WO2020130138A1 - Anticorps anti-pres1 et son utilisation - Google Patents

Anticorps anti-pres1 et son utilisation 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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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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    • 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

La présente invention concerne : un anticorps dirigé contre l'antigène preS1 ou une combinaison de celui-ci; un procédé de mesure d'antigènes preS1 à l'aide de l'anticorps ou d'une combinaison de ceux-ci, et une composition et un kit pour ledit procédé; un procédé d'évaluation d'une infection par le VHB, et une composition et un kit pour celui-ci; et une composition pharmaceutique contenant l'anticorps ou la combinaison de ceux-ci.
PCT/JP2019/050139 2018-12-21 2019-12-20 Anticorps anti-pres1 et son utilisation WO2020130138A1 (fr)

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CN112920277A (zh) * 2021-01-27 2021-06-08 中国农业大学 分析溴氰虫酰胺和氯虫苯甲酰胺残留的vhh-elisa试剂盒及其应用

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JP2018519804A (ja) * 2015-05-22 2018-07-26 フアフイ ヘルス リミテッドＨｕａｈｕｉ Ｈｅａｌｔｈ Ｌｔｄ． 抗Ｐｒｅ−Ｓ１ ＨＢＶ抗体
JP2018533937A (ja) * 2015-10-09 2018-11-22 シャアメン ユニバーシティ Ｂ型肝炎表面抗原に対する抗体及びその使用

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JP2018533937A (ja) * 2015-10-09 2018-11-22 シャアメン ユニバーシティ Ｂ型肝炎表面抗原に対する抗体及びその使用

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Publication number Priority date Publication date Assignee Title
CN112920277A (zh) * 2021-01-27 2021-06-08 中国农业大学 分析溴氰虫酰胺和氯虫苯甲酰胺残留的vhh-elisa试剂盒及其应用

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