WO2020130138A1 - Anti-pres1 antibody and use thereof - Google Patents

Anti-pres1 antibody and use thereof Download PDF

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Publication number
WO2020130138A1
WO2020130138A1 PCT/JP2019/050139 JP2019050139W WO2020130138A1 WO 2020130138 A1 WO2020130138 A1 WO 2020130138A1 JP 2019050139 W JP2019050139 W JP 2019050139W WO 2020130138 A1 WO2020130138 A1 WO 2020130138A1
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amino acid
acid sequence
seq
antibody
variable region
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PCT/JP2019/050139
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French (fr)
Japanese (ja)
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一彰 茶山
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国立大学法人広島大学
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Priority to JP2020561546A priority Critical patent/JP7454855B2/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis

Definitions

  • the present disclosure relates to an antibody against the preS1 antigen of hepatitis B virus (HBV) and its use.
  • HBV hepatitis B virus
  • HBV is a widely existing virus that infects approximately 300 million people worldwide and more than 200 million people are persistently infected. It causes a serious infectious disease that kills more than one million people a year from liver cirrhosis and liver cancer caused by this virus. Most of the viruses become persistent infections due to infection during childhood, but in some cases, even if infected in adults, they become chronic. Some cases of persistent infection develop from chronic hepatitis to cirrhosis, and some cases develop hepatocellular carcinoma. As described above, persistent infection with hepatitis B virus is a serious health problem, and treatment methods therefor, prevention methods for infection, evaluation methods for pathological conditions, and the like are urgently needed.
  • HBV is a virus that has a partially double-stranded circular DNA of 3,200 bases as its genome.
  • HBV particles consist of a core particle that has the HBV gene inside and an outer shell (envelope) that encloses the core particle.
  • HBV antigens present in the outer shell HBc-related antigens present in core particles
  • HBe antigens released into the blood antibodies against these antigens
  • HBV DNA amount, etc. are used as markers for the diagnosis of HBV infection. The combination of these is used to determine the presence or absence of infection, the pathological condition, or the therapeutic effect.
  • HBV antigens When HBV grows in hepatocytes, HBs antigens are overproduced and released into the blood as small spherical particles or rod-shaped particles.Therefore, complete HBV particles and incomplete HBV particles are found in the blood of HBV-infected patients. There are three types of virus particles, rod-shaped particles and small spherical particles, which are various particles. Antibodies against the HBs antigen that are currently used recognize all three types of virus particles. Since HBV infected patients have 10,000 to 100,000 times as many small spherical particles as intact HBV particles in the blood, most of the HBs antigens detected are derived from small spherical particles.
  • HBs antigen has three domains, S, preS1 and preS2, L protein composed of these three domains, lacking the preS1 domain, M protein composed of S domain and preS2 domain, and preS1 domain There is an S protein that lacks the preS2 domain and is composed only of the S domain.
  • the preS1 domain binds to a receptor on hepatocytes and plays an important role in HBV infection.
  • the HBs antigen containing the preS1 domain (also called preS1 antigen) is present only in infectious intact HBV particles and rod-shaped particles, not in small spherical particles.
  • the purpose of the present disclosure is to provide an antibody against the preS1 antigen and its use.
  • the present disclosure provides, in one aspect, an anti-preS1 antibody that binds to an epitope contained in an amino acid sequence selected from SEQ ID NOs: 2, 3 and 4.
  • an anti-preS1 antibody selected from the following (a) to (i): (A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added, CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
  • CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino
  • the present disclosure provides, in a further aspect, a combination of two or more anti-preS1 antibodies independently selected from said antibodies.
  • a method for measuring preS1 antigen which comprises contacting a sample obtained from a subject with the antibody, and detecting a complex of preS1 antigen and the antibody, and Provided are compositions and kits for measuring preS1 antigen, which comprise an antibody or a combination.
  • the present disclosure in a further aspect, comprises a method of assessing HBV infection, comprising measuring preS1 antigen by said measuring method and comparing said measured value with a reference value, and said antibody or combination.
  • a method of assessing HBV infection comprising measuring preS1 antigen by said measuring method and comparing said measured value with a reference value, and said antibody or combination.
  • compositions and kits for assessing HBV infection are provided.
  • the present disclosure provides, in a further aspect, a pharmaceutical composition comprising said antibody or combination.
  • an antibody against preS1 antigen or a combination thereof a method for measuring preS1 antigen using the antibody or a combination thereof and a composition and a kit therefor, a method for evaluating HBV infection and a composition and a kit therefor, and the antibody or the same Pharmaceutical compositions comprising the combination are provided.
  • FIG. 1 shows a partial sequence of the preS1 domain of HBV of each genotype and the binding site of the antibody of the example.
  • FIG. 2A shows the sequences of the light chain variable regions of antibodies 7, 10, 13, 39, 49, 55, 63, 41, and 1A.
  • FIG. 2B shows the heavy chain variable region sequences of antibodies 7, 10, 13, 39, 49, 55, 63, 41, and 1A.
  • FIG. 3 shows the inhibitory effect of antibodies 7, 10, 13, 15, 25, 27, 39, 41, 49, 51, 55 and 63 on HBV infection. The culture supernatant of the antibody-producing hybridoma was used.
  • FIG. 4 shows the inhibitory effect of antibodies 7, 10, 25, 41 and 55 on HBV infection. An antibody purified from the supernatant of the hybridoma after expansion culture was used.
  • FIG. 5 shows the inhibitory effect of antibodies 1A, 1B, 1C, 5A, 5B and 5C on HBV infection.
  • An antibody purified from the supernatant of the hybridoma after expansion culture was used.
  • FIG. 6 shows the HBV infection blocking effect of the combination of antibody 41 and antibody 7 or 10. Culture supernatant of hybridoma producing antibody 41 (0, 0.03, or 0.1 ⁇ g/ml) and culture supernatant of hybridoma producing antibody 7 or 10 (0.003 or 0.01 ⁇ g/ml) were used.
  • FIG. 7 shows the HBV infection inhibitory effect of the combination of antibody 41 and antibody 10 or 55.
  • FIG. 8 shows the HBV infection inhibitory effect of the combination of antibody 49 and antibody 55.
  • Antibody 49 (0, 0.01, or 0.03 ⁇ g/ml) and antibody 55 (0.01 or 0.03 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion culture were used.
  • FIG. 9 shows the HBV infection inhibitory effect of the combination of the antibody 824 having an epitope in the S domain and the antibody 10 or 55.
  • Antibody 824 (0 or 0.03 ⁇ g/ml) and antibody 10 or 55 (0.01 or 0.03 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion culture were used.
  • FIG. 10 shows the preS1 antigen and background absorbance of the antibody used for immobilization. As the immobilized antibody, antibody 41 or antibody 5124A having an epitope in the S domain was used. The results are shown for each group of subject serum divided by the measured values of HBe antigen and HBV-DNA.
  • FIG. 11 shows a calibration curve of preS1 antigen by the positive control serum.
  • FIG. 12 shows comparison of the measured values of the preS1 antigen of the antibody for immobilization (antibody 5124A or 824). Subject No.
  • FIG. 13 shows a comparison of the measured values of the preS1 antigen of the immobilizing antibody (antibody 5124A or 824) in the serum of a subject different from that in FIG. Subject No. and serum dilution are shown in the figure.
  • FIG. 14 shows the measured values of preS1 antigen in serum when the antibody 10, 13, 39 or 55 was used as the detection antibody. Subject No. and serum dilution are shown in the figure.
  • FIG. 15 shows the inhibitory effect of antibodies 41, 1A, and 10 on HBV infection. The antibody (0.01, 0.1, or 1 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion was used. The measurement result of HBs antigen is shown.
  • FIG. 13 shows a comparison of the measured values of the preS1 antigen of the immobilizing antibody (antibody 5124A or 824) in the serum of a subject different from that in FIG. Subject No. and serum dilution are shown in the figure.
  • FIG. 14 shows the measured values
  • FIG. 16 shows the inhibitory effect of antibodies 41, 1A, and 10 on HBV infection.
  • the antibody (0.01, 0.1, or 1 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion was used.
  • the measurement result of HBe antigen is shown.
  • FIG. 17 shows the HBV infection inhibitory effect of the combination of antibody 824 and antibody 41, 1A, or 10.
  • the antibody 824 (0.01 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion culture and the antibody 41, 1A, or 10 (0.01 or 0.1 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion culture were used.
  • the measurement result of HBs antigen is shown.
  • FIG. 18 shows the HBV infection inhibitory effect of the combination of antibody 824 and antibody 41, 1A, or 10.
  • the antibody 824 (0.01 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion culture and the antibody 41, 1A, or 10 (0.01 or 0.1 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion culture were used.
  • the measurement result of HBe antigen is shown.
  • FIG. 19 shows the HBV infection inhibitory effect of two combinations of antibodies 41, 1A, and 10.
  • An antibody (0.01 or 0.1 ⁇ g/ml) purified from the supernatant of the hybridoma after expansion was used.
  • the measurement result of HBs antigen is shown.
  • amino acid residues are represented by the following abbreviations.
  • Ala or A Alanine Arg or R: Arginine Asn or N: Asparagine Asp or D: Aspartic acid Cys or C: Cysteine Gln or Q: Glutamine Glu or E: Glutamic acid Gly or G: Glycine His or H: Histidine Ile or I: Isoleucine Leu or L: Leucine Lys or K: Lysine Met or M: methionine Phe or F: Phenylalanine Pro or P: Proline Ser or S: Serine Thr or T: Threonine Trp or W: Tryptophan Tyr or Y: Tyrosine Val or V: Valine
  • Hepatitis B virus is divided into at least 9 genotypes (Genotype) of A, B, C, D, E, F, G, H, and J.
  • HBV in the present disclosure includes HBV of any genotype.
  • the preS1 antigen means an HBs antigen (Hepatitis B surface antigen, HBsAg) containing a preS1 domain.
  • the HBs antigen in the present disclosure may be a protein of any genotype.
  • a representative amino acid sequence of HBs antigen (Genotype C) (accession No. MH887433) is shown in SEQ ID NO: 1, but the HBs antigen of the present disclosure is not limited to those containing the amino acid sequence of SEQ ID NO: 1.
  • the preS1 domain, preS2 domain, and S domain in SEQ ID NO: 1 are regions 1 to 119, 120 to 174,
  • the anti-preS1 antibody means an antibody that binds to an epitope in the preS1 domain of HBs antigen.
  • the anti-S antibody means an antibody that binds to an epitope in the S domain of HBs antigen
  • the anti-preS2 antibody means an antibody that binds to an epitope in the preS2 domain of HBs antigen.
  • anti-HBs antibody means an antibody against the HBs antigen, and is also meant to include an antibody that binds to an epitope in any of the S domain, preS1 domain and preS2 domain.
  • antibody used herein is meant to include various antibody structures such as polyclonal antibody, monoclonal antibody, multispecific antibody (eg, bispecific antibody), and antibody fragment.
  • Monoclonal antibodies include chimeric antibodies, humanized antibodies and human antibodies.
  • An antibody fragment refers to a molecule that comprises a portion of an antibody as a component, including, but not limited to, antibody heavy and light chain variable regions (V H and V L ), F(ab′) 2 , Fab′, Examples thereof include Fab, Fv, disulphide-linked FV (sdFv), Single-Chain FV (scFV) and polymers thereof.
  • the species of the antibody is not particularly limited, and examples thereof include mouse, rat, rabbit, goat, and human-derived antibodies.
  • the immunoglobulin class of an antibody is determined based on the heavy chain constant region.
  • the immunoglobulin classes include IgA, IgD, IgE, IgG, and IgM, and the corresponding heavy chains are called ⁇ chain, ⁇ chain, ⁇ chain, ⁇ chain, and ⁇ chain, respectively.
  • the immunoglobulin class can be further divided into subclasses (isotypes), eg, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2.
  • the immunoglobulin class of the antibody in the present specification is not particularly limited.
  • the antibody is IgG.
  • the antibody is IgG1 or IgG4.
  • the light chain of an antibody can be divided into a ⁇ chain and a ⁇ chain based on its constant region, but the antibody herein may have either a ⁇ chain or a ⁇ chain.
  • variable region of an antibody is usually composed of three complementarity determining regions (also referred to as CDRs) sandwiched between four framework regions (also referred to as FRs).
  • CDRs complementarity determining regions
  • FRs framework regions
  • abYsis Swindells, MB, Porter, CT, Couch, M., Hurst, J., Abhinandan, KR unless otherwise specified.
  • the binding and binding affinity of antibodies to antigens include enzyme immunoassay (EIA) (including ELISA), radioimmunoassay (RIA), chemiluminescent immunoassay (CIA), fluorescent immunoassay (FIA), etc. Can be confirmed by the immunoassay method described above, BIACORE (registered trademark) surface plasmon resonance assay, or the like.
  • the anti-preS1 antibody is 10 ⁇ 8 M or less, eg, 10 ⁇ 8 M to 10 ⁇ 13 M, 10 ⁇ 8 M to 10 ⁇ 12 M, 10 ⁇ 9 M to 10 ⁇ 12 M, or 10 ⁇ . It has a dissociation constant (Kd) of 9 M to 10 -11 M.
  • binding affinity is measured by BIACORE® surface plasmon resonance assay.
  • the antibody of the present disclosure uses a peptide containing the amino acid sequence of PLGFFPDHQLDPAFG (SEQ ID NO: 2), NSNNPDWDFNPN (SEQ ID NO: 3), or DPAFGANSNN (SEQ ID NO: 4), or a peptide consisting of these amino acid sequences as an immunogen, or To obtain by a general method using a peptide containing the amino acid sequence of NNPDWDFNP (SEQ ID NO: 17), NNPDWDFN (SEQ ID NO: 21), or GFFPDHQLD (SEQ ID NO: 80) or a peptide consisting of these amino acid sequences as an immunogen.
  • the immunogen can be produced by a conventional peptide synthesis method, for example, by a genetic engineering technique or chemical synthesis.
  • the antibody of the present disclosure can also be obtained by preparing an expression vector containing the antibody gene using a genetic engineering technique and expressing it in a cell.
  • Polyclonal antibodies can be prepared by the general methods described in "Antibodies: Laboratory Manual, Lane, H. D. et al. eds., Cold Spring Harbor Laboratory Press, New York, 1989, etc. Specifically, it can be prepared by immunizing a mammal such as rat, mouse, rabbit, goat, or horse with the above immunogen.
  • Monoclonal antibody can be obtained by a known method such as a method for producing a hybridoma producing an antibody, or a method for producing an expression vector containing an antibody gene using a genetic engineering technique and expressing it in a cell.
  • Hybridomas that secrete monoclonal antibodies can be prepared according to the method described in Kohler et al., Nature 256:495, 1975.
  • the immunogen is mixed with an appropriate substance for enhancing antigenicity (eg, keyhole limpet hemocyanin, bovine serum albumin, etc.) and, if necessary, an immunostimulant (Freund's complete or incomplete adjuvant, etc.).
  • an immunostimulant eg. keyhole limpet hemocyanin, bovine serum albumin, etc.
  • immunize a non-human mammal such as rat, mouse, rabbit, goat, or horse.
  • the immunized animal is immunized multiple times at intervals of 3 to 10 days, and 1 to 100 ⁇ g of the peptide as an immunogen is administered.
  • immunocompetent cells are recovered from an immunized animal that has undergone multiple immunizations, and myeloma cells having no autoantibody-producing ability (eg, mouse, rat, guinea pig, hamster, rabbit or Cells derived from mammals such as humans).
  • myeloma cells having no autoantibody-producing ability eg, mouse, rat, guinea pig, hamster, rabbit or Cells derived from mammals such as humans.
  • polyethylene glycol method, electrofusion method, etc. are used for cell fusion.
  • a hybridoma producing a monoclonal antibody is obtained.
  • the monoclonal antibody can be isolated from the culture supernatant obtained by culturing the obtained hybridoma in vitro. It can also be isolated from ascites by culturing in vivo such as ascites of mouse, rat, guinea pig, hamster or rabbit.
  • a monoclonal antibody can also be obtained by preparing an expression vector containing an antibody gene and expressing it in a host cell (PJDelves., ANTIBODY PRODUCTION ESSENTIAL TECHNIQUES., 1997 WILEY; P.Shepherd and C.Dean., Monoclonal Antibodies., 2000 OXFORD UNIVERSITY PRESS; JW Goding., Monoclonal Antibodies:principles and practice., 1993 ACADEMIC PRESS).
  • transgenic animal for example, cow, goat, sheep or pig
  • the gene of the antibody of interest has been incorporated into an endogenous gene
  • transgenic animal production technique for example, cow, goat, sheep or pig
  • a monoclonal antibody derived from the antibody gene can be obtained.
  • the obtained monoclonal antibody is purified by an appropriate combination of methods well known in the art, for example, chromatography with a protein A column, ion exchange chromatography, hydrophobic chromatography, ammonium sulfate salting-out method, gel filtration, affinity chromatography and the like. can do.
  • a chimeric antibody is an antibody containing sequences that are different from each other, and is, for example, an antibody in which variable regions and constant regions that are different from each other are linked.
  • the chimeric antibody is composed of a variable region of an antibody derived from a mammal other than human and a constant region derived from a human antibody.
  • the chimeric antibody is, for example, a polynucleotide encoding a variable region of an antibody derived from a mammal other than human and a polynucleotide encoding a constant region of a human antibody are ligated, and this is incorporated into an expression vector, and the expression vector is used as a host. It can be obtained by introducing and expressing it.
  • CDR is a region that substantially determines the binding specificity of an antibody, and its amino acid sequence is highly diverse. On the other hand, the amino acid sequences constituting FR show high homology even among antibodies having different binding specificities. Thus, CDR grafting allows the binding specificity of one antibody to be transferred to another antibody.
  • a humanized antibody is generally composed of CDRs of an antibody derived from a non-human animal, FRs derived from a human antibody, and constant regions derived from a human antibody.
  • the humanized antibody can be obtained by transplanting the CDR of an antibody derived from a non-human animal into a human antibody.
  • Humanized antibodies can be produced by various methods, but one example is Overlap Extension PCR (Almagro and Fransson, Front.Biosci. 13:1619-1633 (2008)).
  • PCR is performed using an oligonucleotide having a portion overlapping with the end of the CDR of the antibody derived from a non-human animal (for example, a mouse antibody) and the FR of the human antibody as a primer, and derived from a non-human animal.
  • a polynucleotide in which the CDR of the antibody and the FR of the human antibody are linked is synthesized. Then, the obtained polynucleotide is ligated to a polynucleotide encoding a constant region of a human antibody, incorporated into an expression vector, and this expression vector is introduced into a host to be expressed to obtain a humanized antibody. it can.
  • FR selection methods for producing humanized antibodies are known, for example, FR selected by the best fit method (Sims et.al. J. Immunol. 151:2296 (1993)) and the light chain of human antibody. Or FR (Carter et al. Proc. Natl. Acad. Sci. USA 89:4285 (1992); Presta et al. J. Immunol. 151:2623 (derived from a consensus sequence of a specific subgroup of the heavy chain variable region) 1993)) can be used.
  • a human antibody can be obtained, for example, by sensitizing human lymphocytes with a desired antigen in vitro and then fusing the sensitized lymphocytes with human myeloma cells (Japanese Patent Publication No. 1-59878).
  • human myeloma cells that are fusion partners, for example, U266 and the like can be used.
  • human antibodies can also be obtained by immunizing a transgenic animal having the entire repertoire of human antibody genes with a desired antigen (Lonberg, Nat. Biotech. 23: 1117-1125, 2005).
  • variable region of a human antibody is expressed as a single chain antibody (scFv) on the surface of a phage by the phage display method, a phage that binds to the antigen is selected, and the gene of the selected phage is analyzed to obtain the antigen.
  • the DNA sequence encoding the variable region of the human antibody that binds can be determined.
  • this variable region sequence is ligated in frame with the sequence of the human antibody constant region, inserted into an appropriate expression vector, and this expression vector is introduced into a host and expressed to obtain a human antibody. it can.
  • Multispecific antibodies are antibodies that bind to at least two different sites.
  • the multispecific antibody include a bispecific antibody and a trispecific antibody.
  • the multispecific antibody binds the preS1 antigen and one or more other antigens.
  • the multispecific antibody can be produced, for example, by a genetic engineering technique or by binding two or more antibodies having different recognition antigens.
  • the antibody fragment can be obtained, for example, by digesting the antibody with a protease such as papain or pepsin.
  • a protease such as papain or pepsin.
  • it can be obtained by introducing and expressing an expression vector containing a polynucleotide encoding an antibody fragment into a host cell (for example, Co, M. S. et al., J. Immunol. (1994) 152, 2968. -2976; Better, M. and Horwitz, A. H., MethodsEnzymol. (1989)178, 476-496; Pluckthun, A. and Skerra, A., MethodsEnzymol. (1989) 178, 497-515; Lamoyi , E., MethodsEnzymol.
  • an antibody can be obtained by introducing an expression vector containing a polynucleotide encoding the same into a cell and expressing it.
  • an expression vector is constructed so that the sequence encoding the antibody is expressed under an expression control region such as an enhancer and a promoter, and a host cell is transformed with this expression vector to express the antibody.
  • the present disclosure also provides an expression vector containing a polynucleotide encoding an anti-preS1 antibody, and a transformed cell containing a polynucleotide capable of expressing the antibody.
  • an eukaryotic cell such as an animal cell, a plant cell or a fungal cell
  • Animal cells include mammalian cells (eg, CHO, COS, NIH3T3, myeloma, BHK (baby hamster kidney), HeLa, Vero), amphibian cells (eg, Xenopus oocytes), or insect cells (eg, Sf9, Sf21). , Tn5).
  • yeast for example, Saccharomyces genus, for example, Saccharomyces cerevisiae
  • filamentous fungus for example, Aspergillus genus, for example, Aspergillus nigier
  • prokaryotic cells such as E. coli (eg, JM109, DH5 ⁇ , HB101, etc.) and Bacillus subtilis can also be used as host cells.
  • the vector can be introduced into the host cell by a method such as calcium phosphate method, DEAE dextran method, electroporation method, lipofection and the like.
  • the binding of the obtained antibody to the antigen can be confirmed by immunoassay methods such as EIA (including ELISA), RIA, CIA, FIA, and BIACORE (registered trademark) surface plasmon resonance assay.
  • EIA including ELISA
  • RIA including ELISA
  • CIA including CIA
  • FIA FIA
  • BIACORE registered trademark
  • the binding of the antibody to the antigen can also be confirmed by a competitive assay.
  • the antibody is any of the antibodies (a) to (i), any of the antibodies (a′) to (i′), (a′′) to (i′′) described in the present specification. Any of the antibodies of (a''') to (i''), or any of the antibodies of (a''') to (i'''), and preS1 antigen It can be confirmed by examining whether or not there is a competition in binding to.
  • an antibody of the present disclosure is an antibody of any of (a)-(i), an antibody of any of (a′)-(i′), (a′′) described herein.
  • an antibody that competes with the antibody of (1) for binding to the preS1 antigen As used herein, an antibody that competes with a predetermined antibody (that is, a reference antibody) means an antibody that significantly reduces the binding of the reference antibody to the preS1 antigen.
  • the competition experiment can be performed as follows. First, the test antibody and the reference antibody are mixed at various molar ratios.
  • the test antibody is usually used in excess (eg, 1-fold, 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, or 100-fold) with respect to the reference antibody.
  • the reference antibody may be labeled with a suitable label (eg biotin).
  • a detection reagent for example, a secondary antibody labeled with horseradish peroxidase (HRP) or biotinylated
  • HRP-streptavidin is used to measure the amount of labeled reference antibody bound to the preS1 antigen.
  • an antibody of the present disclosure binds a reference antibody to preS1 antigen by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%. % Or more.
  • the antibody of the present disclosure is the antibody of any of (a′) to (i′) and the antibody of (a′′′) to (i′′′) described herein. , Or (a′′′′) to (i′′′′), which competes with the antibody of any of (a′′′′) to (i′′′′) for binding to the preS1 antigen; An antibody that competes with any antibody or any of the antibodies (a′′′) to (i′′′) for binding to the preS1 antigen; or (a′′′) to An antibody that competes with any of the antibodies (i′′′) for binding to the preS1 antigen.
  • the antibody of the present disclosure binds to an epitope contained in an amino acid sequence selected from PLGFFPDHQLDPAFG (SEQ ID NO: 2), NSNNPDWDFNPN (SEQ ID NO: 3), or DPAFGANSNN (SEQ ID NO: 4).
  • the antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO:2 is an antibody that binds to the epitope included in the amino acid sequence of GFFPDHQLD (SEQ ID NO:80).
  • the antibody that binds to the epitope contained in the amino acid sequence of SEQ ID NO:3 is an antibody that binds to the epitope contained in the amino acid sequence of NNPDWDFNP (SEQ ID NO:17) or NNPDWDFN (SEQ ID NO:21). Whether or not it binds to an epitope contained in a given amino acid sequence can be confirmed by investigating whether or not it binds to a peptide consisting of the amino acid sequence, as described in Examples.
  • the antibody of the present disclosure is selected from the following (a)-(i): (A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added, CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
  • CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in
  • the antibodies of the present disclosure are selected from the following (a')-(i'): (A') CDR1 comprising the amino acid sequence of SEQ ID NO: 24, CDR2 containing the amino acid sequence of SEQ ID NO:25, and CDR3 containing the amino acid sequence of SEQ ID NO:26 A light chain variable region containing, and CDR1, which comprises the amino acid sequence of SEQ ID NO: 28, CDR2 containing the amino acid sequence of SEQ ID NO: 29, and CDR3 containing the amino acid sequence of SEQ ID NO: 30
  • An antibody comprising a heavy chain variable region comprising: (B') CDR1 containing the amino acid sequence of SEQ ID NO: 32, CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34 A light chain variable region containing, and CDR1, which comprises the amino acid sequence of SEQ ID NO: 36, CDR2 containing the amino acid sequence of SEQ ID NO:37, and CDR3
  • the antibodies of the present disclosure are selected from the following (a′′)-(i′′): (A'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 23, or the amino acid sequence of SEQ ID NO: 23 or the amino acid sequence of SEQ ID NO: 23 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 27.
  • B'' an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 35.
  • an antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added; (C'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 39.
  • An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added; (D'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 41, or the amino acid sequence of SEQ ID NO: 41 or the amino acid sequence of SEQ ID NO: 41
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues are deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 44.
  • An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added; (E'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 46, or the amino acid sequence of SEQ ID NO: 46 or the amino acid sequence of SEQ ID NO: 46
  • amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 59 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 59 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added; (H′′) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 62.
  • An antibody comprising a heavy chain variable region comprising an amino acid sequence in which ⁇ 5, or 1-3 amino acid residues have been deleted, substituted, or added; and (i′′) the amino acid sequence of SEQ ID NO: 64 and 80% , 85%, 90%, or 95% or more sequence identity, or 1-20, 1-10, 1-5 in the amino acid sequence of SEQ ID NO: 64 or the amino acid sequence of SEQ ID NO: 64 , Or a light chain variable region comprising an amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted, or added, and 80%, 85%, 90%, or 95% or more of the amino acid sequence of SEQ ID NO: 68
  • the antibodies of the present disclosure are selected from the following (a''') to (i'''): (A′′′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27; (B''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35; (C′′′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39; (D′′′) an antibody comprising a light chain variable region containing the amino acid sequence of SEQ ID NO: 41 and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44; (E′′′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:
  • the antibodies of the present disclosure are selected from the following (a′′′′) to (i′′′′): (A′′′′) CDR1, CDR2 in the light chain variable region including the amino acid sequence of SEQ ID NO:23, and CDR1, CDR2 in the heavy chain variable region including the amino acid sequence of SEQ ID NO:27 And an antibody comprising a heavy chain variable region comprising CDR3; (B′′′′) CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:35 and the light chain variable region containing CDR1, CDR2, and CDR3 in the light chain variable region of SEQ ID NO:31 And an antibody comprising a heavy chain variable region comprising CDR3; (C′′′′) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 31 and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 39 And an antibody comprising a heavy chain variable region comprising CDR3; (D′′′′) CDR1,
  • CDR is the definition of Kabat (Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. 1991), the definition of Chothia (Chothia et al., J. Mol. Biol., 1987; 196: 901-917), AdM definition (Martin et al., Proc. Natl. Acad.Sci.USA, 1989; 86: 9268-9272), Contact definition (MacCallum et al., J. Mol. Biol. 1996; 262: 732-745) and the IMGT definition (Lefranc et al., Dev Comp Immunol. 2003; 27(1): 55-77).
  • Kabat Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. 1991
  • Chothia Chothia et al., J. Mol. Biol., 1987; 196: 90
  • an amino acid sequence containing a predetermined amino acid sequence includes an amino acid sequence in which one or more amino acid residues are added to the predetermined amino acid sequence, and a sequence consisting of the predetermined amino acid.
  • a heavy chain variable region or a light chain containing an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of a predetermined heavy chain variable region or light chain variable region A variable region and 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues in the amino acid sequence of a given heavy chain variable region or light chain variable region are deleted or substituted, Alternatively, the heavy chain variable region or the light chain variable region containing the added amino acid sequence includes an antibody in which the CDR in the amino acid sequence of the predetermined heavy chain variable region or light chain variable region is not modified.
  • sequence identity with respect to an amino acid sequence means an amino acid residue that matches between two sequences that are optimally aligned (in a state of maximum matching) over the entire region of the sequence to be compared. Means the ratio of.
  • sequences to be compared may have additions or deletions (eg gaps etc.) in the optimal alignment of the two sequences.
  • Sequence identity can be calculated using a program such as FASTA, BLAST, CLUSTAL W provided in a public database (for example, DDBJ (http://www.ddbj.nig.ac.jp)). Alternatively, it can be determined using commercially available sequence analysis software (eg, Vector NTI (registered trademark) software, GENETYX (registered trademark) ver. 12).
  • variants with improved binding affinity can be obtained by methods based on phage display.
  • the amino acid residue that influences the interaction between the antibody and the antigen is identified by the alanine scanning mutation introduction method, or the crystal structure of the antigen-antibody complex is analyzed to determine the contact point between the antibody and the antigen.
  • the mutagenesis site is determined by identifying A mutant having a desired property can be obtained by preparing a mutant in which the amino acid at this site is modified by error-prone PCR or site-directed mutagenesis, and screening the resulting mutant library. ..
  • the antibody may have a selected immunoglobulin subclass for the regulation of antibody-dependent cellular cytotoxicity (ADCC) activity, complement-dependent cellular cytotoxicity (CDC) activity, or pharmacokinetics, and may have an amino acid sequence of the Fc region or sugar.
  • the chain may be modified.
  • IgG4 can be selected for the purpose of reducing complement activating ability.
  • modifications that reduce or enhance the binding to the Fc receptor or C1q, modifications that increase the binding affinity to FcRn for the purpose of extending the blood half-life, and the like can be performed.
  • the antibody also binds to a polymer such as polyethylene glycol (PEG), polypropylene glycol, polyoxyalkylene or a copolymer of polyethylene glycol and polypropylene glycol, eg, to increase the blood half-life of the antibody or to improve stability. You may have.
  • Antibodies may also be linked to chemotherapeutic agents, toxic peptides, radiochemicals and the like.
  • the antibody combination of the present disclosure is a combination of antibodies that bind to different epitopes. Further, the antibody of the present disclosure may be combined with an anti-HBs antibody that binds to an epitope different from that of the anti-preS1 antibody of the present disclosure. In certain embodiments, anti-preS1 antibodies of the present disclosure are used in combination with anti-S or anti-preS2 antibodies.
  • the antibody of the present disclosure can be used for measurement of preS1 antigen.
  • the methods of the present disclosure include contacting a sample obtained from a subject with an antibody of the present disclosure and detecting a complex of preS1 antigen and the antibody.
  • sample obtained from the subject may be any sample containing the preS1 antigen, and examples thereof include blood, plasma, serum, urine, sweat, milk, colostrum, tears, saliva, semen, vaginal secretions and the like.
  • sample obtained from a subject means not only a sample obtained from a subject itself but also a treatment necessary for forming a complex of preS1 antigen and anti-preS1 antibody in the sample. Used to include later samples.
  • the complex of preS1 antigen and antibody may be measured by any method known in the art.
  • the measurement method include immunoassay methods such as EIA (including ELISA), RIA, CIA, FIA, and a method based on latex agglutination reaction.
  • the antibody may be modified depending on the measuring method.
  • enzymes eg horseradish peroxidase, alkaline phosphatase, luciferase, ⁇ -galactosidase
  • fluorescent substances eg fluorescein, rhodamine
  • radioactive isotopes eg 3 H, 14 C, 32 P, 35 S or 125 I.
  • biotin eg 3 H, 14 C, 32 P, 35 S or 125 I.
  • Immunoassay methods include direct method, indirect method, competitive method, and sandwich method. Each method is usually carried out as follows.
  • the immobilized antigen is brought into contact with a labeled antigen-specific antibody.
  • the indirect method the immobilized antigen is brought into contact with an antigen-specific primary antibody, and subsequently reacted with a labeled secondary antibody specific to the primary antibody.
  • the sandwich method an immobilized antigen-specific antibody is brought into contact with an antigen, and subsequently another labeled antigen-specific antibody (or modified for binding to the label) is bound to the immobilized antibody. Contact with antigen.
  • the competitive method an immobilized antigen-specific antibody is brought into contact with an antigen to be measured and a labeled antigen having a known concentration at the same time. After washing, the antigen-antibody complex can be measured based on the label.
  • the complex of preS1 antigen and antibody is measured by ELISA.
  • the complex of preS1 antigen and antibody is measured by sandwich ELISA.
  • sandwich ELISA an anti-HBs antibody that binds to an epitope different from the anti-preS1 antibody used is immobilized on a solid phase such as a microplate and a sample obtained from the subject is added. After washing, anti-preS1 antibody is added to the solid phase, and the complex of preS1 antigen and anti-preS1 antibody is measured based on the label.
  • the anti-preS1 antibody may be immobilized on a solid phase, and the complex between the antigen and the anti-preS1 antibody may be measured by an anti-HBs antibody that binds to an epitope different from this.
  • the method of the present disclosure uses a combination of the anti-preS1 antibody of the present disclosure and an anti-S antibody or an anti-preS2 antibody.
  • the methods of this disclosure use a combination of anti-preS1 and anti-S antibodies of this disclosure.
  • the anti-preS1 antibody of the present disclosure is used as a detection antibody
  • the anti-S antibody is used as a solid phase antibody
  • the anti-preS1 antibody of the present disclosure is used as a solid phase antibody
  • the anti-S antibody is used as a detection antibody.
  • Accurate measurement can be performed by using an antibody that recognizes an epitope at a site distant from the epitope of the antibody of the present disclosure.
  • anti-S antibody commercially available antibodies such as Hyb-824 and Hyb-5124A, and as the anti-preS2 antibody, Hyb-5520 (both special immunoresearch institute) can be used.
  • anti-S antibody and anti-preS2 antibody can be prepared according to the description of the present disclosure, using a polypeptide having the amino acid sequence of all or part of the S domain or preS2 domain as an immunogen.
  • HBV infection can be evaluated by measuring preS1 antigen.
  • preS1 antigen measurement for HBV infection, a method using an anti-HBs antibody that cannot distinguish infectious HBV particles and small spherical particles, HBe antigen or HBcr antigen or antibody against them, or HBV DNA amount is measured. New information different from the method can be given.
  • the evaluation of HBV infection includes the determination of the presence or absence of infection, the determination of the pathological condition of HBV infection, the determination of the therapeutic effect on HBV infection, the determination of the prognosis of HBV infected subjects, and the like.
  • the results of assessing HBV infection may provide useful information for starting or stopping or discontinuing treatment for HBV infection.
  • Evaluation of HBV infection is performed by comparing the measured value of the target preS1 antigen with the reference value.
  • the reference value is appropriately set according to the purpose of evaluation.
  • the reference value may be a measurement value of another sample that is measured at the same time as the sample obtained from the subject, or may be a value set by a previous measurement. Standard statistical methods can be used for comparison.
  • the reference value is a measurement value of a sample (negative control) that does not contain preS1 antigen.
  • the reference value is a value (for example, an average value) set from the measured values of a plurality of subjects who are not infected with HBV. If the subject's measured value is higher than the reference value (ie, the preS1 antigen is detected), the subject can be determined to be infected with HBV. Also, the higher the subject's measurement, the more severe the HBV infection can be determined to be.
  • the reference value is a previous measurement of the same subject. If the subject's measured value is higher than the reference value, it is determined that the HBV infection is progressing, and if it is lower than the reference value, the HBV infection is determined to be improved.
  • the reference value is a value set from the measured values of a plurality of subjects having a predetermined condition or prognosis of HBV infection. For example, when the measured value of the subject is higher or lower than the reference value, it is determined that the subject has the predetermined disease state or prognosis.
  • the antibody of the present disclosure may be contained in a composition or kit for measuring preS1 antigen, and a composition or kit for evaluating HBV infection.
  • the composition for measurement may contain water, glycerol, a preservative, an antioxidant, a chelating agent, a buffering agent and the like in addition to the antibody of the present disclosure, and the form thereof is not limited, and for example, liquid or solid (Eg freeze-dried powder).
  • the kit comprises an anti-S antibody or an anti-preS2 antibody in addition to the antibody of the present disclosure.
  • the kit comprises an anti-S antibody in addition to the antibodies of this disclosure.
  • the anti-preS1 antibody of the present disclosure as a detection antibody, the anti-S antibody as a solid phase antibody, or the anti-preS1 antibody of the present disclosure as a solid phase antibody, the anti-S antibody as a detection antibody
  • the kit may further include equipment and/or reagents depending on the measuring method.
  • the kit may include a solid phase such as a microplate, preS1 antigen as a standard antigen, a reaction and/or dilution buffer, a coloring reagent, a reaction stopping solution, and an instruction manual.
  • the antibody of the present disclosure or the anti-S antibody or anti-preS2 antibody may be bound to the solid phase in the kit.
  • the antibody of the present disclosure can be used as an active ingredient of a pharmaceutical composition.
  • the antibodies of the present disclosure are used to treat or prevent HBV infection.
  • Treatment of HBV infection includes treatment of symptoms or diseases due to HBV infection, such as treatment of chronic or acute hepatitis B and cirrhosis.
  • Prevention of HBV infection includes prevention of infection after the accident of receiving HBV contaminated needles and prevention of infection after blood transfusion or organ transplantation.
  • the antibody of the present disclosure is administered to a subject in an amount capable of exerting a desired effect (for example, treatment or prevention of HBV infection) (herein, referred to as an effective amount).
  • the dose of the antibody is appropriately selected depending on the administration method, age, weight and health of the subject. For example, 10 ⁇ g/kg to 100 mg/kg, 100 ⁇ g/kg to 10 mg/kg, or 1 mg/kg to 10 mg/kg per day for an adult, consecutive days, days, weeks, or weeks However, the present invention is not limited to this.
  • the administration method of the antibody is also appropriately selected depending on the age, weight, health condition, etc. of the administration subject.
  • the administration method may be oral administration or parenteral administration, but parenteral administration is preferred. Parenteral administration includes subcutaneous administration, intradermal administration, intramuscular administration, intravenous administration and the like, but intravenous administration is preferable.
  • the pharmaceutical composition can be formulated by a conventional method.
  • the pharmaceutical composition includes pharmaceutically acceptable sterilized water, physiological saline, stabilizers, excipients, antioxidants, buffers, preservatives, surfactants, chelating agents, binders and the like. Carrier or additive.
  • the pharmaceutical composition is an antibody that binds to an epitope comprised in the amino acid sequence of SEQ ID NO: 2; or (a), (a'), (a''), (a''') or (a a''').
  • the pharmaceutical composition is an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3; or (b)-(h), (b′)-(h′), (b′′) )-(H''), (b''')-(h''') or (b'''')-(h''').
  • the pharmaceutical composition is an antibody that binds to an epitope comprised in the amino acid sequence of SEQ ID NO: 4; or (i), (i′), (i′′), (i′′′) or (I′′′′) antibody is included.
  • Two or more kinds of the antibodies of the present disclosure may be used in combination as an active ingredient of a pharmaceutical composition.
  • the two or more antibodies of the present disclosure include antibodies that bind different epitopes.
  • the two or more antibodies of the present disclosure bind to an epitope contained in the amino acid sequence of SEQ ID NO: 2 and one antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
  • antibodies or (a), (a'), (a''), (a''') or (a'''') antibody, and (b)-(h), (b) From') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') to (h''') And one or more antibodies of choice.
  • the two or more antibodies of the present disclosure bind to an epitope contained in the amino acid sequence of SEQ ID NO:2, and 1 bind to an epitope contained in the amino acid sequence of SEQ ID NO:4.
  • the two or more antibodies of the present disclosure bind to an epitope contained in the amino acid sequence of SEQ ID NO:3 and one antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO:4.
  • the antibody of the present disclosure may be used in combination with an anti-HBs antibody different from the antibody of the present disclosure, or may be used in combination with other therapeutic agents for HBV infection.
  • Other therapeutic agents include interferons (eg, peginterferon ⁇ -2a) and nucleic acid analogs (eg, lamivudine, adefovir, entecavir, tenofovir).
  • interferons eg, peginterferon ⁇ -2a
  • nucleic acid analogs eg, lamivudine, adefovir, entecavir, tenofovir.
  • the administration schedules of two or more active ingredients may be the same or different.
  • An anti-preS1 antibody which binds to an epitope contained in an amino acid sequence selected from SEQ ID NOs: 2, 3 and 4.
  • the antibody according to 1 above which binds to an epitope contained in the amino acid sequence of SEQ ID NO: 2.
  • [3] 3.
  • the antibody according to 2 above, wherein the antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 2 is an antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 80.
  • the antibody according to 1 above which binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3. [5] 5.
  • the antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 3 is an antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 17 or SEQ ID NO: 21.
  • the antibody according to any one of 1 to 5 above which is selected from the following (a) to (i): (A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added, CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
  • CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence
  • B'' an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 35.
  • an antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added; (C'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 39.
  • An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added; (D'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 41, or the amino acid sequence of SEQ ID NO: 41 or the amino acid sequence of SEQ ID NO: 41
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues are deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 44.
  • An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added; (E'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 46, or the amino acid sequence of SEQ ID NO: 46 or the amino acid sequence of SEQ ID NO: 46
  • amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 59 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 59 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added; (H′′) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57
  • a light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 62.
  • An antibody comprising a heavy chain variable region comprising an amino acid sequence in which ⁇ 5, or 1-3 amino acid residues have been deleted, substituted, or added; and (i′′) the amino acid sequence of SEQ ID NO: 64 and 80% , 85%, 90%, or 95% or more sequence identity, or 1-20, 1-10, 1-5 in the amino acid sequence of SEQ ID NO: 64 or the amino acid sequence of SEQ ID NO: 64 , Or a light chain variable region comprising an amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted, or added, and 80%, 85%, 90%, or 95% or more of the amino acid sequence of SEQ ID NO: 68
  • an antibody comprising a heavy chain variable region comprising a deleted, substituted, or added amino acid sequence comprising a deleted, substituted, or added amino acid sequence.
  • An antibody comprising a light chain variable region comprising, and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:68 comprising the amino acid sequence of SEQ ID NO:68.
  • An anti-preS1 antibody that competes with the antibody according to any one of 7 to 11 above for binding to a preS1 antigen.
  • the anti-preS1 antibody according to the above 12 which competes with the antibody according to the above 8, 10, or 11 for binding to the preS1 antigen.
  • the anti-preS1 antibody according to above 12 which competes with the antibody according to 10 above for binding to a preS1 antigen.
  • Two or more anti-preS1 antibodies One or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3; One or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4; or to the epitope contained in the amino acid sequence of SEQ ID NO:3 One or more antibodies that bind and one or more antibodies that bind to an epitope contained in the amino acid sequence of SEQ ID NO:4, 17.
  • the combination according to 16 above which comprises: [18] Two or more anti-preS1 antibodies 18.
  • the combination according to the above 17, comprising one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 2 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 3.
  • the combination according to the above 17, comprising one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4.
  • Two or more anti-preS1 antibodies (A), (a'), (a''), (a''') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h'''').
  • Antibody (B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h'') or (b''') ) To (h′′′′), and one or more antibodies selected from (i), (i′), (i′′), (i′′′) or (i′′′′) Or (a), (a'), (a''), (a''') or (a'''') antibody, and (i), (i'), (i' 18.
  • Two or more anti-preS1 antibodies (A), (a'), (a''), (a''') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h''''). 19.
  • the combination according to 18 above which comprises the antibody of [22] Two or more anti-preS1 antibodies (B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h'') or (b''') ) To (h′′′′), and one or more antibodies selected from (i), (i′), (i′′), (i′′′) or (i′′′′) 20.
  • the combination according to above 19, which comprises the antibody of [23] Two or more anti-preS1 antibodies (A), (a'), (a''), (a'') or (a'''') antibody and (b), (b'), (b''), (b' )) or (b'''') antibody; (A), (a'), (a''), (a''') or (a'''') antibody and (c), (c'), (c''), (c''), (c''') or (c'''') antibody; (A), (a'), (a''), (a''') or (a'''') antibody and (g), (g'), (g''), (g''') or (g'''') antibody; (A), (a′), (a′′), (a′′′) or (a′′′′) antibody and (i), (i′), (i′′), (i′) '') or (i'''') antibody; or (c), (c
  • a method for measuring preS1 antigen comprising: 16.
  • a method comprising contacting a sample obtained from a subject with the antibody according to any one of 1 to 15 above, and detecting a complex of preS1 antigen and the antibody.
  • 25 The method according to 24 above, wherein the antibody binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
  • the antibody is (b) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b' 25.
  • the method according to 24 above which is an antibody selected from “′′) to (h′′′′). [27] 27.
  • a method for evaluating hepatitis B virus infection comprising: A method comprising measuring the preS1 antigen by the method according to any one of the above 24 to 29, and comparing the measured value with a reference value. [31] 31. The method of claim 30, further comprising administering to the subject an effective amount of the antibody or combination of any of claims 1-23.
  • compositions for measuring preS1 antigen which comprises the antibody or the combination according to any one of 1 to 23 above.
  • a composition for evaluating hepatitis B virus infection which comprises the antibody or the combination according to any one of 1 to 23 above.
  • 34 The composition according to the above 32 or 33, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3. [35] (B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') 34.
  • the composition according to 32 or 33 above which comprises an antibody selected from () to (h′′′′).
  • kits for measuring preS1 antigen which comprises the antibody or the combination according to any one of 1 to 23 above.
  • a kit for evaluating hepatitis B virus infection which comprises the antibody or the combination according to any one of 1 to 23 above.
  • 38. The kit according to 36 or 37 above, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3. [39] (B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') 38.) The kit according to 36 or 37 above, which comprises an antibody selected from (h′′′′). [40] 40.
  • kits according to any of 36 to 39 further comprising an anti-HBs antibody that binds to an epitope different from the antibody according to any of 1 to 15 above.
  • the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody.
  • a pharmaceutical composition comprising the antibody or the combination according to any one of 1 to 23 above.
  • 43. The pharmaceutical composition according to the above 42 which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
  • [49] Includes one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2, and contains one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3 and/or included in the amino acid sequence of SEQ ID NO:4 Used in combination with one or more antibodies that bind to the epitope described above, or that bind to one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3 and/or bind to the epitope contained in the amino acid sequence of SEQ ID NO:4.
  • One or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3, one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2, and/or the amino acid sequence of SEQ ID NO:4 Used in combination with one or more antibodies that bind to the epitope described above, or that bind to one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2 and/or bind to the epitope contained in the amino acid sequence of SEQ ID NO:4.
  • [51] It comprises one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 3 and is used in combination with one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 4, or the amino acid of SEQ ID NO: 4 Further comprising one or more antibodies that bind to the epitope contained in the sequence; or one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4, to the epitope contained in the amino acid sequence of SEQ ID NO:3 Combined with one or more antibodies that bind, or further comprising one or more antibodies that bind to an epitope contained in the amino acid sequence of SEQ ID NO:3, 50.
  • composition according to the above 42. [53] (A), (a'), (a''), (a'') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h'''). Or (b) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''), (b''') to (h''').
  • the pharmaceutical composition according to the above 52 [56] The pharmaceutical composition according to any of 42 to 55, which is used in combination with an anti-HBs antibody that binds to an epitope different from the antibody according to any of 1 to 15 above, or further comprises the anti-HBs antibody. [57] 57. The pharmaceutical composition according to the above 56, wherein the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody. [58] The pharmaceutical composition according to any of the above 42 to 57, for treating or preventing hepatitis B virus infection. [59] The pharmaceutical composition according to any of the above 42 to 58, for the treatment of chronic hepatitis B.
  • [60] 24 A method for treating or preventing hepatitis B virus infection, which method comprises administering to a subject in need thereof an effective amount of an antibody or combination of any of the above 1-23. [61] 61. The method of 60, further comprising assessing a subject for hepatitis B virus infection by the method of 30. [62] 62. The method according to 60 or 61 above, for the treatment of chronic hepatitis B.
  • an antibody was prepared using the following peptides derived from the preS1 domain of the HBs antigen.
  • peptides 21-35, 34-48, and 37-55 were mixed to immunize mice, and hybridomas were obtained by the iliac lymph node method.
  • the peptides Pep1 and Pep2 were mixed to immunize mice to obtain hybridomas.
  • ELISA was performed using the antibody produced by the obtained hybridoma and the peptide used for immunization. Each peptide was immobilized on a 96-well polystyrene plate, and the culture supernatant of the hybridoma was added and incubated. After washing, anti-mouse rabbit antibody labeled with horseradish peroxidase (HRP) was added, and the supernatant showing a positive reaction was made positive, and hybridomas that react with the peptide were selected.
  • HRP horseradish peroxidase
  • Antibodies that react with both 34-48 and 37-55 were considered to have an epitope in the common part (NSNNPDWDFNPN, SEQ ID NO: 3) of both peptides.
  • NSNNPDWDFNPN SEQ ID NO: 3
  • antibodies 7, 10, and 55 were fine-mapped using the peptides shown in Table 3.
  • An amino group-introduced plate (Sumitomo Bakelite) was used as the ELISA plate. 200 ⁇ l of 2% glutaraldehyde solution diluted with bicarbonate buffer (pH 9.6) was added to each well and incubated at room temperature for 2 hours to activate the amino group.
  • PBST PBS containing 0.2% Tween 20, pH 7.4.
  • the antibody purified from the hybridoma supernatant was diluted with blocking buffer to a concentration of 1 ⁇ g/ml, 100 ⁇ l was added to each well, and the mixture was incubated at 37°C for 1 hour. After the incubation, the solution in each well was discarded, and the wells were washed 3 times with 300 ⁇ l of PBST.
  • HRP-labeled Goat anti-mouse IgG H+L
  • HRP conjugate Proteintech Group, Inc
  • blocking buffer 1:2000 (0.5 ⁇ g/ml)
  • 100 ⁇ l was added to each well and incubated at 37°C for 1 hour. Then, the solution in each well was removed, and the wells were washed 3 times with 300 ⁇ l of PBST.
  • 100 ⁇ l of TMB substrate solution was added and reacted at 37° C. for 15 minutes, 100 ⁇ l of 2M sulfuric acid solution was added to stop the reaction, and the absorbance at 450 nm was measured.
  • NNPDWDFN SEQ ID NO: 21
  • NNPDWDFNP SEQ ID NO: 17
  • variable regions of the obtained antibody light and heavy chains and their CDR1, 2 and 3 sequences were analyzed. Identified (FIGS. 2A and B and Tables 4-1 to 4-9). The variable region and CDR were identified by Chothia Numbering Scheme.
  • GFFPDHQLD SEQ ID NO: 80
  • HBV Human hepatocyte chimeric mouse
  • Human primary culture hepatocytes prepared from the liver The effect of HBV infection was investigated.
  • HBV about 20 virus particles/cell
  • culture supernatant of antibody-producing hybridoma or purified antibody were added to human primary hepatocytes (10 5 cells), and after culturing for 13 days, HBs antigen in the culture supernatant was added.
  • HBsAg detection kit Sysmex Corporation
  • the purified antibody was obtained from the culture supernatant of the expanded and produced antibody-producing hybridoma using an antibody purification column (Protenova Co., Ltd.).
  • the antibody concentration in the purified antibody and the culture supernatant was quantified using the Mouse IgG1 ELISA kit.
  • the results are shown as the average (IU/mL) of the measured values in each well.
  • PBS phosphate buffered saline
  • each antibody remarkably suppressed the production of HBs antigen after culturing, and was shown to block HBV infection (FIGS. 3-5).
  • the antibodies 7, 10, 13, 39, 49, 55 and 63 showed a high infection blocking effect (Fig. 3).
  • the inhibitory effect on HBV infection of the combination of antibody 41 and antibody 7 or 10 was examined in human primary culture hepatocytes. It was shown that the infection-inhibiting effect was enhanced by combining the culture supernatant of the hybridoma producing the antibody 41 and the culture supernatant of the hybridoma producing the antibody 7 or 10 (FIG. 6). A synergistic enhancement of the infection-blocking effect was also observed with the combination of the purified antibody, Antibody 41 and Antibody 10 or 55 (FIG. 7). When antibody 49 and antibody 55 were combined, the combined use enhanced the virus growth-inhibitory effect in a dose-dependent manner compared to antibody 49 alone (FIG. 8).
  • an IgG3 antibody of Hyb-824 (Special Immune Research Institute, Inc.) having an epitope in the S domain (also referred to as antibody 824 in the present specification) and antibody 10 or 55 enhances the infection-inhibiting effect. Were observed (Fig. 9).
  • the immobilizing antibody As the immobilizing antibody, the antibody 41 or IgM antibody of hybridoma clone 5124A having an epitope in the S domain (Special Immune Research Institute, Inc.) (also referred to as antibody 5124A in the present specification) was used. Further, in order to observe non-specific adsorption of HBV or enzyme-labeled antibody in serum, a well containing no antibody was prepared, and only 0.2 M phosphate buffer was added while the antibody was immobilized. After immobilization, the antibody solution in the well was discarded, and the well was washed twice with 200 ⁇ l of phosphate buffered saline (PBS, pH 7.4).
  • PBS phosphate buffered saline
  • HRP-labeled antibody 55 diluted with the blocking solution to 1:1000 (3 ⁇ g/ml) was added to each well in an amount of 100 ⁇ l, and incubated at 37° C. for 1 hour. Then, the solution in each well was removed, and the wells were washed 3 times with 300 ⁇ l of PBST. Next, 100 ⁇ l of TMB substrate solution was added and reacted at room temperature for 15 minutes, 100 ⁇ l of 2 M sulfuric acid solution was added to stop the reaction, and the absorbance at 450 nm was measured.
  • the HBc-related antigen was measured by the CLIA method using the hepatitis B virus core-related antigen kit Lumipulse HBcrAg (Fujirebio Inc.).
  • the HBe antibody was measured by CLIA method using Architect i2000SR (Abbott Japan Co., Ltd.) and Architect-HBe antibody reagent. ..
  • the absorbance of HBV-positive subjects was 0.603-3.141. In the absorptiometric method, the practical range of absorbance measurement is said to be 0.05 to 1.5, but when the HBs antigen value (IU/ML) exceeds 1,000, there are many samples with absorbance higher than 1.5 and further dilution is required. It was considered.
  • the correlation of preS1 antigen absorbance with other HBV markers was analyzed using regression analysis, but no correlation was observed with any of the markers (data not shown).
  • antibody 5124A or antibody 824 was used as the immobilized antibody.
  • the antibody solution in the wells was discarded, and the wells were washed 3 times with 200 ⁇ l of phosphate buffered saline (PBS, pH 7.4).
  • 100 ⁇ l of ChonBlockTM Blocking/Sample Dilution Buffer (Chondrex) was added as a blocking solution to each well after washing, and blocking was performed by incubating at room temperature for 1 hour. Then, the blocking solution in the wells was discarded, and the wells were washed 3 times with 200 ⁇ l of PBST (PBS containing 0.2% Tween 20, pH 7.4).
  • HBV antigen is 90,000 IU/ml and HBV-DNA is 9.1 log copies/ml.
  • HBV patient serum is used as positive control (PTC) serum and 50 times diluted solution is prepared using blocking solution, and further 3 times diluted series is prepared. Then, 50 ⁇ l of each of the above antibodies was added to each well in which the antibody was immobilized. Further, with respect to the sera of HBV-positive subjects and negative subjects, a 10-fold dilution series was prepared using a blocking solution, and 50 ⁇ l of each was similarly added to each well. After incubation at 37°C for 1 hour, the solution in each well was discarded after the incubation, and the wells were washed 3 times with 200 ⁇ l of PBST.
  • PTC positive control
  • HRP-labeled antibody 55 diluted to 1:1000 (3 ⁇ g/ml) with ChonBlockTM Detection Antibody Dilution Buffer (Chondrex) was added to each well by 100 ⁇ l, and 37 Incubation was carried out at 0°C for 1 hour. Then, the solution in each well was removed, and the wells were washed 3 times with 200 ⁇ l of PBST. Next, 100 ⁇ l of TMB substrate solution was added and reacted at room temperature for 15 minutes, 100 ⁇ l of 2M sulfuric acid solution was added to stop the reaction, and the absorbance at 450 nm was measured.
  • the absorbances of the antibody 5124A and the antibody 824 were compared at an antibody dilution ratio suitable for each serum, the antibody 824 showed higher detectability than the antibody 5124A (FIG. 12).
  • the absorbance of HBV-negative subjects was low at 0.056 (5124A) and 0.069 (824) even when serum was added at a concentration of 1:10, which was lower than that when PBS containing 3% BSA was used as a blocking solution and a serum diluent. The background absorbance decreased.
  • Example 2 In the same manner as in Example 1, using human primary culture hepatocytes, the inhibitory effects of antibody 41, antibody 1A, and antibody 10 on HBV infection were examined.
  • the amounts of HBs antigen and HBe antigen in the culture supernatant were measured by CLIA method using HBsAg detection kit and HBeAg detection kit (Sysmex Corporation).
  • HBsAg detection kit As a control, a group to which only HBV is added, and IgG1 (OY-TES-1 (G-5), santacruz: sc-390594) or IgG3 (Mouse IgG3 (isotype control), MBL: M078-3 instead of the antibody ) was added to the group. It was shown that each antibody markedly suppressed the production of HBs antigen and HBe antigen after culturing, and blocked HBV infection (FIGS. 15 and 16).
  • the inhibitory effect on HBV infection of the combination of antibody 824 and antibody 41, antibody 1A, or antibody 10 was examined.
  • antibody 824 having an epitope in the S domain with antibody 41, antibody 10 or antibody 1A
  • the inhibitory effect on HBs antigen and HBe antigen production is enhanced in a dose-dependent manner, and a synergistic infection-blocking effect is observed.
  • a synergistic infection blocking effect was also observed by combining the antibody 41, the antibody 10, and the antibody 1A (FIGS. 19 and 20).

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Abstract

The present invention provides: an antibody to the preS1 antigen or a combination thereof; a method for measuring preS1 antigens using the antibody or a combination thereof, and a composition and kit for said method; a method for evaluating HBV infection, and a composition and kit therefor; and a pharmaceutical composition containing the antibody or the combination thereof.

Description

抗ｐｒｅＳ１抗体およびその用途Anti-preS1 antibody and its use
　本出願は、日本国特許出願第２０１８－２３９７１９号について優先権を主張するものであり、ここに参照することによって、その全体が本明細書中へ組み込まれるものとする。
　本開示は、B型肝炎ウイルス（HBV）のpreS1抗原に対する抗体およびその用途に関する。 This application claims priority to Japanese Patent Application No. 2018-239719, which is incorporated herein by reference in its entirety.
The present disclosure relates to an antibody against the preS1 antigen of hepatitis B virus (HBV) and its use.
　HBVは、世界でおよそ3億人が感染し、2億人以上が持続感染している、広く世界中に存在しているウイルスである。年間100万人以上がこのウイルスに由来する肝硬変や肝臓癌により死亡するという、深刻な感染症を引き起こしている。ウイルスは、多くは幼少期の感染により持続感染となるが、成人で感染しても慢性化する症例もある。持続感染となった症例の一部は慢性肝炎から肝硬変へと進展し、また一部の症例は肝細胞癌を発症する。このようにB型肝炎ウイルスの持続感染は深刻な保健上の問題であり、その治療法、感染の予防法、病態の評価方法などが喫緊の課題としてあげられている。 HBV is a widely existing virus that infects approximately 300 million people worldwide and more than 200 million people are persistently infected. It causes a serious infectious disease that kills more than one million people a year from liver cirrhosis and liver cancer caused by this virus. Most of the viruses become persistent infections due to infection during childhood, but in some cases, even if infected in adults, they become chronic. Some cases of persistent infection develop from chronic hepatitis to cirrhosis, and some cases develop hepatocellular carcinoma. As described above, persistent infection with hepatitis B virus is a serious health problem, and treatment methods therefor, prevention methods for infection, evaluation methods for pathological conditions, and the like are urgently needed.
　HBVは、3,200塩基の一部分2本鎖環状DNAをゲノムとして有するウイルスである。HBV粒子は、内部にHBVの遺伝子を持つコア粒子と、これを包む外殻（エンベロープ）からなる。現在、HBV感染の診断には、外殻に存在するHBs抗原、コア粒子に存在するHBc関連抗原、血液中に放出されるHBe抗原、これら抗原に対する抗体や、HBV DNA量などがマーカーとして用いられ、これらを組み合わせて感染の有無や病態または治療効果の判定が行われている。HBVが肝細胞内で増殖する際、HBs抗原が過剰に産生され、小型球形粒子または桿状粒子として血液中に放出されるため、HBV感染患者の血液中には、完全なHBV粒子と、不完全な粒子である桿状粒子および小球形粒子との、3種類のウイルス粒子が存在する。現在用いられているHBs抗原に対する抗体は、これら3種類のウイルス粒子をいずれも認識する。HBV感染患者の血液中には完全なHBV粒子の1万倍～10万倍の小型球形粒子が存在することから、検出されるHBs抗原の大部分は小型球形粒子に由来するものである。 HBV is a virus that has a partially double-stranded circular DNA of 3,200 bases as its genome. HBV particles consist of a core particle that has the HBV gene inside and an outer shell (envelope) that encloses the core particle. Currently, HBV antigens present in the outer shell, HBc-related antigens present in core particles, HBe antigens released into the blood, antibodies against these antigens, HBV DNA amount, etc. are used as markers for the diagnosis of HBV infection. The combination of these is used to determine the presence or absence of infection, the pathological condition, or the therapeutic effect. When HBV grows in hepatocytes, HBs antigens are overproduced and released into the blood as small spherical particles or rod-shaped particles.Therefore, complete HBV particles and incomplete HBV particles are found in the blood of HBV-infected patients. There are three types of virus particles, rod-shaped particles and small spherical particles, which are various particles. Antibodies against the HBs antigen that are currently used recognize all three types of virus particles. Since HBV infected patients have 10,000 to 100,000 times as many small spherical particles as intact HBV particles in the blood, most of the HBs antigens detected are derived from small spherical particles.
　HBs抗原には、S、preS1およびpreS2の3つのドメインが存在し、この3つのドメインから構成されるLタンパク質、preS1ドメインを欠き、SドメインとpreS2ドメインとで構成されるMタンパク質、preS1ドメインとpreS2ドメインとを欠き、Sドメインのみで構成されるSタンパク質がある。preS1ドメインは、肝細胞上の受容体と結合し、HBVの感染に重要な役割を果たす。preS1ドメインを含むHBs抗原（preS1抗原ともいう）は、感染力のある完全なHBV粒子と桿状粒子にのみ存在し、小型球形粒子には存在しない。 HBs antigen has three domains, S, preS1 and preS2, L protein composed of these three domains, lacking the preS1 domain, M protein composed of S domain and preS2 domain, and preS1 domain There is an S protein that lacks the preS2 domain and is composed only of the S domain. The preS1 domain binds to a receptor on hepatocytes and plays an important role in HBV infection. The HBs antigen containing the preS1 domain (also called preS1 antigen) is present only in infectious intact HBV particles and rod-shaped particles, not in small spherical particles.
　本開示の目的は、preS1抗原に対する抗体およびその用途を提供することである。 The purpose of the present disclosure is to provide an antibody against the preS1 antigen and its use.
　本開示は、ある態様において、抗preS1抗体であって、配列番号２、３および４から選択されるアミノ酸配列に含まれるエピトープに結合する抗体を提供する。 The present disclosure provides, in one aspect, an anti-preS1 antibody that binds to an epitope contained in an amino acid sequence selected from SEQ ID NOs: 2, 3 and 4.
　本開示は、さらなる態様において、抗preS1抗体であって、以下の（ａ）～（ｉ）から選択される抗体を提供する：
（ａ）配列番号２４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号２６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号２８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｂ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｃ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｄ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｅ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｆ）配列番号５０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号５４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｇ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｈ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；および
（ｉ）配列番号６５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号６９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号７０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号７１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体。 The present disclosure provides, in a further aspect, an anti-preS1 antibody, selected from the following (a) to (i):
(A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 28 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 30 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(B) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 37 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(C) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(D) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence have been deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 43 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 45 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(E) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and the amino acid sequence of SEQ ID NO: 47 or 1 to 3 in said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(F) CDR1 comprising the amino acid sequence of SEQ ID NO: 50 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 51 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 52 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 54 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:55 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:56 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(G) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 60 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 61 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(H) a CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence;
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:63 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:61 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising: (i) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in said amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 67 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 70 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 71 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
　本開示は、さらなる態様において、前記抗体から独立に選択される２種以上の抗preS1抗体の組み合わせを提供する。 The present disclosure provides, in a further aspect, a combination of two or more anti-preS1 antibodies independently selected from said antibodies.
　本開示は、さらなる態様において、preS1抗原の測定方法であって、対象から得た試料を前記抗体と接触させること、およびpreS1抗原と前記抗体との複合体を検出することを含む方法、並びに前記抗体または組み合わせを含むpreS1抗原の測定用組成物およびキットを提供する。 The present disclosure, in a further aspect, a method for measuring preS1 antigen, which comprises contacting a sample obtained from a subject with the antibody, and detecting a complex of preS1 antigen and the antibody, and Provided are compositions and kits for measuring preS1 antigen, which comprise an antibody or a combination.
　本開示は、さらなる態様において、HBV感染の評価方法であって、前記測定方法によりpreS1抗原を測定すること、および前記測定値を基準値と比較することを含む方法、並びに前記抗体または組み合わせを含むHBV感染の評価用組成物およびキットを提供する。 The present disclosure, in a further aspect, comprises a method of assessing HBV infection, comprising measuring preS1 antigen by said measuring method and comparing said measured value with a reference value, and said antibody or combination. Provided are compositions and kits for assessing HBV infection.
　本開示は、さらなる態様において、前記抗体または組み合わせを含む医薬組成物を提供する。 The present disclosure provides, in a further aspect, a pharmaceutical composition comprising said antibody or combination.
　本開示により、preS1抗原に対する抗体またはその組み合わせ、前記抗体またはその組み合わせを用いるpreS1抗原の測定方法並びにそのための組成物およびキット、HBV感染の評価方法並びにそのための組成物およびキット、並びに前記抗体またはその組み合わせを含む医薬組成物が提供される。 According to the present disclosure, an antibody against preS1 antigen or a combination thereof, a method for measuring preS1 antigen using the antibody or a combination thereof and a composition and a kit therefor, a method for evaluating HBV infection and a composition and a kit therefor, and the antibody or the same Pharmaceutical compositions comprising the combination are provided.
図１は、各遺伝子型（Genotype）のHBVのpreS1ドメインの部分配列および実施例の抗体の結合部位を示す。FIG. 1 shows a partial sequence of the preS1 domain of HBV of each genotype and the binding site of the antibody of the example. 図２Ａは、抗体7、10、13、39、49、55、63、41、および1Aの軽鎖可変領域の配列を示す。FIG. 2A shows the sequences of the light chain variable regions of antibodies 7, 10, 13, 39, 49, 55, 63, 41, and 1A. 図２Ｂは、抗体7、10、13、39、49、55、63、41、および1Aの重鎖可変領域の配列を示す。FIG. 2B shows the heavy chain variable region sequences of antibodies 7, 10, 13, 39, 49, 55, 63, 41, and 1A. 図３は、抗体7、10、13、15、25、27、39、41、49、51、55および63のHBV感染阻止効果を示す。抗体産生ハイブリドーマの培養上清を使用した。FIG. 3 shows the inhibitory effect of antibodies 7, 10, 13, 15, 25, 27, 39, 41, 49, 51, 55 and 63 on HBV infection. The culture supernatant of the antibody-producing hybridoma was used. 図４は、抗体7、10、25、41および55のHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体を使用した。FIG. 4 shows the inhibitory effect of antibodies 7, 10, 25, 41 and 55 on HBV infection. An antibody purified from the supernatant of the hybridoma after expansion culture was used. 図５は、抗体1A、1B、1C、5A、5Bおよび5CのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体を使用した。FIG. 5 shows the inhibitory effect of antibodies 1A, 1B, 1C, 5A, 5B and 5C on HBV infection. An antibody purified from the supernatant of the hybridoma after expansion culture was used. 図６は、抗体41と抗体7または10との組み合わせのHBV感染阻止効果を示す。抗体41を産生するハイブリドーマの培養上清（0、0.03、または0.1 μg/ml）と抗体7または10を産生するハイブリドーマの培養上清（0.003または0.01 μg/ml）を使用した。FIG. 6 shows the HBV infection blocking effect of the combination of antibody 41 and antibody 7 or 10. Culture supernatant of hybridoma producing antibody 41 (0, 0.03, or 0.1 μg/ml) and culture supernatant of hybridoma producing antibody 7 or 10 (0.003 or 0.01 μg/ml) were used. 図７は、抗体41と抗体10または55との組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体41（0、0.03、または0.1 μg/ml）と抗体10または55（0.01または0.03 μg/ml）を使用した。FIG. 7 shows the HBV infection inhibitory effect of the combination of antibody 41 and antibody 10 or 55. Antibody 41 (0, 0.03, or 0.1 μg/ml) and antibody 10 or 55 (0.01 or 0.03 μg/ml) purified from the supernatant of the hybridoma after expansion culture were used. 図８は、抗体49と抗体55との組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体49（0、0.01、または0.03 μg/ml）と抗体55（0.01または0.03 μg/ml）を使用した。FIG. 8 shows the HBV infection inhibitory effect of the combination of antibody 49 and antibody 55. Antibody 49 (0, 0.01, or 0.03 μg/ml) and antibody 55 (0.01 or 0.03 μg/ml) purified from the supernatant of the hybridoma after expansion culture were used. 図９は、Sドメインにエピトープを有する抗体824と、抗体10または55との組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体824（0または0.03 μg/ml）と抗体10または55（0.01または0.03 μg/ml）を使用した。FIG. 9 shows the HBV infection inhibitory effect of the combination of the antibody 824 having an epitope in the S domain and the antibody 10 or 55. Antibody 824 (0 or 0.03 μg/ml) and antibody 10 or 55 (0.01 or 0.03 μg/ml) purified from the supernatant of the hybridoma after expansion culture were used. 図１０は、固相化に使用した抗体のpreS1抗原とバックグラウンドの吸光度を示す。固相化抗体として、抗体41またはSドメインにエピトープを有する抗体5124Aを使用した。被験者血清をHBe抗原およびHBV-DNAの測定値で分けた群ごとに結果を示す。FIG. 10 shows the preS1 antigen and background absorbance of the antibody used for immobilization. As the immobilized antibody, antibody 41 or antibody 5124A having an epitope in the S domain was used. The results are shown for each group of subject serum divided by the measured values of HBe antigen and HBV-DNA. 図１１は、ポジティブコントロール血清によるpreS1抗原の検量線を示す。FIG. 11 shows a calibration curve of preS1 antigen by the positive control serum. 図１２は、固相化用抗体（抗体5124Aまたは824）のpreS1抗原の測定値の比較を示す。被験者No.と血清希釈率を図中に示す。FIG. 12 shows comparison of the measured values of the preS1 antigen of the antibody for immobilization (antibody 5124A or 824). Subject No. and serum dilution are shown in the figure. 図１３は、図１２とは異なる被験者の血清における固相化用抗体（抗体5124Aまたは824）のpreS1抗原の測定値の比較を示す。被験者No.と血清希釈率を図中に示す。FIG. 13 shows a comparison of the measured values of the preS1 antigen of the immobilizing antibody (antibody 5124A or 824) in the serum of a subject different from that in FIG. Subject No. and serum dilution are shown in the figure. 図１４は、抗体10、13、39または55を検出抗体に使用した場合の血清中preS1抗原の測定値を示す。被験者No.と血清希釈率を図中に示す。FIG. 14 shows the measured values of preS1 antigen in serum when the antibody 10, 13, 39 or 55 was used as the detection antibody. Subject No. and serum dilution are shown in the figure. 図１５は、抗体41、1A、および10のHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体（0.01、0.1、または1 μg/ml）を使用した。HBs抗原の測定結果を示す。FIG. 15 shows the inhibitory effect of antibodies 41, 1A, and 10 on HBV infection. The antibody (0.01, 0.1, or 1 μg/ml) purified from the supernatant of the hybridoma after expansion was used. The measurement result of HBs antigen is shown. 図１６は、抗体41、1A、および10のHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体（0.01、0.1、または1 μg/ml）を使用した。HBe抗原の測定結果を示す。FIG. 16 shows the inhibitory effect of antibodies 41, 1A, and 10 on HBV infection. The antibody (0.01, 0.1, or 1 μg/ml) purified from the supernatant of the hybridoma after expansion was used. The measurement result of HBe antigen is shown. 図１７は、抗体824と、抗体41、1A、または10との組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体824（0.01 μg/ml）とハイブリドーマの拡大培養後の上清から精製した抗体41、1A、または10（0.01または0.1 μg/ml）を使用した。HBs抗原の測定結果を示す。FIG. 17 shows the HBV infection inhibitory effect of the combination of antibody 824 and antibody 41, 1A, or 10. The antibody 824 (0.01 μg/ml) purified from the supernatant of the hybridoma after expansion culture and the antibody 41, 1A, or 10 (0.01 or 0.1 μg/ml) purified from the supernatant of the hybridoma after expansion culture were used. The measurement result of HBs antigen is shown. 図１８は、抗体824と、抗体41、1A、または10との組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体824（0.01 μg/ml）とハイブリドーマの拡大培養後の上清から精製した抗体41、1A、または10（0.01または0.1 μg/ml）を使用した。HBe抗原の測定結果を示す。FIG. 18 shows the HBV infection inhibitory effect of the combination of antibody 824 and antibody 41, 1A, or 10. The antibody 824 (0.01 μg/ml) purified from the supernatant of the hybridoma after expansion culture and the antibody 41, 1A, or 10 (0.01 or 0.1 μg/ml) purified from the supernatant of the hybridoma after expansion culture were used. The measurement result of HBe antigen is shown. 図１９は、抗体41、1A、および10のうちの2種の組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体（0.01または0.1 μg/ml）を使用した。HBs抗原の測定結果を示す。FIG. 19 shows the HBV infection inhibitory effect of two combinations of antibodies 41, 1A, and 10. An antibody (0.01 or 0.1 μg/ml) purified from the supernatant of the hybridoma after expansion was used. The measurement result of HBs antigen is shown. 図２０は、抗体41、1A、および10のうちの2種の組み合わせのHBV感染阻止効果を示す。ハイブリドーマの拡大培養後の上清から精製した抗体（0.01または0.1 μg/ml）を使用した。HBe抗原の測定結果を示す。FIG. 20 shows the HBV infection inhibitory effect of two combinations of antibodies 41, 1A, and 10. An antibody (0.01 or 0.1 μg/ml) purified from the supernatant of the hybridoma after expansion was used. The measurement result of HBe antigen is shown.
　本明細書において、アミノ酸残基は以下の略号で表される。
AlaまたはA：アラニン
ArgまたはR：アルギニン
AsnまたはN：アスパラギン
AspまたはD：アスパラギン酸
CysまたはC：システイン
GlnまたはQ：グルタミン
GluまたはE：グルタミン酸
GlyまたはG：グリシン
HisまたはH：ヒスチジン
IleまたはI：イソロイシン
LeuまたはL：ロイシン
LysまたはK：リジン
MetまたはM：メチオニン
PheまたはF：フェニルアラニン
ProまたはP：プロリン
SerまたはS：セリン
ThrまたはT：スレオニン
TrpまたはW：トリプトファン
TyrまたはY：チロシン
ValまたはV：バリン In the present specification, amino acid residues are represented by the following abbreviations.
Ala or A: Alanine
Arg or R: Arginine
Asn or N: Asparagine
Asp or D: Aspartic acid
Cys or C: Cysteine
Gln or Q: Glutamine
Glu or E: Glutamic acid
Gly or G: Glycine
His or H: Histidine
Ile or I: Isoleucine
Leu or L: Leucine
Lys or K: Lysine
Met or M: methionine
Phe or F: Phenylalanine
Pro or P: Proline
Ser or S: Serine
Thr or T: Threonine
Trp or W: Tryptophan
Tyr or Y: Tyrosine
Val or V: Valine
　B型肝炎ウイルス（HBV）は、少なくとも、A、B、C、D、E、F、G、H、Jの9種類の遺伝子型（Genotype）に分けられる。本開示におけるHBVには、いずれの遺伝子型のHBVも含まれる。 Hepatitis B virus (HBV) is divided into at least 9 genotypes (Genotype) of A, B, C, D, E, F, G, H, and J. HBV in the present disclosure includes HBV of any genotype.
　本明細書において、preS1抗原とは、preS1ドメインを含むHBs抗原（Hepatitis B surface antigen, HBsAg）を意味する。本開示におけるHBs抗原は、いずれの遺伝子型のタンパク質であってもよい。HBs抗原（Genotype C）の代表的アミノ酸配列（accession No. MH887433）を配列番号１に示すが、本開示のHBs抗原は配列番号１のアミノ酸配列を含むものに限定されない。

MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPNKDHWPEANQVGAGAFGSGFTPPHGGLLGWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDPAPNMENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI（配列番号１）

　配列番号１におけるpreS1ドメイン、preS2ドメインおよびSドメインは、それぞれ、1番目～119番目、120番目～174番目および175番目～400番目の領域である。当業者は、各遺伝子型のHBs抗原のアミノ酸配列を、GenBankなどのデータベースから容易に取得することができる。 In the present specification, the preS1 antigen means an HBs antigen (Hepatitis B surface antigen, HBsAg) containing a preS1 domain. The HBs antigen in the present disclosure may be a protein of any genotype. A representative amino acid sequence of HBs antigen (Genotype C) (accession No. MH887433) is shown in SEQ ID NO: 1, but the HBs antigen of the present disclosure is not limited to those containing the amino acid sequence of SEQ ID NO: 1.

MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPNKDHWPEANQVGAGAFGSGFTPPHGGLLGWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDPAPNMENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI (SEQ ID NO: 1)

The preS1 domain, preS2 domain, and S domain in SEQ ID NO: 1 are regions 1 to 119, 120 to 174, and 175 to 400, respectively. Those skilled in the art can easily obtain the amino acid sequence of the HBs antigen of each genotype from a database such as GenBank.
　本明細書において、抗preS1抗体とは、HBs抗原のpreS1ドメイン中のエピトープに結合する抗体を意味する。本明細書において、抗S抗体とは、HBs抗原のSドメイン中のエピトープに結合する抗体を意味し、抗preS2抗体とは、HBs抗原のpreS2ドメイン中のエピトープに結合する抗体を意味する。抗HBs抗体なる用語は、HBs抗原に対する抗体を意味し、Sドメイン、preS1ドメインおよびpreS2ドメインのいずれのドメイン中のエピトープに結合する抗体も含む意味で用いられる。 In the present specification, the anti-preS1 antibody means an antibody that binds to an epitope in the preS1 domain of HBs antigen. As used herein, the anti-S antibody means an antibody that binds to an epitope in the S domain of HBs antigen, and the anti-preS2 antibody means an antibody that binds to an epitope in the preS2 domain of HBs antigen. The term anti-HBs antibody means an antibody against the HBs antigen, and is also meant to include an antibody that binds to an epitope in any of the S domain, preS1 domain and preS2 domain.
　本明細書における「抗体」なる用語は、ポリクローナル抗体、モノクローナル抗体、多重特異性抗体（例えば、二重特異性抗体）、抗体断片など、種々の抗体構造を含む意味で用いられる。モノクローナル抗体には、キメラ抗体、ヒト化抗体、ヒト抗体が含まれる。抗体断片は、抗体の一部分を構成要素として含む分子意味し、例えば、限定はされないが、抗体の重鎖および軽鎖可変領域（VHおよびVL）、F(ab')2、Fab'、Fab、Fv、disulphide-linked FV (sdFv) 、Single-Chain FV (scFV) およびこれらの重合体が挙げられる。抗体の種は特に限定されず、例えば、マウス、ラット、ウサギ、ヤギ、ヒト由来の抗体が挙げられる。 The term “antibody” used herein is meant to include various antibody structures such as polyclonal antibody, monoclonal antibody, multispecific antibody (eg, bispecific antibody), and antibody fragment. Monoclonal antibodies include chimeric antibodies, humanized antibodies and human antibodies. An antibody fragment refers to a molecule that comprises a portion of an antibody as a component, including, but not limited to, antibody heavy and light chain variable regions (V H and V L ), F(ab′) 2 , Fab′, Examples thereof include Fab, Fv, disulphide-linked FV (sdFv), Single-Chain FV (scFV) and polymers thereof. The species of the antibody is not particularly limited, and examples thereof include mouse, rat, rabbit, goat, and human-derived antibodies.
　抗体のイムノグロブリンクラスは、重鎖定常領域に基づき決定される。イムノグロブリンクラスとしては、IgA、IgD、IgE、IgG、およびIgMが挙げられ、これらに対応する重鎖はそれぞれ、α鎖、δ鎖、ε鎖、γ鎖、およびμ鎖と呼ばれる。イムノグロブリンクラスは、さらにサブクラス（アイソタイプ）、例えば、IgG1、IgG2、IgG3、IgG4、IgA1およびIgA2に分けることができる。本明細書における抗体のイムノグロブリンクラスは、特に限定はされない。ある実施形態において、抗体は、IgGである。さらなる実施形態において、抗体は、IgG1またはIgG4である。抗体の軽鎖は、その定常領域に基づきκ鎖およびλ鎖に分けることができるが、本明細書における抗体は、κ鎖およびλ鎖のいずれを有していてもよい。 The immunoglobulin class of an antibody is determined based on the heavy chain constant region. The immunoglobulin classes include IgA, IgD, IgE, IgG, and IgM, and the corresponding heavy chains are called α chain, δ chain, ε chain, γ chain, and μ chain, respectively. The immunoglobulin class can be further divided into subclasses (isotypes), eg, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The immunoglobulin class of the antibody in the present specification is not particularly limited. In certain embodiments, the antibody is IgG. In a further embodiment, the antibody is IgG1 or IgG4. The light chain of an antibody can be divided into a κ chain and a λ chain based on its constant region, but the antibody herein may have either a κ chain or a λ chain.
　抗体の可変領域は、通常、４つのフレームワーク領域（framework region）（FRとも記載する）にはさまれた３つの相補性決定領域（complementarity determining region）（CDRとも記載する）で構成される。本明細書において、抗体の可変領域およびCDRは、特に記載する場合を除き、ウェブベースの抗体検索システムのabYsis（Swindells, M.B., Porter, C.T., Couch, M., Hurst, J., Abhinandan, K.R., Nielsen, J.H., Macindoe, G., Hetherington, J. and Martin, A.C., 2017. abYsis: integrated antibody sequence and structure-management, analysis, and prediction. Journal of molecular biology, 429(3), pp.356-364.）において Chothia Numbering Schemeにより特定する。 The variable region of an antibody is usually composed of three complementarity determining regions (also referred to as CDRs) sandwiched between four framework regions (also referred to as FRs). In the present specification, the variable region and CDR of the antibody are abYsis (Swindells, MB, Porter, CT, Couch, M., Hurst, J., Abhinandan, KR unless otherwise specified. , Nielsen, JH, Macindoe, G., Hetherington, J. and Martin, AC, 2017. abYsis: integrated antibody sequence and structure-management, analysis, and prediction. Journal of molecular biology, 429(3), pp.356- 364.) specified by Chothia Numbering Scheme.
　抗体の抗原への結合および結合親和性は、酵素免疫測定法（EIA）（ELISAを含む）、放射免疫測定法（RIA）、化学発光免疫測定法（CIA）、蛍光免疫測定法（FIA）などの免疫測定法や、BIACORE（登録商標）表面プラズモン共鳴アッセイなどにより確認することができる。ある実施形態において、抗preS1抗体は、10-8M以下、例えば10-8M～10-13M、10-8M～10-12M、10-9M～10-12M、または10-9M～10-11Mの解離定数 (Kd) を有する。ある実施形態において、結合親和性は、BIACORE（登録商標）表面プラズモン共鳴アッセイにより測定される。 The binding and binding affinity of antibodies to antigens include enzyme immunoassay (EIA) (including ELISA), radioimmunoassay (RIA), chemiluminescent immunoassay (CIA), fluorescent immunoassay (FIA), etc. Can be confirmed by the immunoassay method described above, BIACORE (registered trademark) surface plasmon resonance assay, or the like. In certain embodiments, the anti-preS1 antibody is 10 −8 M or less, eg, 10 −8 M to 10 −13 M, 10 −8 M to 10 −12 M, 10 −9 M to 10 −12 M, or 10 −. It has a dissociation constant (Kd) of 9 M to 10 -11 M. In certain embodiments, binding affinity is measured by BIACORE® surface plasmon resonance assay.
　本開示の抗体は、PLGFFPDHQLDPAFG（配列番号２）、NSNNPDWDFNPN（配列番号３）、またはDPAFGANSNN（配列番号４）のアミノ酸配列を含むペプチド、またはこれらアミノ酸配列からなるペプチドを免疫原として用いて、あるいは、NNPDWDFNP（配列番号１７）、NNPDWDFN（配列番号２１）、またはGFFPDHQLD（配列番号８０）のアミノ酸配列を含むペプチド、またはこれらアミノ酸配列からなるペプチドを免疫原として用いて、一般的な方法により取得することができる。免疫原は、通常のペプチド合成法により、例えば遺伝子工学的手法または化学合成により、作製することができる。あるいは、本開示の抗体は、遺伝子工学的手法を用いて抗体遺伝子を含む発現ベクターを作製し、細胞で発現させることによっても、得ることができる。 The antibody of the present disclosure uses a peptide containing the amino acid sequence of PLGFFPDHQLDPAFG (SEQ ID NO: 2), NSNNPDWDFNPN (SEQ ID NO: 3), or DPAFGANSNN (SEQ ID NO: 4), or a peptide consisting of these amino acid sequences as an immunogen, or To obtain by a general method using a peptide containing the amino acid sequence of NNPDWDFNP (SEQ ID NO: 17), NNPDWDFN (SEQ ID NO: 21), or GFFPDHQLD (SEQ ID NO: 80) or a peptide consisting of these amino acid sequences as an immunogen. You can The immunogen can be produced by a conventional peptide synthesis method, for example, by a genetic engineering technique or chemical synthesis. Alternatively, the antibody of the present disclosure can also be obtained by preparing an expression vector containing the antibody gene using a genetic engineering technique and expressing it in a cell.
　ポリクローナル抗体は、「Antibodies: A Laboratory Manual, Lane, H. D. et al. eds., Cold Spring Harbor Laboratory Press, New York, 1989」などに記載の一般的な方法により作製することができる。具体的には、前記の免疫原により、ラット、マウス、ウサギ、ヤギ、ウマ等の哺乳動物を免疫することで、作製することができる。 Polyclonal antibodies can be prepared by the general methods described in "Antibodies: Laboratory Manual, Lane, H. D. et al. eds., Cold Spring Harbor Laboratory Press, New York, 1989, etc. Specifically, it can be prepared by immunizing a mammal such as rat, mouse, rabbit, goat, or horse with the above immunogen.
　モノクローナル抗体は、抗体を産生するハイブリドーマを作製する方法や、遺伝子工学的手法を用いて抗体遺伝子を含む発現ベクターを作製して細胞で発現させる方法など、公知の方法により取得することができる。 Monoclonal antibody can be obtained by a known method such as a method for producing a hybridoma producing an antibody, or a method for producing an expression vector containing an antibody gene using a genetic engineering technique and expressing it in a cell.
　モノクローナル抗体を分泌するハイブリドーマは、Kohler et al., Nature 256:495, 1975に記載の方法に準じて作製することができる。まず、免疫原を、抗原性を高めるための適当な物質（例えば、キーホールリンペットヘモシアニンやウシ血清アルブミン等）、および必要に応じて免疫賦活剤（フロイントの完全若しくは不完全アジュバント等）とともに混合し、ラット、マウス、ウサギ、ヤギ、ウマ等の非ヒト哺乳動物に免疫する。通常、免疫動物は3～10日間隔で複数回免疫され、免疫原であるペプチドは1～100μｇが投与される。次いで、複数回の免疫を経た免疫動物から免疫担当細胞（免疫動物において抗体産生能を有する細胞）を回収し、自己抗体産生能のないミエローマ細胞（例えば、マウス、ラット、モルモット、ハムスター、ウサギまたはヒト等の哺乳動物に由来する細胞）と、細胞融合させる。細胞融合には、ポリエチレングリコール法、電気融合法などが用いられる。さらに、融合細胞が有する選択マーカーに基づき細胞融合に成功した細胞を選択し、選択された細胞が産生する抗体の免疫源に対する反応性を、前述の免疫測定法などにより確認することにより、目的のモノクローナル抗体を産生するハイブリドーマが得られる。モノクローナル抗体は、得られたハイブリドーマをインビトロで培養した培養上清から単離することができる。また、マウス、ラット、モルモット、ハムスターまたはウサギ等の腹水中等のインビボで培養し、腹水から単離することもできる。 Hybridomas that secrete monoclonal antibodies can be prepared according to the method described in Kohler et al., Nature 256:495, 1975. First, the immunogen is mixed with an appropriate substance for enhancing antigenicity (eg, keyhole limpet hemocyanin, bovine serum albumin, etc.) and, if necessary, an immunostimulant (Freund's complete or incomplete adjuvant, etc.). And immunize a non-human mammal such as rat, mouse, rabbit, goat, or horse. Usually, the immunized animal is immunized multiple times at intervals of 3 to 10 days, and 1 to 100 μg of the peptide as an immunogen is administered. Then, immunocompetent cells (cells having an antibody-producing ability in the immunized animal) are recovered from an immunized animal that has undergone multiple immunizations, and myeloma cells having no autoantibody-producing ability (eg, mouse, rat, guinea pig, hamster, rabbit or Cells derived from mammals such as humans). For cell fusion, polyethylene glycol method, electrofusion method, etc. are used. Furthermore, by selecting cells that have succeeded in cell fusion based on the selectable marker possessed by the fused cells, and confirming the reactivity of the antibody produced by the selected cells to the immunogen by the immunoassay described above, A hybridoma producing a monoclonal antibody is obtained. The monoclonal antibody can be isolated from the culture supernatant obtained by culturing the obtained hybridoma in vitro. It can also be isolated from ascites by culturing in vivo such as ascites of mouse, rat, guinea pig, hamster or rabbit.
　モノクローナル抗体は、また、抗体遺伝子を含む発現ベクターを作製し宿主細胞で発現させることにより、得ることができる（P.J.Delves., ANTIBODY PRODUCTION ESSENTIAL TECHNIQUES., 1997 WILEY; P.Shepherd and C.Dean., Monoclonal Antibodies., 2000 OXFORD UNIVERSITY PRESS; J.W. Goding., Monoclonal Antibodies:principles and practice., 1993 ACADEMIC PRESS）。さらに、トランスジェニック動物作製技術を用いて、目的とする抗体の遺伝子が内在性遺伝子に組み込まれたトランスジェニック動物（例えば、ウシ、ヤギ、ヒツジまたはブタ）を作製し、そのトランスジェニック動物のミルク中からその抗体遺伝子に由来するモノクローナル抗体を取得することもできる。 A monoclonal antibody can also be obtained by preparing an expression vector containing an antibody gene and expressing it in a host cell (PJDelves., ANTIBODY PRODUCTION ESSENTIAL TECHNIQUES., 1997 WILEY; P.Shepherd and C.Dean., Monoclonal Antibodies., 2000 OXFORD UNIVERSITY PRESS; JW Goding., Monoclonal Antibodies:principles and practice., 1993 ACADEMIC PRESS). Furthermore, a transgenic animal (for example, cow, goat, sheep or pig) in which the gene of the antibody of interest has been incorporated into an endogenous gene is produced by using a transgenic animal production technique, and the transgenic animal's milk Alternatively, a monoclonal antibody derived from the antibody gene can be obtained.
　得られたモノクローナル抗体は、当該分野において周知の方法、例えばプロテインＡカラムによるクロマトグラフィー、イオン交換クロマトグラフィー、疎水クロマトグラフィー、硫安塩析法、ゲル濾過、アフィニティクロマトグラフィー等を適宜組み合わせることにより、精製することができる。 The obtained monoclonal antibody is purified by an appropriate combination of methods well known in the art, for example, chromatography with a protein A column, ion exchange chromatography, hydrophobic chromatography, ammonium sulfate salting-out method, gel filtration, affinity chromatography and the like. can do.
　キメラ抗体は、互いに由来の異なる配列を含む抗体であり、例えば、互いに由来の異なる可変領域と定常領域とを連結した抗体である。ある実施形態において、キメラ抗体は、ヒト以外の哺乳動物由来の抗体の可変領域と、ヒト抗体由来の定常領域から構成される。キメラ抗体は、例えば、ヒト以外の哺乳動物由来の抗体の可変領域をコードするポリヌクレオチドとヒト抗体の定常領域をコードするポリヌクレオチドを連結し、これを発現ベクターに組み込み、この発現ベクターを宿主に導入して発現させることによって、取得することができる。 A chimeric antibody is an antibody containing sequences that are different from each other, and is, for example, an antibody in which variable regions and constant regions that are different from each other are linked. In one embodiment, the chimeric antibody is composed of a variable region of an antibody derived from a mammal other than human and a constant region derived from a human antibody. The chimeric antibody is, for example, a polynucleotide encoding a variable region of an antibody derived from a mammal other than human and a polynucleotide encoding a constant region of a human antibody are ligated, and this is incorporated into an expression vector, and the expression vector is used as a host. It can be obtained by introducing and expressing it.
　CDRは、実質的に抗体の結合特異性を決定する領域であり、そのアミノ酸配列は多様性に富む。一方、FRを構成するアミノ酸配列は、異なる結合特異性を有する抗体の間でも高い相同性を示す。そのため、CDRの移植によって、ある抗体の結合特異性を他の抗体に移植することができる。 CDR is a region that substantially determines the binding specificity of an antibody, and its amino acid sequence is highly diverse. On the other hand, the amino acid sequences constituting FR show high homology even among antibodies having different binding specificities. Thus, CDR grafting allows the binding specificity of one antibody to be transferred to another antibody.
　ヒト化抗体は、一般に、ヒト以外の動物由来の抗体のCDRと、ヒト抗体由来のFRおよびヒト抗体由来の定常領域とから構成される。ヒト化抗体は、ヒト以外の動物由来の抗体のCDRをヒト抗体に移植することにより得ることができる。ヒト化抗体は種々の方法により作製することができるが、一例として、Overlap Extension PCRが挙げられる（Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)）。本方法では、ヒト以外の動物由来の抗体（例えばマウス抗体）のCDRとヒト抗体のFRとの末端にオーバーラップする部分を有するオリゴヌクレオチドをプライマーとして用いてPCRを行い、ヒト以外の動物由来の抗体のCDRとヒト抗体のFRとが連結されたポリヌクレオチドを合成する。次いで、得られたポリヌクレオチドをヒト抗体の定常領域をコードするポリヌクレオチドと連結し、発現ベクターに組み込んで、この発現ベクターを宿主に導入して発現させることにより、ヒト化抗体を取得することができる。 A humanized antibody is generally composed of CDRs of an antibody derived from a non-human animal, FRs derived from a human antibody, and constant regions derived from a human antibody. The humanized antibody can be obtained by transplanting the CDR of an antibody derived from a non-human animal into a human antibody. Humanized antibodies can be produced by various methods, but one example is Overlap Extension PCR (Almagro and Fransson, Front.Biosci. 13:1619-1633 (2008)). In this method, PCR is performed using an oligonucleotide having a portion overlapping with the end of the CDR of the antibody derived from a non-human animal (for example, a mouse antibody) and the FR of the human antibody as a primer, and derived from a non-human animal. A polynucleotide in which the CDR of the antibody and the FR of the human antibody are linked is synthesized. Then, the obtained polynucleotide is ligated to a polynucleotide encoding a constant region of a human antibody, incorporated into an expression vector, and this expression vector is introduced into a host to be expressed to obtain a humanized antibody. it can.
　ヒト化抗体の作製に好適なFRの選択方法は公知であり、例えば、ベストフィット法（Sims et al. J. Immunol. 151:2296 (1993)）により選択されたFRや、ヒト抗体の軽鎖または重鎖可変領域の特定のサブグループのコンセンサス配列に由来するFR（Carter et al. Proc. Natl. Acad. Sci. USA 89:4285 (1992); Presta et al. J. Immunol. 151:2623 (1993)）を用いることができる。 Suitable FR selection methods for producing humanized antibodies are known, for example, FR selected by the best fit method (Sims et.al. J. Immunol. 151:2296 (1993)) and the light chain of human antibody. Or FR (Carter et al. Proc. Natl. Acad. Sci. USA 89:4285 (1992); Presta et al. J. Immunol. 151:2623 (derived from a consensus sequence of a specific subgroup of the heavy chain variable region) 1993)) can be used.
　ヒト抗体は、例えば、ヒトリンパ球をインビトロで所望の抗原で感作し、次いで、感作リンパ球をヒトミエローマ細胞と融合させることによって取得することができる（特公平1-59878号公報）。融合パートナーであるヒトミエローマ細胞には、例えばU266などを利用することができる。また、ヒト抗体は、ヒト抗体遺伝子の全てのレパートリーを有するトランスジェニック動物を所望の抗原で免疫することによっても取得することができる（Lonberg, Nat. Biotech. 23: 1117-1125, 2005)。さらに、ヒト抗体ライブラリーを用いて、パンニングによりヒト抗体を取得する技術も知られている（Antibody Phage Display: Methods and Protocols, Methods in Molecular Biology 178, 2001）。例えば、ヒト抗体の可変領域を一本鎖抗体（scFv）としてファージディスプレイ法によりファージの表面に発現させ、抗原に結合するファージを選択し、選択されたファージの遺伝子を解析することにより、抗原に結合するヒト抗体の可変領域をコードするＤＮＡ配列を決定することができる。次いで、この可変領域配列をヒト抗体定常領域の配列とインフレームで連結し、適当な発現ベクターに挿入して、この発現ベクターを宿主に導入して発現させることにより、ヒト抗体を取得することができる。 A human antibody can be obtained, for example, by sensitizing human lymphocytes with a desired antigen in vitro and then fusing the sensitized lymphocytes with human myeloma cells (Japanese Patent Publication No. 1-59878). For human myeloma cells that are fusion partners, for example, U266 and the like can be used. In addition, human antibodies can also be obtained by immunizing a transgenic animal having the entire repertoire of human antibody genes with a desired antigen (Lonberg, Nat. Biotech. 23: 1117-1125, 2005). Furthermore, a technique for obtaining a human antibody by panning using a human antibody library is also known (AntibodyPhageDisplay: MethodsandProtocols, MethodsinMolecularBiology 178, 2001). For example, the variable region of a human antibody is expressed as a single chain antibody (scFv) on the surface of a phage by the phage display method, a phage that binds to the antigen is selected, and the gene of the selected phage is analyzed to obtain the antigen. The DNA sequence encoding the variable region of the human antibody that binds can be determined. Then, this variable region sequence is ligated in frame with the sequence of the human antibody constant region, inserted into an appropriate expression vector, and this expression vector is introduced into a host and expressed to obtain a human antibody. it can.
　多重特異性抗体は、少なくとも2つの異なる部位に結合する抗体である。多重特異性抗体としては、二重特異性抗体、三重特異性抗体などが挙げられる。ある実施形態において、多重特異性抗体は、preS1抗原と、1以上の他の抗原とに結合する。多重特異性抗体は、例えば、遺伝子工学的手法により、あるいは認識抗原が異なる2つ以上の抗体を結合することにより、作製することができる。 Multispecific antibodies are antibodies that bind to at least two different sites. Examples of the multispecific antibody include a bispecific antibody and a trispecific antibody. In certain embodiments, the multispecific antibody binds the preS1 antigen and one or more other antigens. The multispecific antibody can be produced, for example, by a genetic engineering technique or by binding two or more antibodies having different recognition antigens.
　抗体断片は、例えば、パパイン、ペプシン等のプロテアーゼにより抗体を消化することにより得ることができる。あるいは、抗体断片をコードするポリヌクレオチドを含む発現ベクターを宿主細胞に導入し発現させることで得ることができる（例えば、Co, M. S. et al., J. Immunol. (1994) 152, 2968-2976; Better, M. and Horwitz, A. H., Methods Enzymol. (1989) 178, 476-496; Pluckthun, A. and Skerra, A., Methods Enzymol. (1989) 178, 497-515; Lamoyi, E., Methods Enzymol. (1986) 121, 652-663; Rousseaux, J. et al., Methods Enzymol. (1986) 121, 663-669; Bird, R. E. and Walker, B. W., Trends Biotechnol. (1991) 9, 132-137; Hudson et al., Nat. Med., (2003) 9, 129-134）。 The antibody fragment can be obtained, for example, by digesting the antibody with a protease such as papain or pepsin. Alternatively, it can be obtained by introducing and expressing an expression vector containing a polynucleotide encoding an antibody fragment into a host cell (for example, Co, M. S. et al., J. Immunol. (1994) 152, 2968. -2976; Better, M. and Horwitz, A. H., MethodsEnzymol. (1989)178, 476-496; Pluckthun, A. and Skerra, A., MethodsEnzymol. (1989) 178, 497-515; Lamoyi , E., MethodsEnzymol. (1986) 121, 652-663; Rousseaux, J. et al., Methods Enzymol. (1986)121, 663-669; Bird, R. E. and Walker, B. W., Trends Biotechnol. (1991) 9, 132-137; Hudson et et al., Nat. Med., (2003) 9, 129-134).
　前述のように、抗体は、これをコードするポリヌクレオチドを含む発現ベクターを細胞に導入して発現させることにより、得ることができる。具体的には、抗体をコードする配列がエンハンサーやプロモーターなどの発現制御領域のもとで発現するよう発現ベクターを構築し、この発現ベクターで宿主細胞を形質転換して、抗体を発現させる。 As mentioned above, an antibody can be obtained by introducing an expression vector containing a polynucleotide encoding the same into a cell and expressing it. Specifically, an expression vector is constructed so that the sequence encoding the antibody is expressed under an expression control region such as an enhancer and a promoter, and a host cell is transformed with this expression vector to express the antibody.
　すなわち、本開示はまた、抗preS1抗体をコードするポリヌクレオチドを含む発現ベクター、および前記抗体を発現可能なポリヌクレオチドを含む形質転換細胞を提供する。 That is, the present disclosure also provides an expression vector containing a polynucleotide encoding an anti-preS1 antibody, and a transformed cell containing a polynucleotide capable of expressing the antibody.
　宿主細胞としては、例えば、動物細胞、植物細胞、真菌細胞などの真核細胞を用いることができる。動物細胞としては、哺乳類細胞（例えば、CHO、COS、NIH3T3、ミエローマ、BHK（baby hamster kidney）、HeLa、Vero）、両生類細胞（例えばアフリカツメガエル卵母細胞）、または昆虫細胞（例えば、Sf9、Sf21、Tn5）が挙げられる。真菌細胞としては、酵母（例えば、サッカロミセス（Saccharomyces）属、例えば、サッカロミセス・セレビシエ（Saccharomyces cerevisiae））、糸状菌（例えば、アスペルギルス（Aspergillus）属、例えば、アスペルギルス・ニガー（Aspergillus niger））などが挙げられる。また、大腸菌（E. coli）（例えば、JM109、DH5α、HB101等）、枯草菌などの原核細胞を宿主細胞として用いることもできる。宿主細胞へのベクターの導入は、例えば、リン酸カルシウム法、DEAEデキストラン法、エレクトロポレーション法、リポフェクションなどの方法により行うことができる。 As the host cell, for example, an eukaryotic cell such as an animal cell, a plant cell or a fungal cell can be used. Animal cells include mammalian cells (eg, CHO, COS, NIH3T3, myeloma, BHK (baby hamster kidney), HeLa, Vero), amphibian cells (eg, Xenopus oocytes), or insect cells (eg, Sf9, Sf21). , Tn5). Examples of the fungal cells include yeast (for example, Saccharomyces genus, for example, Saccharomyces cerevisiae), filamentous fungus (for example, Aspergillus genus, for example, Aspergillus nigier), and the like. To be In addition, prokaryotic cells such as E. coli (eg, JM109, DH5α, HB101, etc.) and Bacillus subtilis can also be used as host cells. The vector can be introduced into the host cell by a method such as calcium phosphate method, DEAE dextran method, electroporation method, lipofection and the like.
　得られた抗体の抗原への結合は、EIA（ELISAを含む）、RIA、CIA、FIAなどの免疫測定法や、BIACORE（登録商標）表面プラズモン共鳴アッセイなどにより確認することができる。 The binding of the obtained antibody to the antigen can be confirmed by immunoassay methods such as EIA (including ELISA), RIA, CIA, FIA, and BIACORE (registered trademark) surface plasmon resonance assay.
　抗体の抗原への結合は、競合アッセイにより確認することもできる。例えば、抗体が、本明細書に記載の（ａ）～（ｉ）のいずれかの抗体、（ａ’）～（ｉ’）のいずれかの抗体、（ａ’’）～（ｉ’’）のいずれかの抗体、（ａ’’’）～（ｉ’’’）のいずれかの抗体、または（ａ’’’’）～（ｉ’’’’）のいずれかの抗体と、preS1抗原への結合において競合するか否かを調べることにより、確認することができる。 The binding of the antibody to the antigen can also be confirmed by a competitive assay. For example, the antibody is any of the antibodies (a) to (i), any of the antibodies (a′) to (i′), (a″) to (i″) described in the present specification. Any of the antibodies of (a''') to (i'''), or any of the antibodies of (a'''') to (i''''), and preS1 antigen It can be confirmed by examining whether or not there is a competition in binding to.
　ある実施形態において、本開示の抗体は、本明細書に記載の（ａ）～（ｉ）のいずれかの抗体、（ａ’）～（ｉ’）のいずれかの抗体、（ａ’’）～（ｉ’’）のいずれかの抗体、（ａ’’’）～（ｉ’’’）のいずれかの抗体、または（ａ’’’’）～（ｉ’’’’）のいずれかの抗体と、preS1抗原への結合において競合する抗体である。本明細書において、所定の抗体（すなわち、参照抗体）と競合する抗体とは、参照抗体のpreS1抗原への結合を有意に減少させる抗体を意味する。競合実験は、以下のように行うことができる。まず、試験抗体と参照抗体とを種々のモル比で混合する。試験抗体は、通常、参考抗体に対して過剰量で（例えば、1倍、2倍、5倍、10倍、20倍、50倍、または100倍）用いられる。参照抗体は、適当な標識（例えばビオチン）で標識されていてもよい。次いで、前記混合物を、preS1抗原を固相化したプレートに添加し、反応させた後、プレートを洗浄し、検出試薬（例えば、ホースラディッシュペルオキシダーゼ（HRP）で標識した二次抗体、または、ビオチン化した試験抗体を用いた場合、HRP-ストレプトアビジン）を添加して、preS1抗原に結合した標識参照抗体の量を測定する。ある実施形態において、本開示の抗体は、参照抗体のpreS1抗原への結合を、10％、20％、30％、40％、50％、60％、70％、80％、90％、または95％以上減少させる。 In certain embodiments, an antibody of the present disclosure is an antibody of any of (a)-(i), an antibody of any of (a′)-(i′), (a″) described herein. To (i″) any antibody, (a′″) to (i′″) any antibody, or (a″″) to (i″″) any Antibody that competes with the antibody of (1) for binding to the preS1 antigen. As used herein, an antibody that competes with a predetermined antibody (that is, a reference antibody) means an antibody that significantly reduces the binding of the reference antibody to the preS1 antigen. The competition experiment can be performed as follows. First, the test antibody and the reference antibody are mixed at various molar ratios. The test antibody is usually used in excess (eg, 1-fold, 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, or 100-fold) with respect to the reference antibody. The reference antibody may be labeled with a suitable label (eg biotin). Then, the mixture is added to a plate on which preS1 antigen is immobilized and reacted, then the plate is washed, and a detection reagent (for example, a secondary antibody labeled with horseradish peroxidase (HRP) or biotinylated) is used. HRP-streptavidin) is used to measure the amount of labeled reference antibody bound to the preS1 antigen. In certain embodiments, an antibody of the present disclosure binds a reference antibody to preS1 antigen by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%. % Or more.
　さらなる実施形態において、本開示の抗体は、本明細書に記載の（ａ’）～（ｉ’）のいずれかの抗体、（ａ’’’）～（ｉ’’’）のいずれかの抗体、または（ａ’’’’）～（ｉ’’’’）のいずれかの抗体と、preS1抗原への結合において競合する抗体；本明細書に記載の（ａ’）～（ｉ’）のいずれかの抗体または（ａ’’’）～（ｉ’’’）のいずれかの抗体と、preS1抗原への結合において競合する抗体；または、本明細書に記載の（ａ’’’）～（ｉ’’’）のいずれかの抗体と、preS1抗原への結合において競合する抗体である。 In a further embodiment, the antibody of the present disclosure is the antibody of any of (a′) to (i′) and the antibody of (a′″) to (i″′) described herein. , Or (a″″) to (i″″), which competes with the antibody of any of (a″″) to (i″″) for binding to the preS1 antigen; An antibody that competes with any antibody or any of the antibodies (a′″) to (i′″) for binding to the preS1 antigen; or (a″′) to An antibody that competes with any of the antibodies (i′″) for binding to the preS1 antigen.
　ある実施形態において、本開示の抗体は、PLGFFPDHQLDPAFG（配列番号２）、NSNNPDWDFNPN（配列番号３）、またはDPAFGANSNN（配列番号４）から選択されるアミノ酸配列に含まれるエピトープに結合する。ある実施形態において、配列番号２のアミノ酸配列に含まれるエピトープに結合する抗体は、GFFPDHQLD（配列番号８０）のアミノ酸配列に含まれるエピトープに結合する抗体である。別の実施形態において、配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体は、NNPDWDFNP（配列番号１７）またはNNPDWDFN（配列番号２１）のアミノ酸配列に含まれるエピトープに結合する抗体である。所定のアミノ酸配列に含まれるエピトープに結合するか否かは、実施例に記載するように、当該アミノ酸配列からなるペプチドに結合するか否かを調べることにより、確認することができる。 In one embodiment, the antibody of the present disclosure binds to an epitope contained in an amino acid sequence selected from PLGFFPDHQLDPAFG (SEQ ID NO: 2), NSNNPDWDFNPN (SEQ ID NO: 3), or DPAFGANSNN (SEQ ID NO: 4). In certain embodiments, the antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO:2 is an antibody that binds to the epitope included in the amino acid sequence of GFFPDHQLD (SEQ ID NO:80). In another embodiment, the antibody that binds to the epitope contained in the amino acid sequence of SEQ ID NO:3 is an antibody that binds to the epitope contained in the amino acid sequence of NNPDWDFNP (SEQ ID NO:17) or NNPDWDFN (SEQ ID NO:21). Whether or not it binds to an epitope contained in a given amino acid sequence can be confirmed by investigating whether or not it binds to a peptide consisting of the amino acid sequence, as described in Examples.
　別の実施形態において、本開示の抗体は、以下の（ａ）～（ｉ）から選択される：
（ａ）配列番号２４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号２６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号２８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｂ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｃ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｄ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｅ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｆ）配列番号５０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号５４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｇ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｈ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；および
（ｉ）配列番号６５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号６９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号７０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号７１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体。 In another embodiment, the antibody of the present disclosure is selected from the following (a)-(i):
(A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 28 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 30 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(B) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 37 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(C) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(D) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence have been deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 43 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 45 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(E) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and the amino acid sequence of SEQ ID NO: 47 or 1 to 3 in said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(F) CDR1 comprising the amino acid sequence of SEQ ID NO: 50 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 51 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 52 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 54 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:55 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:56 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(G) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 60 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 61 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(H) a CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence;
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:63 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:61 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising: (i) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in said amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 67 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 70 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 71 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
　さらなる実施形態において、本開示の抗体は、以下の（ａ’）～（ｉ’）から選択される： 
（ａ’）配列番号２４のアミノ酸配列を含むCDR1、
　配列番号２５のアミノ酸配列を含むCDR2、および
　配列番号２６のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号２８のアミノ酸配列を含むCDR1、
　配列番号２９のアミノ酸配列を含むCDR2、および
　配列番号３０のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｂ’）配列番号３２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号３７のアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｃ’）配列番号３２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｄ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号４３のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列を含むCDR2、および
　配列番号４５のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｅ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号４７のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｆ’）配列番号５０のアミノ酸配列を含むCDR1、
　配列番号５１のアミノ酸配列を含むCDR2、および
　配列番号５２のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号５４のアミノ酸配列を含むCDR1、
　配列番号５５のアミノ酸配列を含むCDR2、および
　配列番号５６のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｇ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号６０のアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｈ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号６３のアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；および
（ｉ’）配列番号６５のアミノ酸配列を含むCDR1、
　配列番号６６のアミノ酸配列を含むCDR2、および
　配列番号６７のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号６９のアミノ酸配列を含むCDR1、
　配列番号７０のアミノ酸配列を含むCDR2、および
　配列番号７１のアミノ酸配列を含むCDR3。 In a further embodiment, the antibodies of the present disclosure are selected from the following (a')-(i'):
(A') CDR1 comprising the amino acid sequence of SEQ ID NO: 24,
CDR2 containing the amino acid sequence of SEQ ID NO:25, and CDR3 containing the amino acid sequence of SEQ ID NO:26
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 28,
CDR2 containing the amino acid sequence of SEQ ID NO: 29, and CDR3 containing the amino acid sequence of SEQ ID NO: 30
An antibody comprising a heavy chain variable region comprising:
(B') CDR1 containing the amino acid sequence of SEQ ID NO: 32,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:37, and CDR3 containing the amino acid sequence of SEQ ID NO:38
An antibody comprising a heavy chain variable region comprising:
(C′) CDR1 containing the amino acid sequence of SEQ ID NO: 32,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:38
An antibody comprising a heavy chain variable region comprising:
(D') CDR1 containing the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:43
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:45
An antibody comprising a heavy chain variable region comprising:
(E′) CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:47
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:38
An antibody comprising a heavy chain variable region comprising:
(F') CDR1 comprising the amino acid sequence of SEQ ID NO: 50,
CDR2 containing the amino acid sequence of SEQ ID NO:51 and CDR3 containing the amino acid sequence of SEQ ID NO:52
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 54,
CDR2 containing the amino acid sequence of SEQ ID NO:55, and CDR3 containing the amino acid sequence of SEQ ID NO:56
An antibody comprising a heavy chain variable region comprising:
(G') CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:58
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:60, and CDR3 containing the amino acid sequence of SEQ ID NO:61
An antibody comprising a heavy chain variable region comprising:
(H') CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:58
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:63, and CDR3 containing the amino acid sequence of SEQ ID NO:61
An antibody comprising a heavy chain variable region comprising; and (i′) CDR1 comprising the amino acid sequence of SEQ ID NO:65,
CDR2 containing the amino acid sequence of SEQ ID NO:66, and CDR3 containing the amino acid sequence of SEQ ID NO:67
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 69,
CDR2 containing the amino acid sequence of SEQ ID NO:70, and CDR3 containing the amino acid sequence of SEQ ID NO:71.
　さらなる実施形態において、本開示の抗体は、以下の（ａ’’）～（ｉ’’）から選択される：
（ａ’’）配列番号２３のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号２３のアミノ酸配列もしくは配列番号２３のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号２７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号２７のアミノ酸配列もしくは配列番号２７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｂ’’）配列番号３１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３１のアミノ酸配列もしくは配列番号３１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号３５のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３５のアミノ酸配列もしくは配列番号３５のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｃ’’）配列番号３１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３１のアミノ酸配列もしくは配列番号３１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号３９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３９のアミノ酸配列もしくは配列番号３９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｄ’’）配列番号４１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４１のアミノ酸配列もしくは配列番号４１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号４４のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４４のアミノ酸配列もしくは配列番号４４のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｅ’’）配列番号４６のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４６のアミノ酸配列もしくは配列番号４６のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号４８のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４８のアミノ酸配列もしくは配列番号４８のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｆ’’）配列番号４９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４９のアミノ酸配列もしくは配列番号４９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号５３のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５３のアミノ酸配列もしくは配列番号５３のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｇ’’）配列番号５７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５７のアミノ酸配列もしくは配列番号５７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号５９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５９のアミノ酸配列もしくは配列番号５９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｈ’’）配列番号５７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５７のアミノ酸配列もしくは配列番号５７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号６２のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６２のアミノ酸配列もしくは配列番号６２のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；および
（ｉ’’）配列番号６４のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６４のアミノ酸配列もしくは配列番号６４のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号６８のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６８のアミノ酸配列もしくは配列番号６８のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体。 In a further embodiment, the antibodies of the present disclosure are selected from the following (a″)-(i″):
(A'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 23, or the amino acid sequence of SEQ ID NO: 23 or the amino acid sequence of SEQ ID NO: 23 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 27. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 27 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 27, An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
(B'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 35. Having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 35, or 1-20 in the amino acid sequence of SEQ ID NO: 35, 1-10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
(C'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 39. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 39 or 1 to 20 in the amino acid sequence of SEQ ID NO: 39, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
(D'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 41, or the amino acid sequence of SEQ ID NO: 41 or the amino acid sequence of SEQ ID NO: 41 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues are deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 44. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 44 or 1 to 20 in the amino acid sequence of SEQ ID NO: 44, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(E'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 46, or the amino acid sequence of SEQ ID NO: 46 or the amino acid sequence of SEQ ID NO: 46 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 48 Amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 48 or 1 to 20 in the amino acid sequence of SEQ ID NO: 48, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(F″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 49, or the amino acid sequence of SEQ ID NO: 49 or the amino acid sequence of SEQ ID NO: 49 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 53 With 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 53 or the amino acid sequence of SEQ ID NO: 53 with 1-20, 1-10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(G″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 59. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 59 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 59, An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(H″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 62. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 62 or 1 to 20 in the amino acid sequence of SEQ ID NO: 62, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which ˜5, or 1-3 amino acid residues have been deleted, substituted, or added; and (i″) the amino acid sequence of SEQ ID NO: 64 and 80% , 85%, 90%, or 95% or more sequence identity, or 1-20, 1-10, 1-5 in the amino acid sequence of SEQ ID NO: 64 or the amino acid sequence of SEQ ID NO: 64 , Or a light chain variable region comprising an amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted, or added, and 80%, 85%, 90%, or 95% or more of the amino acid sequence of SEQ ID NO: 68 An amino acid sequence having the sequence identity of, or 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues in the amino acid sequence of SEQ ID NO:68 or the amino acid sequence of SEQ ID NO:68. An antibody comprising a heavy chain variable region comprising a deleted, substituted, or added amino acid sequence.
　さらなる実施形態において、本開示の抗体は、以下の（ａ’’’）～（ｉ’’’）から選択される：
（ａ’’’）配列番号２３のアミノ酸配列を含む軽鎖可変領域、および配列番号２７のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｂ’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域、および配列番号３５のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｃ’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域、および配列番号３９のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｄ’’’）配列番号４１のアミノ酸配列を含む軽鎖可変領域、および配列番号４４のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｅ’’’）配列番号４６のアミノ酸配列を含む軽鎖可変領域、および配列番号４８のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｆ’’’）配列番号４９のアミノ酸配列を含む軽鎖可変領域、および配列番号５３のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｇ’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域、および配列番号５９のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｈ’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域、および配列番号６２のアミノ酸配列を含む重鎖可変領域を含む抗体；および
（ｉ’’’）配列番号６４のアミノ酸配列を含む軽鎖可変領域、および配列番号６８のアミノ酸配列を含む重鎖可変領域を含む抗体。 In a further embodiment, the antibodies of the present disclosure are selected from the following (a''') to (i'''):
(A′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27;
(B''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35;
(C′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39;
(D′″) an antibody comprising a light chain variable region containing the amino acid sequence of SEQ ID NO: 41 and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44;
(E′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:46 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:48;
(F″′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:49 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:53;
(G''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:57 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:59;
(H″′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:57 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:62; and (i″′) an amino acid sequence of SEQ ID NO:64. An antibody comprising a light chain variable region comprising, and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:68.
　ある実施形態において、本開示の抗体は、、以下の（ａ’’’’）～（ｉ’’’’）から選択される：
（ａ’’’’）配列番号２３のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号２７のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｂ’’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号３５のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｃ’’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号３９のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｄ’’’’）配列番号４１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号４４のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｅ’’’’）配列番号４６のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号４８のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｆ’’’’）配列番号４９のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号５３のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｇ’’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号５９のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｈ’’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号６２のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；および
（ｉ’’’’）配列番号６４のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号６８のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体。
　本実施形態において、CDRの特定方法は任意である。好ましくは、CDRは、Kabatの定義 (Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. 1991)、Chothiaの定義(Chothia et al., J. Mol. Biol., 1987; 196: 901-917)、AdMの定義 (Martin et al., Proc.Natl.Acad.Sci.USA, 1989; 86: 9268-9272)、Contactの定義 (MacCallum et al., J. Mol. Biol. 1996; 262: 732-745)、およびIMGTの定義（Lefranc et al., Dev Comp Immunol. 2003; 27(1): 55-77）のいずれかにより特定される。 In certain embodiments, the antibodies of the present disclosure are selected from the following (a″″) to (i″″):
(A″″) CDR1, CDR2 in the light chain variable region including the amino acid sequence of SEQ ID NO:23, and CDR1, CDR2 in the heavy chain variable region including the amino acid sequence of SEQ ID NO:27 And an antibody comprising a heavy chain variable region comprising CDR3;
(B″″) CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:35 and the light chain variable region containing CDR1, CDR2, and CDR3 in the light chain variable region of SEQ ID NO:31 And an antibody comprising a heavy chain variable region comprising CDR3;
(C″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 31 and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 39 And an antibody comprising a heavy chain variable region comprising CDR3;
(D″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 41, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44 And an antibody comprising a heavy chain variable region comprising CDR3;
(E″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO:46, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:48 And an antibody comprising a heavy chain variable region comprising CDR3;
(F″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO:49, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:53 And an antibody comprising a heavy chain variable region comprising CDR3;
(G'''') CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 57, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 59 And an antibody comprising a heavy chain variable region comprising CDR3;
(H″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 57, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 62 And (i″″) a light chain variable region comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 64, and a light chain variable region comprising CDR3, CDR2, and CDR3, and SEQ ID NO: An antibody comprising a heavy chain variable region comprising CDR1, CDR2, and CDR3 in a heavy chain variable region comprising 68 amino acid sequences.
In this embodiment, the method of identifying the CDR is arbitrary. Preferably, CDR is the definition of Kabat (Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. 1991), the definition of Chothia (Chothia et al., J. Mol. Biol., 1987; 196: 901-917), AdM definition (Martin et al., Proc. Natl. Acad.Sci.USA, 1989; 86: 9268-9272), Contact definition (MacCallum et al., J. Mol. Biol. 1996; 262: 732-745) and the IMGT definition (Lefranc et al., Dev Comp Immunol. 2003; 27(1): 55-77).
　本明細書において、所定のアミノ酸配列を含むアミノ酸配列には、その所定のアミノ酸配列に１以上のアミノ酸残基が付加されたアミノ酸配列と、その所定のアミノ酸からなる配列とが包含される。 In the present specification, an amino acid sequence containing a predetermined amino acid sequence includes an amino acid sequence in which one or more amino acid residues are added to the predetermined amino acid sequence, and a sequence consisting of the predetermined amino acid.
　本明細書において、所定の重鎖可変領域または軽鎖可変領域のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列を含む重鎖可変領域または軽鎖可変領域、および、所定の重鎖可変領域または軽鎖可変領域のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域または軽鎖可変領域には、その所定の重鎖可変領域または軽鎖可変領域のアミノ酸配列におけるCDRに改変が生じていない抗体が含まれる。 As used herein, a heavy chain variable region or a light chain containing an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of a predetermined heavy chain variable region or light chain variable region A variable region and 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues in the amino acid sequence of a given heavy chain variable region or light chain variable region are deleted or substituted, Alternatively, the heavy chain variable region or the light chain variable region containing the added amino acid sequence includes an antibody in which the CDR in the amino acid sequence of the predetermined heavy chain variable region or light chain variable region is not modified.
　本明細書における、アミノ酸配列に関する「配列同一性」とは、比較対象の配列の全領域にわたって最適な状態に（一致が最大となる状態に）アラインメントされた２つの配列間で一致するアミノ酸残基の割合を意味する。ここで、比較対象の配列は、２つの配列の最適なアラインメントにおいて、付加または欠失（例えばギャップ等）を有していてもよい。配列同一性は、公共のデータベース（例えば、DDBJ（http://www.ddbj.nig.ac.jp））で提供されるFASTA、BLAST、CLUSTAL W等のプログラムを用いて算出することができる。あるいは、市販の配列解析ソフトウェア（例えば、Vector NTI（登録商標）ソフトウェア、GENETYX（登録商標） ver. 12）を用いて求めることもできる。 As used herein, the term “sequence identity” with respect to an amino acid sequence means an amino acid residue that matches between two sequences that are optimally aligned (in a state of maximum matching) over the entire region of the sequence to be compared. Means the ratio of. Here, the sequences to be compared may have additions or deletions (eg gaps etc.) in the optimal alignment of the two sequences. Sequence identity can be calculated using a program such as FASTA, BLAST, CLUSTAL W provided in a public database (for example, DDBJ (http://www.ddbj.nig.ac.jp)). Alternatively, it can be determined using commercially available sequence analysis software (eg, Vector NTI (registered trademark) software, GENETYX (registered trademark) ver. 12).
　アミノ酸配列を改変し所望の特性を有する抗体を得る方法として、様々な方法が知られている。例えば、結合親和性を改善した変異体は、ファージディスプレイに基づく方法により得ることができる。この方法では、例えば、アラニンスキャニング変異導入法により抗体と抗原の相互作用に影響するアミノ酸残基を同定するか、あるいは、抗原抗体複合体の結晶構造を解析して抗体と抗原の間の接触点を同定するなどして、変異導入部位を決定する。この部位のアミノ酸を改変した変異体をエラープローンPCRや部位特異的変異導入などにより作製し、得られた変異体のライブラリーをスクリーニングすることにより、所望の特性を有する変異体を得ることができる。 Various methods are known as methods for modifying an amino acid sequence to obtain an antibody having desired properties. For example, variants with improved binding affinity can be obtained by methods based on phage display. In this method, for example, the amino acid residue that influences the interaction between the antibody and the antigen is identified by the alanine scanning mutation introduction method, or the crystal structure of the antigen-antibody complex is analyzed to determine the contact point between the antibody and the antigen. The mutagenesis site is determined by identifying A mutant having a desired property can be obtained by preparing a mutant in which the amino acid at this site is modified by error-prone PCR or site-directed mutagenesis, and screening the resulting mutant library. ..
　抗体は、抗体依存性細胞傷害（ADCC）活性や補体依存性細胞傷害（CDC）活性または体内動態の調節のため、イムノグロブリンのサブクラスが選択されていてもよく、Fc領域のアミノ酸配列または糖鎖が改変されていてもよい。例えば、補体活性化能の低減を目的として、IgG4が選択されうる。また、Fc受容体またはC1qへの結合性を低減または増強する改変、血中半減期の延長のためFcRnへの結合親和性を上昇させる改変などが行われうる。 The antibody may have a selected immunoglobulin subclass for the regulation of antibody-dependent cellular cytotoxicity (ADCC) activity, complement-dependent cellular cytotoxicity (CDC) activity, or pharmacokinetics, and may have an amino acid sequence of the Fc region or sugar. The chain may be modified. For example, IgG4 can be selected for the purpose of reducing complement activating ability. In addition, modifications that reduce or enhance the binding to the Fc receptor or C1q, modifications that increase the binding affinity to FcRn for the purpose of extending the blood half-life, and the like can be performed.
　抗体は、また、例えば抗体の血中半減期の延長、または安定性の改善等のため、ポリエチレングリコール（PEG）、ポリプロピレングリコール、ポリオキシアルキレンまたはポリエチレングリコールとポリプロピレングリコールとのコポリマーなどのポリマーと結合していてもよい。抗体は、また、化学療法剤、毒性ペプチド、放射性化学物質などと結合していてもよい。 The antibody also binds to a polymer such as polyethylene glycol (PEG), polypropylene glycol, polyoxyalkylene or a copolymer of polyethylene glycol and polypropylene glycol, eg, to increase the blood half-life of the antibody or to improve stability. You may have. Antibodies may also be linked to chemotherapeutic agents, toxic peptides, radiochemicals and the like.
　2種以上の本開示の抗体を組み合わせて使用することもできる。ある実施形態において、本開示の抗体の組み合わせは、異なるエピトープに結合する抗体の組み合わせである。また、本開示の抗体は、本開示の抗preS1抗体とは異なるエピトープに結合する抗HBs抗体と組み合わせても良い。ある実施形態において、本開示の抗preS1抗体は、抗S抗体または抗preS2抗体と組み合わせて使用される。 It is also possible to use a combination of two or more antibodies of the present disclosure. In certain embodiments, the antibody combination of the present disclosure is a combination of antibodies that bind to different epitopes. Further, the antibody of the present disclosure may be combined with an anti-HBs antibody that binds to an epitope different from that of the anti-preS1 antibody of the present disclosure. In certain embodiments, anti-preS1 antibodies of the present disclosure are used in combination with anti-S or anti-preS2 antibodies.
２．preS1抗原の測定
　本開示の抗体は、preS1抗原の測定に用いることができる。本開示の方法は、対象から得た試料を本開示の抗体と接触させること、およびpreS1抗原と抗体との複合体を検出することを含む。 2. Measurement of preS1 antigen The antibody of the present disclosure can be used for measurement of preS1 antigen. The methods of the present disclosure include contacting a sample obtained from a subject with an antibody of the present disclosure and detecting a complex of preS1 antigen and the antibody.
　対象には、HBVに感染している対象およびHBVへの感染が疑われる対象が含まれる。対象から得る試料は、preS1抗原を含むものであればよく、例えば、血液、血漿、血清、尿、汗、乳汁、初乳、涙液、唾液、精液、膣分泌物などが挙げられる。本明細書における「対象から得た試料」なる用語は、対象から得た試料そのもののみならず、当該試料中のpreS1抗原と抗preS1抗体との複合体の形成のために必要な処理を施した後の試料を包含する意味で用いられる。 ≪Subjects include those infected with HBV and those suspected of being infected with HBV. The sample obtained from the subject may be any sample containing the preS1 antigen, and examples thereof include blood, plasma, serum, urine, sweat, milk, colostrum, tears, saliva, semen, vaginal secretions and the like. In the present specification, the term "sample obtained from a subject" means not only a sample obtained from a subject itself but also a treatment necessary for forming a complex of preS1 antigen and anti-preS1 antibody in the sample. Used to include later samples.
　preS1抗原と抗体との複合体は、当業界に知られるいずれの方法で測定してもよい。測定方法としては、例えば、EIA（ELISAを含む）、RIA、CIA、FIA、ラテックス凝集反応に基づく方法などの免疫測定法が挙げられる。 The complex of preS1 antigen and antibody may be measured by any method known in the art. Examples of the measurement method include immunoassay methods such as EIA (including ELISA), RIA, CIA, FIA, and a method based on latex agglutination reaction.
　抗体は、測定方法に応じて修飾されていてもよい。例えば、酵素（例えば、ホースラディッシュペルオキシダーゼ、アルカリフォスファターゼ、ルシフェラーゼ、β－ガラクトシダーゼ）、蛍光物質（例えば、フルオレセイン、ローダミン）、放射性同位元素（例えば、3H、14C、32P、35Sまたは125I）などで標識されていてもよく、アビジンと結合した標識を結合させるため、ビオチンで修飾されていてもよい。 The antibody may be modified depending on the measuring method. For example, enzymes (eg horseradish peroxidase, alkaline phosphatase, luciferase, β-galactosidase), fluorescent substances (eg fluorescein, rhodamine), radioactive isotopes (eg 3 H, 14 C, 32 P, 35 S or 125 I). ) Or the like, or may be modified with biotin in order to bind a label bound to avidin.
　免疫測定法には、直接法、間接法、競合法、サンドイッチ法などがある。各方法は、通常、以下のように実施される。直接法では、固相化した抗原を、標識した抗原特異的な抗体と接触させる。間接法では、固相化した抗原を、抗原特異的な一次抗体と接触させ、続いて一次抗体に特異的な標識した二次抗体と反応させる。サンドイッチ法では、固相化した抗原特異的な抗体を抗原と接触させ、続いて標識した（または標識との結合のため修飾した）別の抗原特異的な抗体を、固相化抗体に結合した抗原と接触させる。競合法では、固相化した抗原特異的な抗体を、測定対象の抗原および既知濃度の標識抗原と同時に接触させる。洗浄後、標識に基づいて、抗原抗体複合体を測定することができる。 Immunoassay methods include direct method, indirect method, competitive method, and sandwich method. Each method is usually carried out as follows. In the direct method, the immobilized antigen is brought into contact with a labeled antigen-specific antibody. In the indirect method, the immobilized antigen is brought into contact with an antigen-specific primary antibody, and subsequently reacted with a labeled secondary antibody specific to the primary antibody. In the sandwich method, an immobilized antigen-specific antibody is brought into contact with an antigen, and subsequently another labeled antigen-specific antibody (or modified for binding to the label) is bound to the immobilized antibody. Contact with antigen. In the competitive method, an immobilized antigen-specific antibody is brought into contact with an antigen to be measured and a labeled antigen having a known concentration at the same time. After washing, the antigen-antibody complex can be measured based on the label.
　ある実施形態において、preS1抗原と抗体との複合体は、ELISAにより測定される。さらなる実施形態において、preS1抗原と抗体との複合体は、サンドイッチELISAにより測定される。サンドイッチELISAの一実施形態では、使用する抗preS1抗体とは異なるエピトープに結合する抗HBs抗体をマイクロプレートなどの固相に固定し、対象から得た試料を添加する。洗浄後、固相に抗preS1抗体を添加し、標識に基づきpreS1抗原と抗preS1抗体との複合体を測定する。あるいは、抗preS1抗体を固相に固定化し、これとは異なるエピトープに結合する抗HBs抗体によって、抗原と抗preS1抗体との複合体を測定してもよい。 In one embodiment, the complex of preS1 antigen and antibody is measured by ELISA. In a further embodiment, the complex of preS1 antigen and antibody is measured by sandwich ELISA. In one embodiment of the sandwich ELISA, an anti-HBs antibody that binds to an epitope different from the anti-preS1 antibody used is immobilized on a solid phase such as a microplate and a sample obtained from the subject is added. After washing, anti-preS1 antibody is added to the solid phase, and the complex of preS1 antigen and anti-preS1 antibody is measured based on the label. Alternatively, the anti-preS1 antibody may be immobilized on a solid phase, and the complex between the antigen and the anti-preS1 antibody may be measured by an anti-HBs antibody that binds to an epitope different from this.
　ある実施形態において、本開示の方法は、本開示の抗preS1抗体と、抗S抗体または抗preS2抗体との組み合わせを用いる。さらなる実施形態において、本開示の方法は、本開示の抗preS1抗体と抗S抗体との組み合わせを用いる。例えば、本開示の抗preS1抗体を検出用抗体とし、抗S抗体を固相化抗体として、あるいは、本開示の抗preS1抗体を固相化抗体とし、抗S抗体を検出用抗体として、使用することができる。本開示の抗体のエピトープと離れた部位にあるエピトープを認識する抗体を用いることにより、精度のよい測定を行うことができる。抗S抗体としてはHyb-824およびHyb-5124A、抗preS2抗体としてはHyb-5520（いずれも株式会社特殊免疫研究所）などの市販の抗体を用いることができる。また、抗S抗体および抗preS2抗体は、SドメインまたはpreS2ドメインの全体または一部のアミノ酸配列を有するポリペプチドを免疫原として、本開示の記載にしたがい作製することができる。 In one embodiment, the method of the present disclosure uses a combination of the anti-preS1 antibody of the present disclosure and an anti-S antibody or an anti-preS2 antibody. In a further embodiment, the methods of this disclosure use a combination of anti-preS1 and anti-S antibodies of this disclosure. For example, the anti-preS1 antibody of the present disclosure is used as a detection antibody, the anti-S antibody is used as a solid phase antibody, or the anti-preS1 antibody of the present disclosure is used as a solid phase antibody and the anti-S antibody is used as a detection antibody. be able to. Accurate measurement can be performed by using an antibody that recognizes an epitope at a site distant from the epitope of the antibody of the present disclosure. As the anti-S antibody, commercially available antibodies such as Hyb-824 and Hyb-5124A, and as the anti-preS2 antibody, Hyb-5520 (both special immunoresearch institute) can be used. In addition, anti-S antibody and anti-preS2 antibody can be prepared according to the description of the present disclosure, using a polypeptide having the amino acid sequence of all or part of the S domain or preS2 domain as an immunogen.
　preS1抗原を測定することにより、HBV感染を評価することができる。preS1抗原の測定は、HBV感染について、感染性のあるHBV粒子と小型球形粒子とを区別できない抗HBs抗体を用いた方法や、HBe抗原もしくはHBcr抗原またはこれらに対する抗体、またはHBV DNA量を測定する方法とは異なる新たな情報を与えることができる。HBV感染の評価には、感染の有無の判定、HBV感染の病態の判定、HBV感染に対する治療効果の判定、HBV感染対象の予後の判定などが含まれる。HBV感染の評価結果は、HBV感染に対する治療の開始または中止もしくは中断に有用な情報を提供しうる。 HBV infection can be evaluated by measuring preS1 antigen. For preS1 antigen measurement, for HBV infection, a method using an anti-HBs antibody that cannot distinguish infectious HBV particles and small spherical particles, HBe antigen or HBcr antigen or antibody against them, or HBV DNA amount is measured. New information different from the method can be given. The evaluation of HBV infection includes the determination of the presence or absence of infection, the determination of the pathological condition of HBV infection, the determination of the therapeutic effect on HBV infection, the determination of the prognosis of HBV infected subjects, and the like. The results of assessing HBV infection may provide useful information for starting or stopping or discontinuing treatment for HBV infection.
　HBV感染の評価は、対象のpreS1抗原の測定値と基準値とを比較することにより行われる。基準値は、評価の目的に応じて適宜設定される。基準値は、対象から得た試料と同時に測定した別の試料の測定値であってもよく、事前の測定により設定された値であってもよい。比較には、標準的な統計学的方法を用いることができる。 Evaluation of HBV infection is performed by comparing the measured value of the target preS1 antigen with the reference value. The reference value is appropriately set according to the purpose of evaluation. The reference value may be a measurement value of another sample that is measured at the same time as the sample obtained from the subject, or may be a value set by a previous measurement. Standard statistical methods can be used for comparison.
　ある実施形態において、基準値は、preS1抗原を含まない試料（ネガティブコントロール）の測定値である。別の実施形態において、基準値は、HBVに感染していない複数の対象の測定値から設定された値（例えば、平均値）である。対象の測定値が基準値より高い場合（すなわち、preS1抗原が検出された場合）、対象はHBVに感染していると判定することができる。また、対象の測定値が高いほど、HBV感染が重症であると判定することができる。例えば、測定値が高いほど、対象における感染性のHBV粒子が多く、肝内のHBV cccDNA（covalently closed circular DNA）量が多いと判定することができ、治療中止後の再増悪の可能性が高いと判定しうる。あるいは、測定値が高いほど、HBs抗原の消失または持続性感染の消失の見込みが低いと判定しうる。 In one embodiment, the reference value is a measurement value of a sample (negative control) that does not contain preS1 antigen. In another embodiment, the reference value is a value (for example, an average value) set from the measured values of a plurality of subjects who are not infected with HBV. If the subject's measured value is higher than the reference value (ie, the preS1 antigen is detected), the subject can be determined to be infected with HBV. Also, the higher the subject's measurement, the more severe the HBV infection can be determined to be. For example, the higher the measured value, the more infectious HBV particles in the subject, and the greater the amount of HBVcc cDNA (covalently closed closed circular DNA) in the liver, the higher the possibility of re-exacerbation after discontinuation of treatment. Can be determined. Alternatively, the higher the measured value, the lower the likelihood of loss of HBs antigen or loss of persistent infection.
　さらなる実施形態において、基準値は、同じ対象の以前の測定値である。対象の測定値が基準値より高い場合、HBV感染が進行していると判定され、基準値より低い場合、HBV感染が改善していると判定される。 In a further embodiment, the reference value is a previous measurement of the same subject. If the subject's measured value is higher than the reference value, it is determined that the HBV infection is progressing, and if it is lower than the reference value, the HBV infection is determined to be improved.
　さらなる実施形態において、基準値は、HBV感染の所定の病態または予後の複数の対象の測定値から設定された値である。例えば、対象の測定値が基準値より高い場合、または低い場合に、対象が前記所定の病態または予後を有すると判定される。 In a further embodiment, the reference value is a value set from the measured values of a plurality of subjects having a predetermined condition or prognosis of HBV infection. For example, when the measured value of the subject is higher or lower than the reference value, it is determined that the subject has the predetermined disease state or prognosis.
　本開示の抗体は、preS1抗原の測定用組成物またはキット、およびHBV感染の評価用組成物またはキットに含まれていてもよい。測定用組成物は、本開示の抗体に加え、水、グリセロール、防腐剤、酸化防止剤、キレート剤、緩衝剤などを含んでよく、その形態は限定されず、例えば、液体であっても固体（例えば、凍結乾燥粉末）であってもよい。ある実施形態において、キットは、本開示の抗体に加え、抗S抗体または抗preS2抗体を含む。さらなる実施形態において、キットは、本開示の抗体に加え、抗S抗体を含む。例えば、本開示の抗preS1抗体を検出用抗体とし、抗S抗体を固相化抗体として、あるいは、本開示の抗preS1抗体を固相化抗体とし、抗S抗体を検出用抗体として、含むことができる。キットは、さらに、測定方法に応じた機器および／または試薬を含みうる。例えば、キットは、マイクロプレートなどの固相、標準抗原としてのpreS1抗原、反応用および／または希釈用緩衝液、発色試薬、反応停止液および使用説明書などを含んでよい。本開示の抗体または抗S抗体もしくは抗preS2抗体が、キット中の固相に結合していてもよい。 The antibody of the present disclosure may be contained in a composition or kit for measuring preS1 antigen, and a composition or kit for evaluating HBV infection. The composition for measurement may contain water, glycerol, a preservative, an antioxidant, a chelating agent, a buffering agent and the like in addition to the antibody of the present disclosure, and the form thereof is not limited, and for example, liquid or solid (Eg freeze-dried powder). In certain embodiments, the kit comprises an anti-S antibody or an anti-preS2 antibody in addition to the antibody of the present disclosure. In a further embodiment, the kit comprises an anti-S antibody in addition to the antibodies of this disclosure. For example, the anti-preS1 antibody of the present disclosure as a detection antibody, the anti-S antibody as a solid phase antibody, or the anti-preS1 antibody of the present disclosure as a solid phase antibody, the anti-S antibody as a detection antibody, You can The kit may further include equipment and/or reagents depending on the measuring method. For example, the kit may include a solid phase such as a microplate, preS1 antigen as a standard antigen, a reaction and/or dilution buffer, a coloring reagent, a reaction stopping solution, and an instruction manual. The antibody of the present disclosure or the anti-S antibody or anti-preS2 antibody may be bound to the solid phase in the kit.
３．医薬組成物
　本開示の抗体は、医薬組成物の有効成分として使用することができる。ある実施形態において、本開示の抗体は、HBV感染の治療または予防に用いられる。HBV感染の治療には、HBV感染による症状または疾患の治療、例えば慢性または急性B型肝炎および肝硬変の治療が含まれる。HBV感染の予防には、HBVの汚染針の受針事故後の感染予防、輸血または臓器移植の後の感染予防などが含まれる。 3. Pharmaceutical Composition The antibody of the present disclosure can be used as an active ingredient of a pharmaceutical composition. In certain embodiments, the antibodies of the present disclosure are used to treat or prevent HBV infection. Treatment of HBV infection includes treatment of symptoms or diseases due to HBV infection, such as treatment of chronic or acute hepatitis B and cirrhosis. Prevention of HBV infection includes prevention of infection after the accident of receiving HBV contaminated needles and prevention of infection after blood transfusion or organ transplantation.
　本開示の抗体は、所望の効果（例えば、HBV感染の治療または予防）を発揮しうる量（本明細書において、有効量という）で対象に投与される。抗体の投与量は、投与方法、投与対象の年齢、体重、健康状態等によって適宜選択される。例えば、成人１日あたり、10 μg/kg～100 mg/kg、100 μg/kg～10 mg/kg、または1 mg/kg～10 mg/kgを、連日、または数日、１週間もしくは数週間に１回、投与することができるが、これに限定されない。抗体の投与方法も、投与対象の年齢、体重、健康状態等によって適宜選択される。投与方法は、経口投与であっても非経口投与であってもよいが、非経口投与が好ましい。非経口投与としては、皮下投与、皮内投与、筋肉内投与、静脈内投与などが挙げられるが、静脈内投与によることが好ましい。 The antibody of the present disclosure is administered to a subject in an amount capable of exerting a desired effect (for example, treatment or prevention of HBV infection) (herein, referred to as an effective amount). The dose of the antibody is appropriately selected depending on the administration method, age, weight and health of the subject. For example, 10 μg/kg to 100 mg/kg, 100 μg/kg to 10 mg/kg, or 1 mg/kg to 10 mg/kg per day for an adult, consecutive days, days, weeks, or weeks However, the present invention is not limited to this. The administration method of the antibody is also appropriately selected depending on the age, weight, health condition, etc. of the administration subject. The administration method may be oral administration or parenteral administration, but parenteral administration is preferred. Parenteral administration includes subcutaneous administration, intradermal administration, intramuscular administration, intravenous administration and the like, but intravenous administration is preferable.
　医薬組成物は、常法により製剤化することができる。医薬組成物は、抗体に加えて、滅菌水、生理食塩水、安定剤、賦形剤、酸化防止剤、緩衝剤、防腐剤、界面活性剤、キレート剤、結合剤などの、医薬上許容される担体または添加剤を含むことができる。 The pharmaceutical composition can be formulated by a conventional method. In addition to the antibody, the pharmaceutical composition includes pharmaceutically acceptable sterilized water, physiological saline, stabilizers, excipients, antioxidants, buffers, preservatives, surfactants, chelating agents, binders and the like. Carrier or additive.
　ある実施形態において、医薬組成物は、配列番号２のアミノ酸配列に含まれるエピトープに結合する抗体；または、（ａ）、（ａ'）、（ａ’'）、（ａ’’’）または（ａ’’’’）の抗体を含む。別の実施形態において、医薬組成物は、配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体；または、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体を含む。別の実施形態において、医薬組成物は、配列番号４のアミノ酸配列に含まれるエピトープに結合する抗体；または、（ｉ）、（ｉ'）、（ｉ’'）、（ｉ’’’）または（ｉ’’’’）の抗体を含む。 In certain embodiments, the pharmaceutical composition is an antibody that binds to an epitope comprised in the amino acid sequence of SEQ ID NO: 2; or (a), (a'), (a''), (a''') or (a a''''). In another embodiment, the pharmaceutical composition is an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3; or (b)-(h), (b′)-(h′), (b″) )-(H''), (b''')-(h''') or (b'''')-(h''''). In another embodiment, the pharmaceutical composition is an antibody that binds to an epitope comprised in the amino acid sequence of SEQ ID NO: 4; or (i), (i′), (i″), (i″′) or (I″″) antibody is included.
　医薬組成物の有効成分として、2種以上の本開示の抗体を併用してもよい。ある実施形態において、前記2種以上の本開示の抗体は、異なるエピトープに結合する抗体を含む。さらなる実施形態において、前記2種以上の本開示の抗体は、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体；または、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体と、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体とを含む。さらなる実施形態において、前記2種以上の本開示の抗体は、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と、配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体；または、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体と、（ｉ）、（ｉ'）、（ｉ’'）、（ｉ’’’）または（ｉ’’’’）の抗体とを含む。さらなる実施形態において、前記2種以上の本開示の抗体は、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と、配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体；または、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体と、（ｉ）、（ｉ'）、（ｉ’'）、（ｉ’’’）または（ｉ’’’’）の抗体とを含む。本開示の抗体は、本開示の抗体とは異なる抗HBs抗体と併用してもよく、他のHBV感染治療薬と併用してもよい。他の治療薬としては、インターフェロン（例えば、ペグインターフェロンα－２ａ）および核酸アナログ（例えば、ラミブジン、アデホビル、エンテカビル、テノホビル）が挙げられる。本明細書において、2種以上の有効成分を併用する場合、その全てまたは一部の有効成分が同一の組成物に含まれていてもよく、全ての薬剤が別の組成物に含まれていてもよい。また、2種以上の有効成分の投与スケジュールは同じであっても異なっていても良い。 Two or more kinds of the antibodies of the present disclosure may be used in combination as an active ingredient of a pharmaceutical composition. In certain embodiments, the two or more antibodies of the present disclosure include antibodies that bind different epitopes. In a further embodiment, the two or more antibodies of the present disclosure bind to an epitope contained in the amino acid sequence of SEQ ID NO: 2 and one antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3. Or more antibodies; or (a), (a'), (a''), (a''') or (a'''') antibody, and (b)-(h), (b) From') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') to (h'''') And one or more antibodies of choice. In a further embodiment, the two or more antibodies of the present disclosure bind to an epitope contained in the amino acid sequence of SEQ ID NO:2, and 1 bind to an epitope contained in the amino acid sequence of SEQ ID NO:4. Or more than one antibody; or (a), (a'), (a''), (a''') or (a'''') antibody, and (i), (i'), ( i″), (i′″) or (i″″) antibody. In a further embodiment, the two or more antibodies of the present disclosure bind to an epitope contained in the amino acid sequence of SEQ ID NO:3 and one antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO:4. Or more than one antibody; or (b) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') Or at least one antibody selected from (b″″) to (h″″) and (i), (i′), (i″), (i″′) or ( i″″) antibody. The antibody of the present disclosure may be used in combination with an anti-HBs antibody different from the antibody of the present disclosure, or may be used in combination with other therapeutic agents for HBV infection. Other therapeutic agents include interferons (eg, peginterferon α-2a) and nucleic acid analogs (eg, lamivudine, adefovir, entecavir, tenofovir). In the present specification, when two or more active ingredients are used in combination, all or some of the active ingredients may be contained in the same composition, and all drugs are contained in different compositions. Good. In addition, the administration schedules of two or more active ingredients may be the same or different.
　本開示の例示的実施形態を以下に記載する。 Example embodiments of the present disclosure are described below.
［１］
　抗preS1抗体であって、配列番号２、３および４から選択されるアミノ酸配列に含まれるエピトープに結合する、抗体。
［２］
　配列番号２のアミノ酸配列に含まれるエピトープに結合する、前記１に記載の抗体。
［３］
　配列番号２のアミノ酸配列に含まれるエピトープに結合する抗体が、配列番号８０のアミノ酸配列に含まれるエピトープに結合する抗体である、前記２に記載の抗体。
［４］
　配列番号３のアミノ酸配列に含まれるエピトープに結合する、前記１に記載の抗体。
［５］
　配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体が、配列番号１７または配列番号２１のアミノ酸配列に含まれるエピトープに結合する抗体である、前記４に記載の抗体。
［６］
　以下の（ａ）～（ｉ）から選択される、前記１～５のいずれかに記載の抗体：
（ａ）配列番号２４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号２６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号２８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｂ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｃ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｄ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｅ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｆ）配列番号５０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号５４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｇ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｈ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；および
（ｉ）配列番号６５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号６９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号７０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号７１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体。
［７］
　抗preS1抗体であって、以下の（ａ）～（ｉ）から選択される、抗体：
（ａ）配列番号２４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号２６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号２８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号２９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｂ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｃ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｄ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｅ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号４７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｆ）配列番号５０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号５４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号５５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｇ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｈ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；および
（ｉ）配列番号６５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号６６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号６７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号６９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
　配列番号７０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
　配列番号７１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体。
［８］
　抗体が、以下の（ａ’）～（ｉ’）のいずれかの抗体である、前記１～７のいずれかに記載の抗体：
（ａ’）配列番号２４のアミノ酸配列を含むCDR1、
　配列番号２５のアミノ酸配列を含むCDR2、および
　配列番号２６のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号２８のアミノ酸配列を含むCDR1、
　配列番号２９のアミノ酸配列を含むCDR2、および
　配列番号３０のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｂ’）配列番号３２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号３７のアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｃ’）配列番号３２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号３４のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｄ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号４３のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列を含むCDR2、および
　配列番号４５のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｅ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号４７のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号４０のアミノ酸配列を含むCDR2、および
　配列番号３８のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｆ’）配列番号５０のアミノ酸配列を含むCDR1、
　配列番号５１のアミノ酸配列を含むCDR2、および
　配列番号５２のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号５４のアミノ酸配列を含むCDR1、
　配列番号５５のアミノ酸配列を含むCDR2、および
　配列番号５６のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｇ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号６０のアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；
（ｈ’）配列番号４２のアミノ酸配列を含むCDR1、
　配列番号３３のアミノ酸配列を含むCDR2、および
　配列番号５８のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号３６のアミノ酸配列を含むCDR1、
　配列番号６３のアミノ酸配列を含むCDR2、および
　配列番号６１のアミノ酸配列を含むCDR3
　を含む重鎖可変領域を含む抗体；および
（ｉ’）配列番号６５のアミノ酸配列を含むCDR1、
　配列番号６６のアミノ酸配列を含むCDR2、および
　配列番号６７のアミノ酸配列を含むCDR3
　を含む軽鎖可変領域、および、
　配列番号６９のアミノ酸配列を含むCDR1、
　配列番号７０のアミノ酸配列を含むCDR2、および
　配列番号７１のアミノ酸配列を含むCDR3。
［９］
　抗体が、以下の（ａ’’）～（ｉ’’）のいずれかの抗体である、前記１～８のいずれかに記載の抗体：
（ａ’’）配列番号２３のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号２３のアミノ酸配列もしくは配列番号２３のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号２７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号２７のアミノ酸配列もしくは配列番号２７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｂ’’）配列番号３１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３１のアミノ酸配列もしくは配列番号３１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号３５のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３５のアミノ酸配列もしくは配列番号３５のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｃ’’）配列番号３１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３１のアミノ酸配列もしくは配列番号３１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号３９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３９のアミノ酸配列もしくは配列番号３９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｄ’’）配列番号４１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４１のアミノ酸配列もしくは配列番号４１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号４４のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４４のアミノ酸配列もしくは配列番号４４のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｅ’’）配列番号４６のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４６のアミノ酸配列もしくは配列番号４６のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号４８のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４８のアミノ酸配列もしくは配列番号４８のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｆ’’）配列番号４９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４９のアミノ酸配列もしくは配列番号４９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号５３のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５３のアミノ酸配列もしくは配列番号５３のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｇ’’）配列番号５７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５７のアミノ酸配列もしくは配列番号５７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号５９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５９のアミノ酸配列もしくは配列番号５９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｈ’’）配列番号５７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５７のアミノ酸配列もしくは配列番号５７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号６２のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６２のアミノ酸配列もしくは配列番号６２のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；および
（ｉ’’）配列番号６４のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６４のアミノ酸配列もしくは配列番号６４のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
　配列番号６８のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６８のアミノ酸配列もしくは配列番号６８のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体。
［１０］
　抗体が、以下の（ａ’’’）～（ｉ’’’）のいずれかの抗体である、前記１～９のいずれかに記載の抗体：
（ａ’’’）配列番号２３のアミノ酸配列を含む軽鎖可変領域、および配列番号２７のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｂ’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域、および配列番号３５のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｃ’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域、および配列番号３９のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｄ’’’）配列番号４１のアミノ酸配列を含む軽鎖可変領域、および配列番号４４のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｅ’’’）配列番号４６のアミノ酸配列を含む軽鎖可変領域、および配列番号４８のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｆ’’’）配列番号４９のアミノ酸配列を含む軽鎖可変領域、および配列番号５３のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｇ’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域、および配列番号５９のアミノ酸配列を含む重鎖可変領域を含む抗体；
（ｈ’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域、および配列番号６２のアミノ酸配列を含む重鎖可変領域を含む抗体；および
（ｉ’’’）配列番号６４のアミノ酸配列を含む軽鎖可変領域、および配列番号６８のアミノ酸配列を含む重鎖可変領域を含む抗体。
［１１］
　抗体が、以下の（ａ’’’’）～（ｉ’’’’）のいずれかの抗体である、前記１～１０のいずれかに記載の抗体：
（ａ’’’’）配列番号２３のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号２７のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｂ’’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号３５のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｃ’’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号３９のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｄ’’’’）配列番号４１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号４４のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｅ’’’’）配列番号４６のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号４８のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｆ’’’’）配列番号４９のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号５３のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｇ’’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号５９のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
（ｈ’’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号６２のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；および
（ｉ’’’’）配列番号６４のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号６８のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体。
［１２］
　preS1抗原への結合において前記７～１１のいずれかに記載の抗体と競合する、抗preS1抗体。
［１３］
　preS1抗原への結合において前記８、１０、または１１に記載の抗体と競合する、前記１２に記載の抗preS1抗体。
［１４］
　preS1抗原への結合において前記８または１０に記載の抗体と競合する、前記１２に記載の抗preS1抗体。
［１５］
　preS1抗原への結合において前記１０に記載の抗体と競合する、前記１２に記載の抗preS1抗体。 [1]
An anti-preS1 antibody, which binds to an epitope contained in an amino acid sequence selected from SEQ ID NOs: 2, 3 and 4.
[2]
The antibody according to 1 above, which binds to an epitope contained in the amino acid sequence of SEQ ID NO: 2.
[3]
3. The antibody according to 2 above, wherein the antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 2 is an antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 80.
[4]
The antibody according to 1 above, which binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
[5]
5. The antibody according to 4 above, wherein the antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 3 is an antibody that binds to the epitope included in the amino acid sequence of SEQ ID NO: 17 or SEQ ID NO: 21.
[6]
The antibody according to any one of 1 to 5 above, which is selected from the following (a) to (i):
(A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 28 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 30 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(B) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 37 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(C) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(D) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence have been deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 43 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 45 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(E) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and the amino acid sequence of SEQ ID NO: 47 or 1 to 3 in said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(F) CDR1 comprising the amino acid sequence of SEQ ID NO: 50 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 51 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 52 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 54 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:55 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:56 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(G) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 60 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 61 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(H) a CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence;
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:63 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:61 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising: (i) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in said amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 67 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 70 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 71 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
[7]
Anti-preS1 antibody, which is selected from the following (a) to (i):
(A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 28 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 30 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(B) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 37 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(C) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(D) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence have been deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 43 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 45 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(E) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and the amino acid sequence of SEQ ID NO: 47 or 1 to 3 in said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(F) CDR1 comprising the amino acid sequence of SEQ ID NO: 50 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 51 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 52 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 54 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:55 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:56 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(G) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO: 60 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 61 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
(H) a CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence;
CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
CDR2 comprising the amino acid sequence of SEQ ID NO:63 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:61 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising: (i) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in said amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 67 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
CDR2 comprising the amino acid sequence of SEQ ID NO: 70 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 71 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
An antibody comprising a heavy chain variable region comprising:
[8]
The antibody according to any one of 1 to 7 above, wherein the antibody is any one of the following (a′) to (i′):
(A') CDR1 comprising the amino acid sequence of SEQ ID NO: 24,
CDR2 containing the amino acid sequence of SEQ ID NO:25, and CDR3 containing the amino acid sequence of SEQ ID NO:26
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 28,
CDR2 containing the amino acid sequence of SEQ ID NO: 29, and CDR3 containing the amino acid sequence of SEQ ID NO: 30
An antibody comprising a heavy chain variable region comprising:
(B') CDR1 containing the amino acid sequence of SEQ ID NO: 32,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:37, and CDR3 containing the amino acid sequence of SEQ ID NO:38
An antibody comprising a heavy chain variable region comprising:
(C′) CDR1 containing the amino acid sequence of SEQ ID NO: 32,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:38
An antibody comprising a heavy chain variable region comprising:
(D') CDR1 containing the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:43
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:45
An antibody comprising a heavy chain variable region comprising:
(E′) CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:47
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:38
An antibody comprising a heavy chain variable region comprising:
(F') CDR1 comprising the amino acid sequence of SEQ ID NO: 50,
CDR2 containing the amino acid sequence of SEQ ID NO:51 and CDR3 containing the amino acid sequence of SEQ ID NO:52
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 54,
CDR2 containing the amino acid sequence of SEQ ID NO:55, and CDR3 containing the amino acid sequence of SEQ ID NO:56
An antibody comprising a heavy chain variable region comprising:
(G') CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:58
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:60, and CDR3 containing the amino acid sequence of SEQ ID NO:61
An antibody comprising a heavy chain variable region comprising:
(H') CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:58
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
CDR2 containing the amino acid sequence of SEQ ID NO:63, and CDR3 containing the amino acid sequence of SEQ ID NO:61
An antibody comprising a heavy chain variable region comprising; and (i′) CDR1 comprising the amino acid sequence of SEQ ID NO:65,
CDR2 containing the amino acid sequence of SEQ ID NO:66, and CDR3 containing the amino acid sequence of SEQ ID NO:67
A light chain variable region containing, and
CDR1, which comprises the amino acid sequence of SEQ ID NO: 69,
CDR2 containing the amino acid sequence of SEQ ID NO:70, and CDR3 containing the amino acid sequence of SEQ ID NO:71.
[9]
The antibody according to any one of 1 to 8 above, wherein the antibody is any one of the following (a″) to (i″):
(A'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 23, or the amino acid sequence of SEQ ID NO: 23 or the amino acid sequence of SEQ ID NO: 23 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 27. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 27 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 27, An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
(B'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 35. Having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 35, or 1-20 in the amino acid sequence of SEQ ID NO: 35, 1-10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
(C'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 39. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 39 or 1 to 20 in the amino acid sequence of SEQ ID NO: 39, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
(D'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 41, or the amino acid sequence of SEQ ID NO: 41 or the amino acid sequence of SEQ ID NO: 41 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues are deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 44. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 44 or 1 to 20 in the amino acid sequence of SEQ ID NO: 44, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(E'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 46, or the amino acid sequence of SEQ ID NO: 46 or the amino acid sequence of SEQ ID NO: 46 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 48 Amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 48 or 1 to 20 in the amino acid sequence of SEQ ID NO: 48, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(F″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 49, or the amino acid sequence of SEQ ID NO: 49 or the amino acid sequence of SEQ ID NO: 49 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 53 With 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 53 or the amino acid sequence of SEQ ID NO: 53 with 1-20, 1-10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(G″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 59. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 59 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 59, An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
(H″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 62. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 62 or 1 to 20 in the amino acid sequence of SEQ ID NO: 62, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which ˜5, or 1-3 amino acid residues have been deleted, substituted, or added; and (i″) the amino acid sequence of SEQ ID NO: 64 and 80% , 85%, 90%, or 95% or more sequence identity, or 1-20, 1-10, 1-5 in the amino acid sequence of SEQ ID NO: 64 or the amino acid sequence of SEQ ID NO: 64 , Or a light chain variable region comprising an amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted, or added, and 80%, 85%, 90%, or 95% or more of the amino acid sequence of SEQ ID NO: 68 An amino acid sequence having the sequence identity of, or 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues in the amino acid sequence of SEQ ID NO:68 or the amino acid sequence of SEQ ID NO:68. An antibody comprising a heavy chain variable region comprising a deleted, substituted, or added amino acid sequence.
[10]
The antibody according to any one of 1 to 9 above, wherein the antibody is any one of the following (a′″) to (i′″):
(A′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27;
(B''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35;
(C′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39;
(D′″) an antibody comprising a light chain variable region containing the amino acid sequence of SEQ ID NO: 41 and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44;
(E′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:46 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:48;
(F″′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:49 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:53;
(G''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:57 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:59;
(H″′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:57 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:62; and (i″′) an amino acid sequence of SEQ ID NO:64. An antibody comprising a light chain variable region comprising, and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:68.
[11]
The antibody according to any one of 1 to 10 above, wherein the antibody is any of the following (a″″) to (i″″):
(A″″) CDR1, CDR2 in the light chain variable region including the amino acid sequence of SEQ ID NO:23, and CDR1, CDR2 in the heavy chain variable region including the amino acid sequence of SEQ ID NO:27 And an antibody comprising a heavy chain variable region comprising CDR3;
(B″″) CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:35 and the light chain variable region containing CDR1, CDR2, and CDR3 in the light chain variable region of SEQ ID NO:31 And an antibody comprising a heavy chain variable region comprising CDR3;
(C″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 31 and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 39 And an antibody comprising a heavy chain variable region comprising CDR3;
(D″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 41, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44 And an antibody comprising a heavy chain variable region comprising CDR3;
(E″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO:46, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:48 And an antibody comprising a heavy chain variable region comprising CDR3;
(F″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO:49, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:53 And an antibody comprising a heavy chain variable region comprising CDR3;
(G'''') CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 57, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 59 And an antibody comprising a heavy chain variable region comprising CDR3;
(H″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 57, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 62 And (i″″) a light chain variable region comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 64, and a light chain variable region comprising CDR3, CDR2, and CDR3, and SEQ ID NO: An antibody comprising a heavy chain variable region comprising CDR1, CDR2, and CDR3 in a heavy chain variable region comprising 68 amino acid sequences.
[12]
An anti-preS1 antibody that competes with the antibody according to any one of 7 to 11 above for binding to a preS1 antigen.
[13]
The anti-preS1 antibody according to the above 12, which competes with the antibody according to the above 8, 10, or 11 for binding to the preS1 antigen.
[14]
The anti-preS1 antibody according to the above 12, which competes with the antibody according to the above 8 or 10 for binding to a preS1 antigen.
[15]
13. The anti-preS1 antibody according to above 12, which competes with the antibody according to 10 above for binding to a preS1 antigen.
［１６］
　２種以上の抗preS1抗体の組み合わせであって、前記２種以上の抗preS1抗体が前記１～１５のいずれかに記載の抗体から独立に選択される、組み合わせ。
［１７］
　２種以上の抗preS1抗体が、
　配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体；
　配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体；または
　配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体、
　を含む、前記１６に記載の組み合わせ。
［１８］
　２種以上の抗preS1抗体が、
　配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体
　を含む、前記１７に記載の組み合わせ。
［１９］
　２種以上の抗preS1抗体が、
　配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体
　を含む、前記１７に記載の組み合わせ。
［２０］
　２種以上の抗ｐｒｅＳ１抗体が、
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体、および、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体；
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体、および、（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体；または
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体、および、（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体
　を含む、前記１７に記載の組み合わせ。
［２１］
　２種以上の抗ｐｒｅＳ１抗体が、
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体、および、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体
　を含む、前記１８に記載の組み合わせ。
［２２］
　２種以上の抗ｐｒｅＳ１抗体が、
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体、および、（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体
　を含む、前記１９に記載の組み合わせ。
［２３］
　２種以上の抗ｐｒｅＳ１抗体が、
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および（ｂ）、（ｂ’）、（ｂ’’）、（ｂ’’’）または（ｂ’’’’）の抗体；
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体；
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および（ｇ）、（ｇ’）、（ｇ’’）、（ｇ’’’）または（ｇ’’’’）の抗体；
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体；または
　（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体および（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体
　を含む、前記２０に記載の組み合わせ。 [16]
A combination of two or more anti-preS1 antibodies, wherein the two or more anti-preS1 antibodies are independently selected from the antibodies described in any of 1 to 15 above.
[17]
Two or more anti-preS1 antibodies
One or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3;
One or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4; or to the epitope contained in the amino acid sequence of SEQ ID NO:3 One or more antibodies that bind and one or more antibodies that bind to an epitope contained in the amino acid sequence of SEQ ID NO:4,
17. The combination according to 16 above, which comprises:
[18]
Two or more anti-preS1 antibodies
18. The combination according to the above 17, comprising one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 2 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 3.
[19]
Two or more anti-preS1 antibodies
18. The combination according to the above 17, comprising one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3 and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4.
[20]
Two or more anti-preS1 antibodies
(A), (a'), (a''), (a''') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h''''). Antibody;
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') ) To (h″″), and one or more antibodies selected from (i), (i′), (i″), (i′″) or (i″″) Or (a), (a'), (a''), (a''') or (a'''') antibody, and (i), (i'), (i' 18. The combination according to 17 above, which comprises the antibody'), (i''') or (i'''').
[21]
Two or more anti-preS1 antibodies
(A), (a'), (a''), (a''') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h''''). 19. The combination according to 18 above, which comprises the antibody of
[22]
Two or more anti-preS1 antibodies
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') ) To (h″″), and one or more antibodies selected from (i), (i′), (i″), (i′″) or (i″″) 20. The combination according to above 19, which comprises the antibody of
[23]
Two or more anti-preS1 antibodies
(A), (a'), (a''), (a''') or (a'''') antibody and (b), (b'), (b''), (b' )) or (b'''') antibody;
(A), (a'), (a''), (a''') or (a'''') antibody and (c), (c'), (c''), (c''') or (c'''') antibody;
(A), (a'), (a''), (a''') or (a'''') antibody and (g), (g'), (g''), (g''') or (g'''') antibody;
(A), (a′), (a″), (a′″) or (a″″) antibody and (i), (i′), (i″), (i′) '') or (i'''') antibody; or (c), (c'), (c''), (c''') or (c'''') antibody and (i) 21. The combination according to the above 20, which comprises the antibody of (i′), (i″), (i′″) or (i″″).
［２４］
　preS1抗原の測定方法であって、
　対象から得た試料を前記１～１５のいずれかに記載の抗体と接触させること、および
　preS1抗原と前記抗体との複合体を検出すること
　を含む方法。
［２５］
　抗体が、配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体である、前記２４に記載の方法。
［２６］
　抗体が、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される抗体である、前記２４に記載の方法。
［２７］
　preS1抗原と前記抗体との複合体をELISAにより検出する、前記２４～２６のいずれかに記載の方法。
［２８］
　前記１～１５のいずれかに記載の抗体と、これとは異なるエピトープに結合する抗HBs抗体とを用いる、前記２４～２７のいずれかに記載の方法。
［２９］
　抗HBs抗体が、抗S抗体または抗preS2抗体である、前記２８に記載の方法。 [24]
A method for measuring preS1 antigen, comprising:
16. A method comprising contacting a sample obtained from a subject with the antibody according to any one of 1 to 15 above, and detecting a complex of preS1 antigen and the antibody.
[25]
25. The method according to 24 above, wherein the antibody binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
[26]
The antibody is (b) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b' 25. The method according to 24 above, which is an antibody selected from “″) to (h″″).
[27]
27. The method according to any of 24 to 26 above, wherein the complex of preS1 antigen and the antibody is detected by ELISA.
[28]
The method according to any one of 24 to 27 above, wherein the antibody according to any one of 1 to 15 above and an anti-HBs antibody that binds to an epitope different from this antibody are used.
[29]
29. The method according to 28 above, wherein the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody.
［３０］
　B型肝炎ウイルス感染の評価方法であって、
　前記２４～２９のいずれかに記載の方法によりpreS1抗原を測定すること、および
　前記測定値を基準値と比較すること
　を含む方法。
［３１］
　有効量の前記１～２３のいずれかに記載の抗体または組み合わせを対象に投与することをさらに含む、前記３０に記載の方法。 [30]
A method for evaluating hepatitis B virus infection, comprising:
A method comprising measuring the preS1 antigen by the method according to any one of the above 24 to 29, and comparing the measured value with a reference value.
[31]
31. The method of claim 30, further comprising administering to the subject an effective amount of the antibody or combination of any of claims 1-23.
［３２］
　前記１～２３のいずれかに記載の抗体または組み合わせを含む、preS1抗原の測定用組成物。
［３３］
　前記１～２３のいずれかに記載の抗体または組み合わせを含む、B型肝炎ウイルス感染の評価用組成物。
［３４］
　配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体を含む、前記３２または３３に記載の組成物。
［３５］
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される抗体を含む、前記３２または３３に記載の組成物。 [32]
A composition for measuring preS1 antigen, which comprises the antibody or the combination according to any one of 1 to 23 above.
[33]
A composition for evaluating hepatitis B virus infection, which comprises the antibody or the combination according to any one of 1 to 23 above.
[34]
34. The composition according to the above 32 or 33, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
[35]
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') 34. The composition according to 32 or 33 above, which comprises an antibody selected from () to (h″″).
［３６］
　前記１～２３のいずれかに記載の抗体または組み合わせを含む、preS1抗原の測定用キット。
［３７］
　前記１～２３のいずれかに記載の抗体または組み合わせを含む、B型肝炎ウイルス感染の評価用キット。
［３８］
　配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体を含む、前記３６または３７に記載のキット。
［３９］
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される抗体を含む、前記３６または３７に記載のキット。
［４０］
　前記１～１５のいずれかに記載の抗体とは異なるエピトープに結合する抗HBs抗体をさらに含む、前記３６～３９のいずれかに記載のキット。
［４１］
　抗HBs抗体が、抗S抗体または抗preS2抗体である、前記４０に記載のキット。 [36]
A kit for measuring preS1 antigen, which comprises the antibody or the combination according to any one of 1 to 23 above.
[37]
A kit for evaluating hepatitis B virus infection, which comprises the antibody or the combination according to any one of 1 to 23 above.
[38]
38. The kit according to 36 or 37 above, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
[39]
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') 38.) The kit according to 36 or 37 above, which comprises an antibody selected from (h″″).
[40]
40. The kit according to any of 36 to 39, further comprising an anti-HBs antibody that binds to an epitope different from the antibody according to any of 1 to 15 above.
[41]
The kit according to 40, wherein the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody.
［４２］
　前記１～２３のいずれかに記載の抗体または組み合わせを含む、医薬組成物。
［４３］
　配列番号２のアミノ酸配列に含まれるエピトープに結合する抗体を含む、前記４２に記載の医薬組成物。
［４４］
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体を含む、前記４２に記載の医薬組成物。
［４５］
　配列番号３のアミノ酸配列に含まれるエピトープに結合する抗体を含む、前記４２に記載の医薬組成物。
［４６］
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体を含む、前記４２に記載の医薬組成物。
［４７］
　配列番号４のアミノ酸配列に含まれるエピトープに結合する抗体を含む、前記４２に記載の医薬組成物。
［４８］
　（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体を含む、前記４２に記載の医薬組成物。
［４９］
　配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および／または配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および／または配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む、；
　配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および／または配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および／または配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む；または
　配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および／または配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体および／または配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む、
　前記４２に記載の医薬組成物。
［５０］
　配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む；または
　配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号２のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む、
　前記４９に記載の医薬組成物。
［５１］
　配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む；または
　配列番号４のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体を含み、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体と併用されるか、配列番号３のアミノ酸配列に含まれるエピトープに結合する１種以上の抗体をさらに含む、
　前記４９に記載の医薬組成物。
［５２］
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体を含み、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体および／または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体と併用されるか、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体および／または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体をさらに含む；
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および／または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および／または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体をさらに含む；または、
　（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および／または（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体および／または（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体をさらに含む、
　前記４２に記載の医薬組成物。
［５３］
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体を含み、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体と併用されるか、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体をさらに含む；または
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体をさらに含む、
　前記５２に記載の医薬組成物。
［５４］
　（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体を含み、（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体と併用されるか、（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体をさらに含む；または、
　（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体を含み、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体と併用されるか、（ｂ）～（ｈ）、（ｂ’）～（ｈ’）、（ｂ’’）～（ｈ’’）、（ｂ’’’）～（ｈ’’’）または（ｂ’’’’）～（ｈ’’’’）から選択される１種以上の抗体をさらに含む、
　前記５２に記載の医薬組成物。
［５５］
　（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体を含み、（ｂ）、（ｂ’）、（ｂ’’）、（ｂ’’’）または（ｂ’’’’）の抗体、（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体、（ｇ）、（ｇ’）、（ｇ’’）、（ｇ’’’）または（ｇ’’’’）の抗体、または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体と併用されるか、（ｂ）、（ｂ’）、（ｂ’’）、（ｂ’’’）または（ｂ’’’’）の抗体、（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体、（ｇ）、（ｇ’）、（ｇ’’）、（ｇ’’’）または（ｇ’’’’）の抗体、または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体をさらに含む；
　（ｂ）、（ｂ’）、（ｂ’’）、（ｂ’’’）または（ｂ’’’’）の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体をさらに含む；
　（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体または（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体をさらに含む；
　（ｇ）、（ｇ’）、（ｇ’’）、（ｇ’’’）または（ｇ’’’’）の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体をさらに含む；または
　（ｉ）、（ｉ’）、（ｉ’’）、（ｉ’’’）または（ｉ’’’’）の抗体を含み、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体または（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体と併用されるか、（ａ）、（ａ’）、（ａ’’）、（ａ’’’）または（ａ’’’’）の抗体または（ｃ）、（ｃ’）、（ｃ’’）、（ｃ’’’）または（ｃ’’’’）の抗体をさらに含む、
　前記５２に記載の医薬組成物。
［５６］
　前記１～１５のいずれかに記載の抗体とは異なるエピトープに結合する抗HBs抗体と併用されるか、前記抗HBs抗体をさらに含む、前記４２～５５のいずれかに記載の医薬組成物。
［５７］
　抗HBs抗体が、抗S抗体または抗preS2抗体である、前記５６に記載の医薬組成物。
［５８］
　B型肝炎ウイルス感染の治療または予防のための、前記４２～５７のいずれかに記載の医薬組成物。
［５９］
　慢性B型肝炎の治療のための、前記４２～５８のいずれかに記載の医薬組成物。 [42]
A pharmaceutical composition comprising the antibody or the combination according to any one of 1 to 23 above.
[43]
43. The pharmaceutical composition according to the above 42, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 2.
[44]
43. The pharmaceutical composition according to the above 42, which comprises the antibody of (a), (a'), (a''), (a''') or (a'''').
[45]
43. The pharmaceutical composition according to the above 42, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 3.
[46]
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') 43.) The pharmaceutical composition according to the above 42, which comprises one or more antibodies selected from (h″″).
[47]
43. The pharmaceutical composition according to the above 42, which comprises an antibody that binds to an epitope contained in the amino acid sequence of SEQ ID NO: 4.
[48]
43. The pharmaceutical composition according to the above 42, which comprises the antibody of (i), (i'), (i''), (i''') or (i'''').
[49]
Includes one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2, and contains one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3 and/or included in the amino acid sequence of SEQ ID NO:4 Used in combination with one or more antibodies that bind to the epitope described above, or that bind to one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3 and/or bind to the epitope contained in the amino acid sequence of SEQ ID NO:4. Further comprising one or more antibodies;
One or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3, one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2, and/or the amino acid sequence of SEQ ID NO:4 Used in combination with one or more antibodies that bind to the epitope described above, or that bind to one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2 and/or bind to the epitope contained in the amino acid sequence of SEQ ID NO:4. Or one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4, and one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2, and / Or one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 3 or are used in combination with one or more antibodies and/or SEQ ID NO: 3 Further comprising one or more antibodies that bind to an epitope contained in the amino acid sequence of
43. The pharmaceutical composition according to the above 42.
[50]
It contains one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:2, and is used in combination with one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3, or the amino acids of SEQ ID NO:3. Further comprising one or more antibodies that bind to the epitope contained in the sequence; or one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:3, to the epitope contained in the amino acid sequence of SEQ ID NO:2 In combination with one or more antibodies that bind, or further comprising one or more antibodies that bind to an epitope contained in the amino acid sequence of SEQ ID NO:2,
50. The pharmaceutical composition according to the above 49.
[51]
It comprises one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 3 and is used in combination with one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO: 4, or the amino acid of SEQ ID NO: 4 Further comprising one or more antibodies that bind to the epitope contained in the sequence; or one or more antibodies that bind to the epitope contained in the amino acid sequence of SEQ ID NO:4, to the epitope contained in the amino acid sequence of SEQ ID NO:3 Combined with one or more antibodies that bind, or further comprising one or more antibodies that bind to an epitope contained in the amino acid sequence of SEQ ID NO:3,
50. The pharmaceutical composition according to the above 49.
[52]
(A), (a'), (a''), (a''') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h''''). Antibody and/or (i), (i′), (i″), (i′″) or (i″″) antibody, or (b) to (h), (B')-(h'), (b'')-(h''), (b''')-(h''') or (b'''')-(h'''') A) and/or (i), (i′), (i″), (i′″) or (i″″) antibody;
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') ) To (h″″), including one or more antibodies selected from (a), (a′), (a″), (a′″) or (a″″) Antibody and/or (i), (i′), (i″), (i′″) or (i″″) antibody, or (a), (a′) , (A″), (a′″) or (a″″) antibody and/or (i), (i′), (i″), (i″′) or (i '''') further comprising; or
(I), (i′), (i″), (i′″) or (i″″) antibody, and (a), (a′), (a″), ( a″′) or (a″″) antibody and/or (b) to (h), (b′) to (h′), (b″) to (h″), (b Used in combination with one or more antibodies selected from''')-(h''') or (b'''')-(h''''), or (a), (a') , (A″), (a′″) or (a″″) antibody and/or (b) to (h), (b′) to (h′), (b″) to Further comprising one or more antibodies selected from (h″), (b′″) to (h′″) or (b″″) to (h″″),
43. The pharmaceutical composition according to the above 42.
[53]
(A), (a'), (a''), (a''') or (a'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h''''). Or (b) to (h), (b') to (h'), (b'') to (h''), (b''') to (h'''). ) Or (b″″) to (h″″), further containing one or more antibodies selected from the group; or (b) to (h), (b′) to (h′), ( one or more antibodies selected from b″) to (h″), (b′″) to (h′″) or (b″″) to (h″″) And used in combination with the antibody of (a), (a'), (a''), (a''') or (a''''), or (a), (a'), (a) ''), (a''') or (a'''') antibody,
53. The pharmaceutical composition according to the above 52.
[54]
(B) to (h), (b') to (h'), (b'') to (h''), (b''') to (h''') or (b'''') ) To (h″″), including one or more antibodies selected from (i), (i′), (i″), (i″′) or (i″″) The antibody of (i), (i′), (i″), (i′″) or (i″″), or
(I), (i'), (i''), (i''') or (i'''') antibody, and (b) to (h), (b') to (h' ), (b'') to (h''), (b''') to (h''') or (b'''') to (h''''). Or (b) to (h), (b') to (h'), (b'') to (h''), (b''') to (h'''). ) Or (b″″) to (h″″), further comprising one or more antibodies selected from the group consisting of:
53. The pharmaceutical composition according to the above 52.
[55]
(A), (a'), (a''), (a''') or (a'''') antibody is included, and (b), (b'), (b''), ( b''') or (b'''') antibody, (c), (c'), (c''), (c''') or (c'''') antibody, (g ), (g′), (g″), (g′″) or (g″″) antibody, or (i), (i′), (i″), (i″) ') or (i'''') antibody, or (b), (b'), (b''), (b''') or (b'''') antibody, (C), (c'), (c''), (c''') or (c'''') antibody, (g), (g'), (g''), (g' ″) or (g″″) antibody, or (i), (i′), (i″), (i′″) or (i″″) antibody;
(B), (b'), (b''), (b''') or (b'''') antibody is included, and (a), (a'), (a''), ( a)′) or (a″″) antibody, or (a), (a′), (a″), (a′″) or (a″″) Further comprising the antibody of:
(C), (c'), (c''), (c''') or (c'''') antibody is included, and (a), (a'), (a''), ( a″′) or (a″″) antibody or (i), (i′), (i″), (i′″) or (i″″) antibody Or an antibody of (a), (a'), (a''), (a''') or (a'''') or (i), (i'), (i''), (I''') or (i'''') antibody further included;
(G), (g'), (g''), (g''') or (g'''') antibody is included, and (a), (a'), (a''), ( a)′) or (a″″) antibody, or (a), (a′), (a″), (a′″) or (a″″) Or (i), (i′), (i″), (i′″) or (i″″) antibody, (a), (a′), (A''), (a''') or (a'''') antibody or (c), (c'), (c''), (c''') or (c''') ') antibody in combination with (a), (a'), (a''), (a''') or (a'''') antibody or (c), (c') , (C″), (c′″) or (c″″) antibody,
53. The pharmaceutical composition according to the above 52.
[56]
The pharmaceutical composition according to any of 42 to 55, which is used in combination with an anti-HBs antibody that binds to an epitope different from the antibody according to any of 1 to 15 above, or further comprises the anti-HBs antibody.
[57]
57. The pharmaceutical composition according to the above 56, wherein the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody.
[58]
The pharmaceutical composition according to any of the above 42 to 57, for treating or preventing hepatitis B virus infection.
[59]
The pharmaceutical composition according to any of the above 42 to 58, for the treatment of chronic hepatitis B.
［６０］
　B型肝炎ウイルス感染を治療または予防するための方法であって、その処置を必要とする対象に有効量の前記１～２３のいずれかに記載の抗体または組み合わせを投与することを含む方法。
［６１］
　前記３０に記載の方法により対象のB型肝炎ウイルス感染を評価することをさらに含む、前記６０に記載の方法。
［６２］
　慢性B型肝炎の治療のための、前記６０または６１に記載の方法。 [60]
24. A method for treating or preventing hepatitis B virus infection, which method comprises administering to a subject in need thereof an effective amount of an antibody or combination of any of the above 1-23.
[61]
61. The method of 60, further comprising assessing a subject for hepatitis B virus infection by the method of 30.
[62]
62. The method according to 60 or 61 above, for the treatment of chronic hepatitis B.
［６３］
　B型肝炎ウイルス感染の治療または予防に使用するための、前記１～２３のいずれかに記載の抗体または組み合わせ。
［６４］
　慢性B型肝炎の治療に使用するための、前記６３に記載の抗体または組み合わせ。 [63]
The antibody or combination according to any of 1 to 23 above, for use in treating or preventing hepatitis B virus infection.
[64]
64. An antibody or combination according to 63 above, for use in the treatment of chronic hepatitis B.
［６５］
　B型肝炎ウイルス感染を治療または予防するための医薬の製造のための、前記１～２３のいずれかに記載の抗体または組み合わせの使用。
［６６］
　慢性B型肝炎の治療のための医薬の製造のための、前記６５に記載の使用。
[65]
Use of the antibody or combination according to any one of 1 to 23 above for the manufacture of a medicament for treating or preventing hepatitis B virus infection.
[66]
The use according to 65 above, for the manufacture of a medicament for the treatment of chronic hepatitis B.
１．抗体の作製
　HBs抗原のpreS1ドメインに由来する以下のペプチドを用いて、抗体を作製した。

Figure JPOXMLDOC01-appb-T000001
　株式会社アイティーエムにおいて、ペプチド21-35、34-48、および37-55を混合してマウスに免疫し、腸骨リンパ節法によりハイブリドーマを取得した。また、株式会社スクラムにおいて、ペプチドPep1およびPep2を混合してマウスに免疫し、ハイブリドーマを取得した。 1. Preparation of antibody An antibody was prepared using the following peptides derived from the preS1 domain of the HBs antigen.

Figure JPOXMLDOC01-appb-T000001
At ITM Co., Ltd., peptides 21-35, 34-48, and 37-55 were mixed to immunize mice, and hybridomas were obtained by the iliac lymph node method. In Scrum Co., Ltd., the peptides Pep1 and Pep2 were mixed to immunize mice to obtain hybridomas.
　得られたハイブリドーマが産生する抗体と、免疫に使用したペプチドを用いて、ELISAを行った。96ウェルポリスチレンプレートにそれぞれのペプチドを固相化し、ハイブリドーマの培養上清を添加してインキュベートした。洗浄後、ホースラディッシュパーオキシデース（HRP）で標識した抗マウスウサギ抗体を添加し、陽性反応を呈する上清を陽性とし、ペプチドに反応するハイブリドーマを選択した。表２の結果から、21-35、34-48、および37-55を用いて得たハイブリドーマからは、21-35に反応する抗体（15、25、41、および51）と、34-48および37-55の両方に反応する抗体（7、10、13、27、39、49、55、および63）とが得られたことがわかった（表２）。

Figure JPOXMLDOC01-appb-T000002
 ELISA was performed using the antibody produced by the obtained hybridoma and the peptide used for immunization. Each peptide was immobilized on a 96-well polystyrene plate, and the culture supernatant of the hybridoma was added and incubated. After washing, anti-mouse rabbit antibody labeled with horseradish peroxidase (HRP) was added, and the supernatant showing a positive reaction was made positive, and hybridomas that react with the peptide were selected. From the results in Table 2, from the hybridomas obtained using 21-35, 34-48, and 37-55, the antibodies (15, 25, 41, and 51) that react with 21-35, 34-48, and It was found that antibodies reactive with both 37-55 (7, 10, 13, 27, 39, 49, 55, and 63) were obtained (Table 2).

Figure JPOXMLDOC01-appb-T000002
　34-48および37-55の両方に反応する抗体は、両ペプチドの共通部分（NSNNPDWDFNPN、配列番号３）にエピトープが存在すると考えられた。エピトープの決定のため、抗体7、10、および55について、表３に示すペプチドを用いてファインマッピングを行った。ELISAプレートにはアミノ基導入プレート（住友ベークライト）を使用した。重炭酸バッファー（pH9.6）で希釈した2%グルタルアルデヒド溶液200 μlを各ウェルに添加し、室温で2時間インキュベーションして、アミノ基を活性化させた。200 μlの滅菌超純水で各ウェルを2回洗浄後、直ちに、0.2 Mリン酸緩衝液（pH6.0）を用いて100 μMの濃度に希釈した固相化用の各ペプチド溶液100 μlを各ウェルに添加して、4℃で一晩インキュベーションして固相化を行った。固相化後、ウェル中の抗体溶液を廃棄して、200 μlのリン酸緩衝食塩水（PBS, pH 7.4）で3回洗浄した。洗浄後の各ウェルに200 μlのブロッキング溶液（5% スキムミルクと0.05%のTween 20を含むPBS (pH 7.4)）を添加し、37℃で1時間インキュベーションしてブロッキングを行った。その後、ウェル中のブロッキング溶液を廃棄して、200 μlのPBST（0.2% Tween 20を含むPBS, pH7.4）で3回洗浄した。ハイブリドーマの上清より精製された抗体をブロッキングバッファーで1 μg/mlの濃度に希釈し、100 μlずつ各ウェルに添加して、37℃で1時間インキュベーションした。インキュベーション後の各ウェルの溶液は廃棄し、300 μlのPBSTで3回洗浄した。洗浄後、HRPを標識した市販の抗体であるGoat anti-mouse IgG (H+L), HRP conjugate (Proteintech Group, Inc)をブロッキングバッファーで1:2000 (0.5 μｇ/ml)に希釈したものを各ウェルに100 μlずつ添加し、37℃で1時間のインキュベーションを行った。その後、各ウェルの溶液を除去し、300 μlのPBSTで3回洗浄した。次に、100 μlのTMB基質溶液を添加して37℃で15分間反応させ、2Mの硫酸溶液を100 μl添加して反応を止めて、450 nmにおける吸光度を測定した。 Antibodies that react with both 34-48 and 37-55 were considered to have an epitope in the common part (NSNNPDWDFNPN, SEQ ID NO: 3) of both peptides. For epitope determination, antibodies 7, 10, and 55 were fine-mapped using the peptides shown in Table 3. An amino group-introduced plate (Sumitomo Bakelite) was used as the ELISA plate. 200 μl of 2% glutaraldehyde solution diluted with bicarbonate buffer (pH 9.6) was added to each well and incubated at room temperature for 2 hours to activate the amino group. After washing each well twice with 200 μl of sterilized ultrapure water, immediately add 100 μl of each peptide solution for immobilization diluted to a concentration of 100 μM with 0.2 M phosphate buffer (pH 6.0). Immobilization was performed by adding to each well and incubating overnight at 4°C. After immobilization, the antibody solution in the wells was discarded, and the wells were washed 3 times with 200 μl of phosphate buffered saline (PBS, pH 7.4). After washing, 200 μl of blocking solution (PBS (pH 7.4) containing 5% skimmed milk and 0.05% Tween 20) was added to each well and incubated at 37°C for 1 hour to perform blocking. Then, the blocking solution in the wells was discarded, and the wells were washed 3 times with 200 μl of PBST (PBS containing 0.2% Tween 20, pH 7.4). The antibody purified from the hybridoma supernatant was diluted with blocking buffer to a concentration of 1 μg/ml, 100 μl was added to each well, and the mixture was incubated at 37°C for 1 hour. After the incubation, the solution in each well was discarded, and the wells were washed 3 times with 300 μl of PBST. After washing, commercially available HRP-labeled Goat anti-mouse IgG (H+L), HRP conjugate (Proteintech Group, Inc) diluted with blocking buffer to 1:2000 (0.5 μg/ml) was used. 100 μl was added to each well and incubated at 37°C for 1 hour. Then, the solution in each well was removed, and the wells were washed 3 times with 300 μl of PBST. Next, 100 μl of TMB substrate solution was added and reacted at 37° C. for 15 minutes, 100 μl of 2M sulfuric acid solution was added to stop the reaction, and the absorbance at 450 nm was measured.
　表３の結果から、抗体7および10についてはNNPDWDFN（配列番号２１）が、抗体55についてはNNPDWDFNP（配列番号１７）がエピトープであるか、この配列中にエピトープが含まれる可能性が示された。
Figure JPOXMLDOC01-appb-T000003
 From the results in Table 3, it was shown that NNPDWDFN (SEQ ID NO: 21) was the epitope for antibodies 7 and 10 and NNPDWDFNP (SEQ ID NO: 17) was the epitope for antibody 55, or that the epitope may be contained in this sequence. ..
Figure JPOXMLDOC01-appb-T000003
　ペプチドPep1およびPep2を用いて得たハイブリドーマからは、6種類（1A、1B、1C、5A、5B、5C）の抗体が得られた。これら抗体はPep1とPep2の両方のペプチドに反応したことから、両ペプチドの共通部分（DPAFGANSNN、配列番号４）にエピトープが存在すると考えられた。 Six types of antibodies (1A, 1B, 1C, 5A, 5B, 5C) were obtained from the hybridomas obtained using peptides Pep1 and Pep2. Since these antibodies reacted with both Pep1 and Pep2 peptides, it was considered that an epitope exists in the common part (DPAFGANSNN, SEQ ID NO: 4) of both peptides.
　これらのエピトープは各遺伝子型のHBVで配列同一性の高い部分あり（図１）、これら抗体は各遺伝子型のpreS1抗原を認識すると考えられた。 -These epitopes have high sequence identity in HBV of each genotype (Fig. 1), and it was considered that these antibodies recognize preS1 antigen of each genotype.
　抗体の配列解析ツールであるabYsis（http://www.abysis.org/）により、得られた抗体の軽鎖および重鎖の可変領域（VR）、並びにそれらのCDR1、2および3の配列を特定した（図２ＡおよびＢおよび表４－１～４－９）。可変領域およびCDRは、Chothia Numbering Schemeにより特定した。

Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000009
Figure JPOXMLDOC01-appb-T000010
Figure JPOXMLDOC01-appb-T000011
Figure JPOXMLDOC01-appb-T000012
 Using the antibody sequence analysis tool abYsis (http://www.abysis.org/), the variable regions (VR) of the obtained antibody light and heavy chains and their CDR1, 2 and 3 sequences were analyzed. Identified (FIGS. 2A and B and Tables 4-1 to 4-9). The variable region and CDR were identified by Chothia Numbering Scheme.

Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000009
Figure JPOXMLDOC01-appb-T000010
Figure JPOXMLDOC01-appb-T000011
Figure JPOXMLDOC01-appb-T000012
　上記と同様にして、抗体41について、表５に示すペプチドを用いてファインマッピングを行った。表５の結果から、抗体41についてはGFFPDHQLD（配列番号８０）がエピトープであるか、この配列中にエピトープが含まれる可能性が示された。

Figure JPOXMLDOC01-appb-T000013
 Fine mapping was performed on the antibody 41 using the peptides shown in Table 5 in the same manner as described above. From the results in Table 5, it was shown that GFFPDHQLD (SEQ ID NO: 80) is an epitope for antibody 41, or the epitope may be contained in this sequence.

Figure JPOXMLDOC01-appb-T000013
２．抗体によるHBV感染阻止
　ヒト肝細胞キメラマウス（Tateno C, et al., PLoS One. 2015 Nov 4;10(11):e0142145）の肝臓から作製したヒト初代培養肝細胞を用いて、得られた抗体のHBV感染阻止効果を検討した。ヒト初代培養肝細胞（105 細胞）に、HBV（約20ウイルス粒子/細胞）と、抗体産生ハイブリドーマの培養上清または精製抗体とを添加し、13日間培養後、培養上清中のHBs抗原の量を、HBsAg検出キット（シスメックス株式会社）を用いてCLIA法により測定した。精製抗体は、拡大培養した抗体産生ハイブリドーマの培養上清から抗体精製カラム（プロテノバ株式会社）を用いて取得した。精製抗体および培養上清中の抗体濃度は、Mouse IgG1 ELISAキットを用いて定量した。結果は、各ウェルの測定値の平均（IU/mL）で示した。コントロールのリン酸緩衝生理食塩水（PBS）と比較して、各抗体は培養後のHBs抗原の産生を顕著に抑制しており、HBVの感染を阻止することが示された（図３～５）。特に、抗体7、10、13、39、49、55および63は高い感染阻止効果を示した（図３）。 2. Prevention of HBV infection by antibody Human hepatocyte chimeric mouse (Tateno C, et al., PLoS One. 2015 Nov 4;10(11):e0142145) was obtained by using human primary culture hepatocytes prepared from the liver. The effect of HBV infection was investigated. HBV (about 20 virus particles/cell) and culture supernatant of antibody-producing hybridoma or purified antibody were added to human primary hepatocytes (10 5 cells), and after culturing for 13 days, HBs antigen in the culture supernatant was added. Was measured by CLIA method using HBsAg detection kit (Sysmex Corporation). The purified antibody was obtained from the culture supernatant of the expanded and produced antibody-producing hybridoma using an antibody purification column (Protenova Co., Ltd.). The antibody concentration in the purified antibody and the culture supernatant was quantified using the Mouse IgG1 ELISA kit. The results are shown as the average (IU/mL) of the measured values in each well. Compared with the control phosphate buffered saline (PBS), each antibody remarkably suppressed the production of HBs antigen after culturing, and was shown to block HBV infection (FIGS. 3-5). ). In particular, the antibodies 7, 10, 13, 39, 49, 55 and 63 showed a high infection blocking effect (Fig. 3).
　同様に、ヒト初代培養肝細胞において、抗体41と、抗体7または10との組み合わせのHBV感染阻止効果を検討した。抗体41を産生するハイブリドーマの培養上清と、抗体7または10を産生するハイブリドーマの培養上清とを組み合わせることにより、感染阻止効果が増強されることが示された（図６）。精製抗体である抗体41と抗体10または55との組み合わせによっても、相乗的な感染阻止効果の増強が観察された（図７）。抗体49と抗体55とを組み合わせた場合、抗体49単独と比較して併用により用量依存的にウイルスの増殖抑制効果が増強された（図８）。また、Sドメインにエピトープを持つHyb-824（株式会社特殊免疫研究所）のIgG3抗体（本明細書中、抗体824とも記載する）と抗体10または55との組み合わせによっても、感染阻止効果の増強が観察された（図９）。 Similarly, the inhibitory effect on HBV infection of the combination of antibody 41 and antibody 7 or 10 was examined in human primary culture hepatocytes. It was shown that the infection-inhibiting effect was enhanced by combining the culture supernatant of the hybridoma producing the antibody 41 and the culture supernatant of the hybridoma producing the antibody 7 or 10 (FIG. 6). A synergistic enhancement of the infection-blocking effect was also observed with the combination of the purified antibody, Antibody 41 and Antibody 10 or 55 (FIG. 7). When antibody 49 and antibody 55 were combined, the combined use enhanced the virus growth-inhibitory effect in a dose-dependent manner compared to antibody 49 alone (FIG. 8). Further, the combination of an IgG3 antibody of Hyb-824 (Special Immune Research Institute, Inc.) having an epitope in the S domain (also referred to as antibody 824 in the present specification) and antibody 10 or 55 enhances the infection-inhibiting effect. Were observed (Fig. 9).
３．preS1抗原の測定
（１）固相化に使用する抗体の検討（i）
　ELISAプレートにはアミノ基導入プレート（住友ベークライト）を使用した。重炭酸バッファー（pH9.6）で希釈した2%グルタルアルデヒド溶液200 μlを各ウェルに添加し、室温で2時間インキュベーションして、アミノ基を活性化させた。200 μlの滅菌超純水で各ウェルを2回洗浄後、直ちに、0.2 Mリン酸緩衝液（pH6.0）を用いて0.5 μg/mlの濃度に希釈した固相化用抗体の溶液100 μlを各ウェルに添加して、4℃で一晩インキュベーションして固相化を行った。固相化用抗体としては、抗体41またはSドメインにエピトープを持つハイブリドーマクローン5124AのIgM抗体（株式会社特殊免疫研究所）（本明細書中、抗体5124Aとも記載する）を使用した。また、血清中のHBVまたは酵素標識抗体の非特異的な吸着の観察のため、抗体を加えないウェルを準備し、抗体の固相化中は0.2 Mリン酸緩衝液のみ添加した。固相化後、ウェル中の抗体溶液を廃棄し、200 μlのリン酸緩衝食塩水（PBS, pH 7.4）で2回洗浄した。洗浄後の各ウェルに200 μlのブロッキング溶液（3% BSAを含むPBS (pH 7.4）)を添加し、室温で2時間インキュベーションしてブロッキングを行った。その後、ウェル中のブロッキング溶液を廃棄し、HBV陽性被験者血清をブロッキング溶液で10倍に希釈したものを100 μlずつ各ウェルに添加し、37℃で1時間インキュベーションした。ネガティブコントロール（NTC）には血清を添加せず、ブロッキング溶液のみを添加した。インキュベーション後、各ウェルの溶液を廃棄し、300 μlのPBST（0.2% Tween 20を含むPBS, pH7.4)で3回洗浄した。洗浄後、HRPで標識した抗体55を1:1000（3 μg/ml）となるようにブロッキング溶液で希釈したものを各ウェルに100 μlずつ添加し、37℃で1時間のインキュベーションを行った。その後、各ウェルの溶液を除去し、300 μlのPBSTで3回洗浄した。次に、100 μlのTMB基質溶液を添加して室温で15分間反応させ、2 Mの硫酸溶液を100 μl添加して反応を止めて、450 nmにおける吸光度を測定した。 3. Measurement of preS1 antigen (1) Examination of antibody used for immobilization (i)
An amino group-introduced plate (Sumitomo Bakelite) was used as the ELISA plate. 200 μl of a 2% glutaraldehyde solution diluted with bicarbonate buffer (pH 9.6) was added to each well and incubated at room temperature for 2 hours to activate the amino group. After washing each well twice with 200 μl of sterile ultrapure water, immediately immobilize 100 μl of antibody for immobilization antibody diluted to a concentration of 0.5 μg/ml with 0.2 M phosphate buffer (pH 6.0). Was added to each well and incubated at 4° C. overnight for immobilization. As the immobilizing antibody, the antibody 41 or IgM antibody of hybridoma clone 5124A having an epitope in the S domain (Special Immune Research Institute, Inc.) (also referred to as antibody 5124A in the present specification) was used. Further, in order to observe non-specific adsorption of HBV or enzyme-labeled antibody in serum, a well containing no antibody was prepared, and only 0.2 M phosphate buffer was added while the antibody was immobilized. After immobilization, the antibody solution in the well was discarded, and the well was washed twice with 200 μl of phosphate buffered saline (PBS, pH 7.4). After washing, 200 μl of a blocking solution (PBS (pH 7.4) containing 3% BSA) was added to each well and incubated at room temperature for 2 hours for blocking. Then, the blocking solution in the wells was discarded, 100 μl of HBV-positive subject serum diluted 10-fold with the blocking solution was added to each well, and the mixture was incubated at 37° C. for 1 hour. No serum was added to the negative control (NTC), and only the blocking solution was added. After the incubation, the solution in each well was discarded, and the wells were washed 3 times with 300 μl of PBST (PBS containing 0.2% Tween 20, pH 7.4). After washing, HRP-labeled antibody 55 diluted with the blocking solution to 1:1000 (3 μg/ml) was added to each well in an amount of 100 μl, and incubated at 37° C. for 1 hour. Then, the solution in each well was removed, and the wells were washed 3 times with 300 μl of PBST. Next, 100 μl of TMB substrate solution was added and reacted at room temperature for 15 minutes, 100 μl of 2 M sulfuric acid solution was added to stop the reaction, and the absorbance at 450 nm was measured.
　比較した2種類の抗体はいずれもHBs抗原の捕集能を有していたが、抗体5124Aの方がより高い検出能を示した（図１０）。HBVまたは酵素標識抗体の非特異的な吸着による吸光度は、0.128 ± 0.005 (0.087～0.313)であった。 Both of the two kinds of antibodies compared had the ability to collect HBs antigen, but the antibody 5124A showed higher detection ability (Fig. 10). The absorbance due to nonspecific adsorption of HBV or enzyme-labeled antibody was 0.128 ± 0.005 (0.087 to 0.313).
（２）被験者血清中のpreS1抗原の測定
　固相化抗体に抗体5124Aを使用して、上記（１）と同じ方法でHBV陰性被験者10名とHBV陽性被験者50名の血清中のpreS1抗原の測定を行った。被験者血清中のpreS1抗原の吸光度測定値と、既存法によるHBVマーカーの測定値を表６に示す。HBV-DNA値は、リアルタイムPCR法により決定し、HBs抗原およびHBe抗原は、全自動免疫測定装置HISCL（シスメックス株式会社）を使用して、それぞれHBsAg検出キットとHBeAg検出キットを用いてCLIA法で測定した。HBc関連抗原は、Ｂ型肝炎ウイルスコア関連抗原キット　ルミパルスHBcrAg（富士レビオ株式会社）を用いてCLIA法で測定した。HBe抗体は、アーキテクトi2000ＳＲ（アボットジャパン株式会社）とアーキテクト・HBe抗体試薬を用いてCLIA法で測定した。。HBV陽性被験者の吸光度は0.603～3.141であった。吸光光度法では、実用的な吸光度の測定範囲は0.05から1.5と言われているが、HBs抗原値（IU/ML）が1,000を超えると、吸光度1.5を超えるサンプルが多くあり、さらなる希釈が必要と考えられた。preS1抗原の吸光度他のHBVマーカーとの相関を、回帰分析を用いて解析したが、どのマーカーとも相関が見られなかった（データ非提示）。


Figure JPOXMLDOC01-appb-T000014
Figure JPOXMLDOC01-appb-T000015
 (2) Measurement of preS1 antigen in serum of test subjects Using antibody 5124A as the immobilized antibody, measurement of preS1 antigen in serum of 10 HBV-negative subjects and 50 HBV-positive subjects in the same manner as in (1) above. I went. Table 6 shows the measured absorbance of the preS1 antigen in the serum of the subject and the measured value of the HBV marker by the existing method. HBV-DNA value was determined by real-time PCR method, and HBs antigen and HBe antigen were measured by CLIA method using HBsAg detection kit and HBeAg detection kit, respectively, using a fully automated immunoassay device HISCL (Sysmex Corporation). It was measured. The HBc-related antigen was measured by the CLIA method using the hepatitis B virus core-related antigen kit Lumipulse HBcrAg (Fujirebio Inc.). The HBe antibody was measured by CLIA method using Architect i2000SR (Abbott Japan Co., Ltd.) and Architect-HBe antibody reagent. .. The absorbance of HBV-positive subjects was 0.603-3.141. In the absorptiometric method, the practical range of absorbance measurement is said to be 0.05 to 1.5, but when the HBs antigen value (IU/ML) exceeds 1,000, there are many samples with absorbance higher than 1.5 and further dilution is required. It was considered. The correlation of preS1 antigen absorbance with other HBV markers was analyzed using regression analysis, but no correlation was observed with any of the markers (data not shown).


Figure JPOXMLDOC01-appb-T000014
Figure JPOXMLDOC01-appb-T000015
（３）血清希釈率と固相化に使用する抗体の検討（ii）
　ELISAプレートにはアミノ基導入プレート（住友ベークライト）を使用した。重炭酸バッファー（pH9.6）で希釈した2%グルタルアルデヒド溶液200 μlを各ウェルに添加し、室温で2時間インキュベーションして、アミノ基を活性化させた。200 μlの滅菌超純水で各ウェルを2回洗浄後、直ちに、0.2 Mリン酸緩衝液（pH6.0）を用いて0.5 μg/mlの濃度に希釈した固相化用抗体の溶液100 μlを各ウェルに添加して、4℃で一晩インキュベーションを行い、固相化を行った。固相化抗体としては、抗体5124Aまたは抗体824を使用した。固相化後、ウェル中の抗体溶液を廃棄し、200 μlのリン酸緩衝食塩水（PBS, pH 7.4）で3回洗浄した。洗浄後の各ウェルに、ブロッキング溶液としてChonBlock（商標） Blocking/Sample Dilution Buffer (Chondrex)を100 μlずつ添加し、室温で1時間インキュベーションしてブロッキングを行った。その後、ウェル中のブロッキング溶液を廃棄し、200 μlのPBST（0.2% Tween 20を含むPBS, pH7.4)で3回洗浄した。HBs抗原が90,000 IU/mlでHBV-DNAが9.1 Log copies/mlのHBV患者血清をポジティブコントロール（PTC）血清として、ブロッキング溶液を用いて50倍希釈液を作成し、さらに3倍希釈系列を作成して、上記の抗体をそれぞれ固相化したウェルに50 μlずつ添加した。また、HBV陽性被験者および陰性被験者の血清について、ブロッキング溶液を用いて10倍希釈系列を作成し、同じく50 μlずつ各ウェルに添加した。37℃で1時間インキュベーションし、インキュベーション後に各ウェルの溶液を廃棄し、200 μlのPBSTで3回洗浄した。洗浄後、HRPを標識した抗体55を1:1000（3 μg/ml）となるように、ChonBlock（商標） Detection Antibody Dilution Buffer (Chondrex)で希釈したものを各ウェルに100 μlずつ添加し、37℃で1時間のインキュベーションを行った。その後、各ウェルの溶液を除去し、200 μlのPBSTで3回洗浄した。次に、100 μlのTMB基質溶液を添加して室温で15分間反応させ、2Mの硫酸溶液を100 μl添加して反応を止め、450 nmにおける吸光度を測定した。 (3) Examination of serum dilution ratio and antibody used for immobilization (ii)
An amino group-introduced plate (Sumitomo Bakelite) was used as the ELISA plate. 200 μl of a 2% glutaraldehyde solution diluted with bicarbonate buffer (pH 9.6) was added to each well and incubated at room temperature for 2 hours to activate the amino group. After washing each well twice with 200 μl of sterile ultrapure water, immediately immobilize 100 μl of antibody for immobilization antibody diluted to a concentration of 0.5 μg/ml with 0.2 M phosphate buffer (pH 6.0). Was added to each well and incubated overnight at 4° C. to immobilize. As the immobilized antibody, antibody 5124A or antibody 824 was used. After immobilization, the antibody solution in the wells was discarded, and the wells were washed 3 times with 200 μl of phosphate buffered saline (PBS, pH 7.4). 100 μl of ChonBlock™ Blocking/Sample Dilution Buffer (Chondrex) was added as a blocking solution to each well after washing, and blocking was performed by incubating at room temperature for 1 hour. Then, the blocking solution in the wells was discarded, and the wells were washed 3 times with 200 μl of PBST (PBS containing 0.2% Tween 20, pH 7.4). HBV antigen is 90,000 IU/ml and HBV-DNA is 9.1 log copies/ml. HBV patient serum is used as positive control (PTC) serum and 50 times diluted solution is prepared using blocking solution, and further 3 times diluted series is prepared. Then, 50 μl of each of the above antibodies was added to each well in which the antibody was immobilized. Further, with respect to the sera of HBV-positive subjects and negative subjects, a 10-fold dilution series was prepared using a blocking solution, and 50 μl of each was similarly added to each well. After incubation at 37°C for 1 hour, the solution in each well was discarded after the incubation, and the wells were washed 3 times with 200 µl of PBST. After washing, HRP-labeled antibody 55 diluted to 1:1000 (3 μg/ml) with ChonBlock™ Detection Antibody Dilution Buffer (Chondrex) was added to each well by 100 μl, and 37 Incubation was carried out at 0°C for 1 hour. Then, the solution in each well was removed, and the wells were washed 3 times with 200 μl of PBST. Next, 100 μl of TMB substrate solution was added and reacted at room temperature for 15 minutes, 100 μl of 2M sulfuric acid solution was added to stop the reaction, and the absorbance at 450 nm was measured.
　PTC血清の希釈系列を用いてpreS1抗原の検量線を作製したところ、固相化に用いた抗体5124Aは吸光度が2.4、抗体824は2.3付近まで、R2=0.999の近似曲線を作成することが可能であった（表７、図１１）。HBV陽性または陰性被験者血清の各希釈系列のpreS1抗原の吸光度と、既存法で測定したHBs抗原の測定値を表８に示す。吸光度はHBs抗原値が高くなるにつれて高くなった。血清希釈率は10,000 IU/ml以下では1:10、10,000 IU/ml 付近では1:100、100,000 IU/ml付近では1:500～1:1000が良いとみられた。各血清に適した抗体希釈率で抗体5124Aと抗体824の吸光度を比較したところ、抗体824の方が抗体5124Aよりもより高い検出能を示した（図１２）。HBV陰性被験者の吸光度は、血清を1:10の濃度で添加した場合でも0.056 (5124A)、0.069 (824)と低く、ブロッキング溶液と血清希釈液に3% BSAを含むPBSを用いた場合よりもバックグラウンドの吸光度が低下した。

Figure JPOXMLDOC01-appb-T000016
Figure JPOXMLDOC01-appb-T000017
 When a calibration curve of preS1 antigen was prepared using a dilution series of PTC serum, the antibody 5124A used for immobilization had an absorbance of 2.4, the antibody 824 was around 2.3, and an approximate curve of R 2 =0.999 could be created. It was possible (Table 7, Figure 11). Table 8 shows the absorbance of preS1 antigen in each dilution series of HBV positive or negative subject serum and the measured value of HBs antigen measured by the existing method. The absorbance increased as the HBsAg value increased. The serum dilution ratio was considered to be 1:10 below 10,000 IU/ml, 1:100 near 10,000 IU/ml, and 1:500 to 1:1000 around 100,000 IU/ml. When the absorbances of the antibody 5124A and the antibody 824 were compared at an antibody dilution ratio suitable for each serum, the antibody 824 showed higher detectability than the antibody 5124A (FIG. 12). The absorbance of HBV-negative subjects was low at 0.056 (5124A) and 0.069 (824) even when serum was added at a concentration of 1:10, which was lower than that when PBS containing 3% BSA was used as a blocking solution and a serum diluent. The background absorbance decreased.

Figure JPOXMLDOC01-appb-T000016
Figure JPOXMLDOC01-appb-T000017
（４）固相化に使用する抗体の検討（iii）
　上記（３）と同様の方法を用いて、固相化に使用する抗体の比較を、他の被験者の血清を用いて2連で行った。固相化の際の抗体濃度は2 μg/mlで行った。波長450 nmにおける吸光度の平均値を図１３に示す。一部の被験者血清（HBV(+)64および65）では、抗体5124Aの吸光度が高かったが、他の被験者血清では抗体824の吸光度の方が高かった。抗体824の方が抗体5124Aよりも、より多くの被験者血清に対して高い検出能を有することが示唆されたことから、以降の試験では、固相化抗体に抗体824を使用することとした。 (4) Examination of antibodies used for immobilization (iii)
Using the same method as in (3) above, comparison of antibodies used for immobilization was performed in duplicate using sera of other subjects. The antibody concentration during immobilization was 2 μg/ml. The average value of the absorbance at a wavelength of 450 nm is shown in FIG. In some subject sera (HBV(+)64 and 65), antibody 5124A had higher absorbance, while in other subject sera, antibody 824 had higher absorbance. Since it was suggested that the antibody 824 has higher detection ability for a larger amount of subject serum than the antibody 5124A, in the subsequent tests, it was decided to use the antibody 824 as the immobilized antibody.
（５）preS1抗原検出のための検出用抗体の比較
　上記（３）と同様の方法を用い、検出用抗体としてHRP標識した抗体10、13、39または55を用いて、検出能の比較を2連で行った。固相化抗体濃度は0.5μg/ml、検出用抗体濃度は、これまでと同じく1:1000とし、各ウェルに70 μl添加した。波長450 nmにおける吸光度の平均値を図１４に示す。4種類の抗体の検出能はどの血清でも類似した傾向を示し、いずれの抗体もpreS1抗原の測定に使用できることがわかった。測定値は抗体10が最も高く、続いて抗体55、39、13の順で高かった。
(5) Comparison of detection antibodies for detecting preS1 antigen Using the same method as in (3) above, HRP-labeled antibody 10, 13, 39 or 55 was used as the detection antibody to compare the detection capacities. I went in a row. The immobilized antibody concentration was 0.5 μg/ml, and the detection antibody concentration was 1:1000 as before, and 70 μl was added to each well. The average value of absorbance at a wavelength of 450 nm is shown in FIG. It was found that the detectivities of the four types of antibodies showed similar tendencies in all sera, and that all of the antibodies could be used to measure preS1 antigen. The measured value was highest for antibody 10, followed by antibody 55, 39, and 13 in that order.
　実施例１と同様にして、ヒト初代培養肝細胞を用いて、抗体41、抗体1A、および抗体10のHBV感染阻止効果を検討した。培養上清中のHBs抗原およびHBe抗原の量は、HBsAg検出キットおよびHBeAg検出キット（シスメックス株式会社）を用いてCLIA法により測定した。コントロールとして、HBVのみを添加する群、および前記抗体にかえてIgG1（OY-TES-1 (G-5), santacruz: sc-390594）またはIgG3（Mouse IgG3 (isotype control), MBL: M078-3）を添加する群を設定した。各抗体は培養後のHBs抗原およびHBe抗原の産生を顕著に抑制しており、HBVの感染を阻止することが示された（図１５、１６）。 In the same manner as in Example 1, using human primary culture hepatocytes, the inhibitory effects of antibody 41, antibody 1A, and antibody 10 on HBV infection were examined. The amounts of HBs antigen and HBe antigen in the culture supernatant were measured by CLIA method using HBsAg detection kit and HBeAg detection kit (Sysmex Corporation). As a control, a group to which only HBV is added, and IgG1 (OY-TES-1 (G-5), santacruz: sc-390594) or IgG3 (Mouse IgG3 (isotype control), MBL: M078-3 instead of the antibody ) Was added to the group. It was shown that each antibody markedly suppressed the production of HBs antigen and HBe antigen after culturing, and blocked HBV infection (FIGS. 15 and 16).
　同様に、抗体824と、抗体41、抗体1A、または抗体10との組み合わせのHBV感染阻止効果を検討した。Sドメインにエピトープを持つ抗体824と、抗体41、抗体10または抗体1Aとを組み合わせることにより、HBs抗原およびHBe抗原の産生抑制効果が用量依存的に増強され、相乗的な感染阻止効果が観察された（図１７、１８）。また、抗体41、抗体10および抗体1Aをそれぞれ組み合わせることによっても、相乗的な感染阻止効果が観察された（図１９、２０）。 Similarly, the inhibitory effect on HBV infection of the combination of antibody 824 and antibody 41, antibody 1A, or antibody 10 was examined. By combining antibody 824 having an epitope in the S domain with antibody 41, antibody 10 or antibody 1A, the inhibitory effect on HBs antigen and HBe antigen production is enhanced in a dose-dependent manner, and a synergistic infection-blocking effect is observed. (FIGS. 17 and 18). In addition, a synergistic infection blocking effect was also observed by combining the antibody 41, the antibody 10, and the antibody 1A (FIGS. 19 and 20).

Claims (22)

  1. 　抗preS1抗体であって、以下の（ａ）～（ｉ）から選択される、抗体：
    （ａ）配列番号２４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号２５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号２６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号２８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号２９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号３０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｂ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｃ）配列番号３２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号３４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｄ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号４３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号４５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｅ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号４７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号４０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号３８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｆ）配列番号５０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号５１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号５２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号５４のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号５５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号５６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｇ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号６０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｈ）配列番号４２のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号５８のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号６３のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号６１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；および
    （ｉ）配列番号６５のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号６６のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号６７のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号６９のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR1、
    　配列番号７０のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR2、および
    　配列番号７１のアミノ酸配列または前記アミノ酸配列において１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体。     Anti-preS1 antibody, which is selected from the following (a) to (i):
    (A) a CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 25 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 26 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 28 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 29 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 30 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (B) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 containing the amino acid sequence of SEQ ID NO: 37 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (C) a CDR1 comprising the amino acid sequence of SEQ ID NO: 32 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 34 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (D) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence have been deleted, substituted or added,
    CDR2 containing the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 43 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 45 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (E) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and the amino acid sequence of SEQ ID NO: 47 or 1 to 3 in said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 containing the amino acid sequence of SEQ ID NO: 40 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 38 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (F) CDR1 comprising the amino acid sequence of SEQ ID NO: 50 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 51 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 52 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 54 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO:55 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:56 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (G) CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 60 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 61 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
    (H) a CDR1 comprising the amino acid sequence of SEQ ID NO: 42 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence;
    CDR2 comprising the amino acid sequence of SEQ ID NO: 33 or the amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted or added in the amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 58 or the amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1 comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence in which 1 to 3 amino acid residues in the amino acid sequence are deleted, substituted or added,
    CDR2 comprising the amino acid sequence of SEQ ID NO:63 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO:61 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising: (i) a CDR1 comprising the amino acid sequence of SEQ ID NO: 65 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in said amino acid sequence,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 66 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 67 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 69 or an amino acid sequence in which 1 to 3 amino acid residues are deleted, substituted or added in the amino acid sequence,
    CDR2 comprising the amino acid sequence of SEQ ID NO: 70 or the amino acid sequence of 1 to 3 amino acid residues deleted, substituted or added in said amino acid sequence, and 1 to 3 in the amino acid sequence of SEQ ID NO: 71 or said amino acid sequence CDR3 containing amino acid sequence with deletion, substitution, or addition of 4 amino acid residues
    An antibody comprising a heavy chain variable region comprising:
  2. 　抗体が、以下の（ａ’）～（ｉ’）のいずれかの抗体である、請求項１に記載の抗体：
    （ａ’）配列番号２４のアミノ酸配列を含むCDR1、
    　配列番号２５のアミノ酸配列を含むCDR2、および
    　配列番号２６のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号２８のアミノ酸配列を含むCDR1、
    　配列番号２９のアミノ酸配列を含むCDR2、および
    　配列番号３０のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｂ’）配列番号３２のアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列を含むCDR2、および
    　配列番号３４のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列を含むCDR1、
    　配列番号３７のアミノ酸配列を含むCDR2、および
    　配列番号３８のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｃ’）配列番号３２のアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列を含むCDR2、および
    　配列番号３４のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列を含むCDR1、
    　配列番号４０のアミノ酸配列を含むCDR2、および
    　配列番号３８のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｄ’）配列番号４２のアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列を含むCDR2、および
    　配列番号４３のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列を含むCDR1、
    　配列番号４０のアミノ酸配列を含むCDR2、および
    　配列番号４５のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｅ’）配列番号４２のアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列を含むCDR2、および
    　配列番号４７のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列を含むCDR1、
    　配列番号４０のアミノ酸配列を含むCDR2、および
    　配列番号３８のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｆ’）配列番号５０のアミノ酸配列を含むCDR1、
    　配列番号５１のアミノ酸配列を含むCDR2、および
    　配列番号５２のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号５４のアミノ酸配列を含むCDR1、
    　配列番号５５のアミノ酸配列を含むCDR2、および
    　配列番号５６のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｇ’）配列番号４２のアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列を含むCDR2、および
    　配列番号５８のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列を含むCDR1、
    　配列番号６０のアミノ酸配列を含むCDR2、および
    　配列番号６１のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；
    （ｈ’）配列番号４２のアミノ酸配列を含むCDR1、
    　配列番号３３のアミノ酸配列を含むCDR2、および
    　配列番号５８のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号３６のアミノ酸配列を含むCDR1、
    　配列番号６３のアミノ酸配列を含むCDR2、および
    　配列番号６１のアミノ酸配列を含むCDR3
    　を含む重鎖可変領域を含む抗体；および
    （ｉ’）配列番号６５のアミノ酸配列を含むCDR1、
    　配列番号６６のアミノ酸配列を含むCDR2、および
    　配列番号６７のアミノ酸配列を含むCDR3
    　を含む軽鎖可変領域、および、
    　配列番号６９のアミノ酸配列を含むCDR1、
    　配列番号７０のアミノ酸配列を含むCDR2、および
    　配列番号７１のアミノ酸配列を含むCDR3。     The antibody according to claim 1, wherein the antibody is any one of the following (a') to (i'):
    (A') CDR1 comprising the amino acid sequence of SEQ ID NO: 24,
    CDR2 containing the amino acid sequence of SEQ ID NO:25, and CDR3 containing the amino acid sequence of SEQ ID NO:26
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 28,
    CDR2 containing the amino acid sequence of SEQ ID NO: 29, and CDR3 containing the amino acid sequence of SEQ ID NO: 30
    An antibody comprising a heavy chain variable region comprising:
    (B') CDR1 containing the amino acid sequence of SEQ ID NO: 32,
    CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
    CDR2 containing the amino acid sequence of SEQ ID NO:37, and CDR3 containing the amino acid sequence of SEQ ID NO:38
    An antibody comprising a heavy chain variable region comprising:
    (C′) CDR1 containing the amino acid sequence of SEQ ID NO: 32,
    CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:34
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
    CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:38
    An antibody comprising a heavy chain variable region comprising:
    (D') CDR1 containing the amino acid sequence of SEQ ID NO: 42,
    CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:43
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
    CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:45
    An antibody comprising a heavy chain variable region comprising:
    (E′) CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
    CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:47
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
    CDR2 containing the amino acid sequence of SEQ ID NO:40, and CDR3 containing the amino acid sequence of SEQ ID NO:38
    An antibody comprising a heavy chain variable region comprising:
    (F') CDR1 comprising the amino acid sequence of SEQ ID NO: 50,
    CDR2 containing the amino acid sequence of SEQ ID NO:51 and CDR3 containing the amino acid sequence of SEQ ID NO:52
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 54,
    CDR2 containing the amino acid sequence of SEQ ID NO:55, and CDR3 containing the amino acid sequence of SEQ ID NO:56
    An antibody comprising a heavy chain variable region comprising:
    (G') CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
    CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:58
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
    CDR2 containing the amino acid sequence of SEQ ID NO:60, and CDR3 containing the amino acid sequence of SEQ ID NO:61
    An antibody comprising a heavy chain variable region comprising:
    (H') CDR1 comprising the amino acid sequence of SEQ ID NO: 42,
    CDR2 containing the amino acid sequence of SEQ ID NO:33, and CDR3 containing the amino acid sequence of SEQ ID NO:58
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 36,
    CDR2 containing the amino acid sequence of SEQ ID NO:63, and CDR3 containing the amino acid sequence of SEQ ID NO:61
    An antibody comprising a heavy chain variable region comprising; and (i′) CDR1 comprising the amino acid sequence of SEQ ID NO:65,
    CDR2 containing the amino acid sequence of SEQ ID NO:66, and CDR3 containing the amino acid sequence of SEQ ID NO:67
    A light chain variable region containing, and
    CDR1, which comprises the amino acid sequence of SEQ ID NO: 69,
    CDR2 containing the amino acid sequence of SEQ ID NO:70, and CDR3 containing the amino acid sequence of SEQ ID NO:71.
  3. 　抗体が、以下の（ａ’’）～（ｉ’’）のいずれかの抗体である、請求項１または２に記載の抗体：
    （ａ’’）配列番号２３のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号２３のアミノ酸配列もしくは配列番号２３のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号２７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号２７のアミノ酸配列もしくは配列番号２７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｂ’’）配列番号３１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３１のアミノ酸配列もしくは配列番号３１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号３５のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３５のアミノ酸配列もしくは配列番号３５のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｃ’’）配列番号３１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３１のアミノ酸配列もしくは配列番号３１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号３９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号３９のアミノ酸配列もしくは配列番号３９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｄ’’）配列番号４１のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４１のアミノ酸配列もしくは配列番号４１のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号４４のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４４のアミノ酸配列もしくは配列番号４４のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｅ’’）配列番号４６のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４６のアミノ酸配列もしくは配列番号４６のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号４８のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４８のアミノ酸配列もしくは配列番号４８のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｆ’’）配列番号４９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号４９のアミノ酸配列もしくは配列番号４９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号５３のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５３のアミノ酸配列もしくは配列番号５３のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｇ’’）配列番号５７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５７のアミノ酸配列もしくは配列番号５７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号５９のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５９のアミノ酸配列もしくは配列番号５９のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｈ’’）配列番号５７のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号５７のアミノ酸配列もしくは配列番号５７のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号６２のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６２のアミノ酸配列もしくは配列番号６２のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体；および
    （ｉ’’）配列番号６４のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６４のアミノ酸配列もしくは配列番号６４のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む軽鎖可変領域、および
    　配列番号６８のアミノ酸配列と８０％、８５％、９０％、または９５％以上の配列同一性を有するアミノ酸配列、または、配列番号６８のアミノ酸配列もしくは配列番号６８のアミノ酸配列において１～２０個、１～１０個、１～５個、または１～３個のアミノ酸残基が欠失、置換、または付加されたアミノ酸配列を含む重鎖可変領域を含む抗体。     The antibody according to claim 1 or 2, wherein the antibody is any one of the following (a″) to (i″):
    (A'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 23, or the amino acid sequence of SEQ ID NO: 23 or the amino acid sequence of SEQ ID NO: 23 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 27. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 27 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 27, An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
    (B'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 35. Having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 35, or 1-20 in the amino acid sequence of SEQ ID NO: 35, 1-10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
    (C'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 31, or the amino acid sequence of SEQ ID NO: 31 or the amino acid sequence of SEQ ID NO: 31 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 39. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 39 or 1 to 20 in the amino acid sequence of SEQ ID NO: 39, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues are deleted, substituted, or added;
    (D'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 41, or the amino acid sequence of SEQ ID NO: 41 or the amino acid sequence of SEQ ID NO: 41 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues are deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 44. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 44 or 1 to 20 in the amino acid sequence of SEQ ID NO: 44, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
    (E'') an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 46, or the amino acid sequence of SEQ ID NO: 46 or the amino acid sequence of SEQ ID NO: 46 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 48 Amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 48 or 1 to 20 in the amino acid sequence of SEQ ID NO: 48, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
    (F″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 49, or the amino acid sequence of SEQ ID NO: 49 or the amino acid sequence of SEQ ID NO: 49 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 53 With 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 53 or the amino acid sequence of SEQ ID NO: 53 with 1-20, 1-10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
    (G″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 59. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 59 or 1 to 20, 1 to 10 in the amino acid sequence of SEQ ID NO: 59, An antibody comprising a heavy chain variable region comprising an amino acid sequence in which -5, or 1-3 amino acid residues have been deleted, substituted, or added;
    (H″) an amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 57, or the amino acid sequence of SEQ ID NO: 57 or the amino acid sequence of SEQ ID NO: 57 A light chain variable region comprising an amino acid sequence in which 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues have been deleted, substituted, or added, and the amino acid sequence of SEQ ID NO: 62. An amino acid sequence having 80%, 85%, 90%, or 95% or more sequence identity with the amino acid sequence of SEQ ID NO: 62 or 1 to 20 in the amino acid sequence of SEQ ID NO: 62, 1 to 10, 1 An antibody comprising a heavy chain variable region comprising an amino acid sequence in which ˜5, or 1-3 amino acid residues have been deleted, substituted, or added; and (i″) the amino acid sequence of SEQ ID NO: 64 and 80% , 85%, 90%, or 95% or more sequence identity, or 1-20, 1-10, 1-5 in the amino acid sequence of SEQ ID NO: 64 or the amino acid sequence of SEQ ID NO: 64 , Or a light chain variable region comprising an amino acid sequence in which 1 to 3 amino acid residues have been deleted, substituted, or added, and 80%, 85%, 90%, or 95% or more of the amino acid sequence of SEQ ID NO: 68 An amino acid sequence having the sequence identity of, or 1 to 20, 1 to 10, 1 to 5, or 1 to 3 amino acid residues in the amino acid sequence of SEQ ID NO:68 or the amino acid sequence of SEQ ID NO:68. An antibody comprising a heavy chain variable region comprising a deleted, substituted, or added amino acid sequence.
  4. 　抗体が、以下の（ａ’’’）～（ｉ’’’）のいずれかの抗体である、請求項１～３のいずれかに記載の抗体：
    （ａ’’’）配列番号２３のアミノ酸配列を含む軽鎖可変領域、および配列番号２７のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｂ’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域、および配列番号３５のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｃ’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域、および配列番号３９のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｄ’’’）配列番号４１のアミノ酸配列を含む軽鎖可変領域、および配列番号４４のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｅ’’’）配列番号４６のアミノ酸配列を含む軽鎖可変領域、および配列番号４８のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｆ’’’）配列番号４９のアミノ酸配列を含む軽鎖可変領域、および配列番号５３のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｇ’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域、および配列番号５９のアミノ酸配列を含む重鎖可変領域を含む抗体；
    （ｈ’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域、および配列番号６２のアミノ酸配列を含む重鎖可変領域を含む抗体；および
    （ｉ’’’）配列番号６４のアミノ酸配列を含む軽鎖可変領域、および配列番号６８のアミノ酸配列を含む重鎖可変領域を含む抗体。     The antibody according to any one of claims 1 to 3, wherein the antibody is any one of the following (a''') to (i'''):
    (A′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27;
    (B''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:35;
    (C′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:39;
    (D′″) an antibody comprising a light chain variable region containing the amino acid sequence of SEQ ID NO: 41 and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44;
    (E′″) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:46 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:48;
    (F″′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:49 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:53;
    (G''') an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:57 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:59;
    (H″′) an antibody comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:57 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:62; and (i″′) an amino acid sequence of SEQ ID NO:64. An antibody comprising a light chain variable region comprising, and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:68.
  5. 　抗体が、以下の（ａ’’’’）～（ｉ’’’’）のいずれかの抗体である、請求項１～４のいずれかに記載の抗体：
    （ａ’’’’）配列番号２３のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号２７のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｂ’’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号３５のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｃ’’’’）配列番号３１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号３９のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｄ’’’’）配列番号４１のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号４４のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｅ’’’’）配列番号４６のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号４８のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｆ’’’’）配列番号４９のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号５３のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｇ’’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号５９のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；
    （ｈ’’’’）配列番号５７のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号６２のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体；および
    （ｉ’’’’）配列番号６４のアミノ酸配列を含む軽鎖可変領域におけるCDR1、CDR2、およびCDR3を含む軽鎖可変領域、および配列番号６８のアミノ酸配列を含む重鎖可変領域におけるCDR1、CDR2、およびCDR3を含む重鎖可変領域を含む抗体。     The antibody according to any one of claims 1 to 4, wherein the antibody is any one of the following (a'''') to (i''''):
    (A″″) CDR1, CDR2 in the light chain variable region including the amino acid sequence of SEQ ID NO:23, and CDR1, CDR2 in the heavy chain variable region including the amino acid sequence of SEQ ID NO:27 And an antibody comprising a heavy chain variable region comprising CDR3;
    (B″″) CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:35 and the light chain variable region containing CDR1, CDR2, and CDR3 in the light chain variable region of SEQ ID NO:31 And an antibody comprising a heavy chain variable region comprising CDR3;
    (C″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 31 and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 39 And an antibody comprising a heavy chain variable region comprising CDR3;
    (D″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 41, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 44 And an antibody comprising a heavy chain variable region comprising CDR3;
    (E″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO:46, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:48 And an antibody comprising a heavy chain variable region comprising CDR3;
    (F″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO:49, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO:53 And an antibody comprising a heavy chain variable region comprising CDR3;
    (G'''') CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 57, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 59 And an antibody comprising a heavy chain variable region comprising CDR3;
    (H″″) CDR1, CDR2 in the light chain variable region containing the amino acid sequence of SEQ ID NO: 57, and CDR1, CDR2 in the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 62 And (i″″) a light chain variable region comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 64, and a light chain variable region comprising CDR3, CDR2, and CDR3, and SEQ ID NO: An antibody comprising a heavy chain variable region comprising CDR1, CDR2, and CDR3 in a heavy chain variable region comprising 68 amino acid sequences.
  6. 　preS1抗原への結合において請求項１～５のいずれかに記載の抗体と競合する、抗preS1抗体。 An anti-preS1 antibody that competes with the antibody according to any one of claims 1 to 5 for binding to the preS1 antigen.
  7. 　抗preS1抗体であって、配列番号２、３および４から選択されるアミノ酸配列に含まれるエピトープに結合する、抗体。 An anti-preS1 antibody that binds to an epitope contained in the amino acid sequence selected from SEQ ID NOs: 2, 3 and 4.
  8. 　配列番号２のアミノ酸配列に含まれるエピトープに結合する、請求項７に記載の抗体。 The antibody according to claim 7, which binds to an epitope contained in the amino acid sequence of SEQ ID NO: 2.
  9. 　２種以上の抗preS1抗体の組み合わせであって、前記２種以上の抗preS1抗体が請求項１～８のいずれかに記載の抗体から独立に選択される、組み合わせ。 A combination of two or more anti-preS1 antibodies, wherein the two or more anti-preS1 antibodies are independently selected from the antibodies according to any one of claims 1 to 8.
  10. 　preS1抗原の測定方法であって、
    　対象から得た試料を請求項１～８のいずれかに記載の抗体と接触させること、および
    　preS1抗原と前記抗体との複合体を検出すること
    　を含む方法。     A method for measuring preS1 antigen, comprising:
    A method comprising contacting a sample obtained from a subject with an antibody according to any one of claims 1 to 8 and detecting a complex of preS1 antigen and the antibody.
  11. 　請求項１～８のいずれかに記載の抗体と、これとは異なるエピトープに結合する抗HBs抗体とを用いる、請求項１０に記載の方法。 The method according to claim 10, wherein the antibody according to any one of claims 1 to 8 and an anti-HBs antibody that binds to an epitope different from this are used.
  12. 　抗HBs抗体が、抗S抗体または抗preS2抗体である、請求項１１に記載の方法。 The method according to claim 11, wherein the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody.
  13. 　B型肝炎ウイルス感染の評価方法であって、
    　請求項１０～１２のいずれかに記載の方法によりpreS1抗原を測定すること、および
    　前記測定値を基準値と比較すること
    　を含む方法。     A method for evaluating hepatitis B virus infection, comprising:
    A method comprising measuring the preS1 antigen by the method according to claim 10, and comparing the measured value with a reference value.
  14. 　請求項１～９のいずれかに記載の抗体または組み合わせを含む、preS1抗原の測定用組成物。 A composition for measuring preS1 antigen, which comprises the antibody or the combination according to any one of claims 1 to 9.
  15. 　請求項１～９のいずれかに記載の抗体または組み合わせを含む、B型肝炎ウイルス感染の評価用組成物。 A composition for evaluating hepatitis B virus infection, which comprises the antibody or the combination according to any one of claims 1 to 9.
  16. 　請求項１～９のいずれかに記載の抗体または組み合わせを含む、preS1抗原の測定用キット。 A kit for measuring preS1 antigen, which comprises the antibody or the combination according to any one of claims 1 to 9.
  17. 　請求項１～９のいずれかに記載の抗体または組み合わせを含む、B型肝炎ウイルス感染の評価用キット。 A kit for evaluating hepatitis B virus infection, which comprises the antibody or the combination according to any one of claims 1 to 9.
  18. 　請求項１～９のいずれかに記載の抗体とは異なるエピトープに結合する抗HBs抗体をさらに含む、請求項１６または１７に記載のキット。 The kit according to claim 16 or 17, further comprising an anti-HBs antibody that binds to an epitope different from the antibody according to any one of claims 1 to 9.
  19. 　抗HBs抗体が、抗S抗体または抗preS2抗体である、請求項１８に記載のキット。 The kit according to claim 18, wherein the anti-HBs antibody is an anti-S antibody or an anti-preS2 antibody.
  20. 　請求項１～９のいずれかに記載の抗体または組み合わせを含む、医薬組成物。 A pharmaceutical composition comprising the antibody or the combination according to any one of claims 1 to 9.
  21. 　B型肝炎ウイルス感染の治療または予防のための、請求項２０に記載の医薬組成物。 21. The pharmaceutical composition according to claim 20, for treating or preventing hepatitis B virus infection.
  22. 　慢性B型肝炎の治療のための、請求項２０または２１に記載の医薬組成物。 A pharmaceutical composition according to claim 20 or 21 for the treatment of chronic hepatitis B.
PCT/JP2019/050139 2018-12-21 2019-12-20 Anti-pres1 antibody and use thereof WO2020130138A1 (en)

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