. In another embodiment, equivalents may include conservative amino acid substitutions of one or two amino acids within the boundaries of the CDR, e.g., at the amino terminus, carboxy terminus, or both, of the CDR.

[0192] In one aspect, one or more (e.g., any one, two, three, four, five, or six CDRs) of the CDRs of an antibody or fragment thereof disclosed herein are provided. In one embodiment, a set of CDRs is provided that comprises, or alternatively consists essentially of, or even further consists of, one or more (e.g., any one, two, three, four, five, or six CDRs) of the CDRs of an antibody or fragment thereof disclosed herein. In one embodiment, a set of CDRs is provided that comprises, or alternatively consists essentially of, or even further consists of CDRL1, CDRL2, and CDRL3 of the variable regions disclosed herein. In a further embodiment, a set of CDRs is provided that comprises, or alternatively consists essentially of, or even further consists of CDRH1, CDRH2, and CDRH3 of the variable regions disclosed herein. In still further embodiments, a set of CDRs is provided that comprises, or alternatively consists essentially of, or even consists of, CDRL1, CDRL2, and CDRL3 of a variable region disclosed herein, and CDRH1, CDRH2, CDRH3 of another variable region disclosed herein. In certain embodiments, the set of CDRs constitutes a paratope. Additionally or alternatively, the set of CDRs may be a paratope of a DNABII peptide (e.g., but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHIIn yet further embodiments, antibodies, fragments thereof, or equivalents of each thereof are provided that comprise, alternatively consist essentially of, or even consist of, any one or more CDRs disclosed herein. In yet further embodiments, antibodies, fragments thereof, or equivalents of each thereof are provided that comprise, alternatively consist essentially of, or even consist of, a set of CDRs disclosed herein.

[0193] In certain embodiments, the CDRs of SEQ ID NOs: 1-13 are exemplified in the table below. In certain embodiments, CDRH1 of any one of SEQ ID NOs: 1-6, 13, 24, or 26 comprises, consists essentially of, or even consists of amino acids (aa) 50 to 57 of SEQ ID NOs: 1-6, 13, 24, or 26, respectively. In certain embodiments, CDRH2 of any one of SEQ ID NOs: 1-6, 13, 24, or 26 comprises, consists essentially of, or even consists of amino acids (aa) 75 to 82 of SEQ ID NOs: 1-6, 13, 24, or 26, respectively. In certain embodiments, CDRH3 of any one of SEQ ID NOs: 1-6, 13, 24, or 26 comprises, consists essentially of, or even consists of amino acids (aa) 121 to 133 of SEQ ID NOs: 1-6, 13, 24, or 26, respectively. In certain embodiments, CDRL1 of any one of SEQ ID NOs: 7-9, 14, or 25 comprises, consists essentially of, or even consists of amino acids (aa) 47 to aa 57 of SEQ ID NOs: 7-9, 14, or 25, respectively. In certain embodiments, CDRL2 of any one of SEQ ID NOs: 7-9, 14, or 25 comprises, consists essentially of, or even consists of amino acids (aa) 75 to aa 77 of SEQ ID NOs: 7-9, 14, or 25, respectively. In certain embodiments, CDRL3 of any one of SEQ ID NOs: 7-9, 14, or 25 comprises, consists essentially of, or even consists of amino acids (aa) 114 to aa 122 of SEQ ID NOs: 7-9, 14, or 25, respectively. In certain embodiments, CDRL1 of any one of SEQ ID NOs: 10-12 or 27 comprises, consists essentially of, or even consists of amino acids (aa) 47 to aa 52 of SEQ ID NOs: 10-12 or 27, respectively. In certain embodiments, CDRL2 of any one of SEQ ID NOs: 10-12 or 27 comprises, consists essentially of, or even consists of amino acids (aa) 70 to aa 72 of SEQ ID NOs: 10-12 or 27, respectively.In certain embodiments, the CDRL3 of any one of SEQ ID NOs: 10-12 or 27 comprises, consists essentially of, or even consists of amino acids (aa) 109 to aa 116 of SEQ ID NOs: 10-12 or 27, respectively. [Table 10-1] [Table 10-2]

[0194] In certain embodiments, the CDRs identified in the following two tables are provided. [Table 11-1] [Table 11-2] [Table 11-3] [Table 11-4] [Table 11-5]

[0195] In certain embodiments, alternative CDRs are provided that are CDRs identified herein that further comprise an additional 1, 2, 3, 4, 5, 6, 7, or 8 amino acids at the amino or carboxy terminus, or both, of the corresponding variable region sequence. Additionally or alternatively, alternative CDRs are provided that are CDRs identified herein that have 1, 2, 3, 4, 5, 6, 7, or 8 amino acids truncated at the amino or carboxy terminus, or both, of the corresponding variable region sequence. For example, CDR1 of SEQ ID NO: 1 may be amino acids 50 to 57 of SEQ ID NO: 1. However, CDR1 of SEQ ID NO: 1 can also start at amino acid 42, or 43, or 44, or 45, or 46, or 47, or 48, or 49, or 50, or 51, or 52, or 53, or 54, or 55, or 56, or 57, or 58 of SEQ ID NO: 1. Additionally, CDR1 of SEQ ID NO: 1 ends at amino acid 49, or 50, or 51, or 52, or 53, or 54, or 55, or 56, or 57, or 58, or 59, or 60, or 61, or 62, or 63, or 64, or 65 of SEQ ID NO: 1, provided that CDR1 ends after its start. Additionally or alternatively, the CDRs are about 1, or alternatively about 2, or alternatively about 3, or alternatively about 4, or alternatively about 5, or alternatively about 6, or alternatively about 7, or alternatively about 8, or alternatively about 9, or alternatively about 10, or alternatively about 11, or alternatively about 12, or alternatively about 13, or alternatively about 14, or alternatively about 15 amino acids in length.

[0196] In certain embodiments, CDR2 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acid 71 to amino acid 85 of each of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR3 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acid 121 to amino acid 133 of each of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acid 71 to amino acid 81 of each of SEQ ID NOs: 7-9, respectively. In certain embodiments, CDR3 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acid 114 to amino acid 121 of each of SEQ ID NOs: 7-9, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acids 66 to 76 of each of SEQ ID NOs: 10-12, respectively. In certain embodiments, CDR3 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acids 109 to 115 of each of SEQ ID NOs: 10-12, respectively.

[0197] In certain embodiments, CDR1 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acids 50 to 57 of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acids 75 to 82 of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR3 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acids 121 and 122 of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR1 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acids 47 to 57 of SEQ ID NOs: 7-9, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acids 75 to 77 of SEQ ID NOs: 7-9, respectively. In certain embodiments, the CDR3 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acids 114 and 120 of SEQ ID NOs: 7-9, respectively. In certain embodiments, the CDR1 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acids 47 to 52 of SEQ ID NOs: 10-12, respectively. In certain embodiments, the CDR2 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acids 70 to 72 of SEQ ID NOs: 10-12, respectively. In certain embodiments, the CDR3 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acids 109 and 110 of SEQ ID NOs: 10-12, respectively.

[0198] In certain embodiments, CDR1 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acid 47 to amino acid 59 of each of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 1-6 comprises, consists essentially of, or even consists of amino acid 74 to amino acid 83 of each of SEQ ID NOs: 1-6, respectively. In certain embodiments, CDR1 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acid 44 to amino acid 59 of each of SEQ ID NOs: 7-9, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acid 74 to amino acid 81 of each of SEQ ID NOs: 7-9, respectively. In certain embodiments, CDR3 of any one of SEQ ID NOs: 7-9 comprises, consists essentially of, or even consists of amino acid 114 and 122 of each of SEQ ID NOs: 7-9, respectively. In certain embodiments, CDR1 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acid 44 to amino acid 54 of each of SEQ ID NOs: 10-12, respectively. In certain embodiments, CDR2 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acid 79 to amino acid 76 of each of SEQ ID NOs: 10-12, respectively. In certain embodiments, CDR3 of any one of SEQ ID NOs: 10-12 comprises, consists essentially of, or even consists of amino acid 109 and 116 of each of SEQ ID NOs: 10-12, respectively.

[0199] In one aspect, one or more of the variable regions of an antibody or fragment thereof disclosed herein, and / or equivalents of the variable regions thereof, are provided. In further embodiments, antibodies, fragments thereof, or equivalents of each thereof are provided that comprise, alternatively consist essentially of, or even consist of any one or any two or more of the variable regions and / or equivalents of the variable regions disclosed herein. Additionally or alternatively, one or more of the variable regions may be linked to a DNABII peptide (e.g., but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In certain embodiments, the variable region is selected from the following: amino acids (aa) 25 to aa 144 of SEQ ID NOs: 1-6, 13, 24, and 26, aa 21 to aa 132 of SEQ ID NOs: 7-9, 14, and 25, aa 21 to aa 126 of SEQ ID NOs: 10-12 or 27, amino acids 24 to 144 of SEQ ID NOs: 1-6, 13, 24, and 26, amino acids 20 to 132 of SEQ ID NOs: 7-12, 14, 25, and 27, and amino acids 20 to 126 of SEQ ID NOs: 7-12, 14, 25, and 27.

[0200] In further embodiments, the variable region or its equivalent is a variable region disclosed herein having an additional one, two, three, four, five, six, seven, or eight amino acids at its amino terminus, or both, in the sequence corresponding to a SEQ ID NO: provided herein. Additionally or alternatively, the variable region or its equivalent is a variable region disclosed herein having one, two, three, four, five, six, seven, or eight amino acids truncated at its amino terminus, or both, in the sequence corresponding to a SEQ ID NO: provided herein. For example, the variable region of SEQ ID NO: 1 may be amino acids 50 to 57 of SEQ ID NO: 1. However, the variable region for the variable region consisting of amino acids 24 to 144 of SEQ ID NO:1, or its equivalent, can also start at amino acid 16, or 17, or 18, or 19, or 20, or 21, or 22, or 23, or 24, or 25, or 26, or 27, or 28, or 29, or 30, or 31, or 32 of SEQ ID NO:1. Furthermore, the variable region for the variable region consisting of amino acids 24 to 144 of SEQ ID NO:1, or its equivalent, can end at amino acid 136, or 137, or 138, or 139, or 140, or 141, or 142, or 143, or 144, or 145, or 146, or 147, or 148, or 149, or 150, or 151, or 152 of SEQ ID NO:1, provided that the variable region ends after that start.Additionally or alternatively, the variable region is about 90 amino acids in length to about 200 amino acids in length, e.g., about 100 amino acids in length, or alternatively about 110 amino acids in length, or alternatively about 120 amino acids in length, or alternatively about 130 amino acids in length, or alternatively about 140 amino acids in length, or alternatively about 150 amino acids in length, or alternatively about 160 amino acids in length, or alternatively about 170 amino acids in length, or alternatively about 180 amino acids in length, or alternatively about 190 amino acids in length, or alternatively about 200 amino acids in length.

[0201] Additionally or alternatively, an equivalent to an antibody or fragment thereof may contain one or more (e.g., without limitation, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acid differences compared to an antibody or fragment thereof in a region other than the variable domain (referred to herein as a non-VH region). Such non-VH regions include, but are not limited to, the constant region, Fc region, pFc' region, constant heavy chain (CH) domain (e.g., CH1, CH2, CH3, or CH4), constant light chain (CL) domain, or hinge region. It will be understood by those skilled in the art that the antibodies, fragments thereof, or their respective equivalents disclosed herein can be further modified in non-VH regions (e.g., to increase assembly of the heavy chain with the light chain, to be conjugated directly or indirectly to a detectable marker or drug or a purification marker or drug, to increase or decrease complement activation, to enhance or decrease antibody-dependent cellular cytotoxicity (ADCC), or to increase or decrease immune cell activation and recruitment) to provide further equivalents.

[0202] In certain embodiments, equivalents further comprise up to 50, or alternatively up to 30, or alternatively up to 25, or alternatively up to 20, or alternatively up to 15, or alternatively up to 10, or alternatively up to 5 random amino acids at either the amino or carboxy terminus, or both. In certain embodiments, equivalents of an amino acid sequence comprise, consist essentially of, or even consist of an amino acid sequence truncated at the amino or carboxy terminus, or both, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, or 25 amino acids. Such additions and truncations may not alter the three-dimensional arrangement of the CDRs and / or the three-dimensional arrangement of the antibody, fragment thereof, CDR thereof, or CDR-containing peptide.

[0203] In certain embodiments, an antibody or fragment thereof comprises a signal peptide at the amino terminus of the VH and / or the amino terminus of the VL. In one embodiment, the VH signal peptide is different from the VL signal peptide. In another embodiment, the VH signal peptide is the same as the VL signal peptide. In a further embodiment, the signal peptide comprises, consists essentially of, or even consists of the amino acid sequence of amino acids 1-24 of SEQ ID NO: 1. In yet a further embodiment, the signal peptide comprises, consists essentially of, or even consists of the amino acid sequence of amino acids 1-20 of SEQ ID NO: 7. In certain embodiments, an equivalent to an antibody or fragment thereof comprises a signal peptide that is different from the signal peptide of that antibody, provided that the equivalent signal peptide targets the VH and / or VL to the same cellular location as the antibody signal peptide.

[0204] In certain embodiments, an equivalent to an antibody or fragment thereof retains at least 50% (e.g., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%) of the functional activity of the antibody or fragment, or improves one or more thereof. Such functional activities include, but are not limited to, DNABII peptides (e.g., but are not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4, NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI and (ii) a specificity, avidity, and / or affinity for a specific target protein (e.g., a tip region and / or a tail region comprising a nucleotide sequence, ...

[0205] In a further aspect, there is provided an antibody or fragment thereof that competes with the antibody or fragment thereof disclosed herein for binding to an epitope. The antibody or fragment thereof may be a polyclonal antibody, a monoclonal antibody, and / or a humanized antibody.

[0206] In one embodiment, the antibody is a bispecific antibody or a trispecific, tetraspecific, or pentaspecific antibody. In a further embodiment, the antibody is an IgA, IgD, IgE, IgG, or IgM antibody. In another embodiment, the antibody further comprises a constant region selected from an IgA, IgD, IgE, IgG, or IgM constant region. In a specific embodiment, the constant region is an IgG1 constant region. In another embodiment, provided herein are antibodies that compete with the antibodies disclosed herein for binding to an epitope. These can be identified using conventional techniques, such as competitive ELISA.

[0207] The antibodies disclosed herein may be polyclonal, monoclonal, or humanized. In one embodiment, the antibody binds to the "tip" region of a DNABII polypeptide, such as HU or IHF (e.g., IhfA and IhfB). In a further embodiment, the antibody binds to the "tail" region of a DNABII polypeptide, such as HU or IHF (e.g., IhfA and IhfB). As noted above, the present disclosure provides antigen-binding fragments. The antigen-binding fragment is any one of Fab, F(ab')2, Fab', scFv, or Fv, which can be prepared using conventional techniques known to those of skill in the art. In some embodiments of the antibodies provided herein, the antibody is a soluble Fab. In another embodiment, the present disclosure provides a Fab fragment of an antibody disclosed herein, wherein the antibody binds to the tip region of a DNABII peptide (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4). NTHI In one embodiment of the present disclosure, the DNABII is an IHF or HU peptide. In a specific embodiment, the DNABII is an IHF peptide.

[0208] As noted above, the present disclosure provides equivalents to the antibodies and antigen-binding fragments. Equivalents can include polypeptides encoded by polynucleotides that hybridize under high stringency conditions to a polypeptide having at least 80% amino acid identity to the polypeptide, or to the complement of a polynucleotide encoding the polypeptide.

[0209] In some embodiments of the antibodies provided herein, the antibody, fragment thereof, polypeptide, or CDR has a half maximal effective concentration (EC ) of less than 500 ng / mL, or alternatively less than 250 ng / mL, or alternatively less than 200 ng / mL, or alternatively less than 150 ng / mL, or alternatively less than 100 ng / mL, or alternatively less than 90 ng / mL, or alternatively less than 80 ng / mL, or alternatively less than 70 ng / mL, or alternatively less than 65 ng / mL, or alternatively less than 60 ng / mL, or alternatively less than 55 ng / mL, or alternatively less than 50 ng / mL, or alternatively less than 45 ng / mL, or alternatively less than 40 ng / mL, or alternatively less than 35 ng / mL, or alternatively less than 30 ng / mL. 50 ) and a DNABII protein (including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4 NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In a further embodiment, such EC 50 is determined using the ELISA method shown in the Examples.

[0210] In some embodiments of the antibodies provided herein, the antibody, fragment thereof, polypeptide, or CDR is -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, or 10 -12 Equilibrium constant K less than M D and a DNABII protein (including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4 NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI(including In one embodiment, the antibody, fragment thereof, polypeptide or CDR has a K of less than 1000 nM, or alternatively less than 900 nM, or alternatively less than 800 nM, or alternatively less than 700 nM, or alternatively less than 600 nM, or alternatively less than 500 nM, or alternatively less than 400 nM, or alternatively less than 300 nM, or alternatively less than 200 nM, or alternatively less than 100 nM, or alternatively less than 90 nM, or alternatively less than 80 nM, or alternatively less than 70 nM, or alternatively less than 60 nM, or alternatively less than 50 nM, or alternatively less than 40 nM, or alternatively less than 30 nM, or alternatively less than 20 nM, or alternatively less than 15 nM, or alternatively less than 10 nM, or alternatively less than 9 nM, or alternatively less than 8 nM, or alternatively less than 7 nM, or alternatively less than 6 nM, or alternatively less than 5 nM, or alternatively less than 4 nM, or alternatively less than 3 nM, or alternatively less than 2 nM, or alternatively less than 1 nM. D In one embodiment, such a K D is determined using surface plasmon resonance (SPR) methods as shown in the Examples. In some embodiments of the antibodies provided herein, the antigen-binding site specifically binds to a DNABII protein.

[0211] In some embodiments of the antibodies provided herein, the antibody, fragment thereof, polypeptide, or CDR has a molecular weight of 1.0E-02s -1 Less than or equal to 9.0E-03s -1 Less than or equal to 8.0E-03s -1 Less than or equal to 7.0E-03s -1 Less than or equal to 6.0E-03s -1 Less than or equal to 5.0E-03s -1 Less than or equal to 4.0E-03s -1 Less than or equal to 3.0E-03s -1 Less than or equal to 2.0E-03s -1 Less than or equal to 1.0E-03s -1 Less than or equal to 9.0E-04s -1Less than or equal to 8.0E-04s -1 Less than or equal to 7.0E-04s -1 Less than or equal to 6.0E-04s -1 Less than or equal to 5.0E-04s -1 Less than or equal to 4.0E-04s -1 Less than or equal to 3.0E-04s -1 Less than or equal to 2.0E-04s -1 Less than or equal to 1.0E-04s -1 Less than or equal to 9.0E-05s -1 Less than or equal to 8.0E-05s -1 Less than or equal to 7.0E-05s -1 Less than or equal to 6.0E-05s -1 Less than or equal to 5.0E-05s -1 Less than or equal to 4.0E-05s -1 Less than or equal to 3.0E-05s -1 Less than or equal to 2.0E-05s -1 Less than or equal to 1.0E-05s -1 Less than K off and a DNABII protein (including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4 NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, such K off is determined using the surface plasmon resonance (SPR) method shown in the examples.

[0212] In some embodiments of the antibodies provided herein, the antibody, fragment thereof, polypeptide, or CDR has a nucleotide sequence similar to or greater than 9.0E-02M. -1 s -1 Less than or equal to 8.0E-02M -1 s -1 Less than or equal to 7.0E-02M -1 s -1Less than or equal to 6.0E-02M -1 s -1 Less than or equal to 5.0E-02M -1 s -1 Less than or equal to 4.0E-02M -1 s -1 Less than or equal to 3.0E-02M -1 s -1 Less than or equal to 2.0E-02M -1 s -1 Less than or equal to 1.0E-02M -1 s -1 Less than or equal to 9.0E-03M -1 s -1 Less than or equal to 8.0E-03M -1 s -1 Less than or equal to 7.0E-03M -1 s -1 Less than or equal to 6.0E-03M -1 s -1 Less than or equal to 5.0E-03M -1 s -1 Less than or equal to 4.0E-03M -1 s -1 Less than or equal to 3.0E-03M -1 s -1 Less than or equal to 2.0E-03M -1 s -1 Less than or equal to 1.0E-03M -1 s -1 Less than or equal to 9.0E-04M -1 s -1 Less than or equal to 8.0E-04M -1 s -1 Less than or equal to 7.0E-04M -1 s -1 Less than or equal to 6.0E-04M -1 s -1 Less than or equal to 5.0E-04M -1 s -1 Less than or equal to 4.0E-04M -1 s -1 Less than or equal to 3.0E-04M -1 s -1Less than or equal to 2.0E-04M -1 s -1 Less than or equal to 1.0E-04M -1 s -1 Less than or equal to 9.0E-05M -1 s -1 Less than or equal to 8.0E-05M -1 s -1 Less than or equal to 7.0E-05M -1 s -1 Less than or equal to 6.0E-05M -1 s -1 Less than or equal to 5.0E-05M -1 s -1 Less than or equal to 4.0E-05M -1 s -1 Less than or equal to 3.0E-05M -1 s -1 Less than or equal to 2.0E-05M -1 s -1 Less than or equal to 1.0E-05M -1 s -1 Less than K on and a DNABII protein (including but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4 NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, such K on is determined using the surface plasmon resonance (SPR) method shown in the examples.

[0213] In some embodiments of the present invention, IhfA5-mIhfB4 NTHI Binding constant K for chip chimeric peptide (in 1 / M) A is about 3E+05 to about 2E+08. Ais about 3E+05 to about 1E+08, or alternatively about 2E+05 to about 1E+08, or alternatively about 1E+05 to about 1E+08, or alternatively about 1E+06 to about 1E+08, or alternatively about 1E+07 to about 1E+08, or alternatively about 1E+04 to about 1E+09, or alternatively about 1E+05 to about 1E+09, or alternatively about 1E+06 to about 1E+09, or alternatively about 1E+07 to about 1E+09, or alternatively about 1E+08 to about 1E+09, or alternatively about 1E+04 to about 1E+09, or alternatively about 1E+03 to about 1E+10.

[0214] In another embodiment, IhfA5-mIhfB4 NTHI Dissociation constant K for chip chimeric peptide (M) D is about 5E-09 to about 3E-06, or alternatively about 1E-09 to about 1E-06, or alternatively about 1E-08 to about 1E-05, or alternatively about 1E-07 to about 1E-05, or alternatively about 1E-06 to about 1E-05, or alternatively about 1E-09 to about 1E-08, or alternatively about 1E-08 to about 1E-07, or alternatively about 1E-9 to about 1E-08, or alternatively about 1E-10 to about 1E-09, or alternatively about 1E-11 to about 1E-10.

[0215] In one embodiment, IhfA3-IhfB2 NTHI K for tail chimeric peptide (at 1 / M) A is about 7E+06 to about 2E+09, or alternatively about 1E+05 to about 1E+08, or alternatively about 1E+06 to about 1E+08, or alternatively about 1E+07 to about 1E+08, or alternatively about 1E+04 to about 1E+09, or alternatively about 1E+05 to about 1E+09, or alternatively about 1E+06 to about 1E+09, or alternatively about 1E+07 to about 1E+09, or alternatively about It is 1E+08 to about 1E+09, or about 1E+04 to about 1E+09, or alternatively about 1E+03 to about 1E+10, or alternatively about 1E+03 to about 1E+11, or alternatively about 1E+03 to about 1E+12, or alternatively about 1E+09 to about 1E+10, or alternatively about 1E+10 to about 1E+11, or alternatively about 1E+11 to about 1E+12.

[0216] In another embodiment, IhfA3-IhfB2 NTHI K for tail chimeric peptide (M) D is about 6E-10 to about 2E-07, or alternatively about 1E-09 to about 1E-06, or alternatively about 1E-08 to about 1E-05, or alternatively about 1E-07 to about 1E-05, or alternatively about 1E-06 to about 1E-05, or alternatively about 1E-09 to about 1E-08, or alternatively about 1E-08 to about 1E-07, or alternatively about 1E-9 to about 1E-08, or alternatively about 1E-10 to about 1E-09, or alternatively about 1E-11 to about 1E-10, or alternatively about 1E-11 to about 1E-12.

[0217] In some embodiments of the antibodies provided herein, the antibody comprises a chip region of a DNABII protein (including, but not limited to, the chip region of IHF or HU, the chip region of IHFA or IHFB, and / or the chip-chimeric peptide IhfA5-mIhfB4). NTHI In some embodiments of the antibodies provided herein, the antibody, fragment, polypeptide, or CDR thereof that binds to the tail region of a DNABII protein (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2) reduces biofilm biomass in vitro by at least about 10%, or alternatively at least about 15%, or alternatively at least about 20%, or alternatively at least about 25%, or alternatively at least about 30%, or alternatively at least about 35%, or alternatively at least about 40%, or alternatively at least about 45%, or alternatively at least about 50%, or alternatively at least about 55%, or alternatively at least about 60%, or alternatively at least about 65%, or alternatively at least about 70%, or alternatively at least about 75%, or alternatively at least about 80%, or alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 95%. NTHI(including) reduces biofilm biomass in vitro by less than about 1%, or alternatively less than about 2%, or alternatively less than about 3%, or alternatively less than about 4%, or alternatively less than about 5%, or alternatively less than about 6%, or alternatively less than about 7%, or alternatively less than about 8%, or alternatively less than about 9%, or alternatively less than about 10%, or alternatively less than about 12%, or alternatively less than about 15%, or alternatively less than about 20%, or alternatively less than about 25%, or alternatively less than about 30%, or alternatively less than about 35%, or alternatively less than about 40%, or alternatively less than about 45%, or alternatively less than about 50%. In one embodiment, such a change in biomass is determined using the methods set forth in Example 3 or 4.

[0218] In some embodiments of the antibodies provided herein, the antibody comprises a chip region of a DNABII protein (including, but not limited to, the chip region of IHF or HU, the chip region of IHFA or IHFB, and / or the chip-chimeric peptide IhfA5-mIhfB4). NTHIor alternatively at least about 95%, or alternatively at least about 96%, or alternatively at least about 97%, or alternatively at least about 98%, or alternatively at least about 99%, or alternatively at least about 10%, or alternatively at least about 15%, or alternatively at least about 20%, or alternatively at least about 25%, or alternatively at least about 30%, or alternatively at least about 35%, or alternatively at least about 40%, or alternatively at least about 45%, or alternatively at least about 50%, or alternatively at least about 55%, or alternatively at least about 60%, or alternatively at least about 65%, or alternatively at least about 70%, or alternatively at least about 75%, or alternatively at least about 80%, or alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 91%, or alternatively at least about 92%, or alternatively at least about 93%, or alternatively at least about 94%, or alternatively at least about 95%, or alternatively at least about 96%, or alternatively at least about 97%, or alternatively at least about 98%, or alternatively at least about 99%. Some embodiments of the antibodies provided herein comprise a tail region of a DNABII protein (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI (including) reduces bacterial load in a subject by less than about 1%, or alternatively less than about 2%, or alternatively less than about 3%, or alternatively less than about 4%, or alternatively less than about 5%, or alternatively less than about 6%, or alternatively less than about 7%, or alternatively less than about 8%, or alternatively less than about 9%, or alternatively less than about 10%, or alternatively less than about 12%, or alternatively less than about 15%, or alternatively less than about 20%, or alternatively less than about 25%, or alternatively less than about 30%, or alternatively less than about 35%, or alternatively less than about 40%, or alternatively less than about 45%, or alternatively less than about 50%. In one embodiment, such a change in bacterial load is determined using the methods set forth in the Examples.

[0219] In some embodiments of the antibodies provided herein, the antibody comprises a chip region of a DNABII protein (including, but not limited to, the chip region of IHF or HU, the chip region of IHFA or IHFB, and / or the chip-chimeric peptide IhfA5-mIhfB4). NTHI or alternatively at least about 95%, or alternatively at least about 96%, or alternatively at least about 97%, or alternatively at least about 98%, or alternatively at least about 99%, reduces middle ear obstruction in a subject with otitis media (OM) by at least about 10%, or alternatively at least about 15%, or alternatively at least about 20%, or alternatively at least about 25%, or alternatively at least about 30%, or alternatively at least about 35%, or alternatively at least about 40%, or alternatively at least about 45%, or alternatively at least about 50%, or alternatively at least about 55%, or alternatively at least about 60%, or alternatively at least about 65%, or alternatively at least about 70%, or alternatively at least about 75%, or alternatively at least about 80%, or alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 91%, or alternatively at least about 92%, or alternatively at least about 93%, or alternatively at least about 94%, or alternatively at least about 95%, or alternatively at least about 96%, or alternatively at least about 97%, or alternatively at least about 98%, or alternatively at least about 99%. Some embodiments of the antibodies provided herein comprise a tail region of a DNABII protein (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI(including) reduces middle ear obstruction in a subject with otitis media (OM) by less than about 1%, or alternatively less than about 2%, or alternatively less than about 3%, or alternatively less than about 4%, or alternatively less than about 5%, or alternatively less than about 6%, or alternatively less than about 7%, or alternatively less than about 8%, or alternatively less than about 9%, or alternatively less than about 10%, or alternatively less than about 12%, or alternatively less than about 15%, or alternatively less than about 20%, or alternatively less than about 25%, or alternatively less than about 30%, or alternatively less than about 35%, or alternatively less than about 40%, or alternatively less than about 45%, or alternatively less than about 50%. In one embodiment, such changes in middle ear obstruction are determined in an experimental OM model using the methods set forth in the Examples.

[0220] In some embodiments of the antibodies provided herein, the antibody comprises a chip region of a DNABII protein (including, but not limited to, the chip region of IHF or HU, the chip region of IHFA or IHFB, and / or the chip-chimeric peptide IhfA5-mIhfB4). NTHIor at least about 10%, or alternatively at least about 15%, or alternatively at least about 20%, or alternatively at least about 25%, or alternatively at least about 30%, or alternatively at least about 35%, or alternatively at least about 40%, or alternatively at least about 45%, or alternatively at least about 50%, or alternatively at least about 55%, or alternatively at least about 60%, or alternatively at least about 65%, or alternatively at least about 70%, or alternatively at least about 75%, or alternatively or at least about 80%, or alternatively at least about 85%, or alternatively at least about 90%, or alternatively at least about 91%, or alternatively at least about 92%, or alternatively at least about 93%, or alternatively at least about 94%, or alternatively at least about 95%, or alternatively at least about 96%, or alternatively at least about 97%, or alternatively at least about 98%, or alternatively at least about 99%, reducing the relative mucosal biofilm score and / or biomass score in a subject having a mucosal biofilm (e.g., having OM). In one embodiment, the tip region of a DNABII protein (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip-chimeric peptide IhfA5-mIhfB4) is / are used. NTHI In some embodiments of the antibodies provided herein, the antibody, fragment, polypeptide, or CDR that binds to the tail region of a DNABII protein (including, but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2) reduces the relative mucosal biofilm score and / or biomass score in a subject with a mucosal biofilm (e.g., with OM) by at least about 0.5, or alternatively at least about 1, or alternatively at least about 1.5, or alternatively at least about 2, or alternatively at least about 2.5, or alternatively at least about 3, or alternatively at least about 3.5, or alternatively at least about 4. NTHIIn some embodiments of the antibodies provided herein, the antibody, fragment, polypeptide, or CDR thereof that binds to the tail region of a DNABII protein (including but not limited to, the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2) reduces the relative mucosal biofilm score and / or biomass score in a subject with a mucosal biofilm (e.g., with OM) by less than about 1%, or alternatively less than about 2%, or alternatively less than about 3%, or alternatively less than about 4%, or alternatively less than about 5%, or alternatively less than about 6%, or alternatively less than about 7%, or alternatively less than about 8%, or alternatively less than about 9%, or alternatively less than about 10%, or alternatively less than about 12%, or alternatively less than about 15%, or alternatively less than about 20%, or alternatively less than about 25%, or alternatively less than about 30%, or alternatively less than about 35%, or alternatively less than about 40%, or alternatively less than about 45%, or alternatively less than about 50%. NTHI (including) reduces the relative mucosal biofilm score and / or biomass score in a subject having a mucosal biofilm (e.g., having OM) by less than about 0.1, or alternatively less than about 0.2, or alternatively less than about 0.3, or alternatively less than about 0.4, or alternatively less than about 0.5, or alternatively less than about 0.6, or alternatively less than about 0.7, or alternatively less than about 0.8, or alternatively less than about 0.9, or alternatively less than about 1, or alternatively less than about 1.5, or alternatively less than about 2, or alternatively less than about 2.5. In one embodiment, such scores are determined using the methods set forth in the Examples.

[0221] In certain embodiments, the DNABII protein is HU or IHF. In further embodiments, the DNABII protein is IhfA, IhfB, or both. In still further embodiments, the antibody, fragment thereof, polypeptide, or CDR is selected from the tip or tail regions of DNABII proteins (including, but not limited to, the tip region of IHF or HU, the tip region of IHFA or IHFB, the tip-chimeric peptide IhfA5-mIhfB4 NTHI , the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2 NTHI In one embodiment, the antibody, fragment thereof, polypeptide or CDR binds to IhfA5-mIhfB4 NTHI In another embodiment, the antibody, fragment thereof, polypeptide, or CDR binds to the IhfA3-IhfB2 chimeric peptide. NTHI Binds to the tail chimeric peptide.

[0222] In some embodiments of the antibodies provided herein, the antibody is a soluble Fab.

[0223] In some embodiments of the antibodies provided herein, the HC and LC variable domain sequences are components of the same polypeptide chain. In some embodiments of the antibodies provided herein, the HC and LC variable domain sequences are components of different polypeptide chains.

[0224] In some embodiments of the antibodies provided herein, the antibody is a full-length antibody.

[0225] In some embodiments of the antibodies provided herein, the antibodies are chimeric or humanized.

[0226] In some embodiments of the antibodies provided herein, the antibody comprises an Fc domain. In some embodiments of the antibodies provided herein, the antibody is a non-human animal, such as a rat, sheep, cow, dog, cat, or rabbit antibody. In some embodiments of the antibodies provided herein, the antibody is a human or humanized antibody, or is non-immunogenic in humans.

[0227] In some embodiments of the antibodies provided herein, the antibodies comprise human antibody framework regions. Examples of framework regions that can be fused to the LC and HC sequences are known in the art, and examples of such are provided in SEQ ID NOS: 15-23, or their respective equivalents.

[0228] In other embodiments, one or more amino acid residues in the CDRs of the antibodies provided herein are substituted with another amino acid. The substitutions can be "conservative" in the sense that they are substitutions within the same amino acid family. Naturally occurring amino acids can be divided into four families, and conservative substitutions are made within these families: 1) Amino acids with basic side chains: lysine, arginine, histidine. 2) Amino acids with acidic side chains: aspartic acid, glutamic acid 3) Amino acids with uncharged polar side chains: asparagine, glutamine, serine, threonine, tyrosine. 4) Amino acids with nonpolar side chains: glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan, cysteine.

[0229] In another embodiment, one or more amino acid residues are added to or deleted from one or more CDRs of the antibody, such additions or deletions occurring at the N- or C-terminus of the CDR or at a position within the CDR.

[0230] By varying the amino acid sequence of the antibody CDR by adding, deleting, or substituting amino acids, various effects can be achieved, such as increasing the binding affinity to the target antigen.

[0231] It should be appreciated that the antibodies of the present disclosure containing such altered CDR sequences will still bind to DNABII proteins with the same specificity and sensitivity profile as the disclosed antibodies, which can be tested by binding assays such as ELISA or SPR.

[0232] In a further aspect, the antibodies are characterized as being both immunodominant and immunoprotective, as determined using appropriate assays and screens.

[0233] In another embodiment, the antibody can be modified by conventional techniques, which in one embodiment can increase the half-life of the antibody, for example, PEGylation, PEG mimetic, polysialylation, HESylation, or increased glycosylation.

[0234] The antibodies and antigen-binding fragments can further comprise a detectable marker or a purification marker. Antibodies and their derivatives

[0235] The present disclosure provides an antibody that binds to and / or specifically recognizes and binds to an isolated polypeptide for use in the methods disclosed herein. The antibody can be any of the various antibodies described herein, including, but not limited to, polyclonal antibodies, monoclonal antibodies, chimeric antibodies, human antibodies, veneered antibodies, diabodies, humanized antibodies, antibody derivatives, recombinant humanized antibodies, or equivalents (e.g., derivatives) or fragments thereof. In one embodiment, the fragment comprises, or alternatively consists essentially of, or even further consists of, the CDRs of the antibody. In one embodiment, the antibody is detectably labeled or further comprises a detectable label conjugated thereto.

[0236] Hybridoma cell lines producing the monoclonal antibodies disclosed herein are also provided. Further provided herein are compositions comprising, alternatively consisting essentially of, or even consisting of, one or more of the above-described embodiments. Further provided are polynucleotides encoding the amino acid sequences of the antibodies and fragments, as well as methods for recombinantly producing or chemically synthesizing antibody polypeptides and fragments thereof. Antibody polypeptides can be produced in eukaryotic or prokaryotic cells, or by other methods known in the art and described herein.

[0237] Examples of CDR sequences include, but are not limited to, a heavy chain variable region of an antibody or fragment thereof comprising, consisting essentially of, or further consisting of a polypeptide encoded by the following polynucleotide sequence:

[0238] Antibodies can be produced using conventional techniques known in the art and are fully described in the literature.There are several methodologies for producing polyclonal antibodies.For example, polyclonal antibodies are typically produced by immunizing suitable mammals, including but not limited to chickens, goats, guinea pigs, hamsters, horses, mice, rats, and rabbits.Antigens are injected into mammals, and B lymphocytes are induced to produce immunoglobulins specific to the antigen.Immunoglobulins can also be purified from mammalian serum.

[0239] Variations of this methodology include modifications of adjuvants, route and site of administration, injection volume per site, and number of sites per animal for optimal production and humane treatment of animals. For example, adjuvants are typically used to improve or enhance the immune response to antigens. Most adjuvants provide an injection site antigen depot, allowing stow release of antigens to the draining lymph nodes. Other adjuvants include surfactants that promote enrichment of protein antigen molecules over a large surface area and immunostimulatory molecules. Non-limiting examples of adjuvants for polyclonal antibody production include Freund's adjuvant, Ribi adjuvant system, and Titermax. Polyclonal antibodies can be produced using methods known in the art, some of which are described in U.S. Patent Nos. 7,279,559; 7,119,179; 7,060,800; 6,709,659; 6,656,746; 6,322,788; 5,686,073; and 5,670,153.

[0240] Monoclonal antibodies can be produced using conventional hybridoma technology, which is known in the art and well described in the literature. For example, suitable immortal cell lines (e.g., myeloma cell lines including, but not limited to, Sp2 / 0, Sp2 / 0-AG14, NSO, NS1, NS2, AE-1, L.5, P3X63Ag8,653, Sp2 SA3, Sp2 MAI, Sp2 SS1, Sp2 SA5, U397, MIA 144, ACT IV, MOLT4, DA-1, JURKAT, WEHI, K-562, COS, RAJI, NIH 313, HL-60, MLA 144, NAMAIWA, NEURO 2A, CHO, PerC.6, YB2 / O, etc.) or heteromyeloma, its fusion products, or any cells or fusion cells derived therefrom, or any other suitable cell line known in the art (see the following web addresses, e.g., atcc.org, Hybridomas are produced by fusing the cell lines (see cell lines at lifetech.com, last accessed November 26, 2007) with antibody-producing cells, including but not limited to, isolated or cloned spleen, peripheral blood, lymph, tonsil or other immune or B-cell containing cells, or any other cells, expressing heavy or light chain constant or variable or framework or CDR sequences as endogenous or heterologous nucleic acid, recombinant or endogenous, viral, bacterial, algal, prokaryotic, amphibian, insect, reptilian, fish, mammalian, rodent, equine, ovine, caprine, ovine, primate, eukaryotic, genomic DNA, cDNA, rDNA, mitochondrial DNA or RNA, chloroplast DNA or RNA, hnRNA, mRNA, tRNA, single-, double- or triple-stranded, hybridized, etc., or any combination thereof. Antibody-producing cells can also be obtained from the peripheral blood, or in certain embodiments, from the spleen or lymph nodes, of humans or other suitable animals immunized with the antigen of interest, and then screened for the activity of interest. Any other suitable host cells can also be used to express heterologous or endogenous nucleic acid encoding an antibody, specified fragment, or variant thereof, of the present disclosure.The fused cells (hybridomas) or recombinant cells can be isolated using selective culture conditions or other suitable known methods and cloned by limiting dilution or cell sorting, or other known methods.

[0241] Other suitable methods for producing or isolating antibodies of the required specificity can be used, including, but not limited to, recombinant antibody selection from peptide or protein libraries (e.g., but not limited to, bacteriophage, ribosomal, oligonucleotide, cDNA, or other display libraries; e.g., as available from various commercial vendors, such as MorphoSys (Martinsreid / Planegg, Del.), BioInvent (Lund, Sweden), and Affitech (Oslo, Norway)) using methods known in the art. Methods known in the art are described in the patent literature, some of which include U.S. Patent Nos. 4,704,692; 5,723,323; 5,763,192; 5,814,476; 5,817,483; 5,824,514; and 5,976,862. An alternative method involves the production of transgenic animals (e.g., SCID mice, Nguyen et al. (1977) Microbiol. Immunol. 41:901-907 (1997); Sandhu et al. (1996) Crit. Rev. Biotechnol. 16:95-118; Eren et al. (1998) Mumma 1999) capable of producing a repertoire of human antibodies, as known in the art and / or as described herein. 93:154-161). Such techniques include ribosome display (Wanes et al. (1997) Proc. Natl. Acad. Sci. USA 94:4937-4942; Hanes et al. (1998) Proc. Natl. Acad. Sci. USA 95:14130-14135); single-cell antibody production techniques (e.g., selected lymphocyte antibody methods); lymphocyte antibody method) (“SLAM”) (U.S. Patent No. 5,627,052; Wen et al. (1987) J. Immunol 17:887-892; Babcook et al. (1996) Proc. Natl. Acad. Sci. USA 93:7843-7848); gel microdroplets and flow cytometry (Powell et al. (1990) Biotechnol. 8:333-337; One Cell Systems (Cambridge, Mass.); Gray et al. (1995) J. Imm. Meth. 182:155-163; and Kenny et al. (1995) Bio. Technol. 13:787-790); B cell selection (Steenbakkers et al. (1994) Molec. Biol. Reports 19:125-134).

[0242] Antibody derivatives of the present disclosure can also be prepared by delivering a polynucleotide encoding an antibody disclosed herein to a suitable host, e.g., providing a transgenic animal or mammal, such as a goat, cow, horse, sheep, etc., that produces the antibody in its milk. Such methods are known in the art and are described, for example, in U.S. Patent Nos. 5,827,690; 5,849,992; 4,873,316; 5,849,992; 5,994,616; 5,565,362; and 5,304,489.

[0243] The term "antibody derivatives" includes post-translational modifications to the linear polypeptide sequence of an antibody or fragment. For example, U.S. Patent No. 6,602,684 (B1) describes methods for the production of modified glycoforms of antibodies, including whole antibody molecules, antibody fragments, or fusion proteins containing a region equivalent to the Fc region of an immunoglobulin, with enhanced Fe-mediated cytotoxicity, and the glycoproteins thus produced.

[0244] The antibodies disclosed herein also include derivatives that are modified by the covalent attachment of any type of molecule to the antibody, provided that the covalent attachment does not prevent the antibody from generating an anti-idiotypic response. Antibody derivatives include, but are not limited to, antibodies modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, linkage to cellular ligands or other proteins, etc. Moreover, derivatives can contain one or more non-classical amino acids.

[0245] Antibody derivatives can also be prepared by providing transgenic plants and cultured plant cells (e.g., but not limited to, tobacco, corn, and duckweed) that deliver the polynucleotides disclosed herein and produce the antibody, specified portion, or variant in plant parts or cells cultured therefrom. For example, Cramer et al. (1999) Curr. Top. Microbol. Immunol. 240:95-118 and references cited therein describe the production of transgenic tobacco leaves that express large amounts of recombinant proteins, for example, using inducible promoters. Transgenic corn has been used to express mammalian proteins at commercial production levels with biological activity equivalent to that produced in other recombinant systems or purified from natural sources. See, for example, Hood et al. (1999) Adv. Exp. Med. Biol. 464:127-147 and references cited therein. Antibody derivatives have also been produced in large quantities from transgenic plant seeds containing antibody fragments, such as single-chain antibodies (scFv), including tobacco seeds and potato tubers. See, e.g., Conrad et al. (1998) Plant Mol. Biol. 38:101-109 and the references cited therein. Thus, transgenic plants can also be used to produce antibodies according to known methods.

[0246] Antibody derivatives can also be produced, for example, by adding exogenous sequences to modify immunogenicity or to reduce, enhance, or modify binding, affinity, on-rate, off-rate, avidity, specificity, half-life, or any other suitable characteristic. Generally, some or all of the non-human or human CDR sequences are maintained, while the non-human sequences of the variable and constant regions are replaced with variable or constant regions from human or other amino acids or other isotypes.

[0247] In general, the CDR residues are directly and most substantially involved in influencing antigen binding. Antibody humanization or engineering can be performed using any known method, including, but not limited to, those described in U.S. Patent Nos. 5,723,323; 5,976,862; 5,824,514; 5,817,483; 5,814,476; 5,763,192; 5,723,323; 5,766,886; 5,714,352; 6,204,023; 6,180,370; 5,693,762; 5,530,101; 5,585,089; 5,225,539; and 4,816,567.

[0248] The chimeric, humanized, or primatized antibodies of the present disclosure can be prepared based on the sequence of a reference monoclonal antibody prepared using standard molecular biology techniques. Using standard molecular biology techniques, DNA encoding heavy and light chain immunoglobulins can be obtained from the desired hybridoma and engineered to contain non-reference (e.g., human) immunoglobulin sequences. For example, to produce a chimeric antibody, a mouse variable region can be linked to a human constant region using methods known in the art (U.S. Patent No. 4,816,567). To produce a humanized antibody, a mouse CDR region can be inserted into a human framework using methods known in the art (U.S. Patent No. 5,225,539 and U.S. Patent Nos. 5,530,101; 5,585,089; 5,693,762; and 6,180,370). Similarly, to create primatized antibodies, murine CDR regions can be inserted into a primate framework using methods known in the art (WO93 / 02108 and WO99 / 55369).

[0249] Techniques for producing partially to fully human antibodies are known in the art, and any such technique can be used. In one embodiment, fully human antibody sequences are produced in transgenic mice engineered to express human heavy and light chain antibody genes. Several strains of such transgenic mice have been generated that are capable of producing different classes of antibodies. B cells from transgenic mice producing the desired antibody can be fused to generate hybridoma cell lines for sustained production of the desired antibody (see, e.g., Russell et al. (2000) Infection and Immunity April 2000:1820-1826; Gallo et al. (2000) European J. of Immun. 30:534-540; Green (1999) J. of Immun. Methods 231:11-23; Yang et al. (1999A) J. of Leukocyte Biology 66:401-410; Yang (1999B) Cancer Research 59(6):1236-1243; Jakobovits (1998) Advanced Drug Reviews 31:33-42; Green and Jakobovits (1998) J. Exp. Med. 188(3):483-495;Jakobovits (1998) Exp. Opin. Invest. Drugs 7(4):607-614;Tsuda et al. (1997) Genomics 42:413-421;Sherman-Gold (1997) Genetic Engineering News 17(14);Mendez et al. (1997) Nature Genetics 15:146-156;Jakobovits (1996) Weir's Handbook of Experimental Immunology, The Integrated Immune System Vol. IV, 194.1-194.7;Jakobovits (1995) Current Opinion in Biotechnology 6:561-566;Mendez et al. (1995) Genomics 26:294-307; Jakobovits (1994) Current Biology 4(8):761-763; Arbones et al. (1994) Immunity 1(4):247-260; Jakobovits (1993) Nature 362(6417):255-258; Jakobovits et al. (1993) Proc. Natl. Acad. Sci. USA 90(6):2551-2555; and U.S. Patent No. 6,075,181).

[0250] The antibodies disclosed herein can also be modified to produce chimeric antibodies. Chimeric antibodies are antibodies in which various domains of the heavy and light chains of an antibody are encoded by DNA from more than one species. See, for example, U.S. Patent No. 4,816,567.

[0251] Alternatively, the antibodies disclosed herein can be modified to create veneered antibodies. Veneering antibodies are antibodies in which the outer surface amino acid residues of an antibody from one species have been judiciously replaced, or "veneered," with amino acid residues from a second species, so that the antibody from the first species is not immunogenic in the second species, thereby reducing the antibody's immunogenicity. Because the antigenicity of a protein depends primarily on the nature of its surface, replacing exposed residues that differ from those normally found in another mammalian species can reduce the immunogenicity of the antibody. Such judicious outer surface residue replacements should have little or no effect on internal domains or interdomain contacts. Thus, as a result of changes limited to variable region framework residues, ligand binding properties should be unaffected. This process is referred to as "veneering," because only the outer surface, or skin, of the antibody is altered, leaving supporting residues undisturbed.

[0252] Procedures for "veneering" utilize available sequence data for human antibody variable domains compiled by Kabat et al. (1987) Sequences of Proteins of Immunological Interest, 4th ed., Bethesda, Md., National Institutes of Health, updates to this database, and other accessible U.S. and foreign databases (both nucleic acid and protein). Non-limiting examples of methods used to generate veneered antibodies include EP 519596; U.S. Pat. No. 6,797,492; Padlan et al. al. (1991) Mol. Immunol. 28(4-5):489-498.

[0253] The term "antibody derivative" also includes "diabodies," which are small antibody fragments with two antigen-binding sites, comprising a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (see, e.g., EP 404,097; WO 93 / 11161; and Hollinger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448). By using a linker that is too short to allow pairing between the two domains in the same chain, the domains are forced to pair with complementary domains in another chain, creating two antigen-binding sites (see also U.S. Pat. No. 6,632,926, Chen et al., which discloses antibody variants with one or more amino acids inserted into the hypervariable region of the parent antibody and having binding affinity for a target antigen that is at least about two-fold stronger than the binding affinity of the parent antibody for the antigen).

[0254] The term "antibody derivatives" further includes engineered antibody molecules, fragments and single domains, such as scFvs, dAbs, nanobodies, minibodies, unibodies and affibodies (& Hudson (2005) Nature Biotech 23(9):1126-36; U.S. Patent Application Publication No. 2006 / 0211088; PCT International Application Publication No. WO2007 / 059782; U.S. Patent No. 5,831,012).

[0255] The term "antibody derivative" further includes "linear antibodies." Procedures for generating linear antibodies are known in the art and are described in Zapata et al. (1995) Protein Eng. 8(10):1057-1062. Briefly, such antibodies comprise a pair of tandem Ed segments (V) that form a pair of antigen-binding regions. H -C H 1-VH-C H 1) Linear antibodies can be bispecific or monospecific.

[0256] The antibodies disclosed herein can be recovered and purified from recombinant cell culture by known methods, including, but not limited to, protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, and lectin chromatography. High performance liquid chromatography ("HPLC") can also be used for purification.

[0257] The antibodies of the present disclosure include naturally occurring purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from eukaryotic hosts, including, for example, yeast, higher plant, insect, and mammalian cells, or alternatively from prokaryotic hosts, as described above. Several antibody production systems are described in Birch & Radner (2006) Adv. Drug Delivery Rev. 58: 671-685.

[0258] If the antibody being tested binds to a protein or polypeptide, the antibody being tested and the antibody provided by the present disclosure are equivalent. It is also possible to determine, without undue experimentation, whether an antibody has the same specificity as an antibody disclosed herein by determining whether the antibody being tested prevents the antibody disclosed herein from binding to the protein or polypeptide with which it normally reacts. If the antibody being tested competes with the antibody disclosed herein, as indicated by reduced binding by the monoclonal antibody disclosed herein, the two antibodies may bind to the same or closely related epitopes. Alternatively, the antibody disclosed herein can be preincubated with the protein with which it normally reacts to determine whether the antibody being tested is inhibited in its ability to bind to the antigen. If the antibody being tested is inhibited, it almost certainly has the same or closely related epitope specificity as the antibody disclosed herein.

[0259] The term "antibody" is also intended to include antibodies of all immunoglobulin isotypes and subclasses. Monoclonal antibodies of a particular isotype can be prepared directly by selection from the initial fusion, or can be subsequently prepared from parent hybridomas secreting monoclonal antibodies of a different isotype using sib selection techniques to isolate class-switched variants using the procedures described in Steplewski et al. (1985) Proc. Natl. Acad. Sci. USA 82:8653 or Spira et al. (1984) J. Immunol. Methods 74:307. Alternatively, recombinant DNA techniques can be used.

[0260] Isolation of other monoclonal antibodies having the specificity of the monoclonal antibodies described herein can also be achieved by one skilled in the art by raising anti-idiotypic antibodies. Herlyn et al. (1986) Science 232:100. Anti-idiotypic antibodies are antibodies that recognize unique determinants present on the monoclonal antibody of interest.

[0261] In some embodiments disclosed herein, it is useful to detectably or therapeutically label an antibody.Suitable labels are described above.Methods for conjugating antibodies to these agents are known in the art.For illustrative purposes only, antibodies can be labeled with detectable moieties such as radioactive atoms, chromophores, fluorophores, etc.Such labeled antibodies can be used in diagnostic techniques in vivo or in isolated test samples.

[0262] Coupling of an antibody to a low molecular weight hapten can increase the sensitivity of the antibody in an assay. The hapten can then be specifically detected using a second reaction. For example, it is common to use haptens such as biotin, which reacts with avidin, or dinitrophenol, pyridoxal, and fluorescein, which can react with specific anti-hapten antibodies. See Harlow and Lane (1988) supra.

[0263] The variable regions of the antibodies of the present disclosure can be modified by mutating amino acid residues within the VH and / or VL CDR1, CDR2, and / or CDR3 regions to improve one or more binding characteristics (e.g., affinity) of the antibody. Mutations can be introduced by site-directed mutagenesis or PCR-mediated mutagenesis, and the effect on desired antibody binding or other functional properties can be assessed in appropriate in vitro or in vivo assays. In certain embodiments, conservative modifications are introduced, typically altering no more than one, two, three, four, or five residues within the CDR regions. Mutations can be amino acid substitutions, additions, or deletions.

[0264] Framework modifications can be made to the antibody to reduce immunogenicity, for example, by "backmutating" one or more framework residues to the corresponding germline sequence.

[0265] In addition, the antibodies disclosed herein can be engineered to contain modifications within the Fc region to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular cytotoxicity. Such modifications include, but are not limited to, altering the number of cysteine ​​residues in the hinge region to facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody (U.S. Patent No. 5,677,425), and mutating amino acids in the Fc hinge region to decrease the biological half-life of the antibody (U.S. Patent No. 6,165,745).

[0266] Furthermore, the antibodies disclosed herein can be chemically modified. The glycosylation of an antibody can be altered, for example, by modifying one or more glycosylation sites within the antibody sequence to increase the affinity of the antibody for an antigen (U.S. Patent Nos. 5,714,350 and 6,350,861). Alternatively, to increase antibody-dependent cell-mediated cytotoxicity, hypofucosylated antibodies with reduced amounts of fucosyl residues or increased bisected GlcNac structures can be obtained by expressing the antibody in a host cell with altered glycosylation machinery (Shields, RL et al. (2002) J. Biol. Chem. 277:26733-26740; Umana et al. (1999) Nat. Biotech. 17:176-180).

[0267] The antibodies disclosed herein can be pegylated to increase biological half-life by reacting the antibody or fragment thereof with polyethylene glycol (PEG) or a reactive ester or aldehyde derivative of PEG under conditions such that one or more PEG groups become attached to the antibody or antibody fragment. Antibody PEGylation can be carried out by an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any of the PEG forms that have been used to derivatize other proteins, such as mono(C1-C10) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. The antibody to be pegylated can be an aglycosylated antibody. Methods for pegylating proteins are known in the art and can be applied to the antibodies disclosed herein (EP 0154316 and EP 0401384).

[0268] Moreover, antibodies can be chemically modified by conjugating or fusing the antigen-binding region of the antibody to a serum protein such as human serum albumin to increase the half-life of the resulting molecule. Such approaches are described, for example, in EP 0322094 and EP 0486525.

[0269] The antibodies or fragments thereof of the present disclosure can be conjugated to diagnostic agents and used diagnostically, for example, to monitor the onset or progression of disease and determine the effectiveness of a given treatment regimen. Examples of diagnostic agents include enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron-emitting metals using various positron emission tomography techniques, and non-radioactive paramagnetic metal ions. Detectable substances can be coupled or conjugated to antibodies or fragments thereof directly or indirectly via a linker using techniques known in the art. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase. Examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin. An example of a luminescent material includes luminol. Examples of bioluminescent materials include luciferase, luciferin, and aequorin. Examples of suitable radioactive materials include: 125 I, 131I, Indium-111, Lutetium-171, Bismuth-212, Bismuth-213, Astatine-211, Copper-62, Copper-64, Copper-67, Yttrium-90, Iodine-125, Iodine-131, Phosphorus-32, Phosphorus-33, Scandium-47, Silver-111, Gallium-67, Praseodymium-142, Samarium-153, Terbium-161, Dysprosium-166, Holmium-166 , rhenium-186, rhenium-188, rhenium-189, lead-212, radium-223, actinium-225, iron-59, selenium-75, arsenic-77, strontium-89, molybdenum-99, rhodium-1105, palladium-109, praseodymium-143, promethium-149, erbium-169, iridium-194, gold-198, gold-199, and lead-211. Monoclonal antibodies can be indirectly conjugated to radioactive metal ions by use of a bifunctional chelator covalently linked to the antibody. Chelating agents can be attached through amino groups (Meares et al. (1984) Anal. Biochem. 142:68-78); sulfhydral groups of amino acid residues (Koyama (1994) Chem. Abstr. 120:217-262); and carbohydrate groups (Rodwell et al. (1986) PNAS USA 83:2632-2636; Quadri et al. (1993) Nucl. Med. Biol. 20:559-570).

[0270] Additionally, the antibodies or fragments thereof of the present disclosure can be conjugated to a therapeutic agent. Suitable therapeutic agents include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, antimetabolites (methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5-fluorouracil, decarbazine, hydroxyurea, aspartame, thiazolinone ... Laginase, gemcitabine, cladribine, etc.), alkylating agents (mechlorethamine, thioepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, cisplatin, and other platinum derivatives such as carboplatin), antibiotics (dactinomycin (formerly actinomycin), bleomycin, daunorubicin (formerly daunomycin), doxorubicin, idarubicin, mithramycin, mitomycin, mitoxantrone, plicamycin, anthramycin (AMC), etc.), diphtheria toxin and related molecules (diphtheria A chain and its active fragments, and hybrid molecules), ricin toxin (ricin A or deglycosylated Ricin A chain toxin, etc.), cholera toxin, Shiga toxin-like toxins (SLT-I, SLT-II, SLT-IIV), LT toxin, C3 toxin, Shiga toxin, pertussis toxin, tetanus toxin, soybean Bowman-Birk protease inhibitor, Pseudomonas exotoxin, alorin, saporin, modeccin, geranin, abrin A chain, modeccin A chain, alpha-sarcin, Aleuritesfordii protein, dianthin protein, Phytolacca americana proteins (PAPI, PAPII and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrietocin, phenomycin, enomycin toxin and mixed toxins.

[0271] Additional suitable conjugated molecules include ribonucleases (RNases), DNase I, antisense nucleic acids, inhibitory RNA molecules such as siRNA molecules, immunostimulatory nucleic acids, aptamers, ribozymes, triplex-forming molecules, and external guide sequences. Aptamers are small nucleic acids ranging from 15 to 50 bases in length that fold into defined secondary and tertiary structures, such as stem-loops or G-quartets, and can bind small molecules such as ATP (U.S. Patent No. 5,631,146) and theophylline (U.S. Patent No. 5,580,737), as well as large molecules such as reverse transcriptase (U.S. Patent No. 5,786,462) and thrombin (U.S. Patent No. 5,543,293). Ribozymes are nucleic acid molecules that can catalyze chemical reactions, either intramolecularly or intermolecularly. Ribozymes typically cleave nucleic acid substrates by recognizing and binding to the target substrate followed by cleavage. Triplex-forming functional nucleic acid molecules can interact with double-stranded or single-stranded nucleic acids by forming triplexes, where three DNA strands form a complex that relies on both Watson-Crick and Hoogsteen base pairing. Triplex molecules can bind to target regions with high affinity and specificity.

[0272] Functional nucleic acid molecules can act as effectors, inhibitors, modulators, and stimulators of a specific activity possessed by a target molecule, or they can possess a de novo activity independent of any other molecule.

[0273] Therapeutic agents can be linked to antibodies directly or indirectly using any of a number of available methods. For example, agents can be attached at the hinge region of a reduced antibody component via disulfide bond formation using cross-linking agents such as N-succinyl 3-(2-pyridyldithio)propionate (SPDP), or through carbohydrate moieties in the Fc region of the antibody (Yu et al. 1994 Int. J. Cancer 56: 244; Upeslacis et al., "Modification of Antibodies by Chemical Methods," 2004). in Monoclonal antibodies: principles and applications, Birch et al. (eds.), pages 187-230 (Wiley-Liss, Inc. 1995);Price, “Production and Characterization of Synthetic Peptide-Derived Antibodies,” in Monoclonal antibodies: Production, engineering and clinical application, Ritter et al. (eds.), pages 60-84 (Cambridge University Press 1995)).

[0274] Techniques for conjugating therapeutic agents to antibodies are well known (Amon et al. "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy," in Monoclonal Antibodies And Cancer Therapy; Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985);Hellstrom et al. ”Antibodies For Drug Delivery,” in Controlled Drug Delivery (2nd Ed.); Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987);Thorpe ”Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review,” in Monoclonal Antibodies ’84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985);”Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody in Cancer Therapy,” in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985)およびThorpe et al. ”The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates,” (1982) Immunol. Rev. 62:119-58)。

[0275] The antibodies or antigen-binding regions thereof disclosed herein can be linked to another functional molecule, such as another antibody or a receptor ligand, to generate bispecific or multispecific molecules that bind to at least two or more different binding sites or target molecules. Linkage of the antibody to one or more other binding molecules, such as another antibody, antibody fragment, peptide, or binding mimetic, can be achieved, for example, by chemical coupling, genetic fusion, or noncovalent association. Multispecific molecules can further include a third binding specificity in addition to the first and second target epitopes.

[0276] Bispecific and multispecific molecules can be prepared using methods known in the art. For example, each binding unit of a bispecific molecule can be produced separately and then conjugated to each other. When the binding molecule is a protein or peptide, various coupling or cross-linking agents can be used for covalent conjugation. Examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl S-acetyl-thioacetate (SATA), 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-I-carboxylate (sulfo-SMCC) (Karpovsky et al. (1984) J. Exp. Med. 160:1686; Liu et al. (1985) Proc. Natl. Acad. Sci. USA 82:8648). When the binding molecule is an antibody, it can be conjugated via sulfhydryl bonds in the C-terminal hinge regions of the two heavy chains.

[0277] The antibodies of the present disclosure, or fragments thereof, can be linked to a moiety that is toxic to the cells to which the antibody is bound, to form "depleting" antibodies. These antibodies are particularly useful in applications where it is desirable to deplete NK cells.

[0278] The antibodies disclosed herein may also be attached to solid supports, which are particularly useful for immunoassays or purification of the target antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene.

[0279] Antibodies can also be bound to many different carriers. Thus, the present disclosure also provides compositions containing antibodies and other active or inactive substances. Examples of well-known carriers include glass, polystyrene, polypropylene, polyethylene, dextran, nylon, amylase, natural and modified cellulose, polyacrylamide, agarose, and magnetite. The nature of the carrier can be either soluble or insoluble for the purposes disclosed herein. Those skilled in the art will know of other suitable carriers for binding monoclonal antibodies, or can ascertain such using routine experimentation.

[0280] In certain aspects, the present disclosure relates to antibodies or antigen-binding fragments that specifically recognize or bind to the tip or tail domain, tail fragment, or tip fragment of a DNABII protein or fragment thereof. The DNABII protein or fragment thereof may be an IHF or HU polypeptide.

[0281] Functional analysis using antibodies The antibodies disclosed herein can be used to purify the polypeptides disclosed herein and identify biologically equivalent polypeptides and / or polynucleotides.The antibodies disclosed herein can also be used to identify agents that alter the function of the polypeptides disclosed herein.Such antibodies include polyclonal antisera, monoclonal antibodies, and various reagents derived from such preparations, which are familiar to those skilled in the art and described above.

[0282] Antibodies that neutralize the activity of the proteins encoded by the identified genes can also be used in vivo and in vitro to demonstrate function by adding such neutralizing antibodies to in vivo and in vitro test systems, and are also useful as pharmaceutical agents for modulating the activity of the polypeptides disclosed herein.

[0283] Various antibody preparations can also be used in analytical methods such as ELISA assays or Western blots to demonstrate the expression of proteins encoded by the identified genes by test cells in vitro or in vivo. Using appropriate polyclonal antisera and samples derived from experimental samples, fragments of such proteins generated by protease degradation during metabolism can also be identified.

[0284] The antibodies disclosed herein can be used for vaccination or to boost vaccination, either alone or in combination with peptide or protein-based vaccines or dendritic cell-based vaccines.

[0285] Diagnostic and Therapeutic Methods Methods are provided for preventing, inhibiting, or competing for the binding of a DNABII polypeptide or protein to microbial DNA by contacting the DNABII polypeptide or protein, or microbial DNA, with an effective amount of one or more of the above-described agents, such as the antibodies, or antigen-binding fragments thereof, polypeptides, or CDRs disclosed herein, thereby preventing, inhibiting, or competing for the binding of the DNABII protein or polypeptide to the microbial DNA. The DNABII polypeptide can be an IHF or HU peptide. In one aspect, the antibody or antigen-binding fragment thereof binds to the tip region of the DNABII polypeptide (the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip chimeric peptide IhfA5-mIhfB4). NTHIIn one embodiment, an antibody or fragment thereof is provided that comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 24, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 25, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 1, 2, or 3, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 7, 8, or 9, or an equivalent of each thereof. In yet a further embodiment, the contacting is in vivo or in vitro.

[0286] In another embodiment, the antibody or antigen-binding fragment thereof for use in the methods disclosed herein comprises, consists essentially of, or even further consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising a sequence selected from the group of SEQ ID NO: 13, 24, or 26, or an equivalent of each thereof; and / or a light chain (LC) immunoglobulin variable domain sequence comprising a sequence selected from the group of SEQ ID NO: 14, 25, or 27, or an equivalent of each thereof.

[0287] In a further embodiment, one or more of the DNABII polypeptides and / or microbial DNA and / or antibodies or antigen-binding fragments thereof, and / or polypeptides or CDRs disclosed herein are detectably labeled, for example, with a radioisotope or a luminescent molecule that emits a signal when in close contact with each other. The contacting can be performed in vitro or in vivo. These methods can be combined with diagnostic methods, for example, to detect and / or monitor biofilm formation and / or destruction using one or more of the DNABII polypeptides and / or microbial DNA and / or antibodies or antigen-binding fragments thereof, and / or polypeptides or CDRs disclosed herein. In one embodiment, the diagnostic method involves detecting and / or detecting biofilm formation and / or destruction using a tail region or fragment of a DNABII polypeptide (e.g., the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2). NTHIIn a further aspect, the antibody or antigen-binding fragment thereof is detectably labeled. In one embodiment, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 26, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 27, or an equivalent thereof. In further embodiments, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 4, 5, or 6, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 10, 11, or 12, or an equivalent of each thereof.

[0288] In another aspect, a method of disrupting a microbial biofilm is provided by contacting the biofilm with an effective amount of one or more of the above-mentioned agents, such as an antibody or antigen-binding fragment thereof, polypeptide, or CDR, thereby disrupting the microbial biofilm. In one aspect, the antibody or antigen-binding fragment thereof binds to the tip region of a DNABII polypeptide (the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip chimeric peptide IhfA5-mIhfB4). NTHIThe DNABII polypeptide may be an IHF or HU peptide. In one embodiment, an antibody or fragment thereof is provided that comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence of SEQ ID NO: 24 or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence of SEQ ID NO: 25 or an equivalent thereof. In a further embodiment, the antibody or fragment thereof comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence selected from the group of SEQ ID NO: 1, 2, or 3, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence selected from the group of SEQ ID NO: 7, 8, or 9, or an equivalent thereof, In still further embodiments, the contacting is in vivo or in vitro.

[0289] In one aspect, microbial biofilms are produced by microorganisms that export DNABII polypeptides. The DNABII polypeptides can be IHF or HU peptides. In a further aspect, one or more of the antibody, antibody fragment, DNABII polypeptide, and microbial DNA are detectably labeled, for example, with a radioisotope or a luminescent molecule that emits a signal when they come into close contact with each other. The contacting can be carried out in vitro or in vivo. In one aspect, the agents are one or more antibodies and / or antigen-binding fragments that are the same or different from each other. In some embodiments, such antibodies or antigen-binding fragments are administered alone or in combination with each other, or with other agents other than antibodies, or even with further pharmaceutically effective agents, alone or in combination with a pharmaceutically acceptable carrier. These methods can be combined with diagnostic methods to detect and / or monitor the formation and / or destruction of biofilms. In one embodiment, the diagnostic method comprises the step of: NTHI In a further aspect, the antibody or antigen-binding fragment thereof is detectably labeled.

[0290] Also provided are methods of preventing the formation of or disrupting biofilms on a surface, comprising, consisting essentially of, or further consisting of treating a surface prone to or containing a biofilm with an effective amount of one or more of the antibodies or antigen-binding fragments thereof, polypeptides, or CDRs described herein, wherein the antibody or antigen-binding fragment thereof binds to the tip region of a DNABII peptide (the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip chimeric peptide IhfA5-mIhfB4). NTHIAlso provided are methods for binding to DNABII polypeptides, including but not limited to, IHF or HU peptides. In one embodiment, an antibody or fragment thereof is provided that comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence of SEQ ID NO: 26 or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence of SEQ ID NO: 27 or an equivalent thereof. In a further embodiment, the antibody or fragment thereof comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence selected from the group of SEQ ID NO: 4, 5, or 6, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, essentially consists of, or alternatively consists of the amino acid sequence selected from the group of SEQ ID NO: 10, 11, or 12, or an equivalent thereof,

[0291] In one embodiment, the antibody or antigen-binding fragment thereof comprises a detectable label. These methods can be combined with diagnostic methods to detect and / or monitor biofilm formation and / or destruction. In one embodiment, the diagnostic method comprises detecting the tail region or fragment of a DNABII polypeptide (e.g., the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2). NTHIIn a further aspect, the antibody or antigen-binding fragment thereof is detectably labeled. In one embodiment, an antibody or fragment thereof is provided that comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 26, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 27, or an equivalent thereof. In further embodiments, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 4, 5, or 6, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 10, 11, or 12, or an equivalent of each thereof.

[0292] Further provided herein is a method for detecting a biofilm in a subject, comprising, consisting essentially of, or consisting of administering to the subject an effective amount of one or more of the antibodies, or fragments thereof, polypeptides, or CDRs disclosed herein. In one embodiment, the diagnostic method comprises administering to the subject a tail region or fragment of a DNABII polypeptide (e.g., the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2). NTHIThe DNABII polypeptide can be an IHF or HU peptide. In a further aspect, the antibody or antigen-binding fragment thereof is detectably labeled. In one embodiment, an antibody or fragment thereof is provided that comprises a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively further consisting of the amino acid sequence of SEQ ID NO: 26 or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively further consisting of the amino acid sequence of SEQ ID NO: 27 or an equivalent thereof, or consisting essentially of, or alternatively further consisting of: In further embodiments, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 4, 5, or 6, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 10, 11, or 12, or an equivalent of each thereof.

[0293] Methods for preventing or disrupting biofilms in a subject are provided. The methods include administering to the subject a tip region of a DNABII peptide (e.g., a tip region of IHF or HU, a tip region of IHFA or IHFB, and / or a tip chimeric peptide IhfA5-mIhfB4). NTHIIn one aspect, a method of preventing or disrupting biofilms in a subject is provided, comprising, alternatively consisting of, or further consisting of administering to the subject an effective amount of one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein, and / or an effective amount of one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides, or CDRs. The DNABII peptide can be an IHF or HU peptide. In one aspect, the antibody or antigen-binding fragment thereof binds to the tip region of a DNABII polypeptide (the tip region of IHF or HU, the tip region of IHFA or IHFB, and / or the tip chimeric peptide IhfA5-mIhfB4). NTHIIn one embodiment, an antibody or fragment thereof is provided that comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 24, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 25, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 1, 2, or 3, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 7, 8, or 9, or an equivalent of each thereof. In yet a further embodiment, the contacting is in vivo or in vitro. The antibody or antigen-binding fragment thereof may be detectably labeled. These methods may be combined with diagnostic methods to detect and / or monitor biofilm formation and / or destruction. In one embodiment, the diagnostic method may involve the use of a tail region or fragment of a DNABII polypeptide (e.g., the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2). NTHI In a further aspect, the antibody or antigen-binding fragment is detectably labeled.

[0294]

[0013] Methods of treating a condition characterized by biofilm formation in a subject are provided, the methods including administering to the subject a biofilm comprising administering to the subject a tip region of a DNABII polypeptide (e.g., a tip region of IHF or HU, a tip region of IHFA or IHFB, and / or a tip chimeric peptide IhfA5-mIhfB4). NTHI In one aspect, a method is provided for preventing or treating a condition characterized by biofilm formation in a subject by administering to the subject an effective amount of one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein, and / or an effective amount of one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides, or CDRs. In another aspect, a method is provided for inhibiting, preventing, or treating a biofilm-producing microbial infection in a subject, comprising, or alternatively consisting of, or even further consisting of administering to the subject an effective amount of one or more of the antibodies, fragments thereof, polypeptides, or CDRs disclosed herein, and / or an effective amount of one or more polynucleotides or vectors encoding the antibodies, fragments thereof, polypeptides, or CDRs. The DNABII polypeptide can be an IHF or HU peptide. In one embodiment, the antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof that binds to a tip region of a DNABII polypeptide (e.g., a tip region of IHF or HU, a tip region of IHFA or IHFB, and / or a tip chimeric peptide IhfA5-mIhfB4 NTHIIn one embodiment, an antibody or fragment thereof is provided that comprises, essentially consists of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 24, or an equivalent thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of the amino acid sequence of SEQ ID NO: 25, or an equivalent thereof. In a further embodiment, the antibody or fragment thereof comprises, consists essentially of, or alternatively consists of a heavy chain (HC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 1, 2, or 3, or an equivalent of each thereof, and / or a light chain (LC) immunoglobulin variable domain sequence comprising, consisting essentially of, or alternatively consisting of an amino acid sequence selected from the group of SEQ ID NO: 7, 8, or 9, or an equivalent of each thereof. In yet a further embodiment, the contacting is in vivo or in vitro. These methods can be combined with diagnostic methods to detect and / or monitor biofilm formation and / or destruction. In one embodiment, the diagnostic method comprises the step of detecting a tail region or tail fragment of a DNABII polypeptide (e.g., the tail region of IHF or HU, the tail region of IHFA or IHFB, and / or the tail chimeric peptide IhfA3-IhfB2). NTHI In a further aspect, the antibody or antigen-binding fragment is detectably labeled.

[0295] In the above methods, the biofilm is derived from Gram-negative or Gram-positive biofilm-producing bacteria. Non-limiting examples of conditions are selected from the group consisting of chronic non-healing wounds, including venou...

Claims

[Claim 1] The invention described in the specification.

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