Neutralization and use of anti-influenza B antibodies

Abstract

To provide antibodies that have broad neutralizing activity against influenza B viruses, and uses of such antibodies.SOLUTION: The invention provides antibodies and antigen binding fragments thereof that are capable of binding to influenza B virus hemagglutinin (HA) and neutralizing influenza B viruses in two phylogenetically distinct lineages. In one embodiment, the antibody or antigen binding fragment is capable of binding to influenza B virus hemagglutinin and neutralizing influenza B viruses in Yamagata and Victoria lineages.SELECTED DRAWING: Figure 1

Description

[Technical Field] 【0001】 This invention relates to an antibody having broad neutralizing activity against influenza B virus and the use of such an antibody. [Background technology] 【0002】 Influenza viruses pose a significant threat to public health worldwide, causing annual influenza epidemics and occasional global pandemics. Seasonal influenza infections are associated with 200,000 to 500,000 deaths each year, particularly among infants, immunocompromised individuals, and the elderly. Mortality rates typically increase further throughout the season, especially with pandemic influenza outbreaks. There remains a significant unmet medical need for potent antiviral drugs to prevent and treat influenza infections, particularly in underserved populations. 【0003】 There are three types of influenza viruses: A, B, and C. The majority of influenza illnesses are caused by influenza A and B viruses (Thompson et al. (2004) JAMA.292:1333-1340; and Zhou et al. (2012) Clin Infect.Dis.54:1427-1436). The overall structure of influenza A, B, and C viruses is similar, and they include a viral envelope surrounding a central core. The viral envelope contains two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). HA mediates the binding of the virus to and entry into target cells, while NA is involved in the release of progeny viruses from infected cells. 【0004】 The HA protein has a trimer structure and contains three identical copies of a single polypeptide precursor, HA0, which are cleaved during proteolytic maturation into a pH-dependent metastable intermediate having a spherical head (HA1) and a stalk region (HA2) (Wilson et al. (1981) Nature. 289:366-373). The distal spherical head constitutes the majority of the HA1 structure and has a sialic acid binding pocket for viral entry and the major antigen domain. 【0005】 Influenza A viruses can be classified into subtypes based on genetic variations in the hemagglutinin (HA) and neuraminidase (NA) genes. Currently, in seasonal epidemics, the HA subtypes of influenza A H1 and H3 are mainly associated with human disease, while viruses encoding H5, H7, H9, and H10 are associated with sporadic human outbreaks resulting from direct transmission from animals. 【0006】 Unlike influenza A viruses, influenza B viruses were not divided into two subtypes based on surface glycoproteins and were classified as a single homogeneous group until the 1970s. Throughout the 1970s, influenza B viruses began to diverge into two antigenically distinguishable lineages, named Victoria and Yamagata, after their respective first representatives, B / Victoria / 2 / 87 and B / Yamagata / 16 / 88 (Biere et al. (2010) J Clin Microbiol. 48(4): 1425-7; doi: 10.1128 / JCM. 02116-09. Epub 2010 Jan 27). Influenza B viruses are limited to human infection, and both lineages contribute to annual epidemics. The incidence rate caused by influenza B virus is lower than that caused by influenza A H3N2, but higher than that caused by influenza A H1N1 (Zhou et al. (2012) Clin Infect.Dis.54:1427-1436). [Prior art documents] [Non-patent literature] 【0007】 [Non-Patent Document 1] Thompson et al. (2004) JAMA.292:1333-1340 [Non-Patent Document 2] Zhou et al.(2012)Clin Infect.Dis.54:1427-1436 [Non-Patent Document 3] Wilson et al. (1981) Nature.289:366-373 [Non-Patent Document 4] Biere et al. (2010) J Clin Microbiol.48(4):1425-7;doi:10.1128 / JCM.02116-09.Epub 2010 Jan 27 [Overview of the project] [Problems that the invention aims to solve] 【0008】 Neutralizing antibodies induced by influenza virus infection typically target the variable HA1 globular head to block viral receptor binding and are usually strain-specific. Broadly cross-reactive antibodies that neutralize one or more subtypes or lineages are rare. In recent years, several human antibodies capable of neutralizing multiple subtypes of both lineages of influenza B virus have been discovered (Dreyfus et al. (2012) Science.337(6100):1343-8; and Yasugi et al. (2013) PLoS Path.9(2):e1003150). While these antibodies recognize numerous influenza B viruses, they have limited applicability and efficacy, and do not neutralize any influenza A virus strains. To date, no available antibodies broadly neutralize or inhibit all influenza B virus infections or attenuate diseases caused by influenza B viruses. Therefore, there is a need to identify novel antibodies that provide protection against multiple influenza viruses. [Means for solving the problem] 【0009】 The invention described herein provides an isolated antibody or antigen-binding fragment thereof that binds to influenza B virus hemagglutinin (HA) and has the ability to neutralize influenza B virus in two phylogenetically distinct lineages. In one embodiment, the antibody or antigen-binding fragment thereof binds to influenza B virus hemagglutinin and has the ability to neutralize influenza B virus in both Yamagata and Victoria lineages. Examples of Yamagata lineages, but not limited to, include B / AA / 94 (ca B / Ann Arbor / 2 / 94(yamagata)); B / YSI / 98 (ca B / Yamanashi / 166 / 98(yamagata)); B / JHB / 99 (ca B / Johannesburg / 5 / 99(yamagata)); B / SC / 99 (B / Sichuan / 379 / 99(yamagata)); and B / FL / 06 (B / Florida / 4 / 2006(yamagata)). Examples of Victoria bloodlines, though not limited to them, include B / BJ / 97 (ca B / Beijing / 243 / 97(victoria)), B / HK / 01 (B / Hong Kong / 330 / 2001(victoria)), B / MY / 04 (B / Malaysia / 2506 / 2004(victoria)), and B / BNE / 08 (ca B / Brisbane / 60 / 2008(victoria)). 【0010】 In another embodiment, the present invention provides isolated antibodies or antigen-binding fragments thereof that bind to influenza B virus hemagglutinin and have the ability to neutralize influenza B virus in pre-divergent strains. As used herein, the term “pre-divergent” refers to influenza B strains identified before the divergence of influenza B into the Yamagata and Victoria lineages. Examples of pre-divergent influenza B strains include, but are not limited to, B / Lee / 40 (B / Lee / 40); B / AA / 66 (ca B / Ann Arbor / 1 / 66); and B / HK / 72 (B / Hong Kong / 5 / 72). 【0011】 In one embodiment, the antibody or antigen-binding fragment is in an EC range of approximately 1 μg / ml to approximately 50 μg / ml of the antibody. 50 The antibody binds to the influenza B virus. In another embodiment, the antibody or antigen-binding fragment is used to neutralize the influenza B virus in a microneutralization assay at a 50% inhibitory concentration (IC) in the range of approximately 0.001 μg / ml to approximately 5 μg / ml of the antibody, as described in Example 3. 50 It has a neutralizing efficacy expressed as μg / ml. Other microneutralization assays are also described in Example 1. 【0012】 In one embodiment, the antibody has the ability to bind to influenza A virus hemagglutinin. Influenza A virus hemagglutinin includes subtype 1 and subtype 2 hemagglutinin. Subtypes of influenza A virus group 1 include H1, H2, H5, H6, H8, H9, H11, H12, H13, H16 and their variants. Subtypes of influenza A virus group 2 include H3, H4, H7, H10, H14, and H15 and their variants. In one embodiment, the antibody has the ability to bind to one or more subtypes of influenza A virus group 1. In another embodiment, the antibody has the ability to bind to one or more subtypes of influenza A virus group 2. In one embodiment, the antibody has the ability to bind to subtype H9 of influenza A virus group 1. In one embodiment, the present invention provides an isolated antibody or antigen-binding fragment thereof that binds to influenza B virus hemagglutinin (HA) and influenza A virus hemagglutinin (HA) and has the ability to neutralize at least one Yamagata lineage influenza B virus; at least one Victoria lineage influenza B virus; at least one subtype of influenza A virus, or a combination thereof. In one embodiment, the present invention provides an isolated antibody or antigen-binding fragment thereof that binds to influenza B virus hemagglutinin (HA) and hemagglutinin (HA) of one or more subtypes 1 of influenza A virus, and has the ability to neutralize at least one Yamagata lineage influenza B virus or at least one Victoria lineage influenza B virus and at least one subtype 1 of influenza A virus.In one embodiment, the present invention provides an isolated antibody or antigen-binding fragment thereof that binds to hemagglutinin (HA) of influenza B virus and hemagglutinin (HA) of influenza A virus subtype H9, and has the ability to neutralize at least one Yamagata lineage influenza B virus; at least one Victoria lineage influenza B virus; influenza A virus subtype H9, or a combination thereof. 【0013】 In one embodiment, the antibody or antigen-binding fragment is in an EC range of approximately 1 μg / ml to approximately 50 μg / ml of the antibody. 50 In another embodiment, the antibody or antigen-binding fragment neutralizes influenza A virus in a microneutralization assay, with the antibody in an IC50 range of approximately 0.01 μg / ml to approximately 5 μg / ml. 50 It has. 【0014】 In one embodiment, the antibody or fragment thereof of the present invention binds to the spherical head region of HA and neutralizes infection by influenza B virus from two phylogenetically distinct lineages. In another embodiment, the antibody or antigen-binding fragment thereof binds to the spherical head region of HA and neutralizes infection by influenza B virus from both Yamagata and Victoria lineages. The antibody of the present invention is an antibody that binds to the spherical head of anti-influenza B HA and exhibits a broader applicability or superior neutralizing activity against influenza B virus compared to known anti-influenza B antibodies. 【0015】 In one embodiment, the antibody or its antigen-binding fragment is (a) HCDR-1 of SEQ ID NO: 3, HCDR-2 of SEQ ID NO: 4, HCDR-3 of SEQ ID NO: 5, LCDR-1 of SEQ ID NO: 8, LCDR-2 of SEQ ID NO: 9, and LCDR-3 of SEQ ID NO: 10; (b) HCDR-1 of SEQ ID NO: 13, HCDR-2 of SEQ ID NO: 14, HCDR-3 of SEQ ID NO: 15, LCDR-1 of SEQ ID NO: 18, LCDR-2 of SEQ ID NO: 19, LCDR-3 of SEQ ID NO: 20; (c) HCDR-1 of SEQ ID NO: 23, HCDR-2 of SEQ ID NO: 24, HCDR-3 of SEQ ID NO: 25, LCDR-1 of SEQ ID NO: 28, LCDR-2 of SEQ ID NO: 29, and LCDR-3 of SEQ ID NO: 30; (d) HCDR-1 of SEQ ID NO: 33, HCDR-2 of SEQ ID NO: 34, HCDR-3 of SEQ ID NO: 35, LCDR-1 of SEQ ID NO: 38, LCDR-2 of SEQ ID NO: 39, and LCDR-3 of SEQ ID NO: 40; (e) HCDR-1 of SEQ ID NO: 43, HCDR-2 of SEQ ID NO: 44, HCDR-3 of SEQ ID NO: 45, LCDR-1 of SEQ ID NO: 48, LCDR-2 of SEQ ID NO: 49, and LCDR-3 of SEQ ID NO: 50; (f) HCDR-1 of SEQ ID NO: 53, HCDR-2 of SEQ ID NO: 54, HCDR-3 of SEQ ID NO: 55, LCDR-1 of SEQ ID NO: 58, LCDR-2 of SEQ ID NO: 59, and LCDR-3 of SEQ ID NO: 60; (g) HCDR-1 of SEQ ID NO: 63, HCDR-2 of SEQ ID NO: 64, HCDR-3 of SEQ ID NO: 65, LCDR-1 of SEQ ID NO: 68, LCDR-2 of SEQ ID NO: 69, and LCDR-3 of SEQ ID NO: 70; (h) HCDR-1 of SEQ ID NO: 75, HCDR-2 of SEQ ID NO: 76, HCDR-3 of SEQ ID NO: 77, LCDR-1 of SEQ ID NO: 83, LCDR-2 of SEQ ID NO: 84, and LCDR-3 of SEQ ID NO: 85; (i) HCDR-1 of SEQ ID NO: 91, HCDR-2 of SEQ ID NO: 92, HCDR-3 of SEQ ID NO: 93, LCDR-1 of SEQ ID NO: 99, LCDR-2 of SEQ ID NO: 100, and LCDR-3 of SEQ ID NO: 101; (j) HCDR-1 of SEQ ID NO: 107, HCDR-2 of SEQ ID NO: 108, HCDR-3 of SEQ ID NO: 109, LCDR-1 of SEQ ID NO: 115, LCDR-2 of SEQ ID NO: 116, and LCDR-3 of SEQ ID NO: 117; (k) HCDR-1 of SEQ ID NO: 121, HCDR-2 of SEQ ID NO: 122, HCDR-3 of SEQ ID NO: 123, LCDR-1 of SEQ ID NO: 124, LCDR-2 of SEQ ID NO: 125, and LCDR-3 of SEQ ID NO: 126; (l) HCDR-1 of SEQ ID NO: 127, HCDR-2 of SEQ ID NO: 128, HCDR-3 of SEQ ID NO: 129, LCDR-1 of SEQ ID NO: 130, LCDR-2 of SEQ ID NO: 131, and LCDR-3 of SEQ ID NO: 132; (m) HCDR-1 of SEQ ID NO: 133, HCDR-2 of SEQ ID NO: 134, HCDR-3 of SEQ ID NO: 135, LCDR-1 of SEQ ID NO: 136, LCDR-2 of SEQ ID NO: 137, and LCDR-3 of SEQ ID NO: 138; (n) HCDR-1 of SEQ ID NO: 139, HCDR-2 of SEQ ID NO: 140, HCDR-3 of SEQ ID NO: 141, LCDR-1 of SEQ ID NO: 142, LCDR-2 of SEQ ID NO: 143, and LCDR-3 of SEQ ID NO: 144; (o) HCDR-1 of SEQ ID NO: 145, HCDR-2 of SEQ ID NO: 146, HCDR-3 of SEQ ID NO: 147, LCDR-1 of SEQ ID NO: 148, LCDR-2 of SEQ ID NO: 149, and LCDR-3 of SEQ ID NO: 150; (p) HCDR-1 of SEQ ID NO: 78, HCDR-2 of SEQ ID NO: 79, HCDR-3 of SEQ ID NO: 80, LCDR-1 of SEQ ID NO: 86, LCDR-2 of SEQ ID NO: 87, and LCDR-3 of SEQ ID NO: 88; (q) HCDR-1 of SEQ ID NO: 94, HCDR-2 of SEQ ID NO: 95, HCDR-3 of SEQ ID NO: 96, LCDR-1 of SEQ ID NO: 102, LCDR-2 of SEQ ID NO: 103, and LCDR-3 of SEQ ID NO: 104; (r) HCDR-1 of SEQ ID NO: 110, HCDR-2 of SEQ ID NO: 111, HCDR-3 of SEQ ID NO: 112, LCDR-1 of SEQ ID NO: 118, LCDR-2 of SEQ ID NO: 119, and LCDR-3 of SEQ ID NO: 120; and (s) A set of six CDRs as described in any one of (a) to (r), comprising one or more amino acid substitutions, deletions, or insertions. Six CDR sets selected from: HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2, and LCDR-3. 【0016】 In another embodiment, the antibody or its antigen-binding fragment is (a) VH of SEQ ID NO: 2 and VL of SEQ ID NO: 7, (b) VH of sequence number 12 and VL of sequence number 17, (c) VH of SEQ ID NO: 22 and VL of SEQ ID NO: 27, (d) VH of SEQ ID NO: 32 and VL of SEQ ID NO: 37, (e) VH of sequence number 42 and VL of sequence number 47, (f) VH of sequence number 52 and VL of sequence number 57, (g) VH of SEQ ID NO: 62 and VL of SEQ ID NO: 67, (h) VH of sequence number 74 and VL of sequence number 82, (i) VH of SEQ ID NO: 90 and VL of SEQ ID NO: 98, (j) VH of SEQ ID NO: 106 and VL of SEQ ID NO: 114 Each VH and / or VL selected from there has at least 75%, 80%, 85%, 90%, 95%, or 100% identity with the VH and / or VL having at least 75%, 80%, 85%, 90%, 95%, or 100% identity. 【0017】 In further specific embodiments, the antibody or its antigen-binding fragment is (a) VH of SEQ ID NO: 2 and VL of SEQ ID NO: 7, (b) VH of sequence number 12 and VL of sequence number 17, (c) VH of SEQ ID NO: 22 and VL of SEQ ID NO: 27, (d) VH of SEQ ID NO: 32 and VL of SEQ ID NO: 37, (e) VH of sequence number 42 and VL of sequence number 47, (f) VH of sequence number 52 and VL of sequence number 57, (g) VH of SEQ ID NO: 62 and VL of SEQ ID NO: 67, (h) VH of sequence number 74 and VL of sequence number 82, (i) VH of SEQ ID NO: 90 and VL of SEQ ID NO: 98, (j) VH of SEQ ID NO: 106 and VL of SEQ ID NO: 114 Includes VH and VL selected from. 【0018】 In one embodiment, the present invention provides an antibody or an antigen-binding fragment thereof that binds to B-type influenza virus hemagglutinin (HA) and has the ability to neutralize B-type influenza viruses in two phylogenetically distinct lineages. The antibody has the VH amino acid sequence of SEQ ID NO: 71, where Xaa1 of SEQ ID NO: 71 is Val or Glu; Xaa2 of SEQ ID NO: 71 is Leu or Phe; Xaa3 of SEQ ID NO: 71 is Ser or Thr; Xaa4 of SEQ ID NO: 71 is Leu or Ser; Xaa5 of SEQ ID NO: 71 is Ser or Thr; Xaa6 of SEQ ID NO: 71 is Met or Thr; Xaa7 of SEQ ID NO: 71 is Phe or Tyr; Xaa8 of SEQ ID NO: 71 is His or Gln; Xaa9 of SEQ ID NO: 71 is Ser or Asn; Xaa 10 of SEQ ID NO: 71 is Arg or Lys; and Xaa 11 of SEQ ID NO: 71 is Ala or Thr; and has the VL amino acid sequence of SEQ ID NO: 72, where Xaa1 of SEQ ID NO: 72 is Phe or Tyr. In one embodiment, Xaa9 of SEQ ID NO: 71 is Ser. In another embodiment, Xaa4 of SEQ ID NO: 71 is Leu. In yet another embodiment, Xaa1 of SEQ ID NO: 71 is Glu; Xaa5 of SEQ ID NO: 71 is Thr; Xaa6 of SEQ ID NO: 71 is Thr; Xaa7 of SEQ ID NO: 71 is Tyr; Xaa8 of SEQ ID NO: 71 is Gln; Xaa 10 of SEQ ID NO: 71 is Lys; Xaa 11 of SEQ ID NO: 71 is Thr, or a combination thereof. In another embodiment, Xaa1 of SEQ ID NO: 71 is Glu; Xaa5 of SEQ ID NO: 71 is Thr; Xaa6 of SEQ ID NO: 71 is Thr; Xaa7 of SEQ ID NO: 71 is Tyr; Xaa8 of SEQ ID NO: 71 is Gln; Xaa9 of SEQ ID NO: 71 is Ser; Xaa 10 of SEQ ID NO: 71 is Lys; and Xaa 11 of SEQ ID NO: 71 is Thr. 【0019】 In one embodiment, the antibody or its antigen-binding fragment is selected from immunoglobulin molecules, monoclonal antibodies, chimeric antibodies, CDR-transplanted antibodies, humanized antibodies, Fab, Fab', F(ab')2, Fv, disulfide-linked Fv, scFv, single-domain antibodies, diabody antibodies, multispecific antibodies, dual-specific antibodies, and bispecific antibodies. In one embodiment, the antibody or its antigen-binding fragment includes an Fc region. In one embodiment, the antibody or its antigen-binding fragment is IgG1, IgG2, or IgG4 or a fragment thereof. 【0020】 In one embodiment, the present invention provides an antibody or antigen-binding fragment thereof against influenza B virus, which binds to influenza B virus and has the ability to neutralize at least one Yamagata lineage and at least one Victoria lineage of influenza B virus, wherein the antibody or antigen-binding fragment thereof binds to an epitope conserved in at least one Yamagata lineage and at least one Victoria lineage of influenza B virus. In one embodiment, one or more contact residues of the epitope are located within the head region of influenza B HA. In one embodiment, the epitope comprises one or more amino acids selected from 128, 141, 150 and 235 of the sequence of the head region of HA as contact residues (Wang et al. (2008) J. Virol. 82(6):3011-20). 【0021】 In another embodiment, the present invention provides an antibody or antigen-binding fragment thereof against influenza B virus, which binds to influenza B virus hemagglutinin and has the ability to neutralize influenza B virus in two phylogenetically distinct lineages, either by binding to the same epitope as the antibody of the present invention or by competing with the antibody of the present invention for binding to influenza B virus hemagglutinin. In one embodiment, the antibody or antigen-binding fragment is (a) VH of SEQ ID NO: 2 and VL of SEQ ID NO: 7, (b) VH of sequence number 12 and VL of sequence number 17, (c) VH of SEQ ID NO: 22 and VL of SEQ ID NO: 27, (d) VH of SEQ ID NO: 32 and VL of SEQ ID NO: 37, (e) VH of sequence number 42 and VL of sequence number 47, (f) VH of sequence number 52 and VL of sequence number 57, (g) VH of SEQ ID NO: 62 and VL of SEQ ID NO: 67, (h) VH of sequence number 74 and VL of sequence number 82, (i) VH of SEQ ID NO: 90 and VL of SEQ ID NO: 98, (j) VH of SEQ ID NO: 106 and VL of SEQ ID NO: 114 It binds to the same epitope as an antibody having an amino acid sequence selected from the available options, or competes with that antibody for binding to influenza A virus hemagglutinin. 【0022】 The present invention also provides isolated nucleic acids encoding the antibody of the present invention or its antigen-binding fragment, as well as vectors comprising such isolated nucleic acids and host cells comprising such nucleic acids or vectors. In one embodiment, the vector is an expression vector. In another embodiment, the vector is a non-natural recombinant vector. In one embodiment, the vector is a plasmid. In one embodiment, the vector or plasmid comprises a nucleotide sequence encoding the antibody molecule of the present invention or its antigen-binding fragment, the heavy chain or light chain of the antibody molecule of the present invention, the heavy chain or light chain variable domain of the antibody of the present invention, or a portion thereof, or a heavy chain or light chain CDR, operably linked to one or more expression regulatory elements (e.g., promoters, enhancers, transcription terminators, polyadenylation sites, etc.), a selectable marker gene, or a combination thereof. In one embodiment, the vector or plasmid comprises at least one heterologous expression regulatory element, a selectable marker, or a combination thereof. 【0023】 In one embodiment, the present invention provides a method for producing an antibody or an antigen-binding fragment thereof by culturing host cells described herein under conditions suitable for the expression of the antibody or the fragment thereof. In one embodiment, the method includes the step of isolating the antibody or the antigen-binding fragment thereof from a host cell culture. In one embodiment, the host cells are isolated from tissues in which the cells are found naturally. For example, the host cells may be isolated from an organism and maintained in vitro under cell culture. 【0024】 The present invention also provides compositions comprising the antibody of the present invention or its antigen-binding fragment and a pharmaceutically acceptable carrier. In one embodiment, the composition comprises the antibody of the present invention or its antigen-binding fragment and 25 mM His and 0.15 M NaCl at pH 6.0. 【0025】 In one embodiment, the antibody or antigen-binding fragment of the present invention is used in the prevention or treatment of influenza B infection in a subject. In another embodiment, the antibody or antigen-binding fragment is used in the prevention or treatment of influenza A and influenza B infection in a subject. In yet another embodiment, the antibody or antigen-binding fragment of the present invention is used in the manufacture of a drug for the prevention or treatment of influenza B infection in a subject. In yet another embodiment, the antibody or antigen-binding fragment of the present invention is used in the manufacture of a drug for the prevention or treatment of influenza A and influenza B infection in a subject. 【0026】 In one embodiment, the present invention provides a method for the prevention or treatment of influenza B infection in a subject, comprising the step of administering an effective amount of the antibody or antigen-binding fragment thereof to the subject. In another embodiment, the present invention provides a method for the prevention or treatment of influenza A and influenza B infection in a subject, comprising the step of administering an effective amount of the antibody or antigen-binding fragment thereof to the subject. 【0027】 In one embodiment, the antibody or fragment thereof of the present invention is used for in vitro diagnosis of influenza B infection in a subject. In another embodiment, the antibody or fragment thereof of the present invention is used for in vitro diagnosis of influenza A infection in a subject. In yet another embodiment, the antibody or fragment thereof of the present invention is used for in vitro diagnosis of both influenza A and influenza B infection in a subject. [Brief explanation of the drawing] 【0028】 [Figure 1] This shows NK cell activation after incubation with B / HongKong / 330 / 2001 (Victoria strain) and antibody-dependent cytotoxicity (ADCC) as measured by serial dilutions of anti-HA antibodies FBD-94 and FBC-39, as well as mutants lacking Fc effector function (FBD-94 LALA and FBC-39 LALA). [Figure 2] The percentages of surviving animals treated with various concentrations of FBC-39 (A and C), FBD-94 (B and D), and unrelated control antibodies four hours prior to infection with a lethal dose of influenza B / Sichuan / 379 / 99 (Yamagata) (A and B) or influenza B / Hong Kong / 330 / 2001 (Victoria) (C and D) viruses are shown. [Figure 3] This shows the percentage of surviving animals that were infected with a lethal dose of B / Sichuan / 379 / 99(Yamagata)(A and B) or B / Hong Kong / 330 / 2001(Victoria)(C and D) and treated on day 2 post-infection with various doses of FBC-39(A and C) or FBD-94(B and D), or unrelated control antibodies. [Figure 4] This represents the percentage of surviving animals that received a lethal dose of B / Hong Kong / 330 / 2001(Victoria) and were treated with 3 mg / kg of FBC-39(A) or FBD-94(B), or an unrelated control antibody, on day 1, 2, 3, or 4 post-infection. [Figure 5A]The CDRs (within the box) within the VH and VL sequences of the anti-influenza B antibodies FBD-56, FBD-94, FBC-39, FBC-39LSL, FBC-39FSL, FBC-39LTL, FBC-39FTL, FBC-39FSS, FBC-39LTS, and FBC-39FTS, as determined using the Kabat and IMGT numbering systems, are shown. Modified amino acids within the VH and VL sequences are shown in bold and underlined. [Figure 5B] This is a continuation of Figure 5A. [Figure 5C] This is a continuation of Figure 5A. [Figure 6] The heavy chain amino acid sequence (SEQ ID NO: 71) of the anti-influenza B antibody as a general name is shown, based on the heavy chain amino acid sequence (SEQ ID NO: 22) of FBC-39, where Xaa1 may be Val or Glu; Xaa2 may be Leu or Phe; Xaa3 may be Thr or Ser; Xaa4 may be Ser or Leu; Xaa5 may be Thr or Ser; Xaa6 may be Thr or Met; Xaa7 may be Tyr or Phe; Xaa8 may be Gln or His; Xaa9 may be Asn or Ser; Xaa10 may be Lys or Arg; and Xaa11 may be Thr or Ala. [Figure 7] The light chain amino acid sequence (SEQ ID NO: 72) of the anti-influenza B antibody as a general name is shown, based on the light chain amino acid sequence of FBC-39 (SEQ ID NO: 27), where Xaa1 may be Phe or Tyr. [Figure 8] This shows the alignment of HA1 protein from viruses used in the isolation of monoclonal antibody-resistant mutants (MARMs). Amino acid positions found to be contact residues through MARM selection are enclosed in boxes. [Modes for carrying out the invention] 【0029】 introduction The present invention provides human-type antibodies, as well as antigen-binding fragments, derivatives / conjugates, and compositions thereof, which bind to influenza B virus hemagglutinin (HA) and neutralize influenza B viruses in two phylogenetically distinct lineages, as described herein. In one embodiment, the antibody or its antigen-binding fragment binds to influenza B virus hemagglutinin (HA) and neutralizes influenza B viruses in both Yamagata and Victoria lineages, and such anti-influenza B antibodies and fragments thereof are referred to herein as the antibodies of the present invention. In another embodiment, the antibody or its antigen-binding fragment binds to influenza B virus hemagglutinin (HA) and influenza A virus hemagglutinin (HA) and neutralizes at least one Yamagata lineage influenza B virus; at least one Victoria lineage influenza B virus; at least one influenza A virus subtype, or a combination thereof. Such anti-influenza B antibodies and fragments thereof are also referred to herein as the antibodies of the present invention. 【0030】 As used herein, the term “neutralize” means that an antibody or its antigen-binding fragment binds to an infectious pathogen, such as influenza A and / or influenza B virus, and has the biological activity, such as the ability to reduce the pathogenicity of the infectious pathogen. In one embodiment, the antibody or its antigen-binding fragment of the present invention binds immunospecifically to at least one specific epitope or antigenic determinant of influenza A virus; influenza B virus; or a combination thereof. In a further specific embodiment, the antibody or its antigen-binding fragment of the present invention binds immunospecifically to at least one specific epitope or antigenic determinant of influenza B virus hemagglutinin (HA). In another further specific embodiment, the antibody or its binding fragment of the present invention binds immunospecifically to at least one specific epitope or antigenic determinant of the globular head of influenza B virus HA. 【0031】 Antibodies can neutralize the activity of infectious pathogens, such as influenza A and / or influenza B viruses, at various points in the life cycle of the virus. For example, antibodies can interfere with the attachment of the virus to target cells by interfering with the interaction between the virus and one or more cell surface receptors. Alternatively, antibodies can interfere with the post-attachment interaction between the virus and one or more receptors, for example, by interfering with the internal movement of the virus by receptor-mediated endocytosis. 【0032】 As used herein, “antibody” and “antibodies,” also known as immunoglobulins, encompass monoclonal antibodies (including full-length monoclonal antibodies), human antibodies, humanized antibodies, camelid antibodies, chimeric antibodies, single-chain Fvs(scFv), single-chain antibodies, single-domain antibodies, domain antibodies, Fab fragments, F(ab')2 fragments, antibody fragments exhibiting desired biological activity (e.g., antigen-binding moieties), disulfide-bonded Fvs(dsFv), and anti-idiotype (anti-Id) antibodies (e.g., anti-Id antibodies against the antibodies of the present invention), intracellular antibodies, and any of the above epitope-binding fragments. In particular, antibodies include immunoglobulin molecules and immunologically active fragments of immunoglobulin molecules, i.e., molecules containing at least one antigen-binding site. The immunoglobulin molecule may be any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), a subisotype (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or an allotype (e.g., Gm, e.g., G1m(f, z, a, or x), G2m(n), G3m(g, b, or c), Am, Em, and Km(1, 2, or 3)). 【0033】 Human antibodies are typically heterotetrameric glycoproteins with approximately 150,000 daltons, containing two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, although the number of disulfide bonds differs between heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (VH) at one end, followed by several constant domains (CH). Each light chain has a variable domain (VL) at one end and a constant domain (CL) at the other; the constant domain of the light chain aligns with the first constant domain of the heavy chain, and the variable domain of the light chain aligns with the variable domain of the heavy chain. Light chains are classified as either λ-chains or κ-chains based on the amino acid sequence of the light chain constant region. The variable domain of a κ-light chain may also be represented herein as VK. 【0034】 The antibodies of the present invention include full-length antibodies or intact antibodies, antibody fragments containing antigen-binding fragments, natural sequence antibodies or amino acid variants, human antibodies, humanized antibodies, post-translation modified antibodies, chimeric or fusion antibodies, immunoconjugates, and functional fragments thereof. Antibodies can provide desired effector function or serum half-life by modifying the Fc region. As will be discussed in more detail in the following sections, naked antibodies conjugated to the cell surface with an appropriate Fc region can induce cytotoxicity or several other mechanisms, for example, via antibody-dependent cytotoxicity (ADCC), or by recruiting complement in complement-dependent cytotoxicity (CDC), or by expressing one or more effector ligands that recognize the conjugated antibody on influenza A and / or influenza B viruses, and subsequently recruiting nonspecific cytotoxic cells that cause phagocytosis in antibody-dependent cell-mediated phagocytosis (ADCP). Alternatively, specific other Fc regions may be used if it is desirable to eliminate or reduce effector function to minimize side effects or therapeutic complications. Methods for not only enhancing but also reducing or eliminating Fc effector function are described herein. Furthermore, the Fc region of the antibody of the present invention may be modified to enhance its binding affinity to FcRn, thereby increasing its serum half-life. Alternatively, the Fc region may be conjugated to PEG or albumin to increase its serum half-life, or several other conjugations may produce desired effects. 【0035】 In one embodiment, the antibody is useful for diagnosing, preventing, treating and / or mitigating one or more symptoms of influenza B virus infection in mammals. In another embodiment, the antibody is useful for diagnosing, preventing, treating and / or mitigating one or more symptoms of influenza A and influenza B virus infection in animals. As used herein, the term “animal” means mammals, including but not limited to humans, non-human primates, dogs, cats, horses, rabbits, mice, and rats; and non-mammalian species, including but not limited to birds, such as chickens, turkeys, ducks, and quail. 【0036】 The present invention provides compositions comprising the antibody and carrier of the present invention. The composition may be administered to patients in need of treatment for the prevention or treatment of influenza B virus infection. In one embodiment, the composition may be administered to a patient to prevent or treat influenza A virus infection; influenza B virus infection; and combinations thereof. The present invention also provides formulations comprising the antibody and carrier of the present invention. In one embodiment, the formulation is a therapeutic formulation comprising a pharmaceutically acceptable carrier. 【0037】 In certain embodiments, the present invention provides a method useful for preventing or treating influenza B infection in mammals, comprising the step of administering an antibody to a mammal in a therapeutically effective dose. In other embodiments, the present invention provides a method useful for preventing or treating influenza A infection; influenza B infection; and combinations thereof in mammals, comprising the step of administering an antibody to a mammal in a therapeutically effective dose. The antibody therapeutic composition may be administered as directed by a physician, for a short period (acutely), chronically, or intermittently. 【0038】 In certain embodiments, the present invention also provides products, e.g., sterile dosage forms and kits, comprising at least one antibody of the present invention. The kits may be provided to have antibodies intended for in vitro detection and quantification of influenza virus, for example, in ELISA or Western blotting. Such antibodies useful for detection may be provided with labeling, such as fluorescent or radiolabeling. 【0039】 Terminology Before describing the present invention in detail, it should be understood that this invention is not limited to specific compositions or method steps and is itself variable. It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless explicitly indicated otherwise in the context. 【0040】 Unless otherwise defined, all scientific and technical terms used herein have the same meaning as those generally understood by those skilled in the art in the field relating to this invention. For example, *Concise Dictionary of Biomedicine and Molecular Biology*, Juo, Pei-Show (2002) 2nd ed., CRC Press; *The Dictionary of Cell and Molecular Biology*, 3rd ed. (1999) Academic Press; and *Oxford Dictionary of Biochemistry and Molecular Biology*, Revised (2000) Oxford University Press provide many general dictionaries of the terms used in this invention. 【0041】 In this specification, amino acids may be referred to by either their commonly known three-letter or one-letter symbols, as recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Similarly, nucleotides may be referred to by their commonly recognized one-letter codes. 【0042】 Anti-influenza B virus antibodies In certain embodiments, the antibody is an isolated and / or purified antibody and / or a pyrogen-free antibody. The term “purified,” as used herein, refers to other molecules, such as polypeptides and nucleic acid molecules, that have been identified, isolated, and / or recovered from components of their natural environment. Thus, in one embodiment, the antibody of the present invention is a purified antibody isolated from one or more components of its natural environment. The term “isolated antibody,” as used herein, refers to an antibody substantially released from another antibody molecule having different antigen specificity (for example, an isolated antibody that specifically binds to influenza B virus is substantially released from an antibody that specifically binds to antigens other than the influenza B virus HA antibody antigen). Thus, in one embodiment, the antibody of the present invention is an isolated antibody released from an antibody having different specificity. Typically, an isolated antibody is a monoclonal antibody. Furthermore, the isolated antibody of the present invention may also be substantially released from one or more other cellular materials and / or chemical substances, and as used herein, refers to an isolated and purified antibody. In one embodiment of the present invention, a “isolated” monoclonal antibody combination relates to antibodies having different specificity and combined in a well-defined composition. The methods for antibody production and purification / isolation are described in more detail below. 【0043】 The isolated antibody of the present invention comprises an antibody amino acid sequence disclosed herein encoded by any suitable polynucleotide, or any isolated or formulated antibody. 【0044】 The antibody of the present invention immunospecifically binds to at least one specific epitope that is specific to the HA protein of influenza B virus. The term “epitope,” as used herein, refers to a protein determinant that has the ability to bind to an antibody. Epitopes typically consist of chemically active surface groups (groupings) of a molecule, such as amino acid or sugar side chains, and usually possess specific three-dimensional structural properties as well as specific charge properties. Conformal epitopes and non-conformational epitopes are distinguished by the fact that binding to the former, rather than the latter, is lost in the presence of a denaturing solvent. 【0045】 In one embodiment, the antibody or its antigen-binding fragment binds to an epitope present in at least two phylogenetically distinct influenza B strains. In a further specific embodiment, the antibody or its antigen-binding fragment binds to an epitope present in at least one influenza B Yamagata strain and at least one influenza B Victoria strain. In one embodiment, the antibody or its antigen-binding fragment binds to an epitope present in both Yamagata and Victoria strains of influenza B virus. In one embodiment, the antibody or its antigen-binding fragment binds to an epitope conserved in both Yamagata and Victoria strains of influenza B. 【0046】 In one embodiment, the antibody or its antigen-binding fragment is used to target at least one influenza Yamagata strain and at least one influenza Victoria strain, at an effective concentration (EC) of up to half the maximum amount, between approximately 1 ng / ml and approximately 500 ng / ml, or between approximately 1 ng / ml and approximately 250 ng / ml, or between approximately 1 ng / ml and approximately 50 ng / ml, or approximately 500 ng / ml, 250 ng / ml, 100 ng / ml, 50 ng / ml, 40 ng / ml, 30 ng / ml, 20 ng / ml, or less than 15 μg / ml, against at least one influenza Yamagata strain and at least one influenza Victoria strain. 50 In another embodiment, the antibody or its antigen-binding fragment binds to Yamagata and Victoria lineage influenza B viruses at concentrations between approximately 1 ng / ml and 500 ng / ml, or between approximately 1 ng / ml and 250 ng / ml, or between approximately 1 ng / ml and 50 ng / ml, or at concentrations of approximately 500 ng / ml, 250 ng / ml, 100 ng / ml, 50 ng / ml, 40 ng / ml, 30 ng / ml, 20 ng / ml, or less than 15 μg / ml of EC. 50In one embodiment, the antibody or its antigen-binding fragment binds to an epitope present in both Yamagata and Victoria strains of influenza B virus at an EC of approximately 1 ng / ml to approximately 500 ng / ml, or approximately 1 ng / ml to approximately 250 ng / ml, or approximately 1 ng / ml to approximately 50 ng / ml, or approximately 500 ng / ml, 250 ng / ml, 100 ng / ml, 50 ng / ml, 40 ng / ml, 30 ng / ml, 20 ng / ml, or less than 15 μg / ml. 50 They are joined together. 【0047】 In one embodiment, the antibody or its antigen-binding fragment is attached to an epitope present in the Yamagata strain of influenza B with an EC of approximately 1 ng / ml to approximately 100 ng / ml, between 1 ng / ml and approximately 50 ng / ml, between approximately 1 ng / ml and approximately 25 ng / ml, or between approximately 50 ng / ml or less than 25 ng / ml. 50 Furthermore, in the epitopes present in the Victoria lineage of influenza B, EC levels are between approximately 1 ng / ml and 500 ng / ml, or between approximately 1 ng / ml and 250 ng / ml, or between approximately 1 ng / ml and 50 ng / ml, or between approximately 500 ng / ml, 250 ng / ml, 100 ng / ml, or less than 50 ng / ml. 50 They are joined together. 【0048】 In another embodiment, the antibody or its antigen-binding fragment is attached to an epitope present in the Yamagata strain of influenza B with an EC of approximately 1 ng / ml to approximately 100 ng / ml, between 1 ng / ml and approximately 50 ng / ml, between approximately 1 ng / ml and approximately 25 ng / ml, or between approximately 50 ng / ml or less than 25 ng / ml. 50 In the epitopes present in the Victoria lineage of influenza B, EC levels are approximately 1 ng / ml to 500 ng / ml, or between approximately 1 ng / ml and 250 ng / ml, or between approximately 1 ng / ml and 50 ng / ml, or between approximately 500 ng / ml, 250 ng / ml, or less than 100 ng / ml. 50Furthermore, the EC levels on the HA of influenza A are between approximately 1 μg / ml and approximately 50 μg / ml, or approximately 50 μg / ml, 25 μg / ml, 15 μg / ml, or less than 10 μg / ml. 50 In another embodiment, the antibody or its antigen-binding fragment binds to an epitope present in the Yamagata strain of influenza B at an EC of approximately 1 ng / ml to approximately 100 ng / ml, 1 ng / ml to approximately 50 ng / ml, or approximately 1 ng / ml to approximately 25 ng / ml, or approximately 50 ng / ml or less than 25 ng / ml. 50 In the epitopes present in the Victoria lineage of influenza B, EC levels are between approximately 1 ng / ml and 500 ng / ml, or between approximately 1 ng / ml and 250 ng / ml, or between approximately 1 ng / ml and 50 ng / ml, or less than 500 ng / ml, 250 ng / ml, or 100 ng / ml. 50 Furthermore, the EC levels on the H9 HA epitope are between approximately 1 μg / ml and approximately 50 μg / ml, or approximately 50 μg / ml, 25 μg / ml, 15 μg / ml, or less than 10 μg / ml. 50 They are joined together. 【0049】 In one embodiment, the antibody or its antigen-binding fragment recognizes an epitope that is either a linear or a continuous epitope. In another embodiment, the antibody or its antigen-binding fragment recognizes a non-linear or steric epitope. In one embodiment, the epitope is located on the hemagglutinin (HA) glycoprotein of influenza B. In a further specific embodiment, the epitope is located on the head region of the HA glycoprotein of influenza B. In one embodiment, the epitope contains one or more amino acids as contact residues at positions 128, 141, 150, or 235 in the head region of influenza B HA (numbered according to the H3 numbering system described in Wang et al. (2008) J.Virol.82(6):3011-20). In one embodiment, the epitope contains amino acid 128 of the sequence of the head region of influenza B HA as a contact residue. In another embodiment, the epitope includes amino acids 141, 150, and 235 of the head region sequence of influenza B HA as contact residues. 【0050】 One or more epitopes recognized by the antibody or antigen-binding fragment of the present invention may have numerous applications. For example, the purified or synthetic form of the epitope can be used to enhance the immune response (i.e., as a vaccine or to produce antibodies intended for other uses) or to screen serum for antibodies that react with the epitope. In one embodiment, the epitope recognized by the antibody or antigen-binding fragment of the present invention, or an antigen having such an epitope, may be used as a vaccine to enhance the immune response. In another embodiment, the antibody and antigen-binding fragment of the present invention can be used to monitor the quality of a vaccine, for example, by determining whether the antigen in the vaccine contains the correct immunogenic epitope in the correct three-dimensional structure. 【0051】 Variable region As used herein, the term “parent antibody” refers to an antibody encoded by an amino acid sequence used to prepare a variant or derivative, as defined herein. The parent polypeptide may include a naturally occurring antibody sequence (i.e., one that includes naturally occurring allelic variants) or an antibody sequence having existing amino acid sequence modifications (such as other insertions, deletions, and / or substitutions) to a naturally occurring sequence. The parent antibody may be a humanized antibody or a human antibody. In one embodiment, the antibody of the present invention is a variant of the parent antibody. As used herein, the term “variant” refers to an antibody whose amino acid sequence differs from the “parent” antibody amino acid sequence due to the addition, deletion, and / or substitution of one or more amino acid residues in the parent antibody sequence. 【0052】 The antigen-binding moiety of an antibody contains one or more fragments of the antibody that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments contained within the “antigen-binding moiety” of an antibody include: (i) Fab fragments, i.e., monovalent fragments containing the VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, i.e., bivalent fragments containing two Fab fragments linked by a disulfide crosslink at a hinge region; (iii) Fd fragments containing the VH and CH1 domains; (iv) Fv fragments containing the VL and VH domains of a single arm of the antibody; (v) dAb fragments containing the VH domain (Ward et al. (1989) Nature. 341:544-546); and (vi) isolated complementarity-determining regions (CDRs). Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked using recombinant methods with a synthetic linker that allows for the pairing of the VL and VH regions to create a single protein chain that forms a monovalent molecule (known as single-stranded Fv (scFv); see, e.g., Bird et al. (1988) Science. 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-stranded antibodies are also intended to be included within the scope of the antibody term "antigen-binding moiety." These antibody fragments can be obtained using conventional techniques known to those skilled in the art, and the fragments are screened for usefulness in the same manner as for intact antibodies. The antigen-binding moiety can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of intact immunoglobulins. 【0053】 The antibody of the present invention comprises at least one antigen-binding domain, which includes the VH and VL domains described herein. Typical VH and VL domains are shown in Table 1 below. 【0054】 [Table 1] 【0055】 In certain embodiments, the purified antibody comprises a VH and / or VL having a given percentage identity to at least one of the VH and / or VL sequences disclosed herein. As used herein, the term “percent (%) sequence identity” (including “homology”) is defined as the percentage of amino acid residues or nucleotides in a candidate sequence that are identical to an amino acid residue or nucleotide in a reference sequence, e.g., a parent antibody sequence, after the sequences have been aligned and gaps introduced as necessary to obtain the greatest percentage sequence identity (without considering any conservative substitutions as part of sequence identity). The optimal alignment of sequences for comparison can be generated manually, by the local homology algorithm of Smith and Waterman (1981) Ads App.Math.2:482, the local homology algorithm of Neddleman and Wunsch (1970) J.MoI.Biol.48:443, the similarity search method of Pearson and Lipman (1988) Proc.Natl Acad.Sci.USA85:2444, or by computer programs using these algorithms (GAP, BESTFIT, FASTA, BLAST P, BLAST N, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Drive, Madison, Wis.). 【0056】 The antibody of the present invention may contain a VH amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identity with the VH amino acid sequence described herein. In other embodiments, the antibody of the present invention may have a VH amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence of the VH amino acid sequence described herein. 【0057】 The antibodies of the present invention may contain a VL amino acid sequence having at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identity with the VL amino acid sequences described herein. In other embodiments, the antibodies of the present invention may have a VL amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the VL amino acid sequences described herein. 【0058】 Complementarity Determination Area (CDR) While the variable domains (VH and VL) contain antigen-binding regions, their variability is not uniformly distributed throughout the antibody's variable domains. Variability is concentrated within segments called complementarity-determining regions (CDRs) in both the light chain (VL or VK) and heavy chain (VH) variable domains. More highly conserved portions of the variable domains are called framework regions (FRs). Each of the natural heavy and light chain variable domains contains four FRs, primarily in a β-sheet configuration, connected by three CDRs. Loops formed by the CDRs connect these β-sheet structures and, in some cases, form part of them. The CDRs of each chain are held together in close proximity by the FRs and, together with the CDRs from the other chain, contribute to the formation of the antibody's antigen-binding site. The three CDRs of the heavy chain are designated HCDR-1, HCDR-2, and HCDR-3, and the three CDRs of the light chain are designated LCDR-1, LCDR-2, and LCDR-3. 【0059】 In one embodiment, amino acids in the variable domain, complementarity-determining region (CDR), and framework region (FR) of an antibody may be identified according to Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortenings or insertions into the FR or CDR of the variable domain. For example, the heavy chain variable domain may contain a single amino acid insertion after H2 residue 52 (residue 52a according to Kabat) and an inserted residue after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). Kabat numbering of residues may be determined for a given antibody by alignment with a “standard” Kabat numbering sequence in the homology region of the antibody sequence. Maximum alignment of framework residues often requires the insertion of “spacer” residues in the numbering system. Furthermore, the identity of specific individual residues at any given Kabat site number may change from antibody chain to antibody chain due to interspecies or allele differences. 【0060】 According to Kabat et al.'s numbering system, HCDR-1 begins with approximately 31 amino acids (i.e., about 9 residues after the first cysteine ​​residue), contains about 5-7 amino acids, and terminates with the following tyrosine residue. HCDR-2 begins with 15 residues after the terminal of CDR-H1, contains about 16-19 amino acids, and terminates with the following arginine or lysine residue. HCDR-3 begins with approximately 33 amino acid residues after the terminal of HCDR-2, contains 3-25 amino acids, and terminates with the sequence WGXG (where X is any amino acid). LCDR-1 begins with approximately 24 residues (i.e., after the cysteine ​​residue), contains about 10-17 residues, and terminates with the following tyrosine residue. LCDR-2 begins with approximately 16 residues after the terminal of LCDR-1 and contains about 7 residues. LCDR-3 begins approximately 33 residues post-terminus of LCDR-2, contains approximately 7-11 residues, and terminates with the sequence FGXG (where X is any amino acid). Note that the CDR changes significantly from antibody to antibody (and, by definition, does not exhibit homology to the Kabat consensus sequence). The CDR heavy and light chain sequences of the antibodies of the present invention, numbered using the Kabat system, are shown in Tables 2 and 3 below. 【0061】 [Table 2] 【0062】 [Table 3] 【0063】 The Kabat numbering scheme is widely used, but it has several drawbacks. Firstly, because the numbering scheme was developed from sequence data in the absence of structural information, the locations where insertions are made in LCDR-1 and HCDR-1 do not always coincide with the structural insertion locations. Therefore, morphologically equivalent residues within these loops may not be assigned the same number. Secondly, its numbering system is strict and only allows for a limited number of insertions. If there are more residues than the numbering system assigned to insertions allows, there is no standard method for numbering them. 【0064】 In another embodiment, amino acids in the variable domain, complementarity-determining region (CDR), and framework region (FR) of the antibody can be identified using the immunogenetics (IMGT) database (http: / / imgt.cines.fr) (Lefranc et al. (2003) Dev Comp Immunol. 27(1):55-77). The IMGT database was developed using sequence information for immunoglobulin (IgG), T cell receptor (TcR), and major histocompatibility complex (MHC) molecules, unifying numbering across the antibody λ and κ light chains, heavy chain, and T cell receptor chain, and avoiding the use of insertion codes for all but abnormally long insertions. IMGT also considers and combines the definitions of framework (FR) and complementarity-determining region (CDR) from Kabat et al., the characterization of hypervariable loops from Chothia et al., and structural data obtained from X-ray diffraction studies. The CDR heavy and light chain sequences in the antibody of the present invention, numbered using the IMGT system, are shown in Tables 4 and 5 below. Figure 5 shows the alignment of the FBD-56, FBD-94, and FBC-39 sequences, which represent CDR sequences, as identified by Kabat and IMGT. 【0065】 [Table 4] 【0066】 [Table 5] 【0067】 The present invention comprises neutralizing an anti-influenza B antibody containing amino acids in a sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% identical to the amino acid sequence of VH in SEQ ID NO: 2; SEQ ID NO: 12; SEQ ID NO: 22; SEQ ID NO: 32; SEQ ID NO: 42; SEQ ID NO: 52; SEQ ID NO: 62; SEQ ID NO: 74; SEQ ID NO: 90; or SEQ ID NO: 106, and / or at least 75%, 80%, 85%, 90%, 95%, or 100% identical to the amino acid sequence of VL in SEQ ID NO: 7; SEQ ID NO: 17; SEQ ID NO: 27; SEQ ID NO: 37; SEQ ID NO: 47; SEQ ID NO: 57; SEQ ID NO: 67; SEQ ID NO: 82; SEQ ID NO: 98; or SEQ ID NO: 114. In another embodiment, the present invention includes neutralizing an anti-influenza B antibody containing amino acids in a sequence that is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of VH in SEQ ID NO: 2; SEQ ID NO: 12; SEQ ID NO: 22; SEQ ID NO: 32; SEQ ID NO: 42; SEQ ID NO: 52; SEQ ID NO: 62; SEQ ID NO: 74; SEQ ID NO: 90; or SEQ ID NO: 106, and / or at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of VL in SEQ ID NO: 7; SEQ ID NO: 17; SEQ ID NO: 27; SEQ ID NO: 37; SEQ ID NO: 47; SEQ ID NO: 57; SEQ ID NO: 67; SEQ ID NO: 82; SEQ ID NO: 98; or SEQ ID NO: 114. 【0068】 In another embodiment, the present invention provides antibodies and antigen-binding fragments thereof comprising six CDR sets selected from the HCDRs and LCDrs shown in Tables 2-5: HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2, and LCDR-3. In another embodiment, the present invention provides antibodies and antigen-binding fragments thereof comprising six CDR sets containing amino acids in sequences that are at least 75%, 80%, 85%, 90%, 95%, or 100% identical to the amino acid sequences of the HCDRs and LCDrs shown in Tables 2-5: HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2, and LCDR-3. In another embodiment, the present invention provides antibodies and antigen-binding fragments thereof comprising six CDR sets: HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2, and LCDR-3, each containing amino acids in a sequence that is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequences of the HCDRs and LCDRs shown in Tables 2-5. 【0069】 Framework domain Each of the heavy and light chain variable domains contains four framework regions (FR1, FR2, FR3, FR4) which are more highly conserved parts of the variable domain. The four FRs of the heavy chain are named FR-H1, FR-H2, FR-H3, and FR-H4, and the four FRs of the light chain are named FR-L1, FR-L2, FR-L3, and FR-L4. 【0070】 In one embodiment, the Kabat numbering system can be used to identify framework regions. According to Kabat et al., FR-H1 starts at position 1 and terminates at approximately 30 amino acids, FR-H2 is approximately 36–49 amino acids, FR-H3 is approximately 66–94 amino acids, and FR-H4 is approximately 103–113 amino acids. FR-L1 starts at amino acid 1 and terminates at approximately 23 amino acids, FR-L2 is approximately 35–49 amino acids, FR-L3 is approximately 57–88 amino acids, and FR-L4 is approximately 98–107 amino acids. In certain embodiments, the framework region may have substitutions according to the Kabat numbering system, for example, an insertion at 106A in FR-L1. In addition to native substitutions, one or more modifications (e.g., substitutions) of FR residues may also be introduced into the antibody of the present invention, provided that they retain their neutralizing ability. In certain embodiments, these result in an improvement or optimization of the antibody's binding affinity to influenza B virus HA. Examples of framework region residues to be modified include those that directly non-covalently bind to the antigen (Amit et al. (1986) Science. 233:747-753); those that interact with / act on the three-dimensional structure of the CDR (Chothia et al. (1987) J.Mol.Biol. 196:901-917); and / or those involved in the VL-VH interface (U.S. Patent No. 5,225,539). In other embodiments, the framework region may be identified using the IMGT numbering system. 【0071】 In another embodiment, FR may include one or more amino acid changes intended for “germlining.” For example, the heavy and light chain amino acid sequences of a selected antibody can be compared to the heavy and light chain amino acid sequences of the germline, and if certain framework residues of the selected VL and / or VH chains differ from the germline configuration (e.g., as a result of somatic mutation of immunoglobulin), it may be desirable to “reverse mutagenerate” the modified framework residues of the selected antibody with respect to the germline configuration (i.e., modify the framework amino acid sequence of the selected antibody to be the same as the germline framework amino acid sequence) to reduce the chance of immunogenicity, for example. Such “reverse mutagenesis” (or “germlining”) of framework residues can be achieved by standard molecular biological methods for introducing specific mutations (e.g., site-directed mutagenesis; PCR-mediated mutagenesis, etc.). 【0072】 Figure 6 shows the amino acid sequence of the VH domain of the anti-influenza B antibody as a generic name (SEQ ID NO: 71), and the non-germ series residue in FBC-39 (SEQ ID NO: 22) is Xaa 1-11 It was named as such. In one embodiment, the non-reproductive lineage (Xaa 1-11 ) One or more residues are “reverse-mutated” into the germline. In one embodiment, Xaa1 of SEQ ID NO: 71 is Val or Glu; Xaa2 of SEQ ID NO: 71 is Leu or Phe; Xaa3 of SEQ ID NO: 71 is Ser or Thr; Xaa4 of SEQ ID NO: 71 is Leu or Ser; Xaa5 of SEQ ID NO: 71 is Ser or Thr; Xaa6 of SEQ ID NO: 71 is Met or Thr; Xaa7 of SEQ ID NO: 71 is Phe or Tyr; Xaa8 of SEQ ID NO: 71 is His or Gln; Xaa9 of SEQ ID NO: 71 is Ser or Asn; Xaa 10 is Arg or Lys; also Xaa of sequence number 71 11is Ala or Thr. In another embodiment, Xaa1 of SEQ ID NO: 71 is Glu; Xaa5 of SEQ ID NO: 71 is Thr; Xaa6 of SEQ ID NO: 71 is Thr; Xaa7 of SEQ ID NO: 71 is Tyr; Xaa8 of SEQ ID NO: 71 is Gln; Xaa of SEQ ID NO: 71 10 is Lys; Xaa of sequence number 71 11 is Thr, or a combination thereof. In a further specific embodiment, Xaa9 in SEQ ID NO: 71 is Ser. In yet another embodiment, Xaa4 in SEQ ID NO: 71 is Leu. 【0073】 Figure 7 shows the amino acid sequence of the VL domain of the anti-influenza B antibody as a generic name (SEQ ID NO: 72), where the non-germline residue (SEQ ID NO: 27) within FBC-39 is represented by Xaa1. In one embodiment, the non-germline residue (Xaa1) is "reverse-mutated" into the germline. In one embodiment, Xaa1 in SEQ ID NO: 72 is Phe or Tyr. In a further specific embodiment, Xaa1 in SEQ ID NO: 72 is Tyr. 【0074】 The plasmid sequence encoding the antibody of the present invention In addition to the amino acid sequences described above, the present invention also provides nucleotide sequences corresponding to the amino acid sequences and encoding the human antibodies of the present invention. In one embodiment, the present invention provides polynucleotides comprising nucleotide sequences encoding the antibodies or fragments thereof described herein. These include, but are not limited to, nucleotide sequences encoding the amino acid sequences referenced above. Accordingly, the present invention also provides polynucleotide sequences encoding the VH and VL framework regions, including the CDR and FR of the antibodies described herein, and expression vectors for their efficient expression in cells (e.g., mammalian cells). Methods for producing antibodies using polynucleotides are described in more detail below. 【0075】 The present invention also encompasses polynucleotides that hybridize with the polynucleotides encoding the antibodies of the present invention as described herein, under stringent or lower stringency hybridization conditions, as defined herein, for example. The term “stringency,” as used herein, refers to the degree of homology between the probe and the nucleic acid bound to the filter, and the experimental conditions of the hybridization experiment (e.g., temperature and salt concentration), where higher stringency indicates higher percentage homology between the probe and the nucleic acid bound to the filter. 【0076】 Stringent hybridization conditions include, but are not limited to, hybridization with the filter-bound DNA in 6× sodium chloride / sodium citrate (SSC) at approximately 45°C, followed by one or more washes in 0.2× SSC / 0.1% SDS at approximately 50-65°C, highly stringent conditions such as hybridization with the filter-bound DNA in 6× SSC at approximately 45°C, followed by one or more washes in 0.1× SSC / 0.2% SDS at approximately 65°C, or any other stringent hybridization conditions known to those skilled in the art (see, for example, Ausubel et al., eds (1989) Current Protocols in Molecular Biology, vol.1, Green Publishing Associates, Inc. and John Wiley and Sons, Inc., NY, pp. 6.3.1-6.3.6 and 2.10.3). 【0077】 Substantially identical sequences may be polymorphic sequences, i.e., alternative sequences or alleles within a population. The difference between alleles may be as small as that of a single base pair. Substantially identical sequences may also include mutagenic sequences, such as sequences containing silent mutations. Mutations may include changes in one or more residues, deletions of one or more residues, or insertions of one or more additional residues. 【0078】 Polynucleotides may be obtained and their nucleotide sequences determined by any method known in the art. For example, if the nucleotide sequence of an antibody is known, the polynucleotide encoding the antibody may be constructed from chemically synthesized oligonucleotides (as described, for example, in Kutmeier et al. (1994) BioTechniques. 17:242), which involves the synthesis of overlapping oligonucleotides containing a portion of the antibody-encoding sequence, annealing and ligation of the oligonucleotides, and subsequent amplification of the ligated oligonucleotides by PCR. 【0079】 The polynucleotide encoding the antibody may also be prepared from nucleic acids derived from a suitable source. If a clone containing the nucleic acid encoding a particular antibody is not available, but the sequence of the antibody molecule is known, the nucleic acid encoding the immunoglobulin may be obtained by chemical synthesis or from a suitable source (e.g., an antibody cDNA library, or any tissue or cell expressing the antibody, such as a cDNA library prepared from hybridoma cells selected to express the antibody, or nucleic acids isolated therefrom, in one embodiment polyA+RNA) by PCR amplification using synthetic primers that can hybridize to the 3' and 5' ends of the sequence, or by cloning using oligonucleotide probes specific to a particular gene sequence, for example, by identifying a cDNA clone from an antibody-encoding cDNA library. The amplified nucleic acid produced by PCR can then be cloned into a replicable cloning vector using any method well known in the art. 【0080】 Once the nucleotide sequence and corresponding amino acid sequence of an antibody are determined, the nucleotide sequence of the antibody can be manipulated using methods well known in the art for manipulating nucleotide sequences, such as recombinant DNA techniques, site-directed mutagenesis, PCR, etc. (see, for example, Sambrook et al. (1990) Molecular Cloning, A Laboratory Manual, 2d Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY and Ausubel et al., eds. (1998) Current Protocols in Molecular Biology, John Wiley & Sons, NY) to create antibodies with different amino acid sequences, for example, by causing amino acid substitutions, deletions, and / or insertions. 【0081】 Bonding characteristics As described above, the antibody or antigen-binding fragment of the present invention immunospecifically binds to at least one specific epitope or antigenic determinant of the HA protein, peptide, subunit, fragment, moiety, or any combination thereof of influenza B virus, exclusively or preferentially to other polypeptides. In certain embodiments, the epitope or antigenic determinant of the influenza B virus HA protein is a globular head. The terms “epitope” or “antigenic determinant,” as used herein, refer to a protein determinant capable of binding to an antibody. In one embodiment, the term “binding” refers to specific binding as used herein. These protein determinants or epitopes typically involve chemically active surface groupings of molecules such as amino acids or sugar side chains, and usually have specific three-dimensional structural properties as well as specific charge properties. Stereostructural and non-stereostructural epitopes are distinguished in that binding to the former rather than the latter is lost in the presence of a denaturing solvent. The term “discontinuous epitope,” as used herein, refers to a stereostructural epitope on a protein antigen formed from at least two isolated regions within the primary sequence of a protein. 【0082】 The interaction between antigens and antibodies is the same as that of other non-covalent protein-protein interactions. In general, four types of binding interactions exist between antigens and antibodies: (i) hydrogen bonds, (ii) dispersion forces, (iii) electrostatic forces between Lewis acids and Lewis bases, and (iv) hydrophobic interactions. Hydrophobic interactions are the main driving force in antibody-antigen interactions and are based on the repulsion of water by nonpolar groups rather than intermolecular attractive forces (Tanford (1978) Science. 200:1012-8). However, certain physical forces also contribute to antigen-antibody binding, such as the fit or preferential treatment of epitope shapes with different antibody binding sites. Furthermore, other materials and antigens can cross-react with antibodies, thereby competing for available free antibodies. 【0083】 Measuring the affinity constant and specificity of binding between antigen and antibody can help determine the effectiveness of the prophylactic, therapeutic, diagnostic, and research methods using the antibodies of the present invention. “Binding affinity” generally refers to the overall force of the non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, as used herein, “binding affinity” refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y is generally k off / k on This can be expressed by the equilibrium dissociation constant (Kd), which is calculated as a ratio. See, for example, Chen et al. (1999) J.Mol Biol. 293:865-881. Low affinity antibodies generally tend to bind gradually to the antigen and dissociate easily, while high affinity antibodies generally tend to bind more rapidly to the antigen and maintain the bound state for a longer period. Various methods for measuring binding affinity are known in the art, and any of them can be used for the purposes of this invention. 【0084】 One method for determining binding affinity involves measuring the dissociation constant "Kd" by radiolabeled antigen-binding assay (RIA) performed using the Fab version of the antibody of interest and its antigen as described by Chen et al. (1999) J. Mol Biol. 293:865-881. Alternatively, the Kd value may be measured by surface plasmon resonance assay using BIAcore®-2000 or BIAcore®-3000 (BIAcore, Inc., Piscataway, NJ). 6 M -1 If it exceeds S-1, the on-rate can be determined by using fluorescence quenching techniques that measure the increase and decrease in fluorescence emission intensity in the presence of gradually increasing concentrations of antigen. "On-rate" or "rate of association" or "rate of association" or "k on This can also be determined using the same surface plasmon resonance technique as described above. 【0085】 Methods and reagents suitable for determining the binding characteristics of the antibodies of the present invention, or their modified / mutant derivatives, are known in the art and / or commercially available (U.S. Patent Nos. 6,849,425; 6,632,926; 6,294,391; and 6,143,574). Furthermore, instruments and software designed for such dynamical analysis are commercially available (e.g., Biacore® A100 and Biacore® 2000 instruments; Biacore International AB, Uppsala, Sweden). 【0086】 In one embodiment, the antibodies of the present invention (including their antigen-binding fragments or variants) may also be described or specified in terms of their binding affinity to influenza A virus polypeptides; influenza B virus polypeptides; or combinations thereof. Typically, antibodies with high affinity are 10 -7The Kd is less than M. In one embodiment, the antibody or its antigen-binding fragment is 5 × 10⁻¹⁶ of influenza A polypeptide; influenza B polypeptide; its fragment or variant; or a combination thereof. -7 M, 10 -7 M, 5×10 -8 M, 10 -8 M, 5×10 -9 M, 10 -9 M, 5×10 -10 M, 10 -10 M, 5×10 -11 M, 10 -11 M, 5×10 -12 M, 10 -12 M, 5×10 -13 M, 10 -13 M, 5×10 -14 M, 10 -14 M, 5×10 -15 M or 10 -15 It binds with a dissociation constant of M or less, i.e., Kd. In more specific embodiments, the antibody or its antigen-binding fragment binds to influenza A polypeptide; influenza B polypeptide, its fragment or variant; or a combination thereof, with 5 × 10⁻¹⁴ -10 M, 10 -10 M, 5×10 -11 M, 10 -11 M, 5×10 -12 M or 10 -12 The antibody binds with a dissociation constant, i.e., Kd, ​​of M or less. The present invention encompasses antibodies that bind to influenza A polypeptide; influenza B polypeptide; or a combination thereof, with a dissociation constant, i.e., Kd, ​​that falls within the range of any of the individually enumerated values. 【0087】 In another embodiment, the antibody or antigen-binding fragment of the present invention is used with influenza A polypeptide; influenza B polypeptide; fragments or variants thereof; or combinations thereof, for 5 × 10 -2 seconds -1 , 10 -2 seconds -1 , 5×10 -3 seconds -1 Or 10 -3 seconds -1 , 5×10 -4seconds -1 、10 -4 seconds -1 、5×10 -5 seconds -1 、or 10 -5 seconds -1 、5×10 -6 seconds -1 、10 -6 seconds -1 、5×10 -7 seconds -1 or 10 -7 seconds -1 at the following off-rates (k off ). In a more particular embodiment, the antibody or antigen-binding fragment thereof of the present invention binds to an influenza A polypeptide or a fragment or variant thereof at 5×10 -4 seconds -1 、10 -4 seconds -1 、5×10 -5 seconds -1 、or 10 -5 seconds -1 、5×10 -6 seconds -1 、10 -6 seconds -1 、5×10 -7 seconds -1 or 10 -7 seconds -1 at the following off-rates (k off ). The present invention also encompasses antibodies that bind to an influenza A polypeptide; an influenza B polypeptide; or a combination thereof at an off-rate (k off ) within the range between any of the individually recited values. 【0088】 In another embodiment, the antibody or antigen-binding fragment thereof of the present invention binds to an influenza A polypeptide; an influenza B polypeptide; a fragment or variant thereof; or a combination thereof at 10 3 M -1 seconds -1 、5×10 3 M -1 seconds -1 、10 4 M -1 seconds -1 、5×10 4 M-1 seconds -1 , 10 5 M -1 seconds -1 , 5×10 5 M -1 seconds -1 , 10 6 M -1 seconds -1 , 5×10 6 M -1 seconds -1 , 10 7 M -1 seconds -1 , or 5 x 10 7 M -1 seconds -1 The above on-rate (k on ) binds. In more specific embodiments, the antibody or antigen-binding fragment of the present invention binds to influenza A polypeptide; influenza B polypeptide; fragments or variants thereof; or combinations thereof, 10 5 M -1 seconds -1 , 5×10 5 M -1 seconds -1 , 10 6 M -1 seconds -1 , 5×10 6 M -1 seconds -1 , 10 7 M -1 seconds -1 Or 5 x 10 7 M -1 seconds -1 The above on-rate (k on The present invention involves bonding with an on-rate (k) within the range of any of the individually enumerated values ​​of influenza A polypeptide; influenza B polypeptide; or a combination thereof. on It includes antibodies that bind via ). 【0089】 In one embodiment, the binding assay may be carried out as either a direct binding assay or a competitive binding assay. Binding can be detected using a standard ELISA or a standard flow cytometry assay. In a direct binding assay, a candidate antibody is tested for binding to its congener antigen. In a competitive binding assay, on the other hand, the ability of a candidate antibody to compete with known antibodies or other compounds that bind to a specific antigen, such as influenza B virus HA, is evaluated. Generally, any method that enables the binding of an antibody to detectable influenza B virus HA is included within the scope of the present invention for detecting and measuring the binding properties of an antibody. Those skilled in the art will understand these well-known methods, and the reasons for this will not be presented in detail here. These methods are also used to screen a panel of antibodies for those that provide the desired properties. 【0090】 In one embodiment, the antibody of the present invention is immune-specific to influenza B virus HA and has the ability to neutralize influenza B virus infection. In one embodiment, the antibody of the present invention is immune-specific to at least one Yamagata strain influenza B virus and at least one Victoria strain influenza B virus. In another embodiment, the antibody of the present invention is immune-specific to Yamagata strain and Victoria strain influenza B viruses. 【0091】 In another embodiment, the antibody of the present invention is immunospecifically bound to influenza B virus HA and influenza A virus HA, and has the ability to neutralize influenza B virus and influenza A virus infection. In one embodiment, the antibody of the present invention is immunospecifically bound to at least one Yamagata lineage influenza B virus; at least one Victoria lineage influenza B virus and at least one subtype of influenza A virus. 【0092】 The hemagglutinin subtypes of influenza A virus are classified into two major phylogenetic groupings, identified as Group 1, which includes subtypes H1, H2, H5, H6, H8, H9, H11, H12, H13, H16, and H17, and Group 2, which includes subtypes H3, H4, H7, H10, H14, and H15. In one embodiment, the antibody or antigen-binding fragment described in the present invention has the ability to bind to and / or neutralize one or more subtypes of influenza A virus Group 1 selected from H8, H9, H11, H12, H13, H16, and H17 and their variants. In another embodiment, the antibody or antigen-binding fragment described in the present invention has the ability to bind to and / or neutralize one or more subtypes of influenza A virus Group 2 selected from H4, H10, H14, and H15 and their variants. In one embodiment, the antibody of the present invention binds to subtype H9 of influenza A virus Group 1. In one embodiment, the antibody of the present invention binds to and neutralizes subtype H9 of influenza A virus group 1. 【0093】 In one embodiment, the antibody of the present invention is immunospecifically bound to influenza B virus HA and has the ability to neutralize influenza B virus infection. In another embodiment, the antibody of the present invention is immunospecifically bound to influenza A and influenza B virus HA and has the ability to neutralize influenza A and influenza B virus infection. The neutralization assay may be carried out as described in the Examples section of this specification or by other methods known in the art. The term "inhibitory concentration 50%" ("IC") 50 '' (abbreviated as '') represents the concentration of the inhibitor (e.g., the antibody of the present invention) required for 50% neutralization of influenza A virus and / or influenza B virus. A lower IC 50 Those skilled in the art will understand that the values ​​correspond to stronger inhibitors. 【0094】 In one embodiment, the antibody or antigen-binding fragment described in the present invention is used to neutralize influenza B virus in a microneutralization assay with an antibody in the range of approximately 0.001 μg / ml to approximately 5 μg / ml, or in the range of approximately 0.001 μg / ml to approximately 1 μg / ml, or less than 5 μg / ml, less than 2 μg / ml, less than 1 μg / ml, less than 0.5 μg / ml, less than 0.1 μg / ml, less than 0.05 μg / ml, or less than 0.01 μg / ml. 50 It has. 【0095】 In one embodiment, the antibody or antigen-binding fragment described in the present invention is used for IC (Implantable Correction) of influenza B virus in a microneutralization assay at concentrations of approximately 0.001 μg / ml to approximately 5 μg / ml, or approximately 0.001 μg / ml to approximately 1 μg / ml, or less than 5 μg / ml, less than 2 μg / ml, less than 1 μg / ml, less than 0.5 μg / ml, less than 0.1 μg / ml, less than 0.05 μg / ml, or less than 0.01 μg / ml. 50 ; and IC for neutralizing influenza A virus in microneutralization assays, in the range of approximately 0.1 μg / ml to approximately 5 μg / ml, or in the range of approximately 0.1 μg / ml to approximately 2 μg / ml, or less than 5 μg / ml, less than 2 μg / ml, less than 1 μg / ml, or less than 0.5 μg / ml. 50 It has. 【0096】 In one embodiment, the antibody or antigen-binding fragment described in the present invention is used to neutralize influenza B virus in a microneutralization assay at concentrations in the range of approximately 0.001 μg / ml to approximately 50 μg / ml, or in the range of approximately 0.001 μg / ml to approximately 5 μg / ml of the antibody, or in the range of approximately 0.001 μg / ml to approximately 1 μg / ml of the antibody, or less than 10 μg / ml, less than 5 μg / ml, less than 1 μg / ml, less than 0.5 μg / ml, less than 0.1 μg / ml, or less than 0.05 μg / ml or 0.01 μg / ml. 50IC for neutralizing influenza A virus in microneutralization assays, in the range of approximately 0.01 μg / ml to approximately 50 μg / ml, or in the range of approximately 0.05 μg / ml to approximately 5 μg / ml of antibodies, or in the range of approximately 0.1 μg / ml to approximately 2 μg / ml of antibodies, or less than 50 μg / ml, less than 25 μg / ml, less than 10 μg / ml, less than 5 μg / ml, or less than 2 μg / ml. 50 It has. 【0097】 In certain embodiments, the antibodies of the present invention may induce cell death. “Cell death-inducing” antibodies are those that render living cells infertile. Cell death can be measured in vitro in the absence of complement and immunoeffector cells, and antibody-dependent cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC)-induced cell death can be distinguished. Therefore, assays for cell death may be performed using thermo-inactivated serum (i.e., in the absence of complement) and in the absence of immunoeffector cells. To determine whether an antibody can induce cell death, defects in membrane integrity, assessed by methods well known in the art, such as propidium iodide (PI), trypan blue (see Moore et al. (1995) Cytotechnology 17:1-11), 7AAD uptake, or other methods, can be evaluated compared to untreated cells. 【0098】 In specific embodiments, the antibodies of the present invention can induce cell death via apoptosis. “Apoptosis-inducing” antibodies induce programmed cell death, which is determined by annexin V binding, DNA fragmentation, cell contraction, endoplasmic reticulum expansion, cell fragmentation, and / or the formation of membrane vesicles (called apoptotic bodies). Various methods are available for evaluating cellular events associated with apoptosis. For example, phosphatidylserine (PS) translocation can be measured by annexin binding, DNA fragmentation can be evaluated through DNA laddering, and nuclear / chromatin condensation associated with DNA fragmentation can be evaluated by any increase in hypodiploid cells. In one embodiment, the apoptosis-inducing antibody results in approximately 2–50-fold induction of annexin binding in an annexin binding assay compared to untreated cells, approximately 5–50-fold in one embodiment, and approximately 10–50-fold in another embodiment. 【0099】 In another specific embodiment, the antibodies of the present invention may induce cell death via antibody-dependent cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC) and / or antibody-dependent cell-mediated phagocytosis (ADCP). The expression of ADCC and CDC activity of human IgG1 subclass antibodies generally involves the binding of the antibody's Fc region to receptors for antibodies (hereinafter referred to as "FcγR") present on the surface of effector cells such as killer cells, natural killer cells, or activated macrophages. Various complement components may be bound. Regarding this binding, several amino acid residues within the hinge region of the antibody and the second domain of the C region (hereinafter referred to as the "Cγ2 domain") are important (Greenwood et al. (1993) Eur.J.Immunol.23(5):1098-104; Morgan et al. (1995) Immunology.86(2):319-324; Clark, M. (1997) Chemical Immunology.65:88-110), and it has been suggested that the glycans within the Cγ2 domain (Clark, M. (1997) Chemical Immunology.65:88-110) are also important. 【0100】 To evaluate the ADCC activity of the antibody of interest, an in vitro ADCC assay, such as that described in U.S. Patent No. 5,500,362, can be used. This assay may also be performed using a commercially available kit, such as CytoTox96® (Promega). Useful effector cells for such assays include, but are not limited to, peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, and NK cell lines. NK cell lines expressing transgenic Fc receptors (e.g., CD16) and related signaling polypeptides (e.g., FCεRI-γ) can also serve as effector cells (International Publication No. 2006 / 023148). In one embodiment, an NK cell line containing CD16 and possessing luciferase under the NFAT promoter can be used to measure NK cell activation rather than cell lysis or cell death. A similar technique using Jurkat cells instead of NK cells is available from Promega (Promega ADCC Reporter Bioassay #G7010). For example, the ability of any particular antibody to mediate complement activation and / or lysis by ADCC can be assayed. The cells of interest are grown and labeled in vitro, and the antibody is added to the cell culture in combination with immune cells that can be activated by antigen-antibody complexes, i.e., effector cells involved in the ADCC response. The antibody can also be tested for complement activation. In either case, cell lysis is detected by the release of the label from the lysed cells. The degree of cell lysis may also be determined by detecting the release of cytoplasmic proteins (e.g., LDH) into the supernatant. In fact, antibodies can be screened using the patient's own serum as a source of complement and / or immune cells. Antibodies capable of mediating human ADCC in an in vitro test can then be used therapeutically in that particular patient. The ADCC activity of the molecule of interest can also be evaluated in vivo in animal models, such as those disclosed in Clynes et al. (1998) Proc. Natl. Acad. Sci. USA 95:652-656.Furthermore, techniques for modulating (i.e., increasing or decreasing) the levels of ADCC and, optionally, CDC activity of antibodies are well known in the art (e.g., U.S. Patent No. 5,624,821; U.S. Patent No. 6,194,551; U.S. Patent No. 7,317,091). The antibodies of the present invention may have the ability to induce ADCC and / or CDC, or may be modified to have such ability. Assays for determining ADCC function may be performed using human effector cells to evaluate human ADCC function. Such assays may also include those intended to screen for antibodies that induce, mediate, enhance, or block cell death by necrotic and / or apoptotic mechanisms. Such methods, including assays utilizing viable dyes, methods for detecting and analyzing caspases, and assays for measuring DNA breaks, can be used to evaluate apoptotic activity of cells cultured in vitro with the antibody of interest. 【0101】 Antibody production The following describes exemplary techniques for producing antibodies useful in the present invention. 【0102】 Monoclonal antibodies Monoclonal antibodies can be prepared using a wide range of techniques known in the art, including the use of hybridomas (Kohler et al. (1975) Nature. 256:495; Harlow et al. (1988) Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed.); Hammerling, et al. (1981) in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, NY)), recombinant techniques, and phage display techniques, or combinations thereof. The term "monoclonal antibody," as used herein, refers to an antibody obtained from a substantially homogeneous population of antibodies or isolated antibodies, for example, where the individual antibodies constituting the population are identical except for possible naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific and target a single antigenic site. Furthermore, polyclonal antibodies containing different antibodies targeting different determinants (epitopes) can also be used. In contrast to ronal antibody preparations, each monoclonal antibody targets the same determinant on the antigen. In addition to its specificity, monoclonal antibodies have the advantage of being able to be synthesized without contamination by other antibodies. The modifier "monoclonal" should not be interpreted as requiring antibody production by any particular method. The following is a description of typical methods for producing monoclonal antibodies (this description is not intended to be limiting) that can be used, for example, for the production of monoclonal mammalian antibodies, chimeric antibodies, humanized antibodies, human antibodies, domains, diabodies, vaccine bodies, linear antibodies, and multispecific antibodies. 【0103】 Hybridoma Technique Methods for producing and screening specific antibodies using hybridoma technology are routine and well-known in the art. In the hybridoma method, mice or other suitable host animals, such as hamsters, are immunized as described above to produce or induce lymphocytes capable of producing antibodies that can specifically bind to the antigen used for immunization. Alternatively, lymphocytes may be immunized in vitro. After immunization, the lymphocytes are isolated and then fused with myeloma cell lines using a suitable flux or fusion partner, such as polyethylene glycol, to form hybridoma cells (Goding (1986) Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press)). In certain embodiments, the selected myeloma cells are sensitive to a selective medium that efficiently fuses, supports stable and high antibody production by the selected antibody-producing cells, and excludes parent cells that did not fuse. In one embodiment, the myeloma cell lines are mouse myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif., USA, as well as SP-2 and its derivatives, such as X63-Ag8-653 cells, available from the American Type Culture Collection, Rockville, MD., USA. Human myeloma and mouse-human xenomyeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor (1984) J. Immunol., 133:3001; and Brodeur et al. (1987) Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York)). 【0104】 Once hybridoma cells producing antibodies with desired specificity, affinity, and / or activity are identified, the clones can be subcloned using limiting dilution procedures and grown using standard methods (Goding, op. cit.). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, hybridoma cells may be grown in vivo in animals as ascites tumors, for example, by intraperitoneal injection of the cells into mice. 【0105】 The monoclonal antibodies selected by subcloning are preferably separated from culture medium, ascites fluid, or serum by conventional antibody purification procedures, such as affinity chromatography (e.g., Protein A or Protein G Sepharose) or ion exchange chromatography, affinity tagging, hydroxyl apatite chromatography, gel electrophoresis, or dialysis. Exemplary purification methods are described in further detail below. 【0106】 Recombinant DNA techniques Methods for producing and screening specific antibodies using recombinant DNA technology are routine and well-known in the art (e.g., U.S. Patent No. 4,816,567). DNA encoding monoclonal antibodies can be readily isolated and / or sequenced using conventional procedures (e.g., by using oligonucleotide probes that have the ability to specifically bind to the genes encoding the heavy and light chains of mouse antibodies). After isolation, the DNA can be placed into an expression vector, which is then transfected into host cells that do not normally produce antibody proteins, such as Escherichia coli (E. coli) cells, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells, thereby achieving the synthesis of monoclonal antibodies in recombinant host cells. Review articles on recombinant expression of antibody-encoding DNA in bacteria include Skerra et al. (1993) Curr. Opinion in Immunol. 5:256-262 and Pluckthun (1992) Immunol. Revs. 130:151-188. As described below regarding antibodies produced by phage display and the humanization of antibodies, the antibodies of the present invention can be produced by obtaining DNA or genetic material for recombinant antibodies from one or more sources other than hybridomas. 【0107】 Recombinant expression of antibodies or their variants generally requires the construction of an expression vector containing a polynucleotide encoding the antibody. Accordingly, the present invention provides a replicable vector containing a nucleotide sequence encoding an antibody molecule, the heavy or light chain of an antibody, the heavy or light chain variable domain of an antibody or a portion thereof, or the heavy or light chain CDR, operably linked to a promoter. Such a vector may contain a nucleotide sequence encoding the constant region of an antibody molecule (see, for example, U.S. Patent Nos. 5,981,216; 5,591,639; 5,658,759 and 5,122,464), and the variable domain of the antibody may be cloned into such a vector for the expression of the entire heavy chain, the entire light chain, or both the heavy and light chains. 【0108】 In conventional methods, when an expression vector is transferred to a host cell, the transfected cells are then cultured using conventional methods to produce antibodies. Accordingly, the present invention includes a host cell containing the antibody or a fragment thereof, or its heavy chain or light chain, or a portion thereof, or a polynucleotide encoding the single-chain antibody of the present invention, operably linked to a heterologous promoter. In certain embodiments for expressing a double-chain antibody, vectors encoding both the heavy chain and the light chain may be simultaneously expressed in a host cell for the expression of the entire immunoglobulin molecule, as detailed below. 【0109】 Mammalian cell lines available as hosts for recombinant antibody expression are well known in the art and include, but are not limited to, many immortalized cell lines available from the American Type Culture Collection (ATCC), such as Chinese hamster ovary (CHO) cells, Healer cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), human epithelial kidney 293 cells, and numerous other cell lines. Different host cells have characteristic and specific mechanisms for post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be selected to ensure the correct modification and processing of the expressed antibody or a portion thereof. For this purpose, eukaryotic host cells with appropriate cellular mechanisms for primary transcript processing, gene product glycosylation, and phosphorylation may be used. Examples of such mammalian host cells include, but are not limited to, CHO, VERY, BHK, Healer, COS, MDCK, 293, 3T3, W138, BT483, Hs578T, HTB2, BT2O, and T47D, NS0 (a mouse myeloma cell line that does not endogenously produce any functional immunoglobulin chains), SP20, CRL7O3O, and HsS78Bst cells. Monoclonal antibodies can be recombinantly produced using human cell lines developed by immortalizing human lymphocytes. Monoclonal antibodies can be recombinantly produced using the human cell line PER.C6® (Crucell, Netherlands). 【0110】 Further cell lines that may be used as hosts for recombinant antibody expression include, but are not limited to, insect cells (e.g., Sf21 / Sf9, Trichoplusia ni Bti-Tn5b1-4), yeast cells (e.g., S. cerevisiae, Pichia, U.S. Patent No. 7,326,681; etc.), plant cells (U.S. Patent Publication No. 20080066200); and chicken cells (International Publication No. 2008142124). 【0111】 In certain embodiments, the antibody of the present invention is expressed in a cell line exhibiting stable antibody expression. Stable expression can be used for long-term, high-yield production of recombinant proteins. For example, a cell line that stably expresses the antibody molecule may be created. Host cells can be transformed with a appropriately engineered vector containing expression regulatory elements (e.g., promoters, enhancers, transcriptional terminators, polyadenylation sites, etc.) and a selectable marker gene. After introduction of the foreign DNA, the cells may be grown in fortified medium for 1-2 days, then switched to selective medium. The selectable marker in the recombinant plasmid confers resistance to selection, enabling the growth of cells with stably incorporated plasmids into their chromosomes and the formation of foci, which can then be cloned and expanded into a cell line. Methods for producing stable cell lines in high yield are well known in the art, and reagents are generally commercially available. 【0112】 In certain embodiments, the antibody of the present invention is expressed in a cell line exhibiting transient transfection of the antibody. Transient transfection is a process in which nucleic acids introduced into a cell are not integrated into the cell's genome or chromosomal DNA. They are actually maintained in the cell as extrachromosomal elements, such as episomes. The transcription process of the nucleic acids in the episome remains unaffected, and proteins encoded by the nucleic acids in the episome are produced. 【0113】 Cell lines, whether stably or transiently transfected, are maintained in cell culture media and conditions known in the art to result in the expression and production of monoclonal antibodies. In certain embodiments, the mammalian cell culture medium is based on a commercially available culture medium formulation, such as DMEM or Ham F12. In other embodiments, the cell culture medium is modified to support both increased cell growth and increased biological protein expression. As used herein, the terms “cell culture medium,” “culture medium,” and “culture medium formulation” refer to a nutrient solution for maintaining, growing, proliferating, or expanding cells in an artificial in vitro environment outside of a multicellular organism or tissue. Cell culture media may be optimized for specific cell culture applications, including, for example, cell culture growth media formulated to promote cell growth, or cell culture production media formulated to promote recombinant protein production. The terms “nutrient,” “component,” and “constituent” are used herein synonymously to refer to the components that make up a cell culture medium. 【0114】 In one embodiment, the cell line is maintained using a fed-batch method. As used herein, “fed-batch method” refers to a method in which, after being initially incubated in a basal medium, additional nutrients are supplied to the cell culture. For example, a fed-batch method may include adding supplemental medium according to a predetermined replenishment schedule within a given time. Thus, “fed-batch cell culture” refers to a cell culture in which cells, typically mammalian cells, and culture medium are initially supplied to the culture vessel, and additional culture nutrients are continuously or discontinuously and gradually supplied to the culture during culture, with or without periodic cell and / or product retrieval before the end of culture. 【0115】 The cell culture media used and the nutrients contained herein are well known to those skilled in the art. In one embodiment, the cell culture medium comprises a basal medium and at least one hydrolysate, for example, a soy-based hydrolysate, a yeast-based hydrolysate, or a combination of two such hydrolysates, to provide a modified basal medium. In another embodiment, the further nutrients may consist only of the basal medium, such as a concentrated basal medium, or only of the hydrolysates or concentrated hydrolysates. Suitable basal media include, but are not limited to, Dulbecco's Modified Eagle Medium (DMEM), DME / F12, Minimum Essential Medium (MEM), Eagle Basal Medium (BME), RPMI 1640, F-10, F-12, α-Minimum Essential Medium (α-MEM), Glasgow Minimum Essential Medium (G-MEM), PF CHO (see, for example, CHO Protein-Free Medium (Sigma) or Protein-Free EX-CELL™ 325 for CHO Cells, PF CHO Serum-Free Medium (SAFC Bioscience), and Iskov Modified Dulbecco Medium. Other examples of basal media that may be used in the present invention include BME Basal Medium (see also Gibco-Invitrogen; Eagle, H (1965) Proc. Soc. Exp. Biol. Med. 89, 36); Dulbecco's Modified Eagle Medium (DMEM, powder) (Gibco-Invitrogen (#31600); Dulbecco and Examples include Freeman (1959) Virology. 8:396; Smith et al. (1960) Virology. 12:185. See also Tissue Culture Standards Committee, In Vitro 6:2,93); and CMRL 1066 medium (Gibco-Invitrogen (#11530); see also Parker et al. (1957) Special Publications, NYAcademy of Sciences, 5:303). 【0116】 The basal medium may be serum-free, meaning the medium does not contain serum (e.g., fetal bovine serum (FBS), horse serum, goat serum, or any other animal-derived serum known to those skilled in the art), or it may be an animal protein-free medium or a chemically restricted medium. 【0117】 The basal medium may be modified to remove certain non-nutrient components found in standard basal media, such as various inorganic and organic buffers, one or more surfactants, and sodium chloride. By removing such components from the basal cell medium, the concentration of remaining nutrients can be increased, potentially improving overall cell growth and protein expression. In addition, the removed components may be added back to the cell culture medium containing the modified basal cell medium, depending on the requirements of the cell culture conditions. In certain embodiments, the cell culture medium contains the modified basal cell medium and at least one of the following nutrients: an iron source, recombinant growth factors; buffers; surfactants; volumetric osmolality regulators; energy sources; and non-animal hydrolysates. In addition, the modified basal cell medium may optionally contain amino acids, vitamins, or a combination of both amino acids and vitamins. In another embodiment, the modified basal medium further contains glutamine, e.g., L-glutamine, and / or methotrexate. 【0118】 Antibody production can be carried out on a large scale by bioreactor processes using known flow-add, batch, perfusion, or continuous-feed bioreactor methods in the art. Large-scale bioreactors have a capacity of at least 1,000 liters, and in one embodiment, a capacity of about 1,000 to 100,000 liters. Such bioreactors may use agitator impellers to distribute oxygen and nutrients. Small-scale bioreactors generally refer to cell cultures with a volume of about 100 liters or less, and may range from about 1 liter to about 100 liters. Alternatively, single-use bioreactors (SUBs) may be used for both large-scale and small-scale cultures. 【0119】 Temperature, pH, agitation, aeration, and inoculation density may vary depending on the host cells used and the recombinant protein to be expressed. For example, recombinant protein cell cultures may be maintained at a temperature of 30°C to 45°C. The pH of the culture medium may be monitored to maintain an optimal level during the culture process, which may be in the pH range of 6.0 to 8.0 for certain host cells. Mixing in such culture methods may be performed by impeller-driven mixing. The rotation speed of the impeller may be a tip speed of approximately 50 to 200 cm / second, however, depending on the type of host cell being cultured, other airlift or other mixing / aeration systems known in the art may be used. By providing sufficient aeration, a dissolved oxygen concentration of approximately 20% to 80% air saturation in the culture is maintained, again depending on the specific host cell being cultured. Alternatively, air or oxygen may be directly diffused into the culture medium by a bioreactor. Other oxygen supply methods exist, including bubble-free aeration systems using hollow fiber membrane aeration devices. 【0120】 Phage display technique Monoclonal antibodies or antibody fragments can be isolated from antibody phage libraries prepared using techniques described in McCafferty et al. (1990) Nature. 348:552-554, Clackson et al. (1991) Nature (1991) 352:624-628, and Marks et al. (1991) J. Mol. Biol. 222:581-597. In such a method, antibodies can be isolated by screening a recombinant combinatorial antibody library, in one embodiment an scFv phage display library, prepared using human VL and VH cDNA prepared from mRNA derived from human lymphocytes. Methods for preparing and screening such libraries are known in the art. In addition to commercially available kits for preparing phage display libraries (e.g., Pharmacia Recombinant Phage Antibody System, catalog no. 27-9400-01; and Stratagene), commercially available kits for preparing phage display libraries are also available. Examples of methods and reagents particularly suitable for the preparation and screening of antibody display libraries using the SurfZAP (trademark) phage display kit (catalog number 240612) are given in U.S. Patent Nos. 6,248,516; 6,545,142; 6,291,158; 6,291,159; 6,291,160; and 6,291,161. These can be found in the following specifications: 6,680,192; 5,969,108; 6,172,197; 6,806,079; 5,885,793; 6,521,404; 6,544,731; 6,555,313; 6,593,081; 6,582,915; and 7,195,866. Therefore, these techniques are viable alternatives to conventional monoclonal antibody hybridoma techniques for the production and isolation of monoclonal antibodies. 【0121】 In phage display methods, a functional antibody domain is presented on the surface of a phage particle having a polynucleotide sequence encoding it. In detailed embodiments, such phages can be used to present antigen-binding domains expressed from a repertoire or combinatorial antibody library (e.g., human or mouse). Phages expressing antigen-binding domains that bind to a target antigen can be selected or identified by the antigen, for example, using a labeled antigen or an antigen bound to or captured on a solid surface or beads. The phages used in these methods are typically filamentous phages containing fd and M13-binding domains expressed from phages having Fab, Fv, or disulfide-stabilized Fv antibody domains recombinantly fused with either phage gene III or gene VIII protein. 【0122】 As described in the above references, after phage selection, the antibody-coding region can be isolated from the phage and used to create human antibodies, whole antibodies including humanized antibodies, or any other desired antigen-binding fragments, which can then be expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, as described in detail below. For example, recombinant production techniques for Fab, Fab', and F(ab')2 fragments can also be employed using methods known in the art, such as those disclosed in International Publication No. 92 / 22324; Mullinax et al. (1992) BioTechniques. 12(6):864-869; and Better et al. (1988) Science. 240:1041-1043. 【0123】 Examples of techniques that can be used to produce single-chain Fv and antibodies are those described in U.S. Patent No. 4,946,778 and No. 5,258,498. Accordingly, recombinant antibodies can be prepared using the techniques described above and techniques known in the art, in which the binding domain, e.g., ScFv, was isolated from a phage display library. 【0124】 Antibody purification and isolation Following production by recombinant expression or hybridoma expression, the antibody molecule can be purified by any method known in the art for purifying immunoglobulin molecules, for example, by chromatography (e.g., by ion exchange, affinity, particularly affinity for specific antigen protein A or protein G, and size exclusion column chromatography), centrifugation, by solubility differential, or by any other standard protein purification technique. Furthermore, the antibody or fragment thereof of the present invention may be fused to a heterologous polypeptide sequence (referred to herein as a “tag”) known to facilitate purification. 【0125】 When recombinant technology is used, antibodies can be produced intracellularly, in the perimembranous space of the cell membrane, or directly secreted into the culture medium. If antibodies are produced intracellularly, the first step is to remove particulate debris, which is either host cells or lysed fragments, for example by centrifugation or ultrafiltration. Carter et al. (1992) Bio / Technology. 10:163-167 describes a procedure for isolating antibodies secreted into the perimembranous space of Escherichia coli (E. coli). If antibodies are secreted into the culture medium, generally, the supernatant from such an expression system is first concentrated using a commercially available protein concentration filter, such as an Amicon or Millipore Pellicon ultrafiltration unit. Proteolysis may be inhibited in any of the aforementioned steps by including a protease inhibitor such as PMSF, and the growth of exogenous contaminants may be prevented by including antibiotics. 【0126】 Antibody compositions prepared from cells can be purified using, for example, hydroxyl apatite chromatography, hydrophobic interaction chromatography, ion exchange chromatography, gel electrophoresis, dialysis, and / or affinity chromatography, either alone or in combination with other purification steps. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc domain present in the antibody, as will be understood by those skilled in the art. The matrix to which the affinity ligand binds is almost always agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for higher flow rates and shorter processing times that can be achieved with agarose. If the antibody contains a CH3 domain, Bakerbond ABX resin (JTBaker, Phillipsburg, NJ) is useful for purification. Other protein purification techniques, such as fractionation using ion-exchange columns, ethanol precipitation, reverse-phase HPLC, chromatography with silica, chromatography with heparin, Sepharose chromatography using anion or cation exchange resins (such as polyaspartate columns), chromatographic focusing, SDS-PAGE, and ammonium sulfate precipitation, are also available depending on the antibody to be recovered. 【0127】 After any preliminary purification steps, the mixture containing the antibody of interest and contaminants can be subjected to low-pH hydrophobic interaction chromatography, performed using an elution buffer with a pH of approximately 2.5–4.5 and a low salt concentration (e.g., approximately 0–0.25 M salt). 【0128】 Accordingly, in certain embodiments, substantially purified / isolated antibodies of the present invention are provided. In one embodiment, these isolated / purified recombinantly expressed antibodies may be administered to a patient to mediate a prophylactic or therapeutic effect. Prophylactic is a drug therapy or a treatment designed and used to prevent the onset of a disease, disorder, or infection. Therapeutic is specifically related to the treatment of a particular disease, disorder, or infection. Therapeutic dose is the amount required to treat a particular disease, disorder, or infection. In another embodiment, these isolated / purified antibodies may be used to diagnose influenza virus infection, for example, influenza B virus infection, or in other embodiments, influenza A and influenza B virus infection. 【0129】 Human antibodies Human antibodies can be produced using methods well known in the art. Human antibodies avoid some of the problems associated with antibodies that have mouse or rat variable and / or constant regions. The presence of such mouse or rat-derived proteins can lead to rapid clearance of the antibody or to the development of an immune response to the antibody by the patient. 【0130】 Human antibodies can also be obtained by in vitro methods. Suitable examples, though not limited to, include phage display (MedImmune (formerly CAT), Morphosys, Dyax, Biosite / Medarex, Xoma, Symphogen, Alexion (formerly Proliferon), Affimed), ribosome display (MedImmune (formerly CAT)), and yeast display. Phage display technology (see, for example, U.S. Patent No. 5,969,108) allows for the in vitro production of human antibodies or antibody fragments from an immunoglobulin variable (V) domain gene repertoire derived from an unimmunized donor. This technology involves in-frame cloning of antibody V domain genes into major or minor coat protein genes of filamentous bacteriophages such as M13 or fd, and presenting them as functional antibody fragments on the surface of phage particles. Since these filamentous particles contain single-stranded DNA copies of the phage genome, selection based on the functional characteristics of the antibody, as well as the genes encoding antibodies exhibiting those characteristics, can be performed. Therefore, phages mimic some characteristics of B cells. Phage display can be carried out in various formats, as reviewed, for example, in Johnson, Chiswell (1993) Current Opinion in Structural Biology. 3:564-571. Several V gene segment sources can be used for phage display. Clackson et al. (1991) Nature. 352:624-628 isolated diverse arrays of anti-oxazolone antibodies from a small random combinatorial library of V genes derived from the spleen of immunized mice. Essentially, following the techniques described by Marks et al. (1991) J.Mol.Biol. 222:581-597, or Griffith et al. (1993) EMBO J. 12:725-734, a repertoire of V genes derived from non-immunized human donors can be constructed, and antibodies against diverse arrays of antigens (including autoantigens) can be isolated. See also U.S. Patent Nos. 5,565,332 and 5,573,905. 【0131】 As discussed above, human antibodies may also be produced using in vitro activated B cells (see U.S. Patent Nos. 5,567,610 and 5,229,275). 【0132】 Immunoglobulin genes undergo various modifications during the maturation of the immune response, including recombination, isotype switching, and hypermutation between V, D, and J gene segments in the variable region. While recombination and somatic hypermutation are fundamental to antibody diversity and affinity maturation, they can also introduce sequence liabilities, making commercial production of such immunoglobulins as therapeutic agents difficult or increasing the risk of immunogenicity. Generally, mutations in the CDR region are thought to contribute to improved affinity and function, while mutations in the framework region can increase the risk of immunogenicity. This risk can be mitigated by reintroducing framework mutations into the germline while ensuring that antibody activity is not adversely affected. Diversification processes may introduce some structural liabilities, or such structural liabilities may be present in germline sequences contributing to the heavy and light chain variable domains. Regardless of the source, it may be desirable to eliminate potential structural liabilities that can lead to instability, aggregation, product heterogeneity, or increased immunogenicity. Examples of undesirable liabilities include unpaired cysteine ​​(which can lead to scrambling of disulfide bonds or the formation of variable sulfhydryl adducts), N-linked glycosylation sites (resulting in structural and activity heterogeneity), and amidation (e.g., NG, NS), isomerization (DG), oxidation (exposed methionine), and hydrolysis (DP) sites. 【0133】 Therefore, in order to reduce the risk of immunogenicity and improve the pharmaceutical properties, it may be desirable to reintroduce the framework sequence into the germline, reintroduce the CDR into the germline, and / or eliminate structural liabilities. 【0134】 Therefore, in one embodiment, if a particular antibody differs from its respective germline sequence at the amino acid level, the antibody sequence can be reverse-mutated into the germline sequence. Such corrective mutations can occur using standard molecular biological techniques at one, two, three, or more positions, or at any combination of the mutated positions. 【0135】 antibody fragment In certain embodiments, the antibody is an antibody fragment or an antibody containing such fragments. An antibody fragment generally comprises a portion of a full-length antibody that is typically its antigen-binding region or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, Fd, and Fv fragments. Diabodies are linear antibodies (U.S. Patent No. 5,641,870) and single-chain antibody molecules. 【0136】 Traditionally, these fragments were obtained by protein digestion of intact antibodies using techniques known in the art. However, these fragments can now be directly produced by recombinant host cells. Since Fab, Fv, and scFv antibody fragments can all be expressed in and secreted from Escherichia coli (E. coli) cell types, these fragments can be easily mass-produced. In one embodiment, antibody fragments can be isolated from the antibody phage library discussed above. Alternatively, Fab'-SH fragments can be directly recovered from Escherichia coli (E. coli) and chemically coupled to form F(ab')2 fragments (Carter et al. (1992)) Bio / Technology.10:163-167. By another method, F(ab')2 fragments can be directly isolated from recombinant host cell cultures. Other techniques for producing antibody fragments will be apparent to those skilled in the art. In other embodiments, the antibody of choice is a single-chain Fv fragment (scFv). In certain embodiments, the antibody is not a Fab fragment. Fv and scFv are the only species that possess an intact binding site lacking a constant region; therefore, they are suitable for reducing nonspecific binding during in vivo use. scFv fusion proteins may be constructed such that the effector protein fuses to either the amino or carboxyl end of scFv. 【0137】 In certain embodiments, this antibody is a domain antibody, for example, a human antibody with variable heavy chain (V H ) or light chain variable (V L These are antibodies that contain small functional binding units of the antibody corresponding to the ) region. Examples of domain antibodies include, but are not limited to, those of Domantis (see, for example, International Publication No. 04 / 058821; International Publication No. 04 / 081026; International Publication No. 04 / 003019; International Publication No. 03 / 002609; U.S. Patent No. 6,291,158; U.S. Patent No. 6,582,915; U.S. Patent No. 6,696,245; and U.S. Patent No. 6,593,081). 【0138】 In certain embodiments of the present invention, the antibody is a linear antibody. The linear antibody has a pair of tandem Fd segments (V) that form a pair of antigen-binding regions. H -C H1 -V H -C H1 ) includes. See Zapata et al. (1995) Protein Eng. 8(10):1057-1062. 【0139】 Other amino acid sequence modifications In addition to the human antibodies, humanized antibodies and / or chimeric antibodies mentioned above, the present invention also includes variable light chains (V L ) domain and / or variable heavy chain (V H Further modifications of the antibodies of the present invention, including substitutions, additions, and / or deletions of one or more amino acid residues and / or polypeptides in the domain and / or Fc region, and post-translational modifications, as well as variants and fragments thereof. These modifications include antibody conjugates in which an antibody is covalently bound to a portion. Suitable portions for binding to the antibody include, but are not limited to, proteins, peptides, drugs, labels, and cytotoxins. Such modifications to antibodies may be made to alter or fine-tune the (biochemical, binding, and / or functional) properties of the antibody so that it is suitable for the treatment and / or diagnosis of influenza virus infection. Methods for forming conjugates and adding amino acid and / or polypeptide changes and post-translational modifications are known in the art and some of them are detailed below. 【0140】 Amino acid modifications to an antibody inevitably result in a sequence with less than 100% identity to the antibody sequence or parent antibody sequence identified above. In certain embodiments, the antibody may have about 25% to about 95% sequence identity to the amino acid sequence of either the heavy chain or light chain variable domain of the antibody as described herein. Thus, in one embodiment, the modified antibody may have an amino acid sequence having at least 25%, 35%, 45%, 55%, 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity or similarity to the amino acid sequence of either the heavy chain or light chain variable domain of the antibody as described herein. In another embodiment, the modified antibody may have an amino acid sequence having at least 25%, 35%, 45%, 55%, 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity or similarity to the amino acid sequence of the heavy or light chain CDR-1, CDR-2, or CDR-3 of the antibody as described herein. In another embodiment, the modified antibody may have an amino acid sequence having at least 25%, 35%, 45%, 55%, 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity or similarity to the amino acid sequence of the heavy or light chain FR1, FR2, FR3, or FR4 of the antibody as described herein. 【0141】 In certain embodiments, modified antibodies are created by one or more amino acid modifications (e.g., substitutions, deletions, and / or additions) introduced into one or more of the variable regions of the antibody. In other embodiments, amino acid modifications are introduced into the framework region. Modifying one or more framework region residues can result in improved antibody binding affinity to the antigen. This is particularly true when such modifications are made to humanized antibodies in which the framework region may be formed from a different species than the CDR region. Examples of framework region residues to be modified include those that directly and non-covalently bind the antigen (Amit et al. (1986) Science. 233:747-753); those that interact with / cause conformation of the CDR (Chothia et al. (1987) J. Mol. Biol. 196:901-917); and / or V L -V H This includes those involved in the junction (U.S. Patents No. 5,225,539 and No. 6,548,640). In one embodiment, about 1 to about 5 framework residues may be modified. Sometimes this may be sufficient to obtain an antibody variant suitable for use in preclinical trials, even if no hypervariable region residues are modified. However, typically, a modified antibody may contain one or more further hypervariable region modifications. 【0142】 One useful procedure for creating modified antibodies is called "alanine scanning mutagenesis" (Cunningham and Wells (1989) Science. 244:1081-1085). In this method, the amino acid interaction with the target antigen is modified by substituting one or more hypervariable region residues with alanine or polyalanine residues. Then, one or more of these hypervariable region residues that are functionally sensitive to the substitution are refined by introducing an additional or other mutation at the substitution site or for the substitution site. Thus, the site of introduction of the amino acid sequence mutation is predetermined, but the nature of the mutation itself does not need to be predetermined. The Ala mutants produced in this way are screened for their biological activity as described herein. 【0143】 In certain embodiments, substitution mutants involve substituting one or more hypervariable region residues of the parent antibody (e.g., a humanized antibody or a human antibody). Generally, one or more mutants selected for further development may have improved biological properties compared to the parent antibody from which they were created. A convenient method for creating such substitution mutants involves affinity maturation using phage display (Hawkins et al. (1992) J.Mol.Biol.254:889-896 and Lowman et al. (1991) Biochemistry.30(45):10832-10837). In short, by mutating several hypervariable region sites (e.g., 6-7 sites), any possible amino acid substitutions are created at each site. The antibody mutants thus created are presented in a monovalent form from filamentous phage particles as fusions with the M13 gene III product packaged within each particle. Next, the mutants presented by the phage are screened for their biological activity (e.g., binding affinity) as disclosed herein. 【0144】 Mutations in antibody sequences may include substitutions, deletions (including internal deletions), additions (including additions that result in fusion proteins), or conservative substitutions of amino acid residues within and / or adjacent to the amino acid sequence, but which result in "silent" changes in that the change produces a functionally equivalent antibody. Conservative amino acid substitutions may be made based on the similarity of the polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or amphiphilicity of the residues involved. For example, nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine; polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine; positively charged (basic) amino acids include arginine, lysine, and histidine; and negatively charged (acidic) amino acids include aspartic acid and glutamic acid. In addition, glycine and proline are residues that can affect chain orientation. Non-conservative substitutions may involve replacing one member of one of these classes with a member of another. Furthermore, if necessary, non-classical amino acids or chemical amino acid analogs can be introduced into the antibody sequence as substitutions or additions. Non-classical amino acids include, but are not limited to, D-isomers of common amino acids, α-aminoisobutyric acid, 4-aminobutyric acid, Abu, 2-aminobutyric acid, γ-Abu, ε-Ahx, 6-aminohexanoic acid, Aib, 2-aminoisobutyric acid, 3-aminopropionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, β-alanine, fluoroamino acids, designer amino acids such as β-methylamino acids, Cα-methylamino acids, Nα-methylamino acids, and amino acid analogs in general. 【0145】 In another embodiment, any cysteine ​​residues not involved in maintaining the proper conformation of the antibody may also be substituted with serine in general, thereby improving the oxidative stability of the molecule and preventing abnormal crosslinking. Conversely, adding one or more cysteine ​​bonds to the antibody may improve its stability (especially if the antibody is an antibody fragment such as an Fv fragment). 【0146】 Mutant Fc region Mutations in the Fc region (e.g., amino acid substitutions and / or additions and / or deletions) enhance or diminish the effector function of antibodies (e.g., U.S. Patent No. 5,624,821; U.S. Patent No. 5,885,573; U.S. Patent No. 6,538,124; U.S. Patent No. 7,317,091; U.S. Patent No. 5,648,260; U.S. Patent No. 6,538,124; International Publication No. 03 / 074679; International Publication No. 04 / 02920) It is known that the pharmacokinetic properties (e.g., half-life) of the antibody can be modified (see Pamphlet No. 7; International Publication No. 04 / 099249; International Publication No. 99 / 58572; U.S. Patent Application Publication No. 2006 / 0134105; U.S. Patent Application Publication No. 2004 / 0132101; U.S. Patent Application Publication No. 2006 / 0008883), and that the pharmacokinetic properties (e.g., half-life) of the antibody can be modified (see U.S. Patents No. 6,277,375 and No. 7,083,784). Accordingly, in certain embodiments, the antibody of the present invention comprises a modified Fc region (also referred to herein as a “mutated Fc region”), where one or more modifications have been made to the Fc region to alter the functional and / or pharmacokinetic properties of the antibody. Such modifications may result in a decrease or increase in Clq binding and complement-dependent cytotoxicity (CDC) or FcγR binding, and antibody-dependent cytotoxicity (ADCC) or antibody-dependent cell-mediated phagocytosis (ADCP) with respect to IgG. The present invention encompasses antibodies described herein that have mutant Fc regions that have been modified to provide desired effector function through fine-tuning to enhance or diminish effector function. Accordingly, antibodies of the present invention include mutant Fc regions (i.e., Fc regions modified as discussed below). Antibodies of the present invention containing mutant Fc regions are also referred to herein as “Fc mutant antibodies”. As used herein, “natural” refers to an unmodified parent sequence, and antibodies containing a natural Fc region are referred to herein as “natural Fc antibodies”. Fc mutant antibodies can be prepared by a number of methods well known to those skilled in the art. Non-limiting examples include isolating an antibody coding region (e.g., from a hybridoma) and making one or more desired substitutions in the Fc region of the isolated antibody coding region.Alternatively, the antigen-binding portion of the antibody (e.g., the variable region) may be subcloned into a vector encoding the mutant Fc region. In one embodiment, the mutant Fc region exhibits a similar level of effector function induction compared to the natural Fc region. In another embodiment, the mutant Fc region exhibits a higher level of effector function induction compared to the natural Fc region. Several specific embodiments of the mutant Fc region are described in detail below. Methods for measuring effector function are well known in the art. 【0147】 The effector function of an antibody is modified by altering the Fc region, including, but not limited to, amino acid substitution, amino acid addition, amino acid deletion, and post-translational modifications to the Fc amino acid (e.g., glycosylation). By using the methods described below, the effector function of this antibody and the ratio of the binding properties of the Fc region to the FcR (e.g., affinity and specificity) can be fine-tuned to produce a therapeutic antibody with the desired properties. 【0148】 As used herein, the Fc region is understood to include polypeptides that constitute the constant region of an antibody, excluding the first constant region immunoglobulin domain. Thus, Fc refers to the last two constant region immunoglobulin domains of IgA, IgD, and IgG, and the last three constant region immunoglobulin domains of IgE and IgM, and the mobile hinge N-terminus to these domains. With respect to IgA and IgM, Fc may include the J chain. With respect to IgG, Fc may include the immunoglobulin domains C-gamma 2 and C-gamma 3 (Cγ2 and Cγ3), and the hinge between C-gamma 1 (Cγ1) and C-gamma 2 (Cγ2). 【0149】 While the boundaries of the Fc region can vary, the human IgG heavy chain Fc region can be defined as including residues C226 or P230 and their carboxyl ends, where numbering is based on the EU index as shown in Kabat. Fc may refer to this region in isolation, or to this region in the context of an antibody, antibody fragment, or Fc fusion protein. Polymorphisms have been observed at several different Fc positions, including positions 270, 272, 312, 315, 356, and 358 when numbered by the EU index, and therefore slight differences may exist between the presented sequence and the sequence of prior art. 【0150】 In one embodiment, when compared to a natural Fc antibody, the Fc mutant antibody exhibits a modified binding affinity to one or more Fc receptors, including, but not limited to, FcRn, isoforms FcγRIA, FcγRIB, and FcγRIC (FcγRI(CD64); FcγRII(CD32, isoforms FcγRIIA, FcγRIIB, and FcγRIIC); and FcγRIII(CD16, isoforms FcγRIIIA and FcγRIIIB). 【0151】 In one embodiment, the Fc mutant antibody has enhanced binding to one or more Fc ligands compared to the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits an increased or decreased affinity for Fc ligands that is at least 2 times, or at least 3 times, or at least 5 times, or at least 7 times, or at least 10 times, or at least 20 times, or at least 30 times, or at least 40 times, or at least 50 times, or at least 60 times, or at least 70 times, or at least 80 times, or at least 90 times, or at least 100 times, and up to 25 times, or up to 50 times, or up to 75 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times higher or lower than that of the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits an affinity for the Fc ligand that is at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, or at least 5% higher or lower than that of the natural Fc antibody. In a particular embodiment, the Fc mutant antibody has increased affinity for the Fc ligand. In another embodiment, the Fc mutant antibody has decreased affinity for the Fc ligand. 【0152】 In a specific embodiment, the Fc mutant antibody has enhanced binding affinity to the Fc receptor FcγRIIIA. In another specific embodiment, the Fc mutant antibody has enhanced binding affinity to the Fc receptor FcγRIIB. In yet another specific embodiment, the Fc mutant antibody has enhanced binding affinity to both the Fc receptors FcγRIIIA and FcγRIIB. In a particular embodiment, the Fc mutant antibody having enhanced binding affinity to FcγRIIIA does not have the accompanying increase in binding affinity to the FcγRIIB receptor compared to the native Fc antibody. In a specific embodiment, the Fc mutant antibody has reduced binding affinity to the Fc receptor FcγRIIIA. In yet another specific embodiment, the Fc mutant antibody exhibiting modified affinity to FcγRIIIA and / or FcγRIIB has enhanced binding affinity to the Fc receptor FcRn. In yet another specific embodiment, an Fc mutant antibody exhibiting modified affinity for FcγRIIIA and / or FcγRIIB has modified binding affinity for C1q compared to a natural Fc antibody. 【0153】 In one embodiment, the Fc mutant antibody exhibits an affinity for the FcγRIIIA receptor that is at least 2 times, or at least 3 times, or at least 5 times, or at least 7 times, or at least 10 times, or at least 20 times, or at least 30 times, or at least 40 times, or at least 50 times, or at least 60 times, or at least 70 times, or at least 80 times, or at least 90 times, or at least 100 times, or up to 50 times, or up to 60 times, or up to 70 times, or up to 80 times, or up to 90 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times higher or lower than that of the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits an affinity for FcγRIIIA that is at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, or at least 5% higher or lower than that of the natural Fc antibody. 【0154】 In one embodiment, the Fc mutant antibody exhibits an affinity for the FcγRIIB receptor that is at least 2 times, or at least 3 times, or at least 5 times, or at least 7 times, or at least 10 times, or at least 20 times, or at least 30 times, or at least 40 times, or at least 50 times, or at least 60 times, or at least 70 times, or at least 80 times, or at least 90 times, or at least 100 times, or up to 50 times, or up to 60 times, or up to 70 times, or up to 80 times, or up to 90 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times higher or lower than that of the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits an affinity for FcγRIIB that is at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, or at least 5% higher or lower than that of the natural Fc antibody. 【0155】 In one embodiment, the Fc mutant antibody exhibits increased or decreased affinity for C1q compared to the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits at least 2 times, or at least 3 times, or at least 5 times, or at least 7 times, or at least 10 times, or at least 20 times, or at least 30 times, or at least 40 times, or at least 50 times, or at least 60 times, or at least 70 times, or at least 80 times, or at least 90 times, or at least 100 times, or up to 50 times, or up to 60 times, or up to 70 times, or up to 80 times, or up to 90 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times higher or lower affinity for the C1q receptor compared to the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits an affinity for C1q that is at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, or at least 5% higher or lower than that of the natural Fc antibody. In yet another specific embodiment, the Fc mutant antibody exhibiting a modified affinity for Ciq has enhanced binding affinity to the Fc receptor FcRn. In yet another specific embodiment, the Fc mutant antibody exhibiting a modified affinity for C1q has modified binding affinity to FcγRIIIA and / or FcγRIIB compared to that of the natural Fc antibody. 【0156】 It is well known in the art that antibodies have the ability to induce attack and destruction through a series of processes collectively known as antibody effector functions. One of these processes, known as "antibody-dependent cell-mediated cytotoxicity" or "ADCC," refers to a form of cytotoxicity in which secreted Ig bound to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) allows these cytotoxic effector cells to specifically bind to antigen-carrying cells, subsequently killing the cells cytotoxicly. Specific high-affinity IgG antibodies directed towards the cell surface "equip" the cytotoxic cells, and this is essential for such death. Cell lysis is extracellular, requires direct cell-cell contact, and does not involve complements. 【0157】 Another process encompassed by the term "effector function" is complement-dependent cytotoxicity (hereinafter referred to as "CDC"), which refers to the biochemical event of cell destruction by the complement system. The complement system is a complex system of proteins found in normal plasma that, in combination with antibodies, destroys pathogenic bacteria and other foreign cells. 【0158】 Another process encompassed by the term effector function is antibody-dependent cell-mediated phagocytosis (ADCP), which refers to a cell-mediated response in which nonspecific cytotoxic cells expressing one or more effector ligands recognize bound antibodies on the cell, subsequently leading to phagocytosis of the cell. 【0159】 Fc mutant antibodies are intended to be characterized by in vitro functional assays to determine the function of one or more FcγR-mediated effector cells. In certain embodiments, Fc mutant antibodies have similar binding properties and effector cell functions in in vivo models (such as those described and disclosed herein) as in in vitro-based assays. However, the present invention does not exclude Fc mutant antibodies that do not exhibit the desired phenotype in in vitro-based assays but do exhibit the desired phenotype in vivo. 【0160】 In certain embodiments, antibodies containing the Fc variant exhibit enhanced cytotoxicity or phagocytic activity (e.g., ADCC, CDC, and ADCP) compared to antibodies containing the natural Fc region. In specific embodiments, Fc-mutated antibodies have at least 2 times, or at least 3 times, or at least 5 times, or at least 10 times, or at least 50 times, or at least 100 times, or up to 50 times, or up to 75 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times higher cytotoxicity or phagocytic activity compared to natural Fc antibodies. Alternatively, Fc-mutated antibodies exhibit reduced cytotoxicity or phagocytic activity compared to natural Fc antibodies. In specific embodiments, the Fc mutant antibody has at least 2 times, or at least 3 times, or at least 5 times, or at least 10 times, or at least 50 times, or at least 100 times, or up to 50 times, or up to 75 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times lower cytotoxicity or phagocytic activity compared to the natural Fc antibody. 【0161】 In certain embodiments, the Fc mutant antibody exhibits reduced ADCC activity compared to the natural Fc antibody. In other embodiments, the Fc mutant antibody exhibits ADCC activity that is at least 2 times, or at least 3 times, or at least 5 times, or at least 10 times, or at least 50 times, or at least 100 times, or up to 50 times, or up to 75 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times lower than that of the natural Fc antibody. In yet another embodiment, the Fc mutant antibody exhibits ADCC activity that is at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100%, or at least 200%, or at least 300%, or at least 400%, or at least 500% lower than that of the natural Fc antibody. In a particular embodiment, the Fc mutant antibody has undetectable ADCC activity. In a specific embodiment, the reduction and / or ablatement of ADCC activity may be due to the reduced affinity that the Fc mutant antibody exhibits to the Fc ligand and / or receptor. 【0162】 In other embodiments, the Fc mutant antibody exhibits enhanced ADCC activity compared to the natural Fc antibody. In yet another embodiment, the Fc mutant antibody exhibits ADCC activity at least 2 times, or at least 3 times, or at least 5 times, or at least 10 times, or at least 50 times, or at least 100 times, compared to the natural Fc antibody. In yet another embodiment, the Fc mutant antibody exhibits ADCC activity that is at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100%, or at least 200%, or at least 300%, or at least 400%, or at least 500% enhanced compared to the natural Fc antibody. In specific embodiments, the enhanced ADCC activity may be due to the enhanced affinity that the Fc mutant antibody exhibits to the Fc ligand and / or receptor. 【0163】 In certain embodiments, the Fc mutant antibody exhibits enhanced binding to the Fc receptor FcγRIIIA and enhanced ADCC activity compared to the natural Fc antibody. In other embodiments, the Fc mutant antibody has both enhanced ADCC activity and an increased serum half-life compared to the natural Fc antibody. In yet another specific embodiment, the Fc mutant antibody exhibits reduced binding to the Fc receptor FcγRIIIA and decreased ADCC activity compared to the natural Fc antibody. In yet another embodiment, the Fc mutant antibody has both decreased ADCC activity and an increased serum half-life compared to the natural Fc antibody. 【0164】 In certain embodiments, cytotoxicity is mediated by CDC, where the Fc mutant antibody has either enhanced or reduced CDC activity compared to the native Fc antibody. The complement activation pathway is initiated by the binding of a first component of the complement system (C1q) to a molecule, such as an antibody complexed with an alloantigen. To evaluate complement activation, a CDC assay may be performed, for example, as described in Gazzano-Santoro et al. (1996) J. Immunol. Methods, 202:163. 【0165】 In one embodiment, the antibody of the present invention exhibits enhanced CDC activity compared to the natural Fc antibody. In another embodiment, the Fc mutant antibody exhibits CDC activity that is at least 2 times, or at least 3 times, or at least 5 times, or at least 10 times, or at least 50 times, or at least 100 times, or up to 50 times, or up to 75 times, or up to 100 times, or up to 200 times, or 2 to 10 times, or 5 to 50 times, or 25 to 100 times, or 75 to 200 times, or 100 to 200 times higher than that of the natural Fc antibody. In yet another embodiment, the Fc mutant antibody exhibits CDC activity that is at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100%, or at least 200%, or at least 300%, or at least 400%, or at least 500% enhanced compared to the natural Fc antibody. In a specific embodiment, the enhancement of CDC activity may be due to the enhanced affinity that the Fc mutant antibody exhibits for C1q. 【0166】 The antibody of the present invention can exhibit enhanced CDC activity compared to natural Fc antibodies thanks to COMPLEGENT® technology (Kyowa Hakko Kirin Co., Ltd.), which enhances CDC, one of the main mechanisms of action of antibodies. By using a technique called isotype chimerism, in which a portion of IgG3, the antibody isotype, is introduced into the corresponding region of IgG1, the standard isotype in therapeutic antibodies, COMPLEGENT® technology significantly enhances CDC activity compared to either IgG1 or IgG3, while retaining the desired characteristics of IgG1, such as ADCC, PK properties, and protein A binding. Furthermore, COMPLEGENT® technology can be used in combination with POTELLIGENT® technology, thereby creating an even better therapeutic mab (ACCRETAMAB®) with enhanced ADCC and CDC activity. 【0167】 The Fc mutant antibody of the present invention may have enhanced ADCC activity and an increased serum half-life compared to the natural Fc antibody. 【0168】 The Fc mutant antibody of the present invention may have enhanced CDC activity and an increased serum half-life compared to the natural Fc antibody. 【0169】 The Fc mutant antibody of the present invention may have enhanced ADCC activity, enhanced CDC activity, and an increased serum half-life compared to the natural Fc antibody. 【0170】 The serum half-life of proteins containing an Fc region can be increased by increasing the binding affinity of the Fc region to FcRn. The term “antibody half-life,” as used herein, refers to the pharmacokinetic property of an antibody, which is a measure of the mean survival time of an antibody molecule after its administration. Antibody half-life can be expressed as the time required to eliminate 50 percent of a known amount of immunoglobulin from a patient’s body (or other mammal) or a particular compartment therefrom, when measured, for example, in serum (i.e., circulating half-life) or other tissues. Half-life can vary per immunoglobulin or per class of immunoglobulin. Generally, an increase in antibody half-life increases the mean residence time (MRT) in circulation for the administered antibody. 【0171】 Increasing the half-life allows for a reduction in the amount of drug administered to a patient and a reduction in the frequency of administration. To increase the serum half-life of an antibody, salvage receptor-binding epitopes may be incorporated into the antibody (particularly antibody fragments), as described, for example, in U.S. Patent No. 5,739,277. As used herein, the term “salvage receptor-binding epitope” refers to an epitope in the Fc region of an IgG molecule (e.g., IgG1, IgG2, IgG3, or IgG4) that is involved in increasing the in vivo serum half-life of the IgG molecule. 【0172】 Alternatively, antibodies of the present invention with increased half-life may be prepared by modifying amino acid residues identified as being involved in the interaction between the Fc receptor and the FcRn receptor (see, for example, U.S. Patent Nos. 6,821,505 and 7,083,784; and International Publication No. 09 / 058492). In addition, the half-life of antibodies of the present invention may be increased by conjugation with PEG or albumin using techniques widely used in the art. In some embodiments, antibodies having the Fc variant region of the present invention have an increased half-life of about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 80%, about 85%, about 90%, about 95%, about 100%, about 125%, about 150%, or more compared to antibodies having the natural Fc region. In some embodiments, antibodies having an Fc mutation region have an increased half-life of approximately 2x, 3x, 4x, 5x, 10x, 20x, 50x or more compared to antibodies having a natural Fc region, or up to approximately 10x, up to approximately 20x, up to approximately 50x, or 2x to 10x, or 5x to 25x, or 15x to 50x. 【0173】 In one embodiment, the present invention provides an Fc variant in which the Fc region is numbered by the EU index as shown in Kabat as follows: 221, 225, 228, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 250, 251, 252, 254, 255, 256, 257, 262, 263, 264, 265, 266, 267, 268, 269, 279, 280, 284, 292, The amino acids include modifications (e.g., amino acid substitutions, amino acid insertions, amino acid deletions) at one or more positions selected from 296, 297, 298, 299, 305, 308, 313, 316, 318, 320, 322, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 339, 341, 343, 370, 373, 378, 392, 416, 419, 421, 428, 433, 434, 435, 436, 440, and 443. Depending on the circumstances, the Fc area may include modifications in additional and / or alternative positions known to those skilled in the art (e.g., U.S. Patent No. 5,624,821; No. 6,277,375; No. 6,737,056; No. 7,083,784; No. 7,317,091; No. 7,217,797; No. 7,276,585; No. 7,355,008). See also U.S. Patent Application Publication No. 2002 / 0147311; No. 2004 / 0002587; No. 2005 / 0215768; No. 2007 / 0135620; No. 2007 / 0224188; No. 2008 / 0089892; International Publication Brochure No. 94 / 29351; and International Publication Brochure No. 99 / 58572). Furthermore, useful amino acid positions and specific substitutions are illustrated in Tables 2 and 6-10 of U.S. Patent No. 6,737,056; the table presented in Figure 41 of U.S. Patent Application Publication No. 2006 / 024298; the tables presented in Figures 5, 12, and 15 of U.S. Patent Application Publication No. 2006 / 235208; the tables presented in Figures 8, 9, and 10 of U.S. Patent Application Publication No. 2006 / 0173170; and the tables presented in Figures 8-10, 13, and 14 of International Publication No. 09 / 058492. 【0174】 In a specific embodiment, the disclosure provides Fc variants in which the Fc region is numbered by the EU index as shown in Kabat as 221K, 221Y, 225E, 225K, 225W, 228P, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235E, 235F, 236E, 237L, 237M, 237P, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 24 1L, 241Y, 241E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 25 7C, 257M, 257N, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 26 5F, 265V, 265I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 29 6N, 296S, 296T, 296L, 296I, 296H, 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 31 3F, 316D, 318A, 318S, 320A, 320S, 322A, 322S, 325Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 32 7N, 327L, 328S, 328M, 328D, 328E, 328N, 328Q, 328F, 328I, 328V, 328T, 328H,328A, 329F, 329H, 329Q, 330K, 330G, 330T, 330C, 330L, 330Y, 330V, 330I, 330F, 330R, 330H, 331G, 331A, 331L, 331M, 331F, 331W , 331K, 331Q, 331E, 331S, 331V, 331I, 331C, 331Y, 331H, 331R, 331N, 331D, 331T, 332D, 332S, 332W, 332F, 332E, 332N, 332Q, 332 Includes at least one substitution selected from T, 332H, 332Y, 332A, 333A, 333D, 333G, 333Q, 333S, 333V, 334A, 334E, 334H, 334L, 334M, 334Q, 334V, 334Y, 339T, 370E, 370N, 378D, 392T, 396L, 416G, 419H, 421K, 428L, 428F, 433K, 433L, 434A, 424F, 434W, 434Y, 436H, 440Y, and 443W. Depending on the circumstances, the Fc region may include, but is not limited to, additional and / or alternative amino acid substitutions known to those skilled in the art, including, but not limited to, those exemplified in Tables 2 and 6-10 of U.S. Patent No. 6,737,056; the table presented in Figure 41 of U.S. Patent Publication No. 2006 / 024298; the tables presented in Figures 5, 12, and 15 of U.S. Patent Publication No. 2006 / 235208; the tables presented in Figures 8, 9, and 10 of U.S. Patent Publication No. 2006 / 0173170; and the tables presented in Figures 8, 9, and 10 of International Publication No. 09 / 058492. 【0175】 In specific embodiments, the present invention provides an Fc mutant antibody in which the Fc region includes at least one modification (e.g., amino acid substitution, amino acid insertion, amino acid deletion) at one or more positions selected from 228, 234, 235, and 331, as numbered by the EU index as shown in Kabat. In one embodiment, the modification is at least one substitution selected from 228P, 234F, 235E, 235F, 235Y, and 331S, as numbered by the EU index as shown in Kabat. 【0176】 In another specific embodiment, the present invention provides an Fc mutant antibody, wherein the Fc region is an IgG4 Fc region and comprises at least one modification at one or more positions selected from 228 and 235 when numbered according to the EU index as shown in Kabat. In yet another specific embodiment, the Fc region is an IgG4 Fc region, and the non-naturally occurring amino acid is selected from 228P, 235E, and 235Y when numbered according to the EU index as shown in Kabat. 【0177】 In another specific embodiment, the present invention provides an Fc mutant, wherein the Fc region comprises at least one non-naturally occurring amino acid at one or more positions selected from 239, 330, and 332 when numbered according to the EU index as shown in Kabat. In one embodiment, the modification is at least one substitution selected from 239D, 330L, 330Y, and 332E when numbered according to the EU index as shown in Kabat. 【0178】 In a specific embodiment, the present invention provides an Fc mutant antibody, wherein the Fc region comprises at least one non-naturally occurring amino acid at one or more positions selected from 252, 254, and 256 when numbered according to the EU index as shown in Kabat. In one embodiment, the modification is at least one substitution selected from 252Y, 254T, and 256E when numbered according to the EU index as shown in Kabat. In a particularly preferred antibody of the present invention, the modification is three substitutions 252Y, 254T, and 256E (known as "YTE") when numbered according to the EU index as shown in Kabat. See U.S. Patent No. 7,083,784. 【0179】 In certain embodiments, effector function induced by IgG antibodies is strongly dependent on the carbohydrate portion linked to the Fc region of the protein (Ferrara et al. (2006) Biotechnology and Bioengineering. 93:851-861). Therefore, glycosylation of the Fc region can be modified to increase or decrease effector function (e.g., Umana et al. (1999) Nat. Biotechnol. 17:176-180; Davies et al. (2001) Biotechnol Bioeng. 74:288-294; Shields et al. (2002) J Biol Chem 277:26733-26740; Shinkawa et al. (2003) J Biol See Chem.278:3466-3473; U.S. Patent Nos. 6,602,684; 6,946,292; 7,064,191; 7,214,775; 7,393,683; 7,425,446; 7,504,256; U.S. Patent Application Publication No. 2003 / 0157108; 2003 / 0003097; 2009 / 0010921; Potillegent™ technology (Biowa, Inc., Princeton, NJ); GlycoMAb™ glycosylation technology (GLYCART biotechnology AG, Zurich, Switzerland). Accordingly, in one embodiment, the Fc region of the antibody of the present invention includes modified glycosylation of an amino acid residue. In another embodiment, the modified glycosylation of an amino acid residue reduces effector function. In yet another embodiment, the modified glycosylation of an amino acid residue increases effector function. In a specific embodiment, the Fc region is reduced fucosylation. In yet another embodiment, the Fc region is non-fucosylated (see, for example, U.S. Patent Application Publication No. 2005 / 0226867).In one embodiment, these antibodies, which have increased effector function, particularly ADCC, as produced in host cells (e.g., CHO cells, Lemna minor), are engineered to produce highly defucosylated antibodies with more than 100 times higher ADCC compared to antibodies produced by parental cells (Mori et al. (2004) Biotechnol Bioeng. 88:901-908; Cox et al. (2006) Nat Biotechnol. 24:1591-7). 【0180】 The addition of sialic acid to oligosaccharides on IgG molecules can enhance their anti-inflammatory activity and modify their cytotoxicity (Keneko et al. (2006) Science. 313:670-673; Scallon et al. (2007) Mol. Immuno. 44(7):1524-34). The studies referenced above demonstrate that IgG molecules with increased sialylation have anti-inflammatory properties, while IgG molecules with reduced sialylation have increased immunostimulatory properties (e.g., increased ADCC activity). Therefore, antibodies can be modified with sialylation profiles appropriate for specific therapeutic applications (US Patent Application Publication No. 2009 / 0004179 and International Publication No. 2007 / 005786). 【0181】 In one embodiment, the Fc region of the antibody of the present invention includes a modified sialylation profile compared to the natural Fc region. In another embodiment, the Fc region of the antibody of the present invention includes an increased sialylation profile compared to the natural Fc region. In yet another embodiment, the Fc region of the antibody of the present invention includes a decreased sialylation profile compared to the natural Fc region. 【0182】 In one embodiment, the Fc variant of this invention is Ghetie et al. (1997) Nat Biotech. 15:637-40; Duncan et al. (1988) Nature. 332:563-564; Lund et al. (1991) J. Immunol. 147: 2657-2662; al(1992)Mol.Immunol.29:53-59;Alegre et al(1994)Transplantation.57:1537-1543;Hutchins et al.(1995)Proc.Natl.Acad.Sci.USA.92:11980-11984;Jefferis et al(1995)Immunol Lett.44:111-117;Lund et al. al. (1995) Faseb. J. 9:115-119; Jefferis et al (1996) Immunol. Lett. 54:101-104; Lund et al (1996) J. Immunol. 157:4963-4969; Armor et al. (1999) Eur. J. Immunol. 29:2613-2624; Idusogie et al (2000) J. Immunol. 164:4178-4184; et al(2000)J Immunol.164:1925-1933;Xu et al.(2000)Cell Immunol.200:16-26;Idusogie et al(2001)J.Immunol.166:2571-2575;Shields et al.(2001)J.Biol.Chem.276:6591-6604;Jefferis et al. al (2002) Immunol. Lett. 82:57-65; Presta et al.(2002) Biochem Soc Trans 30:487-490); US Patent No. 5,624,821; US ​​Patent No. 5,885,573; US Patent No. 5,677,425; US Patent No. 6,165,745; US Patent No. 6,277,375; US Patent No. 5,869,046; US Patent No. 6,121,022; US Patent No. 5,624,821; US ​​Patent No. 5,648,260; US Patent No. 6,528,624; US Patent No. 6,194,551; US ​​Patent No. 6,737,056; US Patent No. 7,122,6 It may be combined with other known Fc variants, such as those disclosed in Specification 37; Specifications 7,183,387; Specifications 7,332,581; Specifications 7,335,742; Specifications 7,371,826; Specifications 6,821,505; Specifications 6,180,377; Specifications 7,317,091; Specifications 7,355,008; U.S. Patent Application Publication 2004 / 0002587; and International Publication 99 / 58572. Other modifications and / or substitutions and / or additions and / or deletions of the Fc domain will be readily apparent to those skilled in the art. 【0183】 Glycosylation In addition to glycosylation's ability to alter the effector function of an antibody, modified glycosylation in the variable region can alter the antibody's affinity for an antigen. In one embodiment, the glycosylation pattern in the variable region of the antibody is modified. For example, a non-glycosylated antibody can be produced (i.e., this antibody lacks glycosylation). Glycosylation can be modified, for example, to increase the antibody's affinity for an antigen. Such carbohydrate modification can be achieved, for example, by modifying one or more glycosylation sites in the antibody sequence. For example, one or more amino acid substitutions can be produced that result in the removal of one or more glycosylation sites in the variable region framework, thereby removing glycosylation at those sites. Such nonglycosylation can increase the antibody's affinity for an antigen. Such methods are described in more detail in U.S. Patent Nos. 5,714,350 and 6,350,861. One or more amino acid substitutions that result in the removal of glycosylation sites present in the Fc region (e.g., asparagine 297 in IgG) can also be produced. Furthermore, non-glycosylated antibodies can be produced in bacterial cells that lack essential glycosylation mechanisms. 【0184】 Antibody conjugate In certain embodiments, the antibody of the present invention is conjugated or covalently bound to a substance using methods known in the art. In one embodiment, the substance to be bound is a therapeutic agent, a detectable label (also referred to herein as a reporter molecule), or a solid support. Suitable substances for binding to the antibody include, but are not limited to, amino acids, peptides, proteins, polysaccharides, nucleosides, nucleotides, oligonucleotides, nucleic acids, haptens, drugs, hormones, lipids, lipid aggregates, synthetic polymers, polymer microparticles, living cells, viruses, fluorophores, chromophores, dyes, toxins, haptens, enzymes, antibodies, antibody fragments, radioisotopes, solid matrices, semi-solid matrices, and combinations thereof. Methods for conjugating or covalently binding another substance to an antibody are known in the art. 【0185】 In certain embodiments, the antibody of the present invention is conjugated to a solid support. The antibody may be conjugated to a solid support as part of a screening and / or purification and / or manufacturing process. Alternatively, the antibody of the present invention may be conjugated to a solid support as part of a diagnostic method or composition. Solid supports suitable for use in the present invention are typically substantially insoluble in liquid phases. Numerous supports are available and well known to those skilled in the art. Accordingly, solid supports include solid and semi-solid matrices, e.g., aerogels and hydrogels, resins, beads, biochips (including thin-film coated biochips), microfluidic chips, silicon chips, multiwell plates (also known as microtiter plates or microplates), membranes, conductive and non-conductive metals, glass (including microscope slides), and magnetic supports. More specific examples of solid supports include silica gel, polymer films, particles, derivatized plastic films, glass beads, cotton, plastic beads, alumina gel, polysaccharides such as Sepharose, poly(acrylate), polystyrene, poly(acrylamide), polyol, agarose, agar, cellulose, dextran, starch, FICOLL, heparin, glycogen, amylopectin, mannan, inulin, nitrocellulose, diazocellulose, polyvinyl chloride, polypropylene, polyethylene (including poly(ethylene glycol)), nylon, latex beads, magnetic beads, paramagnetic beads, superparamagnetic beads, and starch. 【0186】 In some embodiments, the solid support may include, but is not limited to, reactive functional groups for binding the antibody of the present invention, such as hydroxyl, carboxyl, amino, thiol, aldehyde, halogen, nitro, cyano, amide, urea, carbonate, carbamate, isocyanate, sulfone, sulfonate, sulfonamide, sulfoxide, and the like. 【0187】 A suitable solid support can be selected based on the desired end use and suitability for various synthesis protocols. For example, if amide bond formation is desirable for binding the antibody of the present invention to a solid support, resins generally useful for peptide synthesis can be used, 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 onto polyethylene glycol (TentaGel®, Rapp Polymere, Tubingen, Germany), polydimethylacrylamide resin (available from Milligen / Biosearch, California), or PEGA beads (available from Polymer Laboratories). 【0188】 In certain embodiments, the antibody of the present invention is conjugated with a label for diagnostic and other assays in which the antibody and / or associated ligand can be detected. The label conjugated to the antibody and used in the methods and compositions described herein is any chemical moiety (organic or inorganic) that exhibits maximum absorption at wavelengths greater than 280 nm and retains its spectral characteristics when covalently bound to the antibody. Examples of labels include, but are not limited to, chromophores, fluorophores, fluorescent proteins, phosphorescent dyes, tandem dyes, particles, haptens, enzymes, and radioisotopes. 【0189】 In certain embodiments, the antibody is conjugated to a fluorophore. Thus, the fluorophores used to label the antibody of the present invention include, without limitation: pyrene (including any of the corresponding derivative compounds disclosed in U.S. Patent No. 5,132,432), anthracene, naphthalene, acridine, stilbene, indole or benzindole, oxazole or benzoxazole, thiazole or benzothiazole, 4-amino-7-nitrobenz-2-oxa-1,3-diazole (NBD), cyanine (including any corresponding compound in U.S. Patent Nos. 6,977,305 and 6,974,873), carbocyanine (U.S. Patent Application No. 09 / 557,275; U.S. Patent No. 4,981,977; 5,26 Specification No. 8,486; Specification No. 5,569,587; Specification No. 5,569,766; Specification No. 5,486,616; Specification No. 5,627,027; Specification No. 5, Specification No. 808,044; Specification No. 5,877,310; Specification No. 6,002,003; Specification No. 6,004,536; Specification No. 6,008,373; Specification No. 6 ,043,025; 6,127,134; 6,130,094; 6,133,445; and International Publication Brochures 02 / 26891, 97 / 40104, 99 / 51702, and 01 / 21624; European Patent Application Publication No. 1 065 250 (including any corresponding compound in Specification A1), carbostyryl, porphyrin, salicylate, anthranilate, azulene, perylene, pyridine, quinoline, borapolyazindacene (including any corresponding compound disclosed in U.S. Patent Nos. 4,774,339; 5,187,288; 5,248,782; 5,274,113; and 5,433,896), xanthene (U.S. Patent Nos. 6,162,931; 6,130,101; 6,229,055; 6,339,392; 5,451,343; 5,227,487; 5,442,045; 5,798,276;(including any corresponding compounds disclosed in U.S. Patent No. 5,846,737; U.S. Patent No. 4,945,171; U.S. Patent Application No. 09 / 129,015 and U.S. Patent No. 09 / 922,333), oxazine (including any corresponding compounds disclosed in U.S. Patent No. 4,714,763) or benzoxazine, carbazine (including any corresponding compounds disclosed in U.S. Patent No. 4,810,636), phenalenone, coumarin (U.S. This includes oxazines (including the corresponding compounds disclosed in U.S. Patent Nos. 5,696,157; ​​5,459,276; 5,501,980 and 5,830,912), benzofurans (including the corresponding compounds disclosed in U.S. Patent Nos. 4,603,209 and 4,849,362), and benzphenalenones (including any corresponding compounds disclosed in U.S. Patent No. 4,812,409) and their derivatives. When used herein, oxazines include resolphins (including any corresponding compounds disclosed in U.S. Patent No. 5,242,805), aminooxazinones, diaminooxazines, and their benzo-substituted analogs. 【0190】 In specific embodiments, the fluorophores conjugated to the antibodies described herein include xanthenes (rhodol, rhodamine, fluorescein and their derivatives), coumarins, cyanines, pyrenes, oxazines, and borapolyazindacenes. In other embodiments, such fluorophores are sulfonated xanthenes, fluorinated xanthenes, sulfonated coumarins, fluorinated coumarins, and sulfonated cyanines. Also included are dyes marketed and commonly known under the trademarks ALEXA FLUOR®, DyLight, CY® Dyes, BODIPY®, Oregon Green®, Pacific Blue®, IRDYE®, FAM, FITC, and ROX®. 【0191】 The selection of fluorophores that bind to antibodies can determine the absorption and fluorescence properties of the conjugated antibody. Physical properties of fluorophore labels that can be used on antibodies and antibody-binding ligands include, but are not limited to, spectral properties (absorption, emission, and Stokes shift), fluorescence intensity, lifetime, polarization, and photobleaching rate, or combinations thereof. All of these physical properties can be used to distinguish one fluorophore from another, thus enabling multiplexed analysis. In certain embodiments, the fluorophore has an absorption maximum at wavelengths greater than 480 nm. In other embodiments, the fluorophore absorbs at 488 nm to 514 nm or nearby (particularly suitable for excitation by the output of an argon ion laser excitation source) or near 546 nm (particularly suitable for excitation by a mercury arc lamp). In other embodiments, the fluorophore can emit in the NIR (near-infrared region) for tissue or whole-organism application. Other desirable properties of fluorescence labeling include cell permeability and low toxicity, for example, when antibody labeling is performed on cells or organisms (e.g., living animals). 【0192】 In certain embodiments, an enzyme is the label and is conjugated to the antibody described herein. The enzyme is a desirable label because it can achieve amplification of a detectable signal, resulting in increased assay sensitivity. The enzyme itself does not produce a detectable reaction, but when contacted with a suitable substrate, it degrades the substrate, thereby converting the substrate to produce a fluorescent, colorimetric, or chemiluminescent signal. The enzyme amplifies the detectable signal because one enzyme in the labeling reagent can result in conversion to a detectable signal for multiple substrates. The enzyme substrate is selected so as to yield a preferred measurable product, such as a colorimetric, fluorescent, or chemiluminescent product. Such substrates are widely used in the art and are well known to those skilled in the art. 【0193】 In one embodiment, the combination of a colorimetric substrate or fluorescence-generating substrate and enzyme uses an oxidoreductase such as horseradish peroxidase and a substrate that produces a discernible color (brown and red, respectively) such as 3,3'-diaminobenzidine (DAB) and 3-amino-9-ethylcarbazole (AEC). Other colorimetric oxidoreductase substrates that produce detectable products include, but are not limited to, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), o-phenylenediamine (OPD), 3,3',5,5'-tetramethylbenzidine (TMB), o-dianisidine, 5-aminosalicylic acid, and 4-chloro-1-naphthol. Examples of fluorescence-generating substrates, though not limited to them, include homovanillic acid or 4-hydroxy-3-methoxyphenylacetic acid, reduced phenoxazine and reduced benzothiazine, e.g., Amplex® Red reagent and its variants (U.S. Patent No. 4,384,042), reduced dihydroxanthene, e.g., dihydrofluorescein (U.S. Patent No. 6,162,931), and dihydrorhodamine, e.g., dihydrorhodamine 123. Tyramide peroxidase substrates (U.S. Patent Nos. 5,196,306; 5,583,001 and 5,731,158) may be inherently detectable before the enzyme acts, but they belong to a unique class of peroxidase substrates in that they are "fixed in situ" by the action of peroxidase in the process described as tyramide signal amplification (TSA). These substrates are widely used to label antigens in samples, such as cells, tissues, or arrays, for later detection by microscopy, flow cytometry, optical scanning, and fluorescence quantification. 【0194】 In another embodiment, the combination of a colorimetric (and sometimes fluorescence-generating) substrate and an enzyme is a phosphatase enzyme, such as acid phosphatase, alkaline phosphatase, or a recombinant of such phosphatase, combined with a colorimetric substrate such as 5-bromo-6-chloro-3-indolyl phosphate (BCIP), 6-chloro-3-indolyl phosphate, 5-bromo-6-chloro-3-indolyl phosphate, p-nitrophenyl phosphate, or o-nitrophenyl phosphate, or 4-methylumbelliferyl phosphate, 6,8-difluoro-7-hydroxy-4-methylcoumarinyl phosphate (DiFMUP, U.S. Patent No. 5,830,912), fluorescein diphosphate, 3-O-methylfluorescein phosphate, resolphin phosphate, 9H-(1,3-dichloro-9,9-dimethylacrididine-2-on-7-yl) phosphate (DDAO phosphate), or ELF 97, ELF 39. Used in combination with a fluorescence-generating substrate such as related phosphoric acid (U.S. Patent No. 5,316,906 and U.S. Patent No. 5,443,986). 【0195】 Glycosidases, particularly β-galactosidase, β-glucuronidase, and β-glucosidase, are even more preferred enzymes. Suitable colorimetric substrates include, but are not limited to, 5-bromo-4-chloro-3-indolyl β-D-galactopyranoside (X-gal) and similar indolyl galactosides, glucosides, and glucuronides, o-nitrophenyl β-D-galactopyranoside (ONPG), and p-nitrophenyl β-D-galactopyranoside. In one embodiment, the fluorescence-generating substrate includes resorphin β-D-galactopyranoside, fluorescein digalactoside (FDG), fluorescein diglucuronide and structural variants thereof (US Patent Nos. 5,208,148; 5,242,805; 5,362,628; 5,576,424 and 5,773,236), 4-methylumbelliferyl β-D-galactopyranoside, carboxyumbelliferyl β-D-galactopyranoside, and fluorinated coumarin β-D-galactopyranoside (US Patent No. 5,830,912). 【0196】 Additional enzymes include, but are not limited to, hydrolases such as cholinesterase and peptidase, oxidases such as glucose oxidase and cytochrome oxidase, and reductases for which suitable substrates are known. 【0197】 For some assays, enzymes that produce chemiluminescence and their appropriate substrates are preferred. These include, but are not limited to, luciferase and aequorin in native and recombinant forms. Chemiluminescent substrates for phosphatase, glycosidase and oxidase, such as those containing stable dioxetane, luminol, isoluminol and acridinium ester, are more useful. 【0198】 In another embodiment, haptens such as biotin are also used as labels. Biotin can function to further amplify the detectable signal in the enzyme system and can function as a tag for use in affinity chromatography for isolation purposes, so it is useful. For detection purposes, enzyme conjugates having an affinity for biotin, such as avidin-HRP, are used. Subsequently, when a peroxidase substrate is added, a detectable signal is generated. 【0199】 Haptens also include hormones, naturally occurring and synthetic drugs, pollutants, allergens, effector molecules, growth factors, chemokines, cytokines, lymphokines, amino acids, peptides, chemical intermediates, nucleotides and the like. 【0200】 In certain embodiments, a fluorescent protein may be conjugated to the antibody as a label. Examples of fluorescent proteins include green fluorescent protein (GFP), phycobiliproteins, and their derivatives. Fluorescent proteins, particularly phycobiliproteins, are especially useful for creating labeling reagents labeled with tandem dyes. Such tandem dyes include a fluorescent protein and a fluorophore for the purpose of achieving a larger Stokes shift, where the emission spectrum is shifted further away from the wavelength of the absorption spectrum of the fluorescent protein. This is particularly advantageous for detecting low amounts of antigen in a sample, where the emitted fluorescence light is maximally optimized, or in other words, the emitted light is hardly or never reabsorbed by the fluorescent protein. For this to work, the fluorescent protein and the fluorophore act as an energy transfer pair, where the fluorescent protein emits light at the wavelength absorbed by the fluorophore, and then the fluorophore emits light at a wavelength further away from the fluorescent protein than could be achieved with the fluorescent protein alone. Particularly useful combinations include phycobiliproteins disclosed in U.S. Patent Nos. 4,520,110; 4,859,582; and 5,055,556, and sulfohodamine fluorophores disclosed in U.S. Patent No. 5,798,276, or sulfonated cyanine fluorophores disclosed in U.S. Patent Nos. 6,977,305 and 6,974,873; or sulfonated xanthene derivatives disclosed in U.S. Patent No. 6,130,101, and combinations thereof disclosed in U.S. Patent No. 4,542,104. Alternatively, the fluorophore acts as an energy donor and the fluorescent protein is an energy acceptor. 【0201】 In certain embodiments, the label is a radioactive isotope. Examples of suitable radioactive materials include, but are not limited to, iodine. 121 I, 123 I, 125 I, 131 I), carbon ( 14 C), sulfur ( 35 S), tritium ( 3 H), Indium (111 In, 112 In, 113 mIn, 115 mIn), technetium ( 99 Tc, 99 mTc), thallium ( 201 Ti), Gallium ( 68 Ga, 67 Ga), palladium ( 103 Pd), molybdenum ( 99 Mo), xenon ( 135 Xe), fluorine ( 18 F), 153 Sm, 177 Lu, 159 Gd, 149 PM, 140 La, 175 Yb, 166 Ho, 90 Y, 47 Sc, 186 Re, 188 Re, 142 Pr, 105 Rh and 97 Ru is mentioned. 【0202】 Treatment and use The antibodies and antigen-binding fragments thereof, as well as their variants, of the present invention may be used for the treatment of influenza B virus infection, for the prevention of influenza B virus infection; for the detection, diagnosis, and / or prognosis of influenza B virus infection; or for a combination thereof. In one embodiment, the antibodies and antigen-binding fragments thereof, as well as their variants, of the present invention may be used for the treatment of influenza A and influenza B infection, for the prevention of influenza A and influenza B infection; for the detection, diagnosis, and / or prognosis of influenza A and influenza B infection; or for a combination thereof. 【0203】 The diagnostic method may include contacting an antibody or antibody fragment with a sample. Such a sample may be, for example, a tissue sample taken from the nasal passages, sinus cavities, salivary glands, lungs, liver, pancreas, kidneys, ears, eyes, placenta, gastrointestinal tract, heart, ovaries, pituitary gland, adrenal gland, thyroid gland, brain, or skin. The detection, diagnostic, and / or prognostic methods may also include the detection of an antigen / antibody complex. 【0204】 In one embodiment, the present invention provides a method for treating a subject by administering to the subject an effective amount of the antibody or antigen-binding fragment thereof described in the present invention, or a pharmaceutical composition containing the antibody or antigen-binding fragment thereof. In one embodiment, the antibody or antigen-binding fragment thereof is substantially purified (i.e., substantially freed from substances that limit its effect or cause undesirable side effects). In one embodiment, the antibody or antigen-binding fragment thereof of the present invention is administered after exposure, i.e., after the subject has been exposed to or infected with influenza B virus. In one embodiment, the antibody or antigen-binding fragment thereof of the present invention is administered after exposure, i.e., after the subject has been exposed to or infected with influenza B virus of the Yamagata and / or Victoria lineage. In one embodiment, the antibody or antigen-binding fragment of the present invention is administered after exposure, i.e., after the subject has been exposed to at least one subtype of influenza A virus; influenza B virus of the Yamagata lineage; influenza B virus of the Victoria lineage; or a combination thereof; or after infection with at least one subtype of influenza A virus and / or influenza B virus of the Yamagata and / or Victoria lineage. 【0205】 In another embodiment, the antibody or antigen-binding fragment of the present invention is administered to a subject pre-exposure, i.e., a subject who has not yet been exposed to or infected with influenza B virus. In another embodiment, the antibody or antigen-binding fragment of the present invention is administered to a subject pre-exposure, i.e., a subject who has not yet been exposed to or infected with Yamagata and / or Victoria lineage influenza B virus. In another embodiment, the antibody or antigen-binding fragment of the present invention is administered to a subject pre-exposure, i.e., a subject who has not yet been exposed to or infected with influenza A virus; Yamagata lineage influenza B virus; Victoria lineage influenza virus; or a combination thereof, or a subject who has not yet been infected with influenza A virus; Yamagata lineage influenza B virus; Victoria lineage influenza; or a combination thereof. 【0206】 In one embodiment, the antibody of the present invention or its antigen-binding fragment is administered to a subject who is seronegative to one or more influenza B viruses. In one embodiment, the antibody of the present invention or its antigen-binding fragment is administered to a subject who is seronegative to one or more strains of influenza B viruses. In one embodiment, the antibody of the present invention or its antigen-binding fragment is administered to a subject who is seronegative to one or more subtypes of influenza A and / or one or more influenza B viruses. 【0207】 In another embodiment, the antibody or antigen-conjugated fragment of the present invention is administered to a subject who is seropositive for one or more influenza B viruses. In another embodiment, the antibody or antigen-conjugated fragment of the present invention is administered to a subject who is seropositive for one or more strains of influenza B viruses. In another embodiment, the antibody or antigen-conjugated fragment of the present invention is administered to a subject who is seropositive for one or more subtypes of influenza A viruses and / or one or more influenza B viruses. In one embodiment, the antibody or antigen-conjugated fragment of the present invention is administered to a subject within 1, 2, 3, 4, or 5 days after infection or symptom onset. In another embodiment, the antibody or antigen-conjugated fragment of the present invention may be administered to a subject on 1, 2, 3, 4, 5, 6, or 7 days after infection or symptom onset, and within 2, 3, 4, 5, 6, 7, 8, 9, or 10 days after infection or symptom onset. 【0208】 In one embodiment, the method reduces influenza B virus infection in a subject. In another embodiment, the method reduces influenza A virus infection and / or influenza B virus infection in a subject. In another embodiment, the method prevents or reduces the risk or delay of influenza B virus infection in a subject. In another embodiment, the method prevents or reduces the risk or delay of influenza A and / or influenza B virus infection in a subject. In one embodiment, the subject is an animal. In one embodiment, the subject is a mammal. In further specific embodiments, the subject is a human. In one embodiment, the subject includes, but is not limited to, subjects who are at particular risk of or susceptible to influenza A and / or influenza B virus infection, such as immunocompromised subjects. 【0209】 Treatment can be administered via a single-dose schedule or a multi-dose schedule, and the antibody or antigen-binding fragment of the present invention can be used via passive or active immunization. 【0210】 In one embodiment, the antibody of the present invention or its antigen-binding fragment is administered to a subject in combination with one or more antiviral agents. In another embodiment, the antibody of the present invention or its antigen-binding fragment is administered to a subject in combination with one or more small molecule antiviral agents. The small molecule antiviral agents include neuraminidase inhibitors such as oseltamivir (TAMIFLU®) and zanamivir (RELENZA®), and adamantanes such as amantadine and rimantadine. 【0211】 In another embodiment, the present invention provides compositions to be used as drugs for the prevention or treatment of influenza A and / or influenza B virus infections. In another embodiment, the present invention provides the use of antibodies or antigen-binding fragments thereof and / or proteins comprising target epitopes to which the antibodies or antigen-binding fragments thereof conjugate in the preparation of drugs for the treatment of a subject and / or for diagnosis in a subject. 【0212】 The antibodies and fragments thereof described in the present invention may also be used in kits for the diagnosis of influenza A virus infection; influenza B virus infection; or combinations thereof. Furthermore, epitopes capable of binding to the antibodies of the present invention may be used in kits for monitoring the effectiveness of a vaccination procedure by detecting the presence of protective anti-influenza A and / or influenza B virus antibodies. The antibodies, antibody fragments, or variants and derivatives thereof described in the present invention may also be used in kits for monitoring vaccine production with desired immunogenicity. 【0213】 The present invention also provides a method for preparing a pharmaceutical composition, comprising the step of mixing a monoclonal antibody, which is a monoclonal antibody as described herein, with one or more pharmaceutically acceptable carriers. 【0214】 For administering the antibody or its antigen-binding fragment of the present invention, various delivery systems are known and can be used, but are not limited to those including encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the antibody or antibody fragment, receptor-mediated endocytosis, retroviruses, or other vectors as part of nucleic acids. Delivery methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. In another embodiment, the vaccine may be administered as a DNA vaccine using electroporation techniques, including, but is not limited to, in vivo electroporation. The composition may be administered by any convenient route, for example, by infusion or bolus injection, by absorption through the epithelium or lining of the skin mucosa (e.g., oral mucosa, rectal and intestinal mucosa), or, but is not limited to, administration with other bioactive agents, including small molecule antiviral compositions. Administration may be systemic or topical. Lung administration can also be used, for example, by the use of inhalers or sprayers, and by formulation with aerosolizing agents. In yet another embodiment, the composition may be delivered in a sustained-release system. 【0215】 The present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof of the present invention and a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable,” as used herein, means approved by a federal or state regulatory agency for use in animals, more particularly in humans, or listed in the United States Pharmacopeia or other generally accepted pharmacopoeias. The term “carrier” refers to a diluent, adjuvant, excipient, or medium administered with the therapeutic agent. Such pharmaceutically acceptable carriers may be sterile liquids, such as water, and oils, such as petroleum, animal, plant, or synthetic origins, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, etc. The compositions may also contain small amounts of wetting agents or emulsifiers, or pH buffers, as needed. These compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, etc. Compositions can be formulated as suppositories with conventional binders and carriers such as triglycerides. Oral formulations may contain standard carriers such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. In one embodiment, the pharmaceutical composition contains a therapeutically effective amount of antibody or its antigen-binding fragment, along with an appropriate amount of carrier, preferably in a purified form, to provide a form for appropriate administration to a patient. The formulation should be adapted to the method of administration. 【0216】 Typically, in antibody therapies, the dose administered to a patient is approximately 0.1 mg / kg to 100 mg / kg relative to the patient's body weight. [Examples] 【0217】 Example 1: Construction and optimization of human monoclonal antibodies isolated from memory B cells CD22+IgG+ B cells were selected from donor cryopreserved peripheral blood mononuclear cells (PBMCs) that exhibited high neutralizing titers against both the B / Florida / 4 / 2006 Yamagata lineage (B / FLA / 06) and the B / Brisbane / 60 / 2008 Victoria lineage (B / BNE / 08). These cells were immortalized at a rate of 3 cells / well using Epstein-Barr virus (EBV), CpG oligodeoxyribonucleotide 2006, and supporting cells. The culture supernatant containing antibodies was collected after 14 days and screened using a microneutralization assay (MNA), an improved version of the aforementioned facilitative viral inhibition assay using neuraminidase activity (NA) as readout, to identify antibody clones capable of neutralizing both the Yamagata and Victoria influenza B lineages (Hassantoufighi et al. (2010) Vaccine. 28:790-7). 【0218】 In other words, 10 μl of culture supernatant is used in 400 TCID 50 The cells were incubated with influenza B / BNE / 08 (Victoria strain) or B / FLA / 06 (Yamagata strain) at 37°C for 1 hour. Madin-Darby canine kidney (MDCK) cells were added to plates (20,000 cells / well), incubated for 4 hours, washed twice with medium containing TPCK-trypsin, and then incubated at 37°C for 2 days. After incubation, NA activity was measured by adding methylumbelliferyl-N-acetylneuraminic acid (μ-NANA) (Sigma) at a concentration of 25 μl / well (10 μM) as a fluorescent labeling substrate, and the plates were read using a fluorometer. 【0219】 Three B cell clones (FBC-39, FBD-56, and FBD-94) were found to possess neutralizing activity against both the Victoria and Yamagata strains of influenza B. The VH and VL genes of these clones were sequenced and cloned into IgG1 expression vectors. Recombinant antibodies were produced by transient gene transfer into mammalian cell lines derived from human embryonic kidney (HEK) or Chinese hamster ovary (CHO) cells. The supernatant from the gene-transferred cells was collected 7-10 days after culture, and the antibody (IgG) was purified by protein A chromatography and dialyzed to phosphate-buffered saline (PBS). 【0220】 The Ig BLAST algorithm was used to align mAb sequences against a database of human antibody germline sequences. The VH and VL gene regions in the most recent germline template were identified (Table 6). Non-germlined amino acids were identified by aligning them against these reference sequences. 【0221】 [Table 6] 【0222】 Construction of the FBC-39 mutant In the FBC-39 antibody VL, there was only one non-germline framework residue:F at position 87 in the light chain, where the germline amino acid is Y (i.e., L87F(Y)) according to Kabat numbering (position 103 according to IMGT numbering). The germline sequence is referred to herein as FBC-39-L87Y. The non-germline sequence is referred to as FBC-39-L87F. 【0223】 In the FBC-39 antibody VH, there are 11 Kabat-defined non-germ-sequenced framework residues, namely, according to Kabat numbering, H6V(E), H27L(F), H28S(T), H30L(S), H68S(T), H77M(T), H79F(Y), H81H(Q), H82aS(N), H83R(K), and H93A(T). When using the IMGT definition of framework residues, there are 7 non-germ-sequenced residues: H6V(E), H77S(T), H86M(T), H88F(Y), H90H(Q), H92S(N), and H95R(K). 【0224】 The mutant was constructed in which a total of 12 Kabat-defined non-germline framework residues were converted back to germline amino acids. This antibody construct showed a significant decrease in neutralizing activity and scope against Victoria strain influenza B virus, suggesting that one or more non-germline residues are important for activity. 【0225】 The three non-germ-forming framework residues located at positions H27, H28, and H30 are considered part of HCDR-1 by the IMGT system, but are located within a region defined as VH framework 1 by the Kabat system. Antibody variants were created by restoring all Kabat-defined non-germ-forming framework amino acids to their respective germline residues, with the exception of these three positions: H27L(F), H28S(T), and H30L(S) ("wobbled" between germline and non-germline residues), resulting in seven heavy chain variants: FBC-39LSL, FBC-39FSL; FBC-39LTL; FBC-39FTL; FBC-39- F S S FBC-39-L TS ;FBC-39- FTS (Germline residues are underlined) are FBC-39 FTS The molecule was prepared so that it has three germline amino acids and FBC-39LSL has three wild-type residues at positions H27, H28, and H30. 【0226】 In addition, germline residue N (H92IMGT) at H82 created a promising deamide site (NS) within VH. As a result, this was substituted for wild-type S in FBC-39 in all seven mutants: FBC-39LSL, FBC-39FSL, FBC-39LTL, FBC-39FTL, FBC-39-FSS, FBC-39-LTS, and FBC-39-FTS. Furthermore, all seven FBC-39 mutants share the same light chain sequence (FBC-39-L87Y), which differs from the FBC-39 light chain by only one amino acid. 【0227】 The obtained antibody mutants were expressed, purified, and further characterized as described above. 【0228】 Example 2. Isolated anti-HA antibodies bind to both influenza B HA lineages. To determine the binding and cross-reactivity of isolated antibodies, an HA ELISA binding assay was performed. A 384-well Maxisorb ELISA plate (Nunc) was coated overnight at 4°C with 0.5 μg / ml recombinant HA from Yamagata strain B / FLA / 06 or Victoria strain B / BNE / 08 in PBS. To remove uncoated proteins, the plate was washed with PBS containing 0.1% v / v Tween-20, and a blocking solution containing 1% (w / v) casein (Thermo Scientific) was added at room temperature for 1 hour. The blocking solution was discarded, and 3-fold serial dilutions in PBS of each anti-HA antibody (FBC-39, FBD-56, and FBD-94) were added and incubated at room temperature for 1 hour. The plate was washed three times, and the bound antibodies were detected using peroxidase-conjugated mouse anti-human IgG antibody (Jackson). Antibody binding activity was calculated by measuring the chemiluminescence signal after adding the Supersignal Pico substrate (Thermo Scientific), or by incubating with tetramethylbenzidine (TMB) (available from Kirkegaard and Perry Laboratories, Inc. (KPL), Gaithersburg, MD) as a substrate for one component, stopping the reaction by adding 2N sulfuric acid, and then measuring the color change at 450 nm. 【0229】 [Table 7] 【0230】 Table 7 shows the average EC obtained from three independent experiments. 50 This shows that all three anti-HA IgGs (FBC-39, FBD-56, and FBD-94) bound to recombinant HA from both influenza B lineages. 50 The values ​​were observed across all three antibodies against Yamagata(B / FL / 06)HA. In the case of Victoria(B / BNE / 08), EC was higher against FBD-94 than against either FBD-56 or FBC-39. 50A decrease was observed. 【0231】 The binding activity of germline-linked variants of seven FBC-39 antibodies was tested by ELISA. Table 8 shows the binding results for unpurified anti-HA FBC-39 IgG variants, where FBC-39 FTS has Kabat-defined framework germline-linked amino acids, and FBC-39 LSL has Kabat-defined framework germline-linked amino acids excluding wild-type residues at positions H27, H28, and H30 (Kabat numbering). These results indicate that all variants bound to the Yamagata lineage (B / FL / 06) HA protein, but the variant with an S residue at position H30 lost its binding affinity to the Victoria lineage (B / BNE / 08) HA protein. Four variants, FBC-39LSL, FSL, LTL, and FTL, showed binding affinity to HA proteins from both lineages that was equivalent to or better than FBC-39. 【0232】 [Table 8] 【0233】 Example 3. In vitro cross-reactivity neutralizing activity of anti-influenza B HA IgG against viruses of two different strains. To test the purified mAb activity, a similar microneutralization assay was used, as described in Example 1. Specifically, MDCK cells were cultured in MEM medium (Invitrogen) supplemented with antibiotics, glutamine (complete MEM medium), and 10% (v / v) fetal bovine serum. 60TCID 50 After incubation of the virus (50% tissue culture infectious dose) at room temperature for 30 minutes, it was added to 3-fold diluted antibody in a 384-well plate in a double-well complete MEM medium containing trypsin (Worthington) treated with 0.75 ug / ml TPCK, and 2 × 10⁶ 4Cells were added to the plate in units of 100 cells / well. After incubation for approximately 40 hours in a 33°C, 5% CO2 incubator, NA activity was measured by adding the fluorescently labeled substrate methylumbelliferyl-N-acetylneuraminic acid (M-NANA) (Sigma) to each well, and the plate was incubated at 37°C for 1 hour. Viral replication, as expressed by NA activity, was quantified by reading fluorescence using an Envision Fluorometer (PerkinElmer) with the following settings: excitation 355 nm, emission 460 nm; 10 flashes / well. Neutralization titer (50% inhibitory concentration [IC2]) was measured. 50 ]) is expressed as the final antibody concentration in which the fluorescence signal is reduced by 50% compared to the cell control well. The influenza B virus strains used in Table 9 are as follows: B / Lee / 40 (B / Lee / 40); B / AA / 66 (ca B / Ann Arbor / 1 / 66); B / HK / 72 (B / Hong Kong / 5 / 72); B / BJ / 97 (ca B / Beijing / 243 / 97 (victoria)), B / HK / 01 (B / Hong Kong / 330 / 2001 (victoria)); B / MY / 04 (B / Malaysia / 2506 / 2004 (victoria)); B / BNE / 08 (ca B / Brisbane / 60 / 2008 (victoria)); B / AA / 94 (ca B / Ann Arbor / 2 / 94 (yamagata)); B / YSI / 98 (ca B / Yamanashi / 166 / 98 (yamagata)); B / JHB / 99 (ca B / Johannesburg / 5 / 99(yamagata));B / SC / 99(B / Sichuan / 379 / 99(yamagata));B / FL / 06(B / Florida / 4 / 2006(yamagata)). 【0234】 [Table 9] 【0235】 Table 9 shows the average IC from two independent experiments. 50The results show that anti-HA antibodies neutralized all influenza B viruses tested. FBD-56 and FBD-94 were more potent than FBC-39, but showed some decreased activity against the B / AA / 94 strain. FBC-39 variants, LSL, FSL, LTL, and FTL neutralized all viruses at an IC50 level equivalent to or lower than FBC-39. 50 It neutralized up to that point. 【0236】 Example 4. Binding and neutralization of influenza A H9 virus strain. HA-binding ELISA was performed in the same manner as in Example 2, except that a 384-well plate was coated in PBS with 3 μg / ml recombinant HA derived from influenza A subtype H9 (A / chicken / HK / G9 / 97(H9N2)) at room temperature for 2 hours. The results showed that FBC-39 and germline mutant LTL had similar EC levels of 6.2 and 6.3 μg / ml, respectively. 50 The H9 HA binds to the EC2 at a value of 41.7 and 46.1 μg / ml, and FBC-39LSL and FTL have higher EC2 values ​​of 41.7 and 46.1 μg / ml, respectively. 50 Binding was shown for FBC-39FSL (Table 10). In contrast, FBC-39FSL only weakly bound even at the highest dose tested of 50 μg / ml, and no binding was observed for FBD-56 and FBD-94. 【0237】 [Table 10] 【0238】 To confirm the functional appropriateness of the binding affinity of the influenza A H9 HA protein, a microneutralization assay was performed using the same method as described in Example 3. In this assay, a low-temperature-adapted (ca) attenuated live influenza vaccine virus was prepared by reverse genetics using the same method as described by Jin et al. (2003) Virology. 306:18-24) to possess viral HA and NA genes derived from the A / chicken / Hong Kong / G9 / 97(H9N2) virus, associated with six internal protein genes of the ca A / Ann Arbor / 6 / 60(H2N2) virus. The results of the microneutralization assay are shown in Table 10. Consistent with the binding characteristics, FBC-39 and the variant exhibited biologically appropriate IC. 50 The values ​​indicated that the H9N2 virus was potentially neutralized. FBC-39 and FBC-39 LTL showed IC50 levels of 0.17 and 0.09 μg / ml, respectively. 50 The most potent activity was observed at the highest concentration. As expected, FBD-56, FBD-94, and the control antibody showed no neutralizing activity at the highest concentration tested (50 μg / ml). 【0239】 Example 5. Identification of epitopes by selection of monoclonal antibody-resistant mutants (MARMs). Influenza B virus of the Yamagata lineage (B / Florida / 4 / 2006; B / FLA / 06) and influenza B virus of the Victoria lineage (B / Malyasia / 2506 / 2004; B / MY / 04) were adsorbed onto MDCK cells in a 10×96 well plate at a rate of 30,000 TCID50 / well. FBC-39, FBD-56, and FBD-94 (10×IC) were then used to adsorb the virus and antibody mixture onto MDCK cells. 50 One hour before culturing in the presence of (), high concentrations of FBC-39, FBD-56, and FBD-94 (125 × IC) are administered. 50The cells were incubated with ). For up to 3 days post-infection, putative MARMs exhibiting cytopathic effects (CPE) on infected cells were isolated. The HA gene was amplified by RT-PCR, then sequenced, and resistance in the isolated virus was confirmed by microneutralization assay. MARMs were not isolated from B / FLA / 09 virus when cultured in the presence of FBC-39, FBD-56, or FBD-94. When using Victoria strain (B / MY / 04) virus, MARMs were isolated in the presence of FBD-56 and FBD-94, but in the absence of FBC-39. Sequence analysis showed that two FBD-56 MARMs had a single amino acid substitution at position 128 from glutamic acid (E) to lysine (K) or valine (V) (Table 11). FBD-94 MARMs had a single amino acid substitution from E to K at position 128 (Table 11). A variant of the Yamagata strain B / Florida / 4 / 2006 (B / FLA / 06 G141E) with a single amino acid substitution from glycine (G) to E at position 141 showed an 8-fold reduction in FBC-39 neutralization compared to the wild-type virus (B / FLA / 06). MARM isolation was repeated using only FBC-39 mAbs with this B / FLA / 09 G141E variant, the Yamagata strain virus B / Jiangsu / 10 / 2003 (B / JIN / 03), and a naturally circulating virus with R at position 141 (G141R). One MARM virus was isolated using the B / FLA / 09G141E virus with a single amino acid change from G to arginine (R) at position 235 (Table 11). Two B / JIN / 03 escape mutant viruses were identified, each with a single amino acid substitution: serine (S) to isoleucine (I) at position 150, or E to leucine (L) at position 235 (Table 11).The amino acid substitutions identified in these influenza B MARMs are located within the head region of HA (Wang et al. (2008) J. Virol. 82(6):3011-20), suggesting that FBC-39, FBD-56, and FBD-94 recognize epitopes on the HA head of influenza B virus, that FBD-56 and FBD-94 have important contact at position 128 and share a duplicate epitope, and that FBC-39 has structural epitopes with important contact residues at positions 141, 150, and 235. 【0240】 [Table 11] 【0241】 Example 6. Influenza B anti-HA antibody exhibits Fc effector function. Antibodies have the potential to eliminate virus-infected cells through Fc-effector functions such as antibody-dependent cytotoxicity (ADCC), antibody-dependent phagocytosis, and complement-dependent killing. To confirm that anti-HA antibodies exhibit ADCC activity, we tested their ability to activate NK cells in the presence of influenza B virus using an ADCC bioassay. In this assay, to measure Fc effector activation, we used a human NK cell line (NK92) that has been stably transfected with human FcgIIIA high-affinity receptor and luciferase transgenes under the control of the NFAT promoter. 96-well plates were used, 5.0 × 10⁶ 4 Cells were coated with TCID50 / well of B / Hong Kong / 330 / 2001 (Victoria) virus stock. Serially diluted FBD-94, FBC-39, and Fc effector null mutants having two substitutions L234A and L235A in the Fc region (FBD-94LALA and FBC-39LALA) (Hezareh et al. (2001) J. Virol. 75(24):12161) were applied to the viruses, and then NK cells were incubated at 5.0 × 10⁶. 4Cells were added to individual cells / wells and incubated at 37°C for 4 hours. Luciferase was detected by adding Steady-Glo reagent (Promega) and measured using an Envision plate reader. Figure 1 shows that both FBD-94 and FBC-39 exhibited dose-dependent ADCC activity against influenza B, while the LALA variant did not show activity at the same concentration. 【0242】 Example 7. In vivo prophylactic and therapeutic effects of anti-influenza B IgG in a lethal mouse model of influenza infection. The protective efficacy of monoclonal antibodies that neutralize influenza B was evaluated in a lethal influenza B mouse model. 【0243】 Preventive activity (Figure 2A-D): To test prophylactic efficacy, eight groups of 6-8 week old BALB / c (Harlan Laboratories) mice received a single intraperitoneal (IP) injection of either FBC-39 or FBD-94 antibody in 100 μl volumes at doses of 3, 1, 0.3, or 0.1 mg / kg. In each study, the control group was treated with IP using human isotype-unrelated control IgG at a dose of 3 mg / kg in 100 μl volumes. Four hours after administration, a 15-fold 50 percent lethal dose (15 MLD) was administered intranasally to the mice. 50 ) B / Sichuan / 379 / 99 (Yamagata) (B / Sic / 99) or 10MLD 50Mice were inoculated with 50 μl of B / Hong Kong / 330 / 2001 (Victoria) (B / HK / 01). Mice were weighed on the day of viral loading and monitored daily for weight loss and survival for 14 days (mice with weight loss ≥25% were euthanized). Both FBC-39 and FBD-94 mAbs provided dose-dependent protection. FBC-39 and FBD-94 at doses of 0.3 mg / kg or higher provided 90%–100% protection to animals loaded with B / Sic / 99 (Figures 2A and B) and B / HK / 01 (Figures 2C and D). Lower doses of FBC-39 and FBD-94 at 0.1 mg / kg were also highly protective against B / HK / 01, with survival rates of 90% and 80%, respectively. As expected, none of the mice that received isotype control mAbs at 3 or 30 mg / kg survived loading with B / Sic / 99 or B / HK / 01, respectively. 【0244】 Therapeutic activity (Figures 3A-D and 4A and B): To evaluate the therapeutic effect of the antibody, mice were given 10 ml of dextrose. 50 B / Sic / 99 (Yamagata) or 5MLD 50 Animals were inoculated with B / HK / 01 (Victoria), and on day 2 post-infection (pi), they were injected with 10, 3, 1, or 0.3 mg / kg of FBC-39 or FBD-94. FBC-39 and FBD-94 provided complete protection to B / Sic / 99-loaded animals when administered at doses of 1 mg / kg or higher (Figures 3A and B). In B / HK / 01 infection, FBC-39 and FBD-94 provided complete protection at doses of 0.3 mg / kg and above 0.3 mg / kg (Figures 3C and D). As expected, isotype control mAbs administered at 10 or 30 mg / kg failed to protect mice, resulting in survival rates of 10% and 20%, respectively, against B / Sic / 99 and B / HK / 01 infection. 【0245】 To test the ability of influenza B antibodies to provide protection over time, mice were given 5MLD 50Mice were inoculated with B / HK / 01 and administered 3 mg / kg of FBC-39 or FBD-94 via IP injection (started on day 1, 2, 3, or 4 post-infection). FBC-39 protected 100% of mice when administered on day 1 post-infection, and 80% and 70% on days 2 and 3 post-infection, respectively (Figure 4A). FBD-94 protected 100% of mice when administered on days 1 and 2 post-infection, and 80% and 60% on days 3 and 4 post-infection, respectively (Figure 4B). As expected, mice treated with the same dose of unrelated isotype control antibody had a survival rate of 10% and were not protected. 【0246】 Example 8. Hemagglutination inhibitory activity To determine the possible mechanism of action for the functionality of the antibody of the present invention as an influenza B antibody, hemagglutination inhibition (HAI) assays were performed using a diverse group of influenza B virus strains. In the HAI assay, antibodies that block the association of sialic acid expressed on the cell surface with viral receptors are detected by measuring the inhibition of virus-mediated hemagglutination. Influenza B virus (abbreviated as shown in Table 12 below) was adjusted to 4 HA units determined by incubation with 0.05% turkey erythrocytes (Lampire Biological Laboratories) in the absence of the antibody. FBD-94 and FBC-39 IgG were serially diluted in 2-fold increments in a 96-well U-bottom plate, and the diluted viruses were added to the wells. After incubation for 30-60 minutes, 50 μl of 0.05% turkey erythrocytes were added. The plate was incubated for a further 30-60 minutes, and agglutination was observed. The HAI titer was determined to be the lowest effective concentration (nM) of the antibody that completely inhibited agglutination. Table 12 shows that FBD-94 and FBC39 possessed HAI activity against all influenza B strains tested, providing evidence of binding to the globular head of influenza B HA. Other antibodies of the present invention exhibit similar activity when used in HAI assays. 【0247】 [Table 12] 【0248】 All publications, patents, and patent applications described herein are incorporated herein by reference to the same extent as each individual publication, patent, or patent application is described in detail and individually as is incorporated herein by reference. This specification includes: [1] An isolated antibody or antigen-binding fragment thereof that binds to influenza B virus hemagglutinin (HA) and has the ability to neutralize influenza B virus from two phylogenetically distinct lineages. [2] The isolated antibody or antigen-binding fragment thereof described in [1] above, wherein the antibody binds to influenza B virus hemagglutinin and has the ability to neutralize influenza B virus in both Yamagata and Victoria lineages. [3] The antibody or its antigen-binding fragment is a Yamagata lineage influenza B virus selected from B / AA / 94 (ca B / Ann Arbor / 2 / 94 (yamagata)); B / YSI / 98 (ca B / Yamanashi / 166 / 98 (yamagata)); B / JHB / 99 (ca B / Johannesburg / 5 / 99 (yamagata)); B / SC / 99 (B / Sichuan / 379 / 99 (yamagata)); B / FL / 06 (B / Florida / 4 / 2006 (yamagata)); B / BJ / 97 (ca B / Beijing / 243 / 97 (victoria)), B / HK / 01 (B / Hong Kong / 330 / 2001 (victoria)); B / MY / 04 (B / Malaysia / 2506 / 2004 (victoria)); B / BNE / 08 (ca Influenza B virus of the Victoria lineage selected from B / Brisbane / 60 / 2008(victoria); pre-branching influenza B strains selected from B / Lee / 40(B / Lee / 40); B / AA / 66(ca B / Ann Arbor / 1 / 66); B / HK / 72(B / Hong Kong / 5 / 72); and isolated antibodies or antigen-binding fragments thereof as described in [1] above, which are capable of binding to combinations thereof. [4] The antibody or antigen-binding fragment is in the range of approximately 1 μg / ml to approximately 50 μg / ml of the antibody in terms of EC 50 An antibody or antigen-binding fragment thereof, as described in any of the above [1] to [3], which binds to the influenza B virus. [5] The antibody or antigen-binding fragment is present in the neutralization of influenza B virus in a microneutralization assay at a 50% inhibitory concentration (IC) of the antibody within the range of approximately 0.001 μg / ml to approximately 5 μg / ml. 50 An antibody or antigen-binding fragment thereof, as described in any of [1] to [4] above, having a neutralizing potency expressed as μg / ml. [6] An isolated antibody or antigen-binding fragment thereof according to any of [1] to [5] above, which has the ability to bind to influenza A virus hemagglutinin. [7] The isolated antibody or antigen-binding fragment thereof described in [6] above, wherein the antibody has the ability to bind to hemagglutinin of subtype 1 or subtype 2 of influenza A virus. [8] The isolated antibody or antigen-binding fragment thereof described in [6] above, wherein the antibody has the ability to bind to a subtype of influenza A virus group 1 selected from H8, H9, H11, H12, H13, H16 and their variants. [9] The isolated antibody or antigen-binding fragment thereof described in [6] above, wherein the antibody has the ability to bind to subtype H9 of influenza A virus group 1.

[10] Isolated antibodies or antigen-binding fragments thereof that bind to influenza B virus hemagglutinin (HA) and influenza A virus hemagglutinin (HA) and have the ability to neutralize at least one Yamagata lineage influenza B virus or at least one Victoria lineage influenza B virus and at least one subtype of influenza A virus.

[11] The antibody or antigen-binding fragment is in the range of approximately 1 μg / ml to approximately 50 μg / ml of the antibody in terms of EC 50 An antibody or antigen-binding fragment thereof, as described in any of the above [6] to

[10] , that binds to influenza A HA.

[12] The antibody or antigen-binding fragment neutralizes influenza A virus in a microneutralization assay, with an IC of approximately 0.01 μg / ml to approximately 5 μg / ml. 50 An antibody or antigen-binding fragment thereof according to any of the above [6] to

[10] , having the following characteristics.

[13] The antibody or its antigen-binding fragment (a) HCDR-1 of SEQ ID NO: 3, HCDR-2 of SEQ ID NO: 4, HCDR-3 of SEQ ID NO: 5, LCDR-1 of SEQ ID NO: 8, LCDR-2 of SEQ ID NO: 9, and LCDR-3 of SEQ ID NO: 10; (b) HCDR-1 of SEQ ID NO: 13, HCDR-2 of SEQ ID NO: 14, HCDR-3 of SEQ ID NO: 15, LCDR-1 of SEQ ID NO: 18, LCDR-2 of SEQ ID NO: 19, LCDR-3 of SEQ ID NO: 20; (c) HCDR-1 of SEQ ID NO: 23, HCDR-2 of SEQ ID NO: 24, HCDR-3 of SEQ ID NO: 25, LCDR-1 of SEQ ID NO: 28, LCDR-2 of SEQ ID NO: 29, and LCDR-3 of SEQ ID NO: 30; (d) HCDR-1 of SEQ ID NO: 33, HCDR-2 of SEQ ID NO: 34, HCDR-3 of SEQ ID NO: 35, LCDR-1 of SEQ ID NO: 38, LCDR-2 of SEQ ID NO: 39, and LCDR-3 of SEQ ID NO: 40; (e) HCDR-1 of SEQ ID NO: 43, HCDR-2 of SEQ ID NO: 44, HCDR-3 of SEQ ID NO: 45, LCDR-1 of SEQ ID NO: 48, LCDR-2 of SEQ ID NO: 49, and LCDR-3 of SEQ ID NO: 50; (f) HCDR-1 of SEQ ID NO: 53, HCDR-2 of SEQ ID NO: 54, HCDR-3 of SEQ ID NO: 55, LCDR-1 of SEQ ID NO: 58, LCDR-2 of SEQ ID NO: 59, and LCDR-3 of SEQ ID NO: 60; (g) HCDR-1 of SEQ ID NO: 63, HCDR-2 of SEQ ID NO: 64, HCDR-3 of SEQ ID NO: 65, LCDR-1 of SEQ ID NO: 68, LCDR-2 of SEQ ID NO: 69, and LCDR-3 of SEQ ID NO: 70 (h) HCDR-1 of SEQ ID NO: 75, HCDR-2 of SEQ ID NO: 76, HCDR-3 of SEQ ID NO: 77, LCDR-1 of SEQ ID NO: 83, LCDR-2 of SEQ ID NO: 84, and LCDR-3 of SEQ ID NO: 85; (i) HCDR-1 of SEQ ID NO: 91, HCDR-2 of SEQ ID NO: 92, HCDR-3 of SEQ ID NO: 93, LCDR-1 of SEQ ID NO: 99, LCDR-2 of SEQ ID NO: 100, and LCDR-3 of SEQ ID NO: 101; (j) HCDR-1 of SEQ ID NO: 107, HCDR-2 of SEQ ID NO: 108, HCDR-3 of SEQ ID NO: 109, LCDR-1 of SEQ ID NO: 115, LCDR-2 of SEQ ID NO: 116, and LCDR-3 of SEQ ID NO: 117; (k) HCDR-1 of SEQ ID NO: 121, HCDR-2 of SEQ ID NO: 122, HCDR-3 of SEQ ID NO: 123, LCDR-1 of SEQ ID NO: 124, LCDR-2 of SEQ ID NO: 125, and LCDR-3 of SEQ ID NO: 126; (l) HCDR-1 of SEQ ID NO: 127, HCDR-2 of SEQ ID NO: 128, HCDR-3 of SEQ ID NO: 129, LCDR-1 of SEQ ID NO: 130, LCDR-2 of SEQ ID NO: 131, and LCDR-3 of SEQ ID NO: 132; (m) HCDR-1 of SEQ ID NO: 133, HCDR-2 of SEQ ID NO: 134, HCDR-3 of SEQ ID NO: 135, LCDR-1 of SEQ ID NO: 136, LCDR-2 of SEQ ID NO: 137, and LCDR-3 of SEQ ID NO: 138; (n) HCDR-1 of SEQ ID NO: 139, HCDR-2 of SEQ ID NO: 140, HCDR-3 of SEQ ID NO: 141, LCDR-1 of SEQ ID NO: 142, LCDR-2 of SEQ ID NO: 143, and LCDR-3 of SEQ ID NO: 144; (o) HCDR-1 of SEQ ID NO: 145, HCDR-2 of SEQ ID NO: 146, HCDR-3 of SEQ ID NO: 147, LCDR-1 of SEQ ID NO: 148, LCDR-2 of SEQ ID NO: 149, and LCDR-3 of SEQ ID NO: 150; (p) HCDR-1 of SEQ ID NO: 78, HCDR-2 of SEQ ID NO: 79, HCDR-3 of SEQ ID NO: 80, LCDR-1 of SEQ ID NO: 86, LCDR-2 of SEQ ID NO: 87, and LCDR-3 of SEQ ID NO: 88; (q) HCDR-1 of SEQ ID NO: 94, HCDR-2 of SEQ ID NO: 95, HCDR-3 of SEQ ID NO: 96, LCDR-1 of SEQ ID NO: 102, LCDR-2 of SEQ ID NO: 103, and LCDR-3 of SEQ ID NO: 104; (r) HCDR-1 of SEQ ID NO: 110, HCDR-2 of SEQ ID NO: 111, HCDR-3 of SEQ ID NO: 112, LCDR-1 of SEQ ID NO: 118, LCDR-2 of SEQ ID NO: 119, and LCDR-3 of SEQ ID NO: 120; and (s) A set of six CDRs as described in any one of (a) to (r), comprising one or more amino acid substitutions, deletions, or insertions. Six CDR sets selected from: HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2, LCDR-3, comprising the antibodies or antigen-binding fragments described in any of the above [1] to

[12] .

[14] (a) VH of SEQ ID NO: 2 and VL of SEQ ID NO: 7, (b) VH of sequence number 12 and VL of sequence number 17, (c) VH of SEQ ID NO: 22 and VL of SEQ ID NO: 27, (d) VH of SEQ ID NO: 32 and VL of SEQ ID NO: 37, (e) VH of sequence number 42 and VL of sequence number 47, (f) VH of sequence number 52 and VL of sequence number 57, (g) VH of SEQ ID NO: 62 and VL of SEQ ID NO: 67, (h) VH of sequence number 74 and VL of sequence number 82, (i) VH of SEQ ID NO: 90 and VL of SEQ ID NO: 98, (j) VH of SEQ ID NO: 106 and VL of SEQ ID NO: 114 An antibody or antigen-binding fragment thereof according to any of [1] to

[13] above, comprising a VH having at least 75%, 80%, 85%, 90%, 95%, or 100% identity with a VH and / or a VL having at least 75%, 80%, 85%, 90%, 95%, or 100% identity with a VH and / or VL selected from the above.

[15] (a) VH of sequence number 2 and VL of sequence number 7, (b) VH of sequence number 12 and VL of sequence number 17, (c) VH of SEQ ID NO: 22 and VL of SEQ ID NO: 27, (d) VH of SEQ ID NO: 32 and VL of SEQ ID NO: 37, (e) VH of sequence number 42 and VL of sequence number 47, (f) VH of sequence number 52 and VL of sequence number 57, (g) VH of SEQ ID NO: 62 and VL of SEQ ID NO: 67, (h) VH of sequence number 74 and VL of sequence number 82, (i) VH of SEQ ID NO: 90 and VL of SEQ ID NO: 98, (j) VH of SEQ ID NO: 106 and VL of SEQ ID NO: 114 An antibody or antigen-binding fragment thereof as described in any of [1] to

[14] above, comprising VH and VL selected from [1].

[16] It binds to influenza B virus hemagglutinin (HA) and has the ability to neutralize influenza B viruses in two phylogenetically distinct lineages, and comprises the VH amino acid sequence of SEQ ID NO: 71 and the VL amino acid sequence of SEQ ID NO: 72, where Xaa of SEQ ID NO: 71 1 is Val or Glu; Xaa of SEQ ID NO: 71 2 is Leu or Phe; Xaa of sequence number 71 3 is Ser or Thr; Xaa of sequence number 71 4 is Leu or Ser; Xaa of sequence number 71 5 is Ser or Thr; Xaa of sequence number 71 6 is Met or Thr; Xaa of sequence number 71 7 is Phe or Tyr; Xaa of sequence number 71 8 is His or Gln; Xaa of Sequence ID 71 9 is Ser or Asn; Xaa of sequence number 71 10 is Arg or Lys; and Xaa of sequence number 71 11 is Ala or Thr; here Xaa of sequence number 72 1 Phe or Tyr, an antibody or its antigen-binding fragment.

[17] Xaa of sequence number 71 9 The antibody or antigen-binding fragment thereof described in

[16] above, wherein is Ser.

[18] Xaa of sequence number 71 4 The antibody or antigen-binding fragment thereof as described in

[16] above, wherein is Leu.

[19] Xaa of sequence number 71 1 is Glu; Xaa of sequence number 71 5 is Thr; Xaa of sequence number 71 6 is Thr; Xaa of sequence number 71 7 is Tyr; Xaa of sequence number 71 8 is Gln; Xaa of sequence number 71 10 is Lys; Xaa of sequence number 71 11 An antibody or antigen-binding fragment thereof as described in any of the above [6] to [8], wherein is Thr, or a combination thereof.

[20] Xaa of sequence number 71 1 is Glu; Xaa of sequence number 71 5 is Thr; Xaa of sequence number 71 6 is Thr; Xaa of sequence number 71 7 is Tyr; Xaa of sequence number 71 8 is Gln; Xaa of sequence number 71 9 is Ser; Xaa of sequence number 71 10 is Lys; and Xaa of sequence number 71 11 An antibody or antigen-binding fragment thereof as described in any of the above [[6] to [9]], wherein Thr is present.

[21] Antibodies or antigen-binding fragments include immunoglobulin molecules, monoclonal antibodies, chimeric antibodies, CDR transplant antibodies, humanized antibodies, Fab, Fab', and F(ab'). 2 An antibody or antigen-binding fragment thereof, selected from the group consisting of Fv, disulfide-linked Fv, scFv, single-domain antibody, diabody, multispecific antibody, dual-specific antibody, and bispecific antibody, as described in any of [1] to

[20] above.

[22] An antibody or antigen-binding fragment thereof according to any of [1] to

[21] above, comprising an Fc region.

[23] The antibody or antigen-binding fragment thereof according to any one of [1] to

[22] above, wherein the antibody is IgG1, IgG2, or IgG4 or a fragment thereof.

[24] An antibody against influenza B virus or an antigen-binding fragment thereof, which binds to influenza B virus and has the ability to neutralize at least one Yamagata lineage and at least one Victoria lineage of influenza B virus, and which binds to an epitope conserved in at least one Yamagata lineage and at least one Victoria lineage of influenza B virus.

[25] The antibody or antigen-binding fragment thereof according to

[24] above, wherein one or more contact residues of the epitope are located within the head region of influenza B HA.

[26] The antibody or antigen-binding fragment thereof according to

[24] , wherein the epitope comprises one or more amino acids selected from 128, 141, 150 and 235 of the sequence of the head region of HA as a contact residue.

[27] An antibody against influenza B virus or an antigen-binding fragment thereof that binds to influenza B virus hemagglutinin and has the ability to neutralize influenza B virus in two phylogenetically different lineages, either by binding to the same epitope as the antibody described in any of [1] to

[26] above or by competing with the antibody for binding to influenza B virus hemagglutinin.

[28] (a) VH of SEQ ID NO: 2 and VL of SEQ ID NO: 7, (b) VH of sequence number 12 and VL of sequence number 17, (c) VH of SEQ ID NO: 22 and VL of SEQ ID NO: 27, (d) VH of SEQ ID NO: 32 and VL of SEQ ID NO: 37, (e) VH of sequence number 42 and VL of sequence number 47, (f) VH of sequence number 52 and VL of sequence number 57, (g) VH of SEQ ID NO: 62 and VL of SEQ ID NO: 67, (h) VH of sequence number 74 and VL of sequence number 82, (i) VH of SEQ ID NO: 90 and VL of SEQ ID NO: 98, (j) VH of SEQ ID NO: 106 and VL of SEQ ID NO: 114 An antibody or antigen-binding fragment thereof as described in

[27] above, which binds to the same epitope as an antibody having an amino acid sequence selected from or competes with the antibody for binding to influenza A virus hemagglutinin.

[29] An isolated nucleic acid encoding an antibody or an antigen-binding fragment thereof as described in any of [1] to

[28] above.

[30] A vector containing the isolated nucleic acid described in

[29] above.

[31] A host cell containing the nucleic acid described in

[29] above or the vector described in

[30] above.

[32] A method for producing an antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above, comprising the step of culturing the host cells described in

[31] above under conditions suitable for the expression of the antibody or fragment thereof.

[33] The method according to

[32] , further comprising the step of isolating the antibody or its antigen-binding fragment from the host cell culture.

[34] A composition comprising an antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above and a pharmaceutically acceptable carrier.

[35] A composition comprising an antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above, and 25 mM His and 0.15 M NaCl at pH 6.0.

[36] An antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above, for use in the prevention or treatment of influenza B infection in a subject.

[37] An antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above, for use in the prevention or treatment of influenza A and influenza B infection in subjects.

[38] Use of an antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above in the manufacture of a drug for the prevention or treatment of influenza B infection in a subject.

[39] Use of an antibody or antigen-binding fragment thereof as described in any of [1] to

[28] above in the manufacture of a drug for the prevention or treatment of influenza A and influenza B infection in a subject.

[40] A method for the prevention or treatment of influenza B infection in a subject, comprising the step of administering an effective amount of an antibody or antigen-binding fragment thereof described in any of [1] to

[28] above to the subject.

[41] A method for the prevention or treatment of influenza A and influenza B infection in a subject, comprising the step of administering to the subject an effective amount of an antibody or antigen-binding fragment thereof described in any of [1] to

[28] above.

[42] Use of any antibody or fragment thereof described in any of [1] to

[28] above for in vitro diagnosis of influenza B infection in a subject. 【0249】 Sequence listing information Sequence ID 1 (FBD-56 VH DNA) [ka] Sequence ID 2 (FBD-56 VH protein) [ka] Sequence ID 3 (FBD-56 HCDR-1-Kabat): DYAMN Sequence ID 4 (FBD-56 HCDR-2-Kabat):VISWDSGRIGYADSVKG Sequence ID 5 (FBD-56 HCDR-3-Kabat): DMLAYYSDNSGKKYNVYGMDV Sequence ID 6 (FBD-56 VL DNA) [ka] Sequence ID 7 (FBD-56 VL protein) [ka] Sequence ID 8 (FBD-56 LCDR-1-Kabat): ASQSVSTFLA Sequence ID 9 (FBD-56 LCDR-2-Kabat): DASNRAT Sequence ID 10 (FBD-56 LCDR-3-Kabat): QQRSHWPPI Sequence ID 11 (FBD-94 VH DNA) [ka] Sequence ID No. 12 (FBD-94 VH protein) [ka] Sequence ID 13 (FBD-94 HCDR-1-Kabat): DYAIN Sequence ID 14 (FBD-94 HCDR-2-Kabat): IISWDSGRIGYADSVRG Sequence ID 15 (FBD-94 HCDR-3-Kabat):DMLAYYYDGSGIRYNLYGMDV Sequence ID No. 16 (FBD-94 VL DNA) [ka] Sequence ID No. 17 (FBD-94 VL protein) [ka] Sequence ID 18 (FBD-94 LCDR-1-Kabat):RASRSITTFLA Sequence ID 19 (FBD-94 LCDR-2-Kabat): DASNRAT Sequence ID 20 (FBD-94 LCDR-3-Kabat): QQRDHWPPI Sequence ID 21 (FBC-39 VH DNA) [ka] Sequence ID No. 22 (FBC-39 VH protein) [ka] Sequence ID 23 (FBC-39 HCDR-1-Kabat): NAWMS Sequence ID 24 (FBC-39 HCDR-2-Kabat):RIKSNTDGGTTDYAAPVKG Sequence ID 25 (FBC-39 HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID No. 26 (FBC-39 VL DNA) [ka] Sequence ID No. 27 (FBC-39 VL protein) [ka] Sequence ID 28 (FBC-39 LCDR-1-Kabat):RASQDISTWLA Sequence ID 29 (FBC-39 LCDR-2-Kabat): AASSLQS Sequence ID 30 (FBC-39 LCDR-3-Kabat): QQANSFPPT Sequence ID 31 (FBC-39 LSL VH DNA) [ka] Sequence ID 32 (FBC-39 LSL VH protein) [ka] Sequence ID 33 (FBC-39 LSL HCDR-1-Kabat): NAWMS Sequence ID 34 (FBC-39 LSL HCDR-2-Kabat):RIKSNTDGGTTDYAAPVKG Sequence ID 35 (FBC-39 LSL HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID 36 (FBC-39 LSL VL DNA) [ka] Sequence ID 37 (FBC-39 LSL VL protein) [ka] Sequence ID 38 (FBC-39 LSL LCDR-1-Kabat):RASQDISTWLA Sequence ID 39 (FBC-39 LSL LCDR-2-Kabat): AASSLQS Sequence ID 40 (FBC-39 LSL LCDR-3-Kabat): QQANSFPPT Sequence ID No. 41 (FBC-39 FSL VH DNA) [ka] Sequence ID 42 (FBC-39 FSL VH protein) [ka] Sequence ID 43 (FBC-39 FSL HCDR-1-Kabat): NAWMS Sequence ID 44 (FBC-39 FSL HCDR-2-Kabat):RIKSNTDGGTTDYAAPVKG Sequence ID 45 (FBC-39 FSL HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID No. 46 (FBC-39 FSL VL DNA) [ka] Sequence ID No. 47 (FBC-39 FSL VL protein) [ka] Sequence ID 48 (FBC-39 FSL LCDR-1-Kabat):RASQDISTWLA Sequence ID 49 (FBC-39 FSL LCDR-2-Kabat): AASSLQS Sequence ID 50 (FBC-39 FSL LCDR-3-Kabat): QQANSFPPT Sequence ID 51 (FBC-39 LTL VH DNA) [ka] Sequence ID 52 (FBC-39 LTL VH protein) [ka] Sequence ID 53 (FBC-39 LTL HCDR-1-Kabat): NAWMS Sequence ID 54 (FBC-39 LTL HCDR-2-Kabat):RIKSNTDGGTTDYAAPVKG Sequence ID 55 (FBC-39 LTL HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID 56 (FBC-39 LTL VL DNA) [ka] Sequence ID No. 57 (FBC-39 LTL VL protein) [ka] Sequence ID 58 (FBC-39 LTL LCDR-1-Kabat):RASQDISTWLA Sequence ID 59 (FBC-39 LTL LCDR-2-Kabat): AASSLQS Sequence ID 60 (FBC-39 LTL LCDR-3-Kabat): QQANSFPPT Sequence ID 61 (FBC-39 FTL VH DNA) [ka] Sequence ID 62 (FBC-39 FTL VH protein) [ka] Sequence ID 63 (FBC-39 FTL HCDR-1-Kabat): NAWMS Sequence ID 64 (FBC-39 FTL HCDR-2-Kabat):RIKSNTDGGTTDYAAPVKG Sequence ID 65 (FBC-39 FTL HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID 66 (FBC-39 FTL VL DNA) [ka] Sequence ID 67 (FBC-39 FTL VL protein) [ka] Sequence ID 68 (FBC-39 FTL LCDR-1-Kabat):RASQDISTWLA Sequence ID 69 (FBC-39 FTL LCDR-2-Kabat): AASSLQS Sequence ID 70 (FBC-39 FTL LCDR-3-Kabat): QQANSFPPT Sequence ID No. 71 (FBC-39 VH protein - with variable amino acids) (See Figure 6) Sequence ID No. 72 (FBC-39 VL protein - with variable amino acids) (See Figure 7) Sequence ID 73 (FBC-39 FSS VH DNA) [ka] Sequence ID No. 74 (FBC-39 FSS VH protein) [ka] Sequence ID 75 (FBC-39 FSS HCDR-1-Kabat):RASQDISTWLA Sequence ID 76 (FBC-39 FSS HCDR-2-Kabat):RIKSNTDGGTTDYAAPVKG Sequence ID 77 (FBC-39 FSS HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID 78 (FBC-39 FSS HCDR-1-IMGT): GFSFSNAW Sequence ID 79 (FBC-39 FSS HCDR-2-IMGT): IKSNTDGGTT Sequence ID 80 (FBC-39 FSS HCDR-3-IMGT):TTDGPYSDDFRSGYAARYRYFGMDV Sequence ID No. 81 (FBC-39 FSS VL DNA) [ka] Sequence ID No. 82 (FBC-39 FSS VL protein) [ka] Sequence ID 83 (FBC-39 FSS LCDR-1-Kabat):RASQDISTWLA Sequence ID 84 (FBC-39 FSS LCDR-2-Kabat):AASSLQS Sequence ID 85 (FBC-39 FSS LCDR-3-Kabat): QQANSFPPT Sequence ID 86 (FBC-39 FSS LCDR-1-IMGT): QDISTW Sequence ID 87 (FBC-39 FSS LCDR-2-IMGT): AAS Sequence ID 88 (FBC-39 FSS LCDR-3-IMGT): QQANSFPPT Sequence ID 89 (FBC-39 LTS VH DNA): [ka] Sequence ID 90 (FBC-39 LTS VH protein): [ka] Sequence ID 91 (FBC-39 LTS HCDR-1-Kabat):RASQDISTWLA Sequence ID 92 (FBC-39 LTS HCDR-2-Kabat): RIKSNTDGGTTDYAAPVKG Sequence ID 93 (FBC-39 LTS HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID 94 (FBC-39 LTS HCDR-1-IMGT): GLTFSNAW Sequence ID 95 (FBC-39 LTS HCDR-2-IMGT): IKSNTDGGTT Sequence ID 96 (FBC-39 LTS HCDR-3-IMGT):TTDGPYSDDFRSGYAARYRYFGMDV Sequence ID 97 (FBC-39 LTS VL DNA) [ka] Sequence ID 98 (FBC-39 LTS VL protein) [ka] Sequence ID 99 (FBC-39 LTS LCDR-1-Kabat):RASQDISTWLA Sequence ID 100 (FBC-39 LTS LCDR-2-Kabat): AASSLQS Sequence ID 101 (FBC-39 LTS LCDR-3-Kabat): QQANSFPPT Sequence ID 102 (FBC-39 LTS LCDR-1-IMGT): QDISTW Sequence ID 103 (FBC-39 LTS LCDR-2-IMGT): AAS Sequence ID 104 (FBC-39 LTS LCDR-3-IMGT): QQANSFPPT Sequence ID 105 (FBC-39 FTS VH DNA): [ka] Sequence ID 106 (FBC-39 FTS VH protein): [ka] Sequence ID 107 (FBC-39 FTS HCDR-1-Kabat):RASQDISTWLA Sequence ID 108 (FBC-39 FTS HCDR-2-Kabat): RIKSNTDGGTTDYAAPVKG Sequence ID 109 (FBC-39 FTS HCDR-3-Kabat):DGPYSDDFRSGYAARYRYFGMDV Sequence ID 110 (FBC-39 FTS HCDR-1-IMGT): GFTFSNAW Sequence ID 111 (FBC-39 FTS HCDR-2-IMGT): IKSNTDGGTT Sequence ID 112 (FBC-39 FTS HCDR-3-IMGT):TDGPYSDDFRSGYAARYRYFGMDV Sequence ID 113 (FBC-39 FTS VL DNA) [ka] Sequence ID No. 114 (FBC-39 FTS VL protein) [ka] Sequence ID 115 (FBC-39 FTS LCDR-1-Kabat):RASQDISTWLA Sequence ID 116 (FBC-39 FTS LCDR-2-Kabat): AASSLQS Sequence ID 117 (FBC-39 FTS LCDR-3-Kabat): QQANSFPPT Sequence ID 118 (FBC-39 FTS LCDR-1-IMGT): QDISTW Sequence ID 119 (FBC-39 FTS LCDR-2-IMGT): AAS Sequence ID 120 (FBC-39 FTS LCDR-3-IMGT): QQANSFPPT Sequence ID 121 (FBC-39 HCDR-1-IMGT): GLSFLNAW Sequence ID 122 (FBC-39 HCDR-2-IMGT): IKSNTDGGTT Sequence ID 123 (FBC-39 HCDR-3-IMGT):TDGPYSDDFRSGYAARYRYFGMDVW Sequence ID 124 (FBC-39 LCDR-1-IMGT): QDISTW Sequence ID 125 (FBC-39 LCDR-2-IMGT): AAS Sequence ID 126 (FBC-39 LCDR-3-IMGT): QQANSFPPT Sequence ID 127 (FBC-39 LSL HCDR-1-IMGT): GLSFLNAW Sequence ID 128 (FBC-39 LSL HCDR-2-IMGT): IKSNTDGGTT Sequence ID 129 (FBC-39 LSL HCDR-3-IMGT):TTDGPYSDDFRSGYAARYRYFGMDV Sequence ID 130 (FBC-39 LSL LCDR-1-IMGT): QDISTW Sequence ID 131 (FBC-39 LSL LCDR-2-IMGT): AAS Sequence ID 132 (FBC-39 LSL LCDR-3-IMGT): QQANSFPPT Sequence ID 133 (FBC-39 FSL HCDR-1-IMGT): GFSFLNAW Sequence ID 134 (FBC-39 FSL HCDR-2-IMGT): IKSNTDGGTT Sequence ID 135 (FBC-39 LSL HCDR-3-IMGT):TTDGPYSDDFRSGYAARYRYFGMDV Sequence ID 136 (FBC-39 FSL LCDR-1-IMGT): QDISTW Sequence ID 137 (FBC-39 FSL LCDR-2-IMGT): AAS Sequence ID 138 (FBC-39 FSL LCDR-3-IMGT): QQANSFPPT Sequence ID 139 (FBC-39 LTL HCDR-1-IMGT): GLTFLNAW Sequence ID 140 (FBC-39 LTL HCDR-2-IMGT): IKSNTDGGTT Sequence ID 141 (FBC-39 LTL HCDR-3-IMGT):TTDGPYSDDFRSGYAARYRYFGMDV Sequence ID 142 (FBC-39 LTL LCDR-1-IMGT): QDISTW Sequence ID 143 (FBC-39 LTL LCDR-2-IMGT): AAS Sequence ID 144 (FBC-39 LTL LCDR-3-IMGT): QQANSFPPT Sequence ID 145 (FBC-39 FTL HCDR-1-IMGT): GFTFLNAW Sequence ID 146 (FBC-39 FTL HCDR-2-IMGT): IKSNTDGGTT Sequence ID 147 (FBC-39 FTL HCDR-3-IMGT):TTDGPYSDDFRSGYAARYRYFGMDV Sequence ID 148 (FBC-39 FTL LCDR-1-IMGT): QDISTW Sequence ID 149 (FBC-39 FTL LCDR-2-IMGT): AAS Accession Number 150 (FBC-39 FTL LCDR-3-IMGT): QQANSFPPT <Sequence Listing> <110> MEDIMMUNE, LLC HUMABS BIOMED SA <120> NEUTRALIZING ANTI-INFLUENZA B ANTIBODIES AND USES THEREOF <130> PA24-048 <150> US62 / 024,804 <151> 2014-07-15 <160> 154 <170> PatentIn version 3.5 <210> 1 <211> 391 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 1 gaagtgcagc tggtggagtc tgggggacac ttggtgcagc ctggcaggtc cctgagactc 60 tcctgtgcag cctctggatt cacctttgag gattatgcca tgaattgggt ccggcaagct 120 ccagggaagg gcctggagtg ggtctcagtc attagttggg acagtggtag gataggctat 180 gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctcctcgtat 240 ctgcaaatga acagtctgag acctgaggac actgccttgt attattgtgt aagagatatg 300 ttggcttatt attctgacaa tagtggcaaa aaatacaacg tctacggtat ggacgtctgg 360 ggccaaggga ccacggtcac cgtctcctca g 391 <210> 2 <211> 130 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 2 Glu Val Gln Leu Val Glu Ser Gly Gly His Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Glu Asp Tyr 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser Trp Asp Ser Gly Arg Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Ser Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Val Arg Asp Met Leu Ala Tyr Tyr Ser Asp Asn Ser Gly Lys Lys Tyr 100 105 110 Asn Val Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val 115 120 125 Ser Ser 130 <210> 3 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 3 Asp Tyr Ala Met Asn 1 5 <210> 4 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 4 Val Ile Ser Trp Asp Ser Gly Arg Ile Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 5 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 5 Asp Met Leu Ala Tyr Tyr Ser Asp Asn Ser Gly Lys Lys Tyr Asn Val 1 5 10 15 Tyr Gly Met Asp Val 20 <210> 6 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 6 gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca gggccagtca gagtgtttcc accttcttag cctggtacca acagaaacct 120 ggccaggctc ccaggctcct catgtatgat gcatccaaca gggccactgg catcccagcc 180 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagaacct 240 gaagattttg caatttacta ctgtcagcag cgtagccact ggcctcctat cttcggccaa 300 gggacacgac tggagattaa ac 322 <210> 7 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 7 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Thr Phe 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Met 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Arg Ser His Trp Pro Pro 85 90 95 Ile Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 8 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 8 Ala Ser Gln Ser Val Ser Thr Phe Leu Ala 1 5 10 <210> 9 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 9 Asp Ala Ser Asn Arg Ala Thr 1 5 <210> 10 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 10 Gln Gln Arg Ser His Trp Pro Pro Ile 1 5 <210> 11 <211> 391 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 11 gaagtgcagc tggtggagtc tgggggaggc ttggtgcaac ctggcaggtc cctgagactc 60 tcctgtgcag tttctggatt catctttgaa gattatgcca taaactgggt ccggcaagct 120 ccagggaagg gcctggagtg ggtctcaatt attagttggg acagtggtag gataggctac 180 gcggactctg tgaggggccg attcaccatc tccagagaca acgccaagaa ctcctcgttt 240 ctgcaaatga acagtctgag acccgaagac acggccgtgt attattgtgt aaaagatatg 300 ttggcgtatt attatgatgg tagcggcatc aggtacaacc tctacggtat ggacgtctgg 360 ggccaaggga ccacggtcac cgtctcctca g 391 <210> 12 <211> 130 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 12 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Ile Phe Glu Asp Tyr 20 25 30 Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ile Ile Ser Trp Asp Ser Gly Arg Ile Gly Tyr Ala Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Ser Phe 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Lys Asp Met Leu Ala Tyr Tyr Tyr Asp Gly Ser Gly Ile Arg Tyr 100 105 110 Asn Leu Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val 115 120 125 Looking Looking 130 <210> 13 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 13 Asp Tyr Ala Ile Asn 1 5 <210> 14 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 14 Ile Ile Ser Trp Asp Ser Gly Arg Ile Gly Tyr Ala Asp Ser Val Arg 1 5 10 15 Gly <210> 15 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 15 Asp Met Leu Ala Tyr Tyr Tyr Asp Gly Ser Gly Ile Arg Tyr Asn Leu 1 5 10 15 Tyr Gly Met Asp Val 20 <210> 16 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 16 gaaattgtgt tgacacagtc tccagccact ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca gggccagtcg gagtattacc accttcttag cctggtacca acaaaaacct 120 ggccaggctc ccaggctcct catctacgat gcatccaaca gggccactgg cgtcccagcc 180 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcaacag cctagagcct 240 gacgattttg caatttatta ctgtcagcag cgtgaccact ggcctccgat cttcggccaa 300 gggacacgac tggagattaa ac 322 <210> 17 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 17 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Arg Ser Ile Thr Thr Phe 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Glu Pro 65 70 75 80 Asp Asp Phe Ala Ile Tyr Tyr Cys Gln Gln Arg Asp His Trp Pro Pro 85 90 95 Ile Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 18 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 18 Arg Ala Ser Arg Ser Ile Thr Thr Phe Leu Ala 1 5 10 <210> 19 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 19 Asp Ala Ser Asn Arg Ala Thr 1 5 <210> 20 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 20 Gln Gln Arg Asp His Trp Pro Pro Ile 1 5 <210> 21 <211> 403 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 21 gaggtgcagc tggtggtgtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggact cagtttcctt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc agcatctcaa gagacgattc aaagaacatg 240 ctgtttctgc atatgagcag cctgagaacc gaggacacag ccgtctatta ctgcgccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct cag 403 <210> 22 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 22 Glu Val Gln Leu Val Val Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Ser Phe Leu Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Ser Ile Ser Arg Asp Asp Ser Lys Asn Met 65 70 75 80 Leu Phe Leu His Met Ser Ser Leu Arg Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Ala Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 23 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 23 Asn Ala Trp Met Ser 1 5 <210> 24 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 24 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 25 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 25 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 26 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 26 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttactt ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 27 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 27 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 28 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 28 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 29 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 29 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 30 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 30 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 31 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 31 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggact ctctttcctt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 32 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 32 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Ser Phe Leu Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 33 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 33 Asn Ala Trp Met Ser 1 5 <210> 34 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 34 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 35 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 35 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 36 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 36 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 37 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 37 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 38 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 38 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 39 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 39 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 40 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 40 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 41 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 41 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggatt ctctttcctt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 42 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 42 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Leu Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 43 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 43 Asn Ala Trp Met Ser 1 5 <210> 44 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 44 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 45 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 45 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 46 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 46 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 47 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 47 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 48 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 48 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 49 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 49 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 50 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 50 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 51 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 51 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggact cactttcctt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 52 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 52 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Leu Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 53 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 53 Asn Ala Trp Met Ser 1 5 <210> 54 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 54 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 55 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 55 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 56 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 56 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 57 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 57 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 58 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 58 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 59 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 59 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 60 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 60 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 61 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 61 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggatt cactttcctt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 62 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 62 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Leu Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 63 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 63 Asn Ala Trp Met Ser 1 5 <210> 64 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 64 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 65 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 65 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 66 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 66 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 67 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 67 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 68 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 68 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 69 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 69 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 70 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 70 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 71 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <220> <221> MOD_RES <222> (6)..(6) <223> Val or Glu <220> <221> MOD_RES <222> (27)..(27) <223> Leu or Phe <220> <221> MOD_RES <222> (28)..(28) <223> Thr or Ser <220> <221> MOD_RES <222> (30)..(30) <223> Ser or Leu <220> <221> MOD_RES <222> (71)..(71) <223> Thr or Ser <220> <221> MOD_RES <222> (80)..(80) <223> Thr or Met <220> <221> MOD_RES <222> (82)..(82) <223> Tyr or Phe <220> <221> MOD_RES <222> (84)..(84) <223> Gln or His <220> <221> MOD_RES <222> (86)..(86) <223> Asn or Ser <220> <221> MOD_RES <222> (89)..(89) <223> Lys or Arg <220> <221> MOD_RES <222> (99)..(99) <223> Thr or Ala <400> 71 Glu Val Gln Leu Val Xaa Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Xaa Xaa Phe Xaa Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Xaa Ile Ser Arg Asp Asp Ser Lys Asn Xaa 65 70 75 80 Leu Xaa Leu Xaa Met Xaa Ser Leu Xaa Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Xaa Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 72 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <220> <221> MOD_RES <222> (87)..(87) <223> Phe or Tyr <400> 72 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Xaa Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 73 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 73 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggatt ctctttcagt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 74 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 74 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 75 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 75 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 76 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 76 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 77 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 77 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 78 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 78 Gly Phe Ser Phe Ser Asn Ala Trp 1 5 <210> 79 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 79 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 80 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 80 Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala 1 5 10 15 Arg Tyr Arg Tyr Phe Gly Met Asp Val 20 25 <210> 81 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 81 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 82 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 82 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 83 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 83 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 84 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 84 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 85 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 85 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 86 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 86 Gln Asp Ile Ser Thr Trp 1 5 <210> 87 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 87 Ala Ala Ser 1 <210> 88 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 88 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 89 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 89 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggact cactttcagt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 90 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 90 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Ser Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 91 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 91 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 92 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 92 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 93 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 93 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 94 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 94 Gly Leu Thr Phe Ser Asn Ala Trp 1 5 <210> 95 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 95 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 96 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 96 Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala 1 5 10 15 Arg Tyr Arg Tyr Phe Gly Met Asp Val 20 25 <210> 97 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 97 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 98 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 98 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 99 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 99 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 100 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 100 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 101 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 101 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 102 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 102 Gln Asp Ile Ser Thr Trp 1 5 <210> 103 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 103 Ala Ala Ser 1 <210> 104 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 104 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 105 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 105 gaggtgcagc tggtggagtc tgggggaggc ttggtaaagc ctggggggtc ccttagactc 60 tcctgtgcag cctctggatt cactttcagt aacgcctgga tgagctgggt ccgccaggct 120 ccagggaagg gcctggagtg ggttggccgt attaaaagta atactgatgg tgggacaaca 180 gactacgccg cacccgtgaa aggcagattc accatctcaa gagacgattc aaagaacacg 240 ctgtatctgc aaatgagcag cctgaaaacc gaggacacag ccgtctatta ctgcaccaca 300 gatggacctt actctgacga ttttagaagt ggttatgccg cacgctaccg ttatttcgga 360 atggacgtct ggggccaagg gaccacggtc accgtctcct ca 402 <210> 106 <211> 134 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 106 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Ala 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Gly Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60 Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Ser Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr 100 105 110 Ala Ala Arg Tyr Arg Tyr Phe Gly Met Asp Val Trp Gly Gln Gly Thr 115 120 125 Thr Val Thr Val Ser Ser 130 <210> 107 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 107 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 108 <211> 19 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 108 Arg Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro 1 5 10 15 Val Lys Gly <210> 109 <211> 23 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 109 Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg Tyr 1 5 10 15 Arg Tyr Phe Gly Met Asp Val 20 <210> 110 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 110 Gly Phe Thr Phe Ser Asn Ala Trp 1 5 <210> 111 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 111 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 112 <211> 24 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 112 Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg 1 5 10 15 Tyr Arg Tyr Phe Gly Met Asp Val 20 <210> 113 <211> 322 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polynucleotide <400> 113 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc 60 atcacttgtc gggcgagtca ggatattagc acctggttag cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180 agattcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcagcag gctaacagtt tccctccgac ttttggccag 300 gggaccaagc tggagatcaa ac 322 <210> 114 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 114 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 115 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 115 Arg Ala Ser Gln Asp Ile Ser Thr Trp Leu Ala 1 5 10 <210> 116 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 116 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 117 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 117 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 118 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 118 Gln Asp Ile Ser Thr Trp 1 5 <210> 119 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 119 Ala Ala Ser 1 <210> 120 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 120 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 121 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 121 Gly Leu Ser Phe Leu Asn Ala Trp 1 5 <210> 122 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 122 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 123 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 123 Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala Arg 1 5 10 15 Tyr Arg Tyr Phe Gly Met Asp Val Trp 20 25 <210> 124 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 124 Gln Asp Ile Ser Thr Trp 1 5 <210> 125 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 125 Ala Ala Ser 1 <210> 126 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 126 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 127 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 127 Gly Leu Ser Phe Leu Asn Ala Trp 1 5 <210> 128 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 128 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 129 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 129 Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala 1 5 10 15 Arg Tyr Arg Tyr Phe Gly Met Asp Val 20 25 <210> 130 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 130 Gln Asp Ile Ser Thr Trp 1 5 <210> 131 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 131 Ala Ala Ser 1 <210> 132 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 132 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 133 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 133 Gly Phe Ser Phe Leu Asn Ala Trp 1 5 <210> 134 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 134 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 135 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 135 Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala 1 5 10 15 Arg Tyr Arg Tyr Phe Gly Met Asp Val 20 25 <210> 136 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 136 Gln Asp Ile Ser Thr Trp 1 5 <210> 137 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 137 Ala Ala Ser 1 <210> 138 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 138 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 139 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 139 Gly Leu Thr Phe Leu Asn Ala Trp 1 5 <210> 140 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 140 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 141 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 141 Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala 1 5 10 15 Arg Tyr Arg Tyr Phe Gly Met Asp Val 20 25 <210> 142 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 142 Gln Asp Ile Ser Thr Trp 1 5 <210> 143 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 143 Ala Ala Ser 1 <210> 144 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 144 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 145 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 145 Gly Phe Thr Phe Leu Asn Ala Trp 1 5 <210> 146 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 146 Ile Lys Ser Asn Thr Asp Gly Gly Thr Thr 1 5 10 <210> 147 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 147 Thr Thr Asp Gly Pro Tyr Ser Asp Asp Phe Arg Ser Gly Tyr Ala Ala 1 5 10 15 Arg Tyr Arg Tyr Phe Gly Met Asp Val 20 25 <210> 148 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 148 Gln Asp Ile Ser Thr Trp 1 5 <210> 149 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 149 Ala Ala Ser 1 <210> 150 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 150 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 151 <211> 347 <212> PRT <213> Influenza B virus <400> 151 Asp Arg Ile Cys Thr Gly Ile Thr Ser Ser Asn Ser Pro His Val Val 1 5 10 15 Lys Thr Ala Thr Gln Gly Glu Val Asn Val Thr Gly Val Ile Pro Leu 20 25 30 Thr Thr Thr Pro Thr Lys Ser His Phe Ala Asn Leu Lys Gly Thr Glu 35 40 45 Thr Arg Gly Lys Leu Cys Pro Lys Cys Leu Asn Cys Thr ...

Claims

[Claim 1] An isolated antibody or its antigen-binding fragment that binds to the spherical head region of influenza B virus hemagglutinin (HA), binds to influenza A virus HA, neutralizes influenza B viruses in two phylogenetically distinct lineages, and has the ability to neutralize at least one influenza A virus subtype, An isolated antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a set of six CDRs, including HCDR-1 of SEQ ID NO: 139, HCDR-2 of SEQ ID NO: 140, HCDR-3 of SEQ ID NO: 141, LCDR-1 of SEQ ID NO: 142, LCDR-2 consisting of the amino acid sequence AAS, and LCDR-3 of SEQ ID NO:

144. [Claim 2] An isolated antibody or antigen-binding fragment thereof according to claim 1, comprising VH having at least 90% or 95% identity with the amino acid sequence of SEQ ID NO: 52, and VL having at least 90% or 95% identity with the amino acid sequence of SEQ ID NO:

57. [Claim 3] The aforementioned antibody or its antigen-binding fragment: (a) Having the ability to bind to influenza B virus hemagglutinin and neutralize influenza B virus in both Yamagata and Victoria lineages, and / or (b) Yamagata lineage influenza B virus selected from B / AA / 94 (ca B / Ann Arbor / 2 / 94 (yamagata)); B / YSI / 98 (ca B / Yamanashi / 166 / 98 (yamagata)); B / JHB / 99 (ca B / Johannesburg / 5 / 99 (yamagata)); B / SC / 99 (B / Sichuan / 379 / 99 (yamagata)); B / FL / 06 (B / Florida / 4 / 2006 (yamagata)); B / BJ / 97 (ca Influenza B viruses of the Victoria lineage selected from B / Beijing / 243 / 97 (victoria)), B / HK / 01 (B / Hong Kong / 330 / 2001 (victoria)); B / MY / 04 (B / Malaysia / 2506 / 2004 (victoria)); B / BNE / 08 (ca B / Brisbane / 60 / 2008 (victoria)); pre-branching influenza B strains selected from B / Lee / 40 (B / Lee / 40); B / AA / 66 (ca B / Ann Arbor / 1 / 66); B / HK / 72 (B / Hong Kong / 5 / 72); and combinations thereof capable of binding. The isolated antibody or antigen-binding fragment thereof according to claim 1. [Claim 4] The isolated antibody or antigen-binding fragment according to claim 1, wherein the antibody or antigen-binding fragment comprises a mutated Fc region. [Claim 5] The isolated antibody or antigen-binding fragment according to claim 4, wherein the antibody or antigen-binding fragment thereof has an increased serum half-life compared to that of a natural Fc antibody. [Claim 6] The variant Fc regions, when numbered according to the EU index as shown in Kabat, are 221, 225, 228, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 250, 251, 252, 254, 255, 256, 257, 262, 263, 264, 265, 266, 267, 268, 269, 279, 280, 284, 292, 296, 297, 298, An isolated antibody or antigen-binding fragment thereof according to claim 4, comprising modifications at one or more positions selected from 299, 305, 308, 313, 316, 318, 320, 322, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 339, 341, 343, 370, 373, 378, 392, 416, 419, 421, 428, 433, 434, 435, 436, 440, and 443. [Claim 7] The isolated antibody or antigen-binding fragment thereof according to claim 6, wherein the modification is selected from substitution, insertion, and deletion. [Claim 8] When the mutated Fc region is numbered according to the EU index as shown in Kabat, it is 221K, 221Y, 225E, 225K, 225W, 228P, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235E, 235F, 236E, 237L, 237M, 237P, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W, 243L, 24 3Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 257C, 257M, 257N, 262I, 262A, 262T, 26 2E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 265F, 265V, 265I, 265L, 265H, 265T, 26 6I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 29 6G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 313F, 316D, 318A, 318S, 320A, 320S, 32 2A, 322S, 325Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 327N, 327L, 328S, 328M, 328D, 328E, 32 8N, 328Q, 328F, 328I, 328V, 328T, 328H, 328A, 329F, 329H, 329Q, 330K, 330G,330T, 330C, 330L, 330Y, 330V, 330I, 330F, 330R, 330H, 331G, 331A, 331L, 331M, 331F, 331W, 331K, 331Q, 331E, 331S, 331V, 331 I, 331C, 331Y, 331H, 331R, 331N, 331D, 331T, 332D, 332S, 332W, 332F, 332E, 332N, 332Q, 332T, 332H, 332Y, 332A, 333A, 333D, 3 An isolated antibody or antigen-binding fragment thereof according to claim 4, comprising at least one substitution selected from 33G, 333Q, 333S, 333V, 334A, 334E, 334H, 334L, 334M, 334Q, 334V, 334Y, 339T, 370E, 370N, 378D, 392T, 396L, 416G, 419H, 421K, 428L, 428F, 433K, 433L, 434A, 424F, 434W, 434Y, 436H, 440Y, and 443W. [Claim 9] The mutated Fc region is: (a) At least one qualification of one or more positions selected from 228, 234, 235 and 331 when numbered by the EU index as shown in Kabat, (b) At least one qualification of one or more positions selected from 228 and 235 when numbered by the EU index as shown in Kabat, (c) At least one qualification of one or more positions selected from 239, 330 and 332 when numbered by the EU index as shown in Kabat, or (d) At least one qualification of one or more positions selected from 252, 254, and 256 when numbered by the EU index as shown in Kabat, The isolated antibody or antigen-binding fragment thereof according to claim 4, comprising: [Claim 10] The mutated Fc region is: (a) One or more substitutions selected from 228P, 234F, 235E, 235F, 235Y, and 331S when numbered by the EU index as shown in Kabat, (b) One or more substitutions selected from 228P, 235E and 235Y when numbered by the EU index as shown in Kabat, (c) One or more substitutions selected from 239D, 330L, 330Y, and 332E when numbered by the EU index as shown in Kabat, (d) One or more substitutions selected from 252Y, 254T and 256E when numbered by the EU index as shown in Kabat, or (e) Three substitutions of 252Y, 254T and 256E when numbered by the EU index as shown in Kabat, The isolated antibody or antigen-binding fragment thereof according to claim 4, comprising: [Claim 11] The isolated antibody or antigen-binding fragment thereof according to claim 4, wherein the mutated Fc region includes an IgG4 Fc region. [Claim 12] An isolated nucleic acid encoding an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 11. [Claim 13] A vector for producing an isolated antibody or antigen-binding fragment thereof that binds to influenza B virus hemagglutinin (HA) and has the ability to neutralize influenza B virus in two phylogenetically distinct lineages, comprising the isolated nucleic acid described in claim 12. [Claim 14] A host cell capable of expressing an isolated antibody or antigen-binding fragment thereof that binds to influenza B virus hemagglutinin (HA) and has the ability to neutralize influenza B viruses from two phylogenetically different lineages, comprising the nucleic acid described in claim 12. [Claim 15] A method for producing an antibody or antigen-binding fragment thereof according to any one of claims 1 to 11, comprising the step of culturing a host cell containing the nucleic acid or a vector containing the nucleic acid according to claim 12 under conditions suitable for the expression of the antibody or fragment thereof. [Claim 16] A composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 11, and a pharmaceutically acceptable carrier. [Claim 17] A composition comprising an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 11, (a) For use in the prevention or treatment of influenza B infection in subjects, (b) For use in the prevention or treatment of influenza A and influenza B infections in subjects, and / or (c) For in vitro diagnosis of influenza B infection in subjects, composition. [Claim 18] An antibody or antigen-binding fragment thereof according to any one of claims 1 to 11, (a) EC in the range of 1 μg / ml to 50 μg / ml of antibody ５０ Binds to influenza A HA; and / or (b) IC for neutralization of influenza A virus in a microneutralization assay, with antibody concentration in the range of 0.01 μg / ml to 5 μg / ml. ５０ Having, An antibody or its antigen-binding fragment.

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