US20190076521A1 - Soybean allergy antigen - Google Patents

Soybean allergy antigen Download PDF

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Publication number
US20190076521A1
US20190076521A1 US16/081,032 US201716081032A US2019076521A1 US 20190076521 A1 US20190076521 A1 US 20190076521A1 US 201716081032 A US201716081032 A US 201716081032A US 2019076521 A1 US2019076521 A1 US 2019076521A1
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Prior art keywords
amino acid
protein
seq
acid sequence
nucleotide sequence
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US16/081,032
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Inventor
Kayoko Matsunaga
Akiko Yagami
Naoshi Shimojo
Masashi Nakamura
Nayu Sato
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Hoyu Co Ltd
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Hoyu Co Ltd
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Assigned to HOYU CO., LTD. reassignment HOYU CO., LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: MATSUNAGA, Kayoko, NAKAMURA, MASASHI, SATO, Nayu, SHIMOJO, Naoshi, YAGAMI, Akiko
Publication of US20190076521A1 publication Critical patent/US20190076521A1/en
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    • 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
    • G01N33/564—Immunoassay; Biospecific binding assay; Materials therefor for pre-existing immune complex or autoimmune disease, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/30—Removing undesirable substances, e.g. bitter substances
    • 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
    • A61K39/35—Allergens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A61K49/0006—Skin tests, e.g. intradermal testing, test strips, delayed hypersensitivity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K4/00—Peptides having up to 20 amino acids in an undefined or only partially defined sequence; Derivatives thereof
    • C07K4/10—Peptides having up to 20 amino acids in an undefined or only partially defined sequence; Derivatives thereof from plants
    • 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
    • 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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/24—Immunology or allergic disorders

Definitions

  • the present invention relates to a novel antigen of an allergy to soybean.
  • the present invention also relates to a kit, a composition, and a method for diagnosing allergy to soybean.
  • the present invention also relates to a pharmaceutical composition comprising such an antigen and soybean or processed products of soybean in which such an antigen is eliminated or reduced.
  • the present invention further relates to a tester for determining the presence or absence of a soybean antigen in an object of interest.
  • IgE antibodies specific to particular antigens are produced. Physiological consequences caused by interaction between such IgE antibodies and such particular antigens elicit allergic reactions.
  • antigen reagents are commonly prepared simply by grinding a candidate allergenic food, material or the like (Patent Literature 1).
  • conventional antigen reagents do not necessarily contain only a particular antigenic protein inducing an allergic reaction (allergen component), and rather contain different types of protein components.
  • conventional antigen reagents contain varied amounts of allergen components.
  • allergen component a particular antigenic protein inducing an allergic reaction
  • allergen component a particular protein acting as an allergen component
  • diagnosis efficiency was not sufficiently high when using a conventional allergy testing agent in patients possessing an IgE antibody binding to an allergen component present in small amounts in an allergen such as food.
  • the severity and symptoms of an allergic reaction do not necessarily correlate with the content of an allergen component. Even when a patient's IgE antibody reacts with an allergen component present in trace amounts in a candidate allergic food, material or the like, the allergic reaction may develop allergic symptoms or may affect the severity of those symptoms.
  • Electric charge-based purification methods can be exemplified by column chromatography using ion exchange resins, and isoelectric focusing. Purifications based on molecular weight difference can be exemplified by centrifugal separation, molecular-sieve column chromatography, and SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).
  • the present invention provides novel antigens of an allergy to soybean.
  • the present invention also provides methods and kits for diagnosing allergy to soybean.
  • the present invention also provides pharmaceutical compositions comprising such an antigen and soybean or processed products of soybean in which such an antigen is eliminated or reduced.
  • the present invention further provides testers for determining the presence or absence of a soybean antigen in an object of interest.
  • the present inventors had made intensive studies to identify causative antigens of an allergy to soybean. As a result, the inventors succeeded in identifying novel antigens to which an IgE antibody in the serum of a soybean-allergic patient specifically binds. The present invention has been completed based on this finding.
  • the present invention can be as defined below.
  • kits for diagnosing an allergy to a soybean comprising, as an antigen, at least one of proteins defined below in any of the following (1) to (8):
  • (1A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 2;
  • (1A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 2;
  • (1A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 1;
  • (1A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 1;
  • (1A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1; or
  • (2A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 3;
  • (2A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 3;
  • (2A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 3; or
  • (3A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 6;
  • (3A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 6;
  • (3A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 5;
  • (3A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 5; or
  • (3A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 5; or
  • (4A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 8;
  • (4A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 8;
  • (4A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 7;
  • (4A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 7; or
  • (4A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 7; or
  • (5A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 9;
  • (5A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 9; or
  • (6A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 11;
  • (6A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 11; or
  • (6A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 11; or
  • (7A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 14;
  • (7A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 14;
  • (7A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 13;
  • (7A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 13; or
  • (7A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 13; or
  • (8A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 15;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 15; or
  • (8A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 15; or
  • composition for diagnosing an allergy to a soybean comprising, as an antigen, at least one of proteins as defined above in any of (1) to (8) of [1].
  • a method for providing an indicator for diagnosing an allergy to a soybean in a subject comprising the steps of:
  • a pharmaceutical composition comprising at least one of proteins as defined above in any of (1) to (8) of [1].
  • a tester for determining the presence or absence of a soybean antigen in an object of interest comprising an antibody that binds to at least one of proteins as defined above in any of (1) to (8) of [1].
  • a soybean-derived antigen which is at least one of proteins as defined above in any of (1) to (8) of [1] and is causative of an allergy to soybean.
  • kits for diagnosing an allergy to a soybean comprising, as an antigen, at least one of proteins defined below in any one of (1 ⁇ ) to (55 ⁇ ):
  • (1 ⁇ A1-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 70;
  • (1 ⁇ A1-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 70;
  • (1 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 69;
  • (1 ⁇ A1-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 69; or
  • (1 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 69;
  • (1 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 71-87,
  • (1 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 89;
  • (1 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 89;
  • (1 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 88;
  • (1 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 88; or
  • (1 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 88; or
  • (1 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 90-104;
  • (2 ⁇ A1-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 106;
  • (2 ⁇ A1-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 106;
  • (2 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 105;
  • (2 ⁇ A1-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 105; or
  • (2 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 105; or
  • (2 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 107-120;
  • (2 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 122;
  • (2 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 122;
  • (2 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 121;
  • (2 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 121; or
  • (2 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 121; or
  • (2 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 123-137;
  • (2 ⁇ A3-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 139;
  • (2 ⁇ A3-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 139;
  • (2 ⁇ A3-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 138;
  • (2 ⁇ A3-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 138; or
  • (2 ⁇ A3-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 138; or
  • (2 ⁇ B3) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 140-153;
  • (4 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 155;
  • (4 ⁇ A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 155;
  • (4 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 154;
  • (4 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 154; or
  • (4 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 154; or
  • (4 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 156 and 157;
  • (5 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 159;
  • (5 ⁇ A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 159;
  • (5 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 158;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 158; or
  • (5 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 158; or
  • (5 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 160 and 161;
  • (6 ⁇ ) (6 ⁇ A1) aconitate hydratase 1 or a variant thereof, which is defined below in any of (6 ⁇ A1-a) to (6 ⁇ A1-e):
  • (6 ⁇ A1-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 163;
  • (6 ⁇ A1-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 163;
  • (6 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 162;
  • (6 ⁇ A1-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 162; or
  • (6 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 162;
  • (6 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 164-167; or
  • (6 ⁇ A2) aconitate hydratase 1 or a variant thereof, which is defined below in any of (6 ⁇ A2-a) to (6 ⁇ A2-e):
  • (6 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 169;
  • (6 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 169;
  • (6 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 168;
  • (6 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 168; or
  • (6 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 168; or
  • (6 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 170-172;
  • (7 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 174;
  • (7 ⁇ A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 174;
  • (7 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 173;
  • (7 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 173; or
  • (7 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 173; or
  • (7 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 175-187;
  • (8 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 188;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 188; or
  • (8 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 188; or
  • (8 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 190-199;
  • (9 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 201;
  • (9 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 200;
  • (9 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 200; or
  • (9 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 200; or
  • (10 ⁇ A1-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 204;
  • (10 ⁇ A1-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 204;
  • (10 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 203;
  • (10 ⁇ A1-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 203; or
  • (10 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 203;
  • (10 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 205-218;
  • (10 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 220;
  • (10 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 220;
  • (10 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 219;
  • (10 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 219; or
  • (10 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 219; or
  • (10 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 221-231;
  • (11 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 233;
  • (11 ⁇ A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 233;
  • (11 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 232;
  • (11 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 232; or
  • (11 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 232; or
  • (11 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 234-237;
  • (12 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 239;
  • (12 ⁇ A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 239;
  • (12 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 238;
  • (12 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 238; or
  • (12 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 238; or
  • (12 ⁇ B) a protein comprising an amino acid sequence of SEQ ID NO: 240;
  • V-type proton ATPase subunit B 2 or a variant thereof, which is defined below in any of (14 ⁇ A1-a) to (14 ⁇ A1-e):
  • (14 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 279;
  • (14 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 279;
  • V-type proton ATPase subunit B 2 or a variant thereof, which is defined below in any of (14 ⁇ A2-a) to (14 ⁇ A2-e):
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 290;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 290; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 290;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 305; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 305; or
  • (17 ⁇ A1-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 320;
  • (17 ⁇ A1-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 320;
  • (17 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 319;
  • (17 ⁇ A1-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 319; or
  • (17 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 319;
  • (17 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 321-325;
  • (17 ⁇ A2) rab GDP dissociation inhibitor alpha-like or a variant thereof, which is defined below in any of (17 ⁇ A2-a) to (17 ⁇ A2-e):
  • (17 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 327;
  • (17 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 327;
  • (17 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 326;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 326; or
  • (17 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 326; or
  • (17 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 328-332;
  • (18 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 334;
  • (18 ⁇ A-b) a protein comprising an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 334;
  • (18 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 333;
  • (18 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 333; or
  • (18 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 333; or
  • (18 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 335-339;
  • sucrose binding protein homolog S-64 or a variant thereof, which is defined below in any of (19 ⁇ A-a) to (19 ⁇ A-e):
  • (19 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 340;
  • (19 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 340; or
  • (19 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 340; or
  • (25 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 347;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 347; or
  • (25 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 347;
  • (25 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 349-356;
  • (25 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 357;
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 357; or
  • (26 ⁇ ) (26 ⁇ A1) phosphoglycerate kinase, cytosolic or a variant thereof, which is defined below in any of (26 ⁇ A1-a) to (26 ⁇ A1-e):
  • (26 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 366;
  • (26 ⁇ A1-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 366; or
  • (26 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 366;
  • (26 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 368-382;
  • (26 ⁇ A2) phosphoglycerate kinase, cytosolic or a variant thereof, which is defined below in any of (26 ⁇ A2-a) to (26 ⁇ A2-e):
  • (26 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 383;
  • (26 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 383;
  • (26 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 385-392;
  • (26 ⁇ A3-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 393;
  • (26 ⁇ A3-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 393; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 421; or
  • (28 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 428;
  • (28 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 428; or
  • (34 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 452;
  • (34 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 452;
  • (34 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 455;
  • (34 ⁇ A2-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 455; or
  • (35 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 459;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 459; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 459;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 467; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 467;
  • (35 ⁇ A3-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 472;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 472; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 472; or
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 478; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 478;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 484; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 484;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 488; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 488; or
  • (38 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 494;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 494; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 494; or
  • (39 ⁇ ) (39 ⁇ A) ATP synthase subunit O mitochondrial-like or a variant thereof, which is defined below in any of (39 ⁇ A-a) to (39 ⁇ A-e):
  • (39 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 514;
  • (39 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 514; or
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 517; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 517; or
  • (41 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 522;
  • (41 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 521;
  • (41 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 521; or
  • (41 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 521; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 524;
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 532; or
  • (45 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 540;
  • (45 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ 1D NO: 540;
  • (45 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 542-545;
  • (46 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 552;
  • (46 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 552; or
  • (46 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 552; or
  • (48 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 564;
  • (48 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 564; or
  • (50 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 571;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 571; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 571; or
  • (51 ⁇ A-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 578;
  • (51 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 577;
  • (51 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 577; or
  • (51 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 577; or (51 ⁇ B) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 579-582;
  • (52 ⁇ A-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 583;
  • (52 ⁇ A-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 583; or
  • (52 ⁇ A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 583; or
  • triosephosphate isomerase isoform X1 or a variant thereof, which is defined below in any of (53 ⁇ A1-a) to (53 ⁇ A1-e):
  • (53 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 589;
  • (53 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 589;
  • (53 ⁇ A2) triosephosphate isomerase, cytosolic or a variant thereof, which is defined below in any of (53 ⁇ A2-a) to (53 ⁇ A2-e):
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 598; or
  • (53 ⁇ A1-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 605;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 605; or
  • (54 ⁇ A1-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 605;
  • (54 ⁇ B1) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 607-611;
  • (54 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 612;
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 612; or
  • (54 ⁇ B2) a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 614-616; or
  • (55 ⁇ ) (55 ⁇ A1) 51 kDa seed maturation protein precursor or a variant thereof, which is defined below in any of (55 ⁇ A1-a) to (55 ⁇ A1-e):
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 617;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 617; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 617;
  • a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 636; or
  • a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 636; or
  • composition for diagnosing an allergy to a soybean comprising, as an antigen, at least one of proteins as defined above in any of (1 ⁇ ) to (55 ⁇ ) of [9].
  • a method for providing an indicator for diagnosing an allergy to a soybean in a subject comprising the steps of:
  • a pharmaceutical composition comprising at least one of proteins as defined above in any of (1 ⁇ ) to (55 ⁇ ) of [9].
  • a tester for determining the presence or absence of a soybean antigen in an object of interest comprising an antibody that binds to at least one of proteins as defined above in any of (1 ⁇ ) to (55 ⁇ ) of [9].
  • a soybean-derived antigen which is at least one of proteins as defined above in any of (1 ⁇ ) to (55 ⁇ ) of [9] and is causative of an allergy to soybean.
  • the present invention can provide novel antigens of an allergy to soybean. Since the novel antigens (allergen components) that trigger a soybean allergy were identified according to this invention, this invention can provide highly sensitive methods and kits for diagnosing an allergy to soybean, pharmaceutical compositions comprising such an antigen, soybean or processed products of soybean in which such an antigen is eliminated or reduced.
  • FIG. 1 is a photograph of a gel showing a protein electrophoretic pattern in two-dimensional electrophoresis of proteins contained in soybean.
  • the bands at the left of the photograph are bands of molecular weight markers.
  • FIG. 2 is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of a healthy subject.
  • FIG. 3A is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 1.
  • the arrows indicate spots where the serum of the soybean-allergic patient specifically reacted.
  • FIG. 3B is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 2.
  • the arrows indicate spots where the serum of the soybean-allergic patient specifically reacted.
  • FIG. 3C is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 3.
  • the arrows indicate spots where the serum of the soybean-allergic patient specifically reacted.
  • FIG. 3D is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 4.
  • the arrows indicate spots where the serum of the soybean-allergic patient specifically reacted.
  • FIG. 4 is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of a healthy subject.
  • FIG. 5A is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 1 ⁇ .
  • the spots indicated with a white line or enclosed in a square are spot where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5B is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 2 ⁇ .
  • the spots indicated with a white line or enclosed in a square are spot where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5C is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 3 ⁇ .
  • the spots indicated with a white line or enclosed in a square are spot where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5D is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 4 ⁇ .
  • the spots indicated with a while line are spots where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5E is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 5 ⁇ .
  • the spots indicated with a while line are spots where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5F is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 6 ⁇ .
  • the spots indicated with a white line or enclosed in a square are spot where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5G is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 7 ⁇ .
  • the spots enclosed in a square are spots where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5H is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 8 ⁇ .
  • the spots indicated with a white line or enclosed in a square are spot where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • FIG. 5I is a photograph of an immunoblot of a two-dimensional electrophoretic pattern of proteins contained in soybean stained with serum of Soybean-allergic patient 9 ⁇ .
  • the spots indicated with a white line or enclosed in a square are spot where the serum of the soybean-allergic patient specifically reacted.
  • the numbers (n) indicated are the numbers of the spots and correspond to the spots indicated with “n ⁇ ” herein.
  • Amino acid sequences herein are represented by one-letter amino acid symbols well known to those skilled in the art and the left end corresponds to the amino terminal and the right end corresponds to the carboxy terminal.
  • the “allergy” refers to the state in which, when a certain antigen enters the body of a living individual sensitized to said antigen, the living individual shows a hypersensitive reaction detrimental to him/her.
  • IgE antibodies specific to antigens are produced. IgE antibodies bind to mast cells or basophils.
  • an antigen specific to such an IgE antibody enters again the body of a patient with an allergic disease, said antigen combines with the IgE antibody bound to mast cells or basophils, and the IgE antibody crosslinks said antigen on the cell surface, resulting in physiological effects of IgE antibody-antigen interaction.
  • physiological effects include release of histamine, serotonin, heparin, eosinophil chemotactic factors, leucotrienes, or the like. These released substances provoke an allergic reaction resulting from the combination of an IgE antibody with particular antigens. Such allergic reactions caused by particular antigens occur through the aforementioned pathway.
  • the “allergy to soybean” refers to the state in which an individual has an allergic reaction caused by proteins, etc. present in soybean which act as an antigen.
  • the allergy to soybean can produce an allergic reaction upon contact with, or consumption of, an antigen present in soybean.
  • allergic reactions caused by consumption of foods are particularly referred to as “food allergies”.
  • the allergy to soybean may be a food allergy.
  • the “antigen” refers to a substance that provokes an allergic reaction, and is also referred to as an “allergen component”.
  • the antigen is preferably a protein.
  • the “protein” refers to a molecule having a structure in which naturally occurring amino acids are joined together by peptide bond.
  • the number of amino acids present in a protein is not particularly limited, but proteins having about 2 to 50 amino acids joined together by peptide bond are in some cases called “peptides”. In the case where amino acids can form different enantiomers, the amino acids are understood to form an L-enantiomer, unless otherwise indicated.
  • the amino acid sequences of proteins or peptides as used herein are represented by one-letter symbols of amino acids in accordance with standard usage and the notational convention commonly used in the art. The leftward direction represents the amino-terminal direction, and the rightward direction represents the carboxy-terminal direction.
  • soybean proteins contained in soybean were analyzed by the aforementioned technique to identify causative antigens of an allergy to soybean. To be specific, soybean proteins were subjected to two-dimensional electrophoresis under the conditions described below.
  • the electrophoresis in the first dimension was isoelectric focusing, which was performed using isoelectric focusing gels with a gel-strip length of 5 to 10 cm and a gel pH range of 3 to 10.
  • the isoelectric focusing was performed using the IPG gels, Immobiline Drystrip (pH3-10NL), produced by GE Healthcare Bio-Sciences Corporation (hereinafter abbreviated as “GE”).
  • the electrophoresis system used was IPGphor produced by GE.
  • the maximum current of the electrophoresis system was limited to 75 ⁇ A per gel strip.
  • the voltage program adopted to perform the first-dimensional isoelectric focusing was as follows: (1) a constant voltage step was performed at a constant voltage of 300 V until the volt-hours reached 750 Vhr (the current variation width during electrophoresis for 30 minutes before the end of this step was 5 ⁇ A); (2) the voltage was increased gradually to 1000 V for 300 Vhr; (3) the voltage was further increased gradually to 5000 V for 4500 Vhr; and then (4) the voltage was held at a constant voltage of 5000 V until the total Vhr reached 12000.
  • the electrophoresis in the second dimension was SDS-PAGE, which was performed using polyacrylamide gels whose gel concentration at the distal end in the direction of electrophoresis was set to 3 to 6% and whose gel concentration at the proximal end was set to a higher value than that at the distal end. More specifically, the SDS-PAGE was performed using NuPAGE 4-12% Bris-Tris Gels (IPG well, Mini, 1 mm) produced by Life Technologies. The electrophoresis system used was XCell SureLock Mini-Cell produced by Life Technologies. The electrophoresis was run at a constant voltage of 200 V for about 45 minutes using an electrophoresis buffer composed of 50 mM MOPS, 50 mM Tris base, 0.1% (w/v) SDS and 1 mM EDTA.
  • Spot 1 Molecular weight 1 to 30 kDa, pI 3.0 to 7.0 Spot 2: Molecular weight 30 to 75 kDa, pI 4.0 to 8.5 Spot 3: Molecular weight 20 to 50 kDa, pI 7.0 to 12.0 Spot 4: Molecular weight 20 to 50 kDa, pI 6.0 to 10.0 Spot 5: Molecular weight 2 to 30 kDa, pI 4.0 to 9.0 Spot 6: Molecular weight 50 to 100 kDa, pI 5.0 to 10.0 Spot 7: Molecular weight 5 to 37 kDa, pI 3.0 to 7.0 Spot 8: Molecular weight 75 to 200 kDa, pI 4.0 to 6.0 Spot 1 ⁇ : Molecular weight 70 to 120 kDa, pI 2.5 to 6.5 Spot 2 ⁇ : Molecular weight 50 to 120 kDa, pI 2.5 to 6.5 Spot 4 ⁇ : Molecular weight 70 to 120 kDa, pI 3.5 to 7.5 Spo
  • the mass spectroscopic data obtained for spot 1 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Bowman-Birk type proteinase inhibitor D-II (amino acid sequence: SEQ ID NO: 2, encoding nucleotide sequence: SEQ ID NO: 1).
  • the antigen in spot 1 can be any of (1A-a) to (1A-e) and (1B) as defined below:
  • (1A-e) a protein comprising an amino acid sequence encoded by a nucleic acid that hybridizes under stringent conditions with a nucleic acid having a nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1;
  • a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 17-20, preferably a protein comprising at least 2 or 3 or all sequences of the amino acid sequences; more preferably a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 17-19, preferably a protein comprising at least 2 or all sequences of the amino acid sequences.
  • the amino acid sequence of any of SEQ ID NOs: 17-20 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the proteins of (1A-a) to (1A-e) and (1B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (1A-a) to (1A-e) and (1B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 1 to 30 kDa, preferably around 2 to 25 kDa, more preferably around 5 to 20 kDa and an isoelectric point of 3.0 to 7.0, preferably 4.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 1 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 2 (Bowman-Birk type proteinase inhibitor D-II) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 2 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Ribulose bisphosphate carboxylase large chain (amino acid sequence: SEQ ID NO: 4, encoding nucleotide sequence: SEQ ID NO: 3).
  • the antigen in spot 2 can be any of (2A-a) to (2A-e) and (2B) as defined below:
  • the proteins of (2A-a) to (2A-e) and (2B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (2A-a) to (2A-e) and (2B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 30 to 75 kDa, preferably around 37 to 70 kDa, more preferably around 40 to 60 kDa and an isoelectric point of 4.0 to 8.5, preferably 5.5 to 7.5 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 2 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 4 (Ribulose bisphosphate carboxylase large chain) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 3 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Gamma-glutamyl hydrolase (amino acid sequence: SEQ ID NO: 6, encoding nucleotide sequence: SEQ ID NO: 5).
  • the antigen in spot 3 can be any of (3A-a) to (3A-e) and (3B) as defined below:
  • the amino acid sequence(s) of SEQ ID NOs: 29 and/or 30 may be an amino acid sequence(s) derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the proteins of (3A-a) to (3A-e) and (3B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (3A-a) to (3A-e) and (3B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 20 to 50 kDa, preferably around 25 to 40 kDa, more preferably around 25 to 37 kDa and an isoelectric point of 7.0 to 12.0, preferably 8.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 3 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 6 (Gamma-glutamyl hydrolase) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 4 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Guanine nucleotide-binding protein subunit beta-like protein (amino acid sequence: SEQ ID NO: 8, encoding nucleotide sequence: SEQ ID NO: 7).
  • the antigen in spot 4 can be any of (4-a) to (4-e) and (4B) as defined below:
  • the proteins of (4A-a) to (4A-e) and (4B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (4A-a) to (4A-e) and (4B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 20 to 50 kDa, preferably around 25 to 40 kDa, more preferably around 25 to 37 kDa and an isoelectric point of 6.0 to 10.0, preferably 7.0 to 9.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 4 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 8 (Guanine nucleotide-binding protein subunit beta-like protein) or causative of an allergy to soybean.
  • Protein SLE3 amino acid sequence: SEQ ID NO: 10, encoding nucleotide sequence: SEQ ID NO: 9.
  • the antigen in spot 5 can be any of (5A-a) to (5A-e) and (5B) as defined below:
  • the proteins of (5A-a) to (5A-e) and (5B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (5A-a) to (5A-e) and (5B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 2 to 30 kDa, preferably around 3 to 25 kDa, more preferably around 5 to 20 kDa and an isoelectric point of 4.0 to 9.0, preferably 5.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 5 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 10 (Protein SLE3) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 6 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Seed biotin-containing protein SBP65 (amino acid sequence: SEQ ID NO: 12, encoding nucleotide sequence: SEQ ID NO: 11).
  • the antigen in spot 6 can be any of (6A-a) to (6A-e) and (6B) as defined below:
  • the proteins of (6A-a) to (6A-e) and (6B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (6A-a) to (6A-e) and (6B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 50 to 100 kDa, preferably around 55 to 90 kDa, more preferably around 60 to 75 kDa and an isoelectric point of 5.0 to 10.0, preferably 6.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 6 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 12 (Seed biotin-containing protein SBP65) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 7 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Ferritin-2 (amino acid sequence: SEQ ID NO: 14, encoding nucleotide sequence: SEQ ID NO: 13).
  • the antigen in spot 7 can be any of (7A-a) to (7A-e) and (7B) as defined below:
  • the proteins of (7A-a) to (7A-e) and (7B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (7A-a) to (7A-e) and (7B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 5 to 37 kDa, preferably around 10 to 30 kDa, more preferably around 15 to 25 kDa and an isoelectric point of 3.0 to 7.0, preferably 4.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 7 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 14 (Ferritin-2) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 8 on a mass spectrometer was analyzed by comparing the data against the Uniprot protein data, and as a result, the antigen in question was identified as Cell division cycle protein 48 homolog (amino acid sequence: SEQ ID NO: 16, encoding nucleotide sequence: SEQ ID NO: 15).
  • the antigen in spot 8 can be any of (8A-a) to (8A-e) and (8B) as defined below:
  • the proteins of (8A-a) to (8A-e) and (8B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins of (8A-a) to (8A-e) and (8B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 75 to 200 kDa, preferably around 100 to 170 kDa, more preferably around 100 to 150 kDa and an isoelectric point of 3.0 to 7.0, preferably 4.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 8 has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 16 (Cell division cycle protein 48 homolog) or causative of an allergy to soybean.
  • endoplasmin homolog isoform X2 (amino acid sequence: SEQ ID NO: 70, encoding nucleotide sequence: SEQ ID NO: 69) was identified as a protein comprising any of SEQ ID NOs: 71 to 87 and endoplasmin homolog isoform X2 (amino acid sequence: SEQ ID NO: 89, encoding nucleotide sequence: SEQ ID NO: 88) was identified as a protein comprising any of SEQ ID NOs: 90 to 104.
  • the antigen in spot 1 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (1 ⁇ A1-a) to (1 ⁇ A1-e), (1 ⁇ B1), (1 ⁇ A2-a) to (1 ⁇ A2-e), and (1 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 71 to 87 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids;
  • the protein selected from the group consisting of the proteins as defined above in (1 ⁇ A1-a) to (1 ⁇ A1-e), (1 ⁇ B1), (1 ⁇ A2-a) to (1 ⁇ A2-e), and (1 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (1 ⁇ A1-a) to (1 ⁇ A1-e), (1 ⁇ B1), (1 ⁇ A2-a) to (1 ⁇ A2-e), and (1 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 70 to 120 kDa, preferably around 80 to 110 kDa, and an isoelectric point of 2.5 to 6.5, preferably 3.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 1 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 70 or 89 (endoplasmin homolog isoform X2) or causative of an allergy to soybean.
  • heat shock cognate protein 80 (amino acid sequence: SEQ ID NO: 106, encoding nucleotide sequence: SEQ ID NO: 105) was identified as a protein comprising any of SEQ ID NOs: 107 to 120
  • heat shock protein 90-1 (amino acid sequence: SEQ ID NO: 122, encoding nucleotide sequence: SEQ ID NO: 121) was identified as a protein comprising any of SEQ ID NOs: 123 to 137
  • hypothetical protein GLYMA_02G302500 was identified as a protein comprising any of SEQ ID NOs: 140 to 153.
  • the antigen in spot 1 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (2 ⁇ A1-a) to (2 ⁇ A1-e), (2 ⁇ B1), (2 ⁇ A2-a) to (2 ⁇ A2-e), (2 ⁇ B2), (2 ⁇ A3-a) to (2 ⁇ A3-e), and (2 ⁇ B3):
  • the amino acid sequence of any of SEQ ID NOs: 107 to 120 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (2 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 122; (2 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 122; (2 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 121; (2 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably at least 75%,
  • the amino acid sequence of any of SEQ ID NOs: 123 to 137 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the protein selected from the group consisting of the proteins as defined above in (2 ⁇ A1-a) to (2 ⁇ A1-e), (2 ⁇ B1), (2 ⁇ A2-a) to (2 ⁇ A2-e), (2 ⁇ B2), (2 ⁇ A3-a) to (2 ⁇ A3-e), and (2 ⁇ B3) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (2 ⁇ A1-a) to (2 ⁇ A1-e), (2 ⁇ B1), (2 ⁇ A2-a) to (2 ⁇ A2-e), (2 ⁇ B2), (2 ⁇ A3-a) to (2 ⁇ A3-e), and (2 ⁇ B3) can be proteins that are found in a protein spot with a molecular weight of around 50 to 120 kDa, preferably around 60 to 110 kDa, and an isoelectric point of 2.5 to 6.5, preferably 3.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 2 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 106 (heat shock cognate protein 80), the protein having the amino acid sequence of SEQ ID NO: 122 (heat shock protein 90-1), or the protein having the amino acid sequence of SEQ ID NO: 139 (hypothetical protein GLYMA_02G302500), or causative of an allergy to soybean.
  • embryo-specific urease isoform X1 (amino acid sequence: SEQ ID NO: 155, encoding nucleotide sequence: SEQ ID NO: 154) was identified as a protein comprising SEQ ID NO: 156 or 157.
  • the antigens in spot 4 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (4 ⁇ A-a) to (4 ⁇ A-e) and (4 ⁇ B):
  • the proteins selected from the group consisting of (4 ⁇ A-a) to (4 ⁇ A-e) and (4 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (4 ⁇ A-a) to (4 ⁇ A-e) and (4 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 70 to 120 kDa, preferably around 80 to 110 kDa and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 4 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 155 (embryo-specific urease isoform X1) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 5 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, alpha-1,4 glucan phosphorylase L isozyme, chloroplastic/amyloplastic (amino acid sequence: SEQ ID NO: 159, encoding nucleotide sequence: SEQ ID NO: 158) was identified as a protein comprising SEQ ID NO: 160 or 161.
  • the antigens in spot 5 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (5 ⁇ A-a) to (5 ⁇ A-e) and (5 ⁇ B):
  • the proteins selected from the group consisting of (5 ⁇ A-a) to (5 ⁇ A-e) and (5 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (5 ⁇ A-a) to (5 ⁇ A-e) and (5 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 70 to 170 kDa, preferably around 80 to 160 kDa and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 5 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 159 (alpha-1,4 glucan phosphorylase L isozyme, chloroplastic/amyloplastic) or causative of an allergy to soybean.
  • aconitate hydratase 1 (amino acid sequence: SEQ ID NO: 163, encoding nucleotide sequence: SEQ ID NO: 162) was identified as a protein comprising any of SEQ ID NOs: 164 to 167 and aconitate hydratase 1 (amino acid sequence: SEQ ID NO: 169, encoding nucleotide sequence: SEQ ID NO: 168) was identified as a protein comprising any of SEQ ID NOs: 170 to 172.
  • the antigen in spot 6 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (6 ⁇ A1-a) to (6 ⁇ A1-e), (6 ⁇ B1), (6 ⁇ A2-a) to (6 ⁇ A2-e), and (6 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 164 to 167 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (6 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 169; (6 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 169; (6 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 168; (6 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably at least 75%
  • the protein selected from the group consisting of the proteins as defined above in (6 ⁇ A1-a) to (6 ⁇ A1-e), (6 ⁇ B1), (6 ⁇ A2-a) to (6 ⁇ A2-e), and (6 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (6 ⁇ A1-a) to (6 ⁇ A1-e), (6 ⁇ B1), (6 ⁇ A2-a) to (6 ⁇ A2-e), and (6 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 70 to 120 kDa, preferably around 80 to 110 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 6 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 163 or 169 (aconitate hydratase 1) or causative of an allergy to soybean.
  • SEQ ID NO: 175 AGITVIQIDEAALREGLPLRK (SEQ ID NO: 176) ALGVDTVPVLVGPVTYLLLSKPAK (SEQ ID NO: 177) DEAFFSGNAAALASRK (SEQ ID NO: 178) DEVEDLEK (SEQ ID NO: 179) ISEEEYVK (SEQ ID NO: 180) IVEVNALAK (SEQ ID NO: 181) LIRNELAK (SEQ ID NO: 182) LLSVFREGVK (SEQ ID NO: 183) LVVSTSSSLLHTAVDLVNETK (SEQ ID NO: 184) SFSLLSLLPK (SEQ ID NO: 185) SSPRVTNEAVQK (SEQ ID NO: 186) TLTSLNGVTAYGFDLVRGTNTLDLIK (SEQ ID NO: 187) YGAGIGPGVYDIHSPRIPPTEEIADRINK
  • methionine synthase (amino acid sequence: SEQ ID NO: 174, encoding nucleotide sequence: SEQ ID NO: 173) was identified as a protein comprising any of SEQ ID NOs: 175 to 187.
  • the antigens in spot 7 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (7 ⁇ A-a) to (7 ⁇ A-e) and (7 ⁇ B):
  • the proteins selected from the group consisting of (7 ⁇ A-a) to (7 ⁇ A-e) and (7 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (7 ⁇ A-a) to (7 ⁇ A-e) and (7 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 50 to 120 kDa, preferably around 60 to 110 kDa and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 7 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 174 (methionine synthase) or causative of an allergy to soybean.
  • AIGIDRFGASAPAGRIYK (SEQ ID NO: 191) ALPTYTPESPADATRNLSQTNLNALAK (SEQ ID NO: 192) ANSYSVHGSALGAK (SEQ ID NO: 193) ATADAALVEK (SEQ ID NO: 194) DKPTLIK (SEQ ID NO: 195) GGYTISDNSTGNKPDVILIGTGSELEIAAK (SEQ ID NO: 196) LPQLPGTSIEGVEK (SEQ ID NO: 197) NGNNGYDDIRAAIK (SEQ ID NO: 198) SVNTIRFLAIDAVEK (SEQ ID NO: 199) VTTTIGYGSPNK
  • the mass spectroscopic data obtained for spot 8 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, transketolase, chloroplastic (amino acid sequence: SEQ ID NO: 189, encoding nucleotide sequence: SEQ ID NO: 188) was identified as a protein comprising any of SEQ ID NOs: 190 to 199.
  • the antigens in spot 8 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (8 ⁇ A-a) to (8 ⁇ A-e) and (8 ⁇ B):
  • the proteins selected from the group consisting of (8 ⁇ A-a) to (8 ⁇ A-e) and (8 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (8 ⁇ A-a) to (8 ⁇ A-e) and (8 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 50 to 120 kDa, preferably around 60 to 110 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 8 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 189 (transketolase, chloroplastic) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 9 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, lysine-tRNA ligase, cytoplasmic-like isoform X2 (amino acid sequence: SEQ ID NO: 201, encoding nucleotide sequence: SEQ ID NO: 200) was identified as a protein comprising SEQ ID NO: 202.
  • the antigens in spot 9 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (9 ⁇ A-a) to (9 ⁇ A-e) and (9 ⁇ B):
  • the proteins selected from the group consisting of (9 ⁇ A-a) to (9 ⁇ A-e) and (9 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (9 ⁇ A-a) to (9 ⁇ A-e) and (9 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 50 to 120 kDa, preferably around 60 to 110 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 9 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 201 (lysine-tRNA ligase, cytoplasmic-like isoform X2) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 10 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, chaperonin CPN60-2, mitochondrial (amino acid sequence: SEQ ID NO: 204, encoding nucleotide sequence: SEQ ID NO: 203) was identified as a protein comprising any of SEQ ID NOs: 205 to 218 and chaperonin CPN60-2, mitochondrial-like isoform X1 (amino acid sequence: SEQ ID NO: 220, encoding nucleotide sequence: SEQ ID NO: 219) was identified as a protein comprising any of SEQ ID NOs: 221 to 231.
  • the amino acid sequence of any of SEQ ID NOs: 205 to 218 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (10 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 220; (10 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 220; (10 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 219; (10 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably at least 75%
  • the protein selected from the group consisting of the proteins as defined above in (10 ⁇ A1-a) to (10 ⁇ A1-e), (10 ⁇ B1), (10 ⁇ A2-a) to (10 ⁇ A2-e), and (10 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (10 ⁇ A1-a) to (10 ⁇ A1-e), (10 ⁇ B1), (10 ⁇ A2-a) to (10 ⁇ A2-e), and (10 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 10 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 204 (haperonin CPN60-2, mitochondrial) or the protein having the amino acid sequence of SEQ ID NO: 220 (chaperonin CPN60-2, mitochondrial-like isoform X1) or causative of an allergy to soybean.
  • AIDVLNFTPLNGK (SEQ ID NO: 235)
  • FNNVFVK (SEQ ID NO: 236)
  • GFGFVNFANVDDAAK (SEQ ID NO: 237)
  • polyadenylate-binding protein 8 isoform X3 (amino acid sequence: SEQ ID NO: 233, encoding nucleotide sequence: SEQ ID NO: 232) was identified as a protein comprising any of SEQ ID NOs: 234 to 237.
  • the antigens in spot 11 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (11 ⁇ A-a) to (11 ⁇ A-e) and (11 ⁇ B):
  • the proteins selected from the group consisting of (11 ⁇ A-a) to (11 ⁇ A-e) and (11 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (11 ⁇ A-a) to (11 ⁇ A-e) and (11 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 11 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 233 (polyadenylate-binding protein 8 isoform X3) or causative of an allergy to soybean.
  • the proteins selected from the group consisting of (12 ⁇ A-a) to (12 ⁇ A-e) and (12 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (12 ⁇ A-a) to (12 ⁇ A-e) and (12 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 12 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 239 (pyrophosphate-fructose 6-phosphate 1-phosphotransferase subunit alpha-like) or causative of an allergy to soybean.
  • protein disulfide isomerase-like protein precursor (amino acid sequence: SEQ ID NO: 242, encoding nucleotide sequence: SEQ ID NO: 241) was identified as a protein comprising any of SEQ ID NOs: 243 to 252 and protein disulfide-isomerase (amino acid sequence: SEQ ID NO: 254, encoding nucleotide sequence: SEQ ID NO: 253) was identified as a protein comprising any of SEQ ID NOs: 255 to 278.
  • the antigen in spot 13 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (13 ⁇ A1-a) to (13 ⁇ A1-e), (13 ⁇ B1), (13 ⁇ A2-a) to (13 ⁇ A2-e), and (13 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 205 to 218 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (13 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 254; (13 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 254; (13 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 253; (13 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the protein selected from the group consisting of the proteins as defined above in (13 ⁇ A1-a) to (13 ⁇ A1-e), (13 ⁇ B1), (13 ⁇ A2-a) to (13 ⁇ A2-e), and (13 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (13 ⁇ A1-a) to (13 ⁇ A1-e), (13 ⁇ B1), (13 ⁇ A2-a) to (13 ⁇ A2-e), and (13 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 13 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 242 (protein disulfide isomerase-like protein precursor) or the protein having the amino acid sequence of SEQ ID NO: 254 (protein disulfide-isomerase) or causative of an allergy to soybean.
  • V-type proton ATPase subunit B2 (amino acid sequence: SEQ ID NO: 280, encoding nucleotide sequence: SEQ ID NO: 279) was identified as a protein comprising any of SEQ ID NOs: 281 to 284 and V-type proton ATPase subunit B2 (amino acid sequence: SEQ ID NO: 286, encoding nucleotide sequence: SEQ ID NO: 285) was identified as a protein comprising any of SEQ ID NOs: 287 to 289.
  • the antigen in spot 14 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (14 ⁇ A1-a) to (14 ⁇ A1-e), (14 ⁇ B1), (14 ⁇ A2-a) to (14 ⁇ A2-e), and (14 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 281 to 284 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (14 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 286; (14 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 286; (14 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 285; (14 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the protein selected from the group consisting of the proteins as defined above in (14 ⁇ A1-a) to (14 ⁇ A1-e), (14 ⁇ B1), (14 ⁇ A2-a) to (14 ⁇ A2-e), and (14 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (14 ⁇ A1-a) to (14 ⁇ A1-e), (14 ⁇ B1), (14 ⁇ A2-a) to (14 ⁇ A2-e), and (14 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa, and an isoelectric point of 2.5 to 6.5, preferably 3.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 14 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 280 or 286 (V-type proton ATPase subunit B2) or causative of an allergy to soybean.
  • the antigen in spot 15 ⁇ , 16 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (15, 16 ⁇ A1-a) to (15, 16 ⁇ A1-e), (15, 16 ⁇ B1), (15, 16 ⁇ A2-a) to (15, 16 ⁇ A2-e), and (15, 16 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 292 to 304 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids;
  • 15, 16 ⁇ A2-a a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 306;
  • 15, 16 ⁇ A2-b a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 306;
  • 15, 16 ⁇ A2-c a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 305;
  • 15, 16 ⁇ A2-d a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least
  • the protein selected from the group consisting of the proteins as defined above in (15, 16 ⁇ A1-a) to (15, 16 ⁇ A1-e), (15, 16 ⁇ B1), (15, 16 ⁇ A2-a) to (15, 16 ⁇ A2-e), and (15, 16 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (15, 16 ⁇ A1-a) to (15, 16 ⁇ A1-e), (15, 16 ⁇ B1), (15, 16 ⁇ A2-a) to (15, 16 ⁇ A2-e), and (15, 16 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 20 to 90 kDa, preferably around 30 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 15 ⁇ or 16 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 291 (UTP-glucose-1-phosphate uridylyltransferase) or the protein having the amino acid sequence of SEQ ID NO: 306 (hypothetical protein GLYMA_02G241100) or causative of an allergy to soybean.
  • guanosine nucleotide diphosphate dissociation inhibitor 2 (amino acid sequence: SEQ ID NO: 320, encoding nucleotide sequence: SEQ ID NO: 319) was identified as a protein comprising any of SEQ ID NOs: 321 to 325 and rab GDP dissociation inhibitor alpha-like (amino acid sequence: SEQ ID NO: 327, encoding nucleotide sequence: SEQ ID NO: 326) was identified as a protein comprising any of SEQ ID NOs: 328 to 332.
  • the antigen in spot 17 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (17 ⁇ A1-a) to (17 ⁇ A1-e), (17 ⁇ B1), (17 ⁇ A2-a) to (17 ⁇ A2-e), and (17 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 321 to 325 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the protein selected from the group consisting of the proteins as defined above in (17 ⁇ A1-a) to (17 ⁇ A1-e), (17 ⁇ B1), (17 ⁇ A2-a) to (17 ⁇ A2-e), and (17 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (17 ⁇ A1-a) to (17 ⁇ A1-e), (17 ⁇ B1), (17 ⁇ A2-a) to (17 ⁇ A2-e), and (17 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 20 to 90 kDa, preferably around 30 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 17 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 320 (guanosine nucleotide diphosphate dissociation inhibitor 2) or the protein having the amino acid sequence of SEQ ID NO: 327 (rab GDP dissociation inhibitor alpha-like) or causative of an allergy to soybean.
  • DFETRVETEGGRIRVLK (SEQ ID NO: 336) DFETRVETEGGRIRVLK (SEQ ID NO: 336) DNIVSSLDNVAK (SEQ ID NO: 337) GRAVLGLVSESETEK (SEQ ID NO: 338) LFDQQNEGSIFAISREQVRALAPTK (SEQ ID NO: 339) RPTISNGYGRLTEVGPDDDEK
  • the mass spectroscopic data obtained for spot 18 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, hypothetical protein GLYMA_10G028300 (amino acid sequence: SEQ ID NO: 334, encoding nucleotide sequence: SEQ ID NO: 333) was identified as a protein comprising any of SEQ ID NOs: 335 to 339.
  • the antigens in spot 18 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (18 ⁇ A-a) to (18 ⁇ A-e) and (18 ⁇ B):
  • the proteins selected from the group consisting of (18 ⁇ A-a) to (18 ⁇ A-e) and (18 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (18 ⁇ A-a) to (18 ⁇ A-e) and (18 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 18 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 334 (hypothetical protein GLYMA_10G028300) or causative of an allergy to soybean.
  • sucrose binding protein homolog S-64 amino acid sequence: SEQ ID NO: 341, encoding nucleotide sequence: SEQ ID NO: 340
  • sucrose binding protein homolog S-64 amino acid sequence: SEQ ID NO: 341, encoding nucleotide sequence: SEQ ID NO: 340
  • the antigens in spot 19 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (19 ⁇ A-a) to (19 ⁇ A-e) and (19 ⁇ B):
  • the proteins selected from the group consisting of (19 ⁇ A-a) to (19 ⁇ A-e) and (19 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (19 ⁇ A-a) to (19 ⁇ A-e) and (19 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 30 to 90 kDa, preferably around 40 to 80 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 19 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 341 (sucrose binding protein homolog S-64) or causative of an allergy to soybean.
  • DLFPNENELVVK (SEQ ID NO: 350) EIFSLRDTTQEDPREVTAAK (SEQ ID NO: 351) LHGGTPANFLDVGGNASENQVVEAFK (SEQ ID NO: 352) LNFDDNAAYRQK (SEQ ID NO: 353) SQILAGGRGLGTFK (SEQ ID NO: 354) TDQVEDIAGK (SEQ ID NO: 355) VPVVVRLEGTNVDQGK (SEQ ID NO: 356) VVDGLALK (SEQ ID NO: 359) DLFPNENELVVK (SEQ ID NO: 360) EIFALRDTTQEDPREVTAAK (SEQ ID NO: 361) LHGGTPANFLDVGGNASEGQVVEAFK (SEQ ID NO: 362) LNFDDNAAYRQK (SEQ ID NO: 363) SQILAGGRGLGTFK (SEQ ID NO: 364) TDQVEDIAGK (SEQ ID NO: 365) VVDG
  • succinyl-CoA ligase [ADP-forming] subunit beta mitochondrial (amino acid sequence: SEQ ID NO: 348, encoding nucleotide sequence: SEQ ID NO: 347) was identified as a protein comprising any of SEQ ID NOs: 349 to 356 and succinyl-CoA ligase [ADP-forming] subunit beta, mitochondrial (amino acid sequence: SEQ ID NO: 358, encoding nucleotide sequence: SEQ ID NO: 357) was identified as a protein comprising any of SEQ ID NOs: 359 to 365.
  • the antigen in spot 25 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (25 ⁇ A1-a) to (25 ⁇ A1-e), (25 ⁇ B1), (25 ⁇ A2-a) to (25 ⁇ A2-e), and (25 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 349 to 356 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (25 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 358; (25 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 358; (25 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 357; (25 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the protein selected from the group consisting of the proteins as defined above in (25 ⁇ A1-a) to (25 ⁇ A1-e), (25 ⁇ B1), (25 ⁇ A2-a) to (25 ⁇ A2-e), and (25 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (25 ⁇ A1-a) to (25 ⁇ A1-e), (25 ⁇ B1), (25 ⁇ A2-a) to (25 ⁇ A2-e), and (25 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 20 to 90 kDa, preferably around 30 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 25 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 348 or 358 (succinyl-CoA ligase [ADP-forming] subunit beta, mitochondrial) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 26 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, phosphoglycerate kinase, cytosolic (amino acid sequence: SEQ ID NO: 367, encoding nucleotide sequence: SEQ ID NO: 366) was identified as a protein comprising any of SEQ ID NOs: 368 to 382, phosphoglycerate kinase, cytosolic (amino acid sequence: SEQ ID NO: 384, encoding nucleotide sequence: SEQ ID NO: 383) was identified as a protein comprising any of SEQ ID NOs: 385 to 392, and phosphoglycerate kinase, cytosolic (amino acid sequence: SEQ ID NO: 394, encoding nucleotide sequence: SEQ ID NO: 393) was identified as a protein comprising any of SEQ ID NOs: 395 to 409.
  • the antigen in spot 26 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (26 ⁇ A1-a) to (26 ⁇ A1-e), (26 ⁇ B1), (26 ⁇ A2-a) to (26 ⁇ A2-e), (26 ⁇ B2), (26 ⁇ A3-a) to (26 ⁇ A3-e), and (26 ⁇ B3):
  • the amino acid sequence of any of SEQ ID NOs: 368 to 382 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (26 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 384; (26 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 384; (26 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 383; (26 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the amino acid sequence of any of SEQ ID NOs: 385 to 392 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the protein selected from the group consisting of the proteins as defined above in (26 ⁇ A1-a) to (26 ⁇ A1-e), (26 ⁇ B1), (26 ⁇ A2-a) to (26 ⁇ A2-e), (26 ⁇ B2), (26 ⁇ A3-a) to (26 ⁇ A3-e), and (26 ⁇ B3) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (26 ⁇ A1-a) to (26 ⁇ A1-e), (26 ⁇ B1), (26 ⁇ A2-a) to (26 ⁇ A2-e), (26 ⁇ B2), (26 ⁇ A3-a) to (26 ⁇ A3-e), and (26 ⁇ B3) can be proteins that are found in a protein spot with a molecular weight of around 20 to 90 kDa, preferably around 30 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 25 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 367, 384, or 394 (phosphoglycerate kinase, cytosolic) or causative of an allergy to soybean.
  • alcohol dehydrogenase 1 (amino acid sequence: SEQ ID NO: 411, encoding nucleotide sequence: SEQ ID NO: 410) was identified as a protein comprising any of SEQ ID NOs: 412 to 416
  • alcohol dehydrogenase 1 (amino acid sequence: SEQ ID NO: 418, encoding nucleotide sequence: SEQ ID NO: 417) was identified as a protein comprising any of SEQ ID NOs: 419 and 420
  • alcohol dehydrogenase 1-like (amino acid sequence: SEQ ID NO: 422, encoding nucleotide sequence: SEQ ID NO: 421) was identified as a protein comprising any of SEQ ID NOs: 423 to 427.
  • the antigen in spot 27 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (27 ⁇ A1-a) to (27 ⁇ A1-e), (27 ⁇ B1), (27 ⁇ A2-a) to (27 ⁇ A2-e), (27 ⁇ B2), (27 ⁇ A3-a) to (27 ⁇ A3-e), and (27 ⁇ B3):
  • the amino acid sequence of any of SEQ ID NOs: 412 to 416 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids;
  • the amino acid sequence of any of SEQ ID NOs: 419, 420 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the amino acid sequence of any of SEQ ID NOs: 423 to 427 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the protein selected from the group consisting of the proteins as defined above in (27 ⁇ A1-a) to (27 ⁇ A1-e), (27 ⁇ B1), (27 ⁇ A2-a) to (27 ⁇ A2-e), (27 ⁇ B2), (27 ⁇ A3-a) to (27 ⁇ A3-e), and (27 ⁇ B3) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (27 ⁇ A1-a) to (27 ⁇ A1-e), (27 ⁇ B1), (27 ⁇ A2-a) to (27 ⁇ A2-e), (27 ⁇ B2), (27 ⁇ A3-a) to (27 ⁇ A3-e), and (27 ⁇ B3) can be proteins that are found in a protein spot with a molecular weight of around 20 to 90 kDa, preferably around 30 to 80 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 27 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 411 or 418 (alcohol dehydrogenase 1) or the protein having the amino acid sequence of SEQ ID NO: 422 (alcohol dehydrogenase 1-like) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 28 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, 35 kDa seed maturation protein isoform X1 (amino acid sequence: SEQ ID NO: 429, encoding nucleotide sequence: SEQ ID NO: 428) was identified as a protein comprising any of SEQ ID NO: 430 to 433.
  • the antigens in spot 28 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (28 ⁇ A-a) to (28 ⁇ A-e) and (28 ⁇ B):
  • proteins selected from the group consisting of (28 ⁇ A-a) to (28 ⁇ A-e) and (28 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (28 ⁇ A-a) to (28 ⁇ A-e) and (28 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 10 to 70 kDa, preferably around 20 to 60 kDa and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 28 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 422 (35 kDa seed maturation protein isoform X1) or causative of an allergy to soybean.
  • glyceraldehyde-3-phosphate dehydrogenase isoform X1 (amino acid sequence: SEQ ID NO: 435, encoding nucleotide sequence: SEQ ID NO: 434) was identified as a protein comprising any of SEQ ID NO: 436 to 439, glyceraldehyde-3-phosphate dehydrogenase, cytosolic (amino acid sequence: SEQ ID NO: 441, encoding nucleotide sequence: SEQ ID NO: 440) was identified as a protein comprising any of SEQ ID NO: 442 to 445, and uncharacterized protein LOC100782924 (amino acid sequence: SEQ ID NO: 447, encoding nucleotide sequence: SEQ ID NO: 446) was identified as a protein comprising any of SEQ
  • the antigen in spot 29 ⁇ to 33 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (29-33 ⁇ A1-a) to (29-33 ⁇ A1-e), (29-33 ⁇ B1), (29-33 ⁇ A2-a) to (29-33 ⁇ A2-e), (29-33 ⁇ B2), (29-33 ⁇ A3-a) to (29-33 ⁇ A3-e), and (29-33 ⁇ B3):
  • the amino acid sequence of any of SEQ ID NOs: 436 to 439 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids;
  • the amino acid sequence of any of SEQ ID NOs: 442 to 445 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the protein selected from the group consisting of the proteins as defined above in (29-33 ⁇ A1-a) to (29-33 ⁇ A1-e), (29-33 ⁇ B1), (29-33 ⁇ A2-a) to (29-33 ⁇ A2-e), (29-33 ⁇ B2), (29-33 ⁇ A3-a) to (29-33 ⁇ A3-e), and (29-33 ⁇ B3) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (29-33 ⁇ A1-a) to (29-33 ⁇ A1-e), (29-33 ⁇ B1), (29-33 ⁇ A2-a) to (29-33 ⁇ A2-e), (29-33 ⁇ B2), (29-33 ⁇ A3-a) to (29-33 ⁇ A3-e), and (29-33 ⁇ B3) can be proteins that are found in a protein spot with a molecular weight of around 10 to 70 kDa, preferably around 20 to 60 kDa, and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in any of spots 29 ⁇ to 33 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 435 (glyceraldehyde-3-phosphate dehydrogenase isoform X1), the protein having the amino acid sequence of SEQ ID NO: 441 (glyceraldehyde-3-phosphate dehydrogenase, cytosolic), or the protein having the amino acid sequence of SEQ ID NO: 447 (uncharacterized protein LOC100782924) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 34 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, bifunctional UDP-glucose 4-epimerase and UDP-xylose 4-epimerase 1-like (amino acid sequence: SEQ ID NO: 453, encoding nucleotide sequence: SEQ ID NO: 452) was identified as a protein comprising SEQ ID NO: 454 and uncharacterized protein LOC100803483 (amino acid sequence: SEQ ID NO: 456, encoding nucleotide sequence: SEQ ID NO: 455) was identified as a protein comprising SEQ ID NO: 457 or 458.
  • the antigen in spot 34 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (34 ⁇ A1-a) to (34 ⁇ A1-e), (34 ⁇ B1), (34 ⁇ A2-a) to (34 ⁇ A2-e), and (34 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 464 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (34 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 456; (34 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 456; (34 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 455; (34 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably at least 7
  • the protein selected from the group consisting of the proteins as defined above in (34 ⁇ A1-a) to (34 ⁇ A1-e), (34 ⁇ B1), (34 ⁇ A2-a) to (34 ⁇ A2-e), and (34 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (34 ⁇ A1-a) to (34 ⁇ A1-e), (34 ⁇ B1), (34 ⁇ A2-a) to (34 ⁇ A2-e), and (34 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 10 to 70 kDa, preferably around 20 to 60 kDa, and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 34 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 453 (bifunctional UDP-glucose 4-epimerase and UDP-xylose 4-epimerase 1-like) or the protein having the amino acid sequence of SEQ ID NO: 456 (uncharacterized protein LOC100803483) or causative of an allergy to soybean.
  • elongation factor 1-alpha (amino acid sequence: SEQ ID NO: 460, encoding nucleotide sequence: SEQ ID NO: 459) was identified as a protein comprising any of SEQ ID NOs: 461 to 466
  • elongation factor 1-alpha (amino acid sequence: SEQ ID NO: 468, encoding nucleotide sequence: SEQ ID NO: 467) was identified as a protein comprising any of SEQ ID NOs: 469 to 471
  • elongation factor-1A isoform X1 (amino acid sequence: SEQ ID NO: 473, encoding nucleotide sequence: SEQ ID NO: 472) was identified as a protein comprising any of SEQ ID NOs: 474 to 477.
  • the antigen in spot 35 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (35 ⁇ A1-a) to (35 ⁇ A1-e), (35 ⁇ B1), (35 ⁇ A2-a) to (35 ⁇ A2-e), (35 ⁇ B2), (35 ⁇ A3-a) to (35 ⁇ A3-e), and (35 ⁇ B3):
  • the amino acid sequence of any of SEQ ID NOs: 412 to 416 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (35 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 468; (35 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 468; (35 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 467; (35 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the amino acid sequence of any of SEQ ID NOs: 469 to 471 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • 35 ⁇ A3-a a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 473;
  • 35 ⁇ A3-b a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 473;
  • 35 ⁇ A3-c a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 472;
  • 35 ⁇ A3-d a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least
  • the protein selected from the group consisting of the proteins as defined above in (35 ⁇ A1-a) to (35 ⁇ A1-e), (35 ⁇ B1), (35 ⁇ A2-a) to (35 ⁇ A2-e), (35 ⁇ B2), (35 ⁇ A3-a) to (35 ⁇ A3-e), and (35 ⁇ B3) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (35 ⁇ A1-a) to (35 ⁇ A1-e), (35 ⁇ B1), (35 ⁇ A2-a) to (35 ⁇ A2-e), (35 ⁇ B2), (35 ⁇ A3-a) to (35 ⁇ A3-e), and (35 ⁇ B3) can be proteins that are found in a protein spot with a molecular weight of around 20 to 90 kDa, preferably around 30 to 80 kDa, and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 35 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 460 or 468 (elongation factor 1-alpha) or the protein having the amino acid sequence of SEQ ID NO: 473 (elongation factor-1A isoform X1) or causative of an allergy to soybean.
  • SEQ ID NO: 480 EGSSIINTTSVNAYK (SEQ ID NO: 481) FPPQQQTQPGK (SEQ ID NO: 482) GAIVAYTRGLALQLVSK (SEQ ID NO: 483) LLDYTSTK (SEQ ID NO: 486) EGATVAFTYVK (SEQ ID NO: 487) QVIDLVVK (SEQ ID NO: 490) EGSSIINTTSVNAYK (SEQ ID NO: 491) FPPQQQTQPGK (SEQ ID NO: 492) GAIVAYTRGLALQLVSK (SEQ ID NO: 493) LLDYTSTK
  • seed maturation protein PM34 (amino acid sequence: SEQ ID NO: 479, encoding nucleotide sequence: SEQ ID NO: 478) was identified as a protein comprising any of SEQ ID NOs: 480 to 483, uncharacterized protein LOC100809384 (amino acid sequence: SEQ ID NO: 485, encoding nucleotide sequence: SEQ ID NO: 484) was identified as a protein comprising SEQ ID NO: 486 or 487, and hypothetical protein GLYMA_10G155300 (amino acid sequence: SEQ ID NO: 489, encoding nucleotide sequence: SEQ ID NO: 488) was identified as a protein comprising any of SEQ ID NOs: 4490 to 493.
  • the antigen in spot 36 ⁇ , 37 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (36, 37 ⁇ A1-a) to (36, 37 ⁇ A1-e), (36, 37 ⁇ B1), (36, 37 ⁇ A2-a) to (36, 37 ⁇ A2-e), (36, 37 ⁇ B2), (36, 37 ⁇ A3-a) to (36, 37 ⁇ A3-e), and (36, 37 ⁇ B3):
  • the amino acid sequence of any of SEQ ID NOs: 480 to 483 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (36, 37 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 485; (36, 37 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 485; (36, 37 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 484; (36, 37 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide
  • a protein comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 486 and 487, preferably a protein comprising both the amino acid sequences.
  • the amino acid sequence of any of SEQ ID NOs: 486, 487 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids.
  • the protein selected from the group consisting of the proteins as defined above in (36, 37 ⁇ A1-a) to (36, 37 ⁇ A1-e), (36, 37 ⁇ B1), (36, 37 ⁇ A2-a) to (36, 37 ⁇ A2-e), (36, 37 ⁇ B2), (36, 37 ⁇ A3-a) to (36, 37 ⁇ A3-e), and (36, 37 ⁇ B3) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (36, 37 ⁇ A1-a) to (36, 37 ⁇ A1-e), (36, 37 ⁇ B1), (36, 37 ⁇ A2-a) to (36, 37 ⁇ A2-e), (36, 37 ⁇ B2), (36, 37 ⁇ A3-a) to (36, 37 ⁇ A3-e), and (36, 37 ⁇ B3) can be proteins that are found in a protein spot with a molecular weight of around 10 to 70 kDa, preferably around 20 to 60 kDa, and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 36 ⁇ or 37 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 479 (seed maturation protein PM34), the protein having the amino acid sequence of SEQ ID NO: 485 (uncharacterized protein LOC100809384), or the protein having the amino acid sequence of SEQ ID NO: 489 (hypothetical protein GLYMA_10G155300) or causative of an allergy to soybean.
  • SEQ ID NO: 496 AAGSTTAQYVGEK (SEQ ID NO: 497) ALATGWSAAHYSTEITVEGTK (SEQ ID NO: 498) AQVAGSTTVQYAGEK (SEQ ID NO: 499) DTIGRVDNITTPLAK (SEQ ID NO: 500) DTIGSVDHITTPLAEK (SEQ ID NO: 501) ERELASDRARSGNIVK (SEQ ID NO: 502) ERYERANQATSEAAK (SEQ ID NO: 503) GGGEEQGRRREEIGEENK (SEQ ID NO: 504) GLAAAAAGENAK (SEQ ID NO: 505) GRVVIESEK (SEQ ID NO: 506) GVAEYAGQK (SEQ ID NO: 507) LGSAAHTLVK (SEQ ID NO: 508) QSSLEEIFK (SEQ ID NO: 509) QSSLEEISK (SEQ ID NO: 510) REQKPLGSIIGESFQGVQEK (SEQ ID NO: 511)
  • the mass spectroscopic data obtained for spot 38 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, seed biotin-containing protein SBP65-like isoform X1 (amino acid sequence: SEQ ID NO: 495, encoding nucleotide sequence: SEQ ID NO: 494) was identified as a protein comprising any of SEQ ID NOs: 496 to 513.
  • the antigens in spot 38 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (38 ⁇ A-a) to (38 ⁇ A-e) and (38 ⁇ B):
  • proteins selected from the group consisting of (38 ⁇ A-a) to (38 ⁇ A-e) and (38 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (38 ⁇ A-a) to (38 ⁇ A-e) and (38 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 70 to 170 kDa, preferably around 80 to 160 kDa and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 38 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 495 (seed biotin-containing protein SBP65-like isoform X1) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 39 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, ATP synthase subunit O, mitochondrial-like (amino acid sequence: SEQ ID NO: 515, encoding nucleotide sequence: SEQ ID NO: 514) was identified as a protein comprising SEQ ID NO: 516.
  • the antigens in spot 39 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (39 ⁇ A-a) to (39 ⁇ A-e) and (39 ⁇ B):
  • the proteins selected from the group consisting of (39 ⁇ A-a) to (39 ⁇ A-e) and (39 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (39 ⁇ A-a) to (39 ⁇ A-e) and (39 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 5 to 60 kDa, preferably around 10 to 50 kDa and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 39 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 515 (ATP synthase subunit O, mitochondrial-like) or causative of an allergy to soybean.
  • the antigens in spot 40 ⁇ , 56 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (40, 56 ⁇ A-a) to (40, 56 ⁇ A-e) and (40, 56 ⁇ B):
  • the proteins selected from the group consisting of (40, 56 ⁇ A-a) to (40, 56 ⁇ A-e) and (40, 56 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (40, 56 ⁇ A-a) to (40, 56 ⁇ A-e) and (40, 56 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 5 to 60 kDa, preferably around 10 to 50 kDa and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 40 ⁇ or 56 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 518 (P24 oleosin isoform A) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 41 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, uncharacterized protein At3g49720-like isoform X2 (amino acid sequence: SEQ ID NO: 522, encoding nucleotide sequence: SEQ ID NO: 521) was identified as a protein comprising SEQ ID NO: 523.
  • the antigens in spot 41 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (41 ⁇ A-a) to (41 ⁇ A-e) and (41 ⁇ B):
  • the proteins selected from the group consisting of (41 ⁇ A-a) to (41 ⁇ A-e) and (41 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (41 ⁇ A-a) to (41 ⁇ A-e) and (41 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 5 to 60 kDa, preferably around 10 to 50 kDa and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 41 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 515 (ATP synthase subunit O, mitochondrial-like) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spots 42 ⁇ and 43 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, superoxide dismutase [Mn], mitochondrial (amino acid sequence: SEQ ID NO: 525, encoding nucleotide sequence: SEQ ID NO: 524) was identified as a protein comprising any of SEQ ID NOs: 526 to 531 and hypothetical protein GLYMA_04G221300 (amino acid sequence: SEQ ID NO: 533, encoding nucleotide sequence: SEQ ID NO: 532) was identified as a protein comprising any of SEQ ID NOs: 534 to 539.
  • Mn superoxide dismutase
  • the antigen in spot 42 ⁇ , 43 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (42, 43 ⁇ A1-a) to (42, 43 ⁇ A1-e), (42, 43 ⁇ B1), (42, 43 ⁇ A2-a) to (42, 43 ⁇ A2-e), and (42, 43 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 526 to 531 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (42, 43 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 533; (42, 43 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 533; (42, 43 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 532; (42, 43 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide
  • the protein selected from the group consisting of the proteins as defined above in (42, 43 ⁇ A1-a) to (42, 43 ⁇ A1-e), (42, 43 ⁇ B1), (42, 43 ⁇ A2-a) to (42, 43 ⁇ A2-e), and (42, 43 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (42, 43 ⁇ A1-a) to (42, 43 ⁇ A1-e), (42, 43 ⁇ B1), (42, 43 ⁇ A2-a) to (42, 43 ⁇ A2-e), and (42, 43 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 5 to 60 kDa, preferably around 10 to 50 kDa, and an isoelectric point of 5.5 to 10.5, preferably 6.0 to 10.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 42 ⁇ or 43 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 525 (superoxide dismutase [Mn], mitochondrial) or the protein having the amino acid sequence of SEQ ID NO: 533 (hypothetical protein GLYMA_04G221300) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 45 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, uncharacterized protein LOC100499771 (amino acid sequence: SEQ ID NO: 541, encoding nucleotide sequence: SEQ ID NO: 540) was identified as a protein comprising any of SEQ ID NOs: 542 to 545 and hypothetical protein GLYMA_09G192800 (amino acid sequence: SEQ ID NO: 547, encoding nucleotide sequence: SEQ ID NO: 546) was identified as a protein comprising any of SEQ ID NOs: 548 to 551.
  • the antigen in spot 45 ⁇ in the present invention can be any of proteins selected from the group consisting of the proteins as defined below in (45 ⁇ A1-a) to (45 ⁇ A1-e), (45 ⁇ B1), (45 ⁇ A2-a) to (45 ⁇ A2e), and (45 ⁇ B2):
  • the amino acid sequence of any of SEQ ID NOs: 542 to 545 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (45 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 547; (45 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 547; (45 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 546; (45 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the protein selected from the group consisting of the proteins as defined above in (45 ⁇ A1-a) to (45 ⁇ A1-e), (45 ⁇ B1), (45 ⁇ A2-a) to (45 ⁇ A2-e), and (45 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (45 ⁇ A1-a) to (45 ⁇ A1-e), (45 ⁇ B1), (45 ⁇ A2-a) to (45 ⁇ A2-e), and (45 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 2 to 50 kDa, preferably around 5 to 40 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 45 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 541 (uncharacterized protein LOC100499771) or the protein having the amino acid sequence of SEQ ID NO: 547 (hypothetical protein GLYMA_09G192800) or causative of an allergy to soybean.
  • seed maturation protein PM22 (amino acid sequence: SEQ ID NO: 553, encoding nucleotide sequence: SEQ ID NO: 552) was identified as a protein comprising SEQ ID NO: 554.
  • the antigens in spot 46 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (46 ⁇ A-a) to (46 ⁇ A-e) and (46 ⁇ B):
  • the proteins selected from the group consisting of (46 ⁇ A-a) to (46 ⁇ A-e) and (46 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (46 ⁇ A-a) to (46 ⁇ A-e) and (46 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 2 to 50 kDa, preferably around 5 to 40 kDa and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 46 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 553 (seed maturation protein PM22) or causative of an allergy to soybean.
  • eukaryotic translation initiation factor 5A-2 (amino acid sequence: SEQ ID NO: 556, encoding nucleotide sequence: SEQ ID NO: 555) was identified as a protein comprising SEQ ID NO: 557 or 558 and uncharacterized protein LOC100305510 (amino acid sequence: SEQ ID NO: 560, encoding nucleotide sequence: SEQ ID NO: 559) was identified as a protein comprising any of SEQ ID NOs: 561 to 563.
  • the amino acid sequence of any of SEQ ID NOs: 557, 558 may be an amino acid sequence derived therefrom by deletion, substitution, insertion or addition of one or several amino acids; (47 ⁇ A2-a) a protein comprising an amino acid sequence with deletion, substitution, insertion or addition of one or several amino acids in SEQ ID NO: 560; (47 ⁇ A2-b) a protein comprising an amino acid sequence having at least 70%, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% identity to the amino acid sequence of SEQ ID NO: 560; (47 ⁇ A2-c) a protein comprising an amino acid sequence encoded by a nucleotide sequence with deletion, substitution, insertion or addition of one or several nucleotides in SEQ ID NO: 559; (47 ⁇ A2-d) a protein comprising an amino acid sequence encoded by a nucleotide sequence having at least 70%, preferably
  • the protein selected from the group consisting of the proteins as defined above in (47 ⁇ A1-a) to (47 ⁇ A1-e), (47 ⁇ B1), (47 ⁇ A2-a) to (47 ⁇ A2-e), and (47 ⁇ B2) also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the protein selected from the group consisting of the proteins as defined above in (47 ⁇ A1-a) to (47 ⁇ A1-e), (47 ⁇ B1), (47 ⁇ A2-a) to (47 ⁇ A2-e), and (47 ⁇ B2) can be proteins that are found in a protein spot with a molecular weight of around 2 to 50 kDa, preferably around 5 to 40 kDa, and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 47 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 556 (eukaryotic translation initiation factor 5A-2) or the protein having the amino acid sequence of SEQ ID NO: 560 (uncharacterized protein LOC100305510) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 48 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, uncharacterized protein LOC100306570 (amino acid sequence: SEQ ID NO: 565, encoding nucleotide sequence: SEQ ID NO: 564) was identified as a protein comprising any of SEQ ID NOs: 566 to 570.
  • the antigens in spot 48 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (48 ⁇ A-a) to (48 ⁇ A-e) and (48 ⁇ B):
  • the proteins selected from the group consisting of (48 ⁇ A-a) to (48 ⁇ A-e) and (48 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (48 ⁇ A-a) to (48 ⁇ A-e) and (48 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 2 to 50 kDa, preferably around 5 to 40 kDa and an isoelectric point of 3.5 to 8.5, preferably 4.0 to 8.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 48 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 565 (uncharacterized protein LOC100306570) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 50 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, uncharacterized protein LOC100813859 (amino acid sequence: SEQ ID NO: 572, encoding nucleotide sequence: SEQ ID NO: 571) was identified as a protein comprising any of SEQ ID NOs: 573 to 576.
  • the antigens in spot 50 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (50 ⁇ A-a) to (50 ⁇ A-e) and (50 ⁇ B):
  • the proteins selected from the group consisting of (50 ⁇ A-a) to (50 ⁇ A-e) and (50 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (50 ⁇ A-a) to (50 ⁇ A-e) and (50 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 2 to 50 kDa, preferably around 5 to 40 kDa and an isoelectric point of 2.5 to 6.5, preferably 3.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 50 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 572 (uncharacterized protein LOC100813859) or causative of an allergy to soybean.
  • EAAESTLSAYK (SEQ ID NO: 580) LLDSRLIPSASSGDSK (SEQ ID NO: 581) QAFDEAIAELDTLGEESYK (SEQ ID NO: 582) VSAAADNEELTVEERNLLSVAYK
  • the mass spectroscopic data obtained for spot 51 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, 14-3-3-like protein (amino acid sequence: SEQ ID NO: 578, encoding nucleotide sequence: SEQ ID NO: 577) was identified as a protein comprising any of SEQ ID NOs: 579 to 582.
  • the antigens in spot 51 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (51 ⁇ A-a) to (51 ⁇ A-e) and (51 ⁇ B):
  • the proteins selected from the group consisting of (51 ⁇ A-a) to (51 ⁇ A-e) and (51 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (51 ⁇ A-a) to (51 ⁇ A-e) and (51 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 5 to 60 kDa, preferably around 10 to 50 kDa and an isoelectric point of 2.5 to 6.5, preferably 3.0 to 6.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.
  • a protein that is an antigen in spot 51 ⁇ has activity equivalent to that of the protein having the amino acid sequence of SEQ ID NO: 578 (14-3-3-like protein) or causative of an allergy to soybean.
  • the mass spectroscopic data obtained for spot 52 ⁇ on a mass spectrometer was analyzed by comparing the data against the NCBI protein data, and as a result, probable fructokinase-4 (amino acid sequence: SEQ ID NO: 584, encoding nucleotide sequence: SEQ ID NO: 583) was identified as a protein comprising any of SEQ ID NOs: 585 to 588.
  • the antigens in spot 52 ⁇ in the present invention can be any protein selected from the group consisting of the proteins as defined below in (52 ⁇ A-a) to (52 ⁇ A-e) and (52 ⁇ B):
  • the proteins selected from the group consisting of (52 ⁇ A-a) to (52 ⁇ A-e) and (52 ⁇ B) as defined above also include those proteins whose amino acid residues are modified by phosphorylation, sugar chain modification, aminoacylation, ring-opening, deamination or the like.
  • the proteins selected from the group consisting of (52 ⁇ A-a) to (52 ⁇ A-e) and (52 ⁇ B) as defined above can be proteins that are found in a protein spot with a molecular weight of around 5 to 60 kDa, preferably around 10 to 50 kDa and an isoelectric point of 3.5 to 7.5, preferably 4.0 to 7.0 on gels used in the two-dimensional electrophoresis performed under the conditions described above in the subsection titled “Identification of antigens”.

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