Anti-allergen antibodies and their uses

Abstract

The present invention provides antibodies that bind to birch pollen allergens, particularly the Bet v 1 allergen, and also to Bet v 1 homologous allergens. The present invention also provides compositions and kits comprising distinct antibodies that bind to distinct non-overlapping epitopes of Bet v 1 and Bet v 1 homologous proteins. Furthermore, the present invention relates to the use of such antibodies, compositions, and kits for, for example, preventing or treating tree pollen allergies and food-related allergies.

Description

Detailed description of the invention

[0001] The present invention relates to antibodies that bind to birch pollen allergens and related tree pollen allergens and related food allergens, particularly to Bet v 1 and Bet v 1 homologous allergen / protein. The present invention also relates to compositions and kits comprising distinct antibodies that bind to distinct non-overlapping epitopes of Bet v 1 and Bet v 1 homologous protein. Furthermore, the present invention relates to the use of the antibodies, compositions and kits for preventing or treating, for example, tree pollen allergies and food allergies, particularly birch pollen allergies and related tree pollen allergies, as well as food-related allergies (such as apple allergies, hazelnut allergies, and walnut allergies).

[0002] Allergies are disorders caused by hypersensitivity of the immune system. When exposed to an allergen, allergen-binding immunoglobulin E (IgE) antibodies are produced. These antibodies are pre-bound to FcεRI receptors on mast cells and basophils, triggering the release of inflammatory compounds (such as histamine, leukotrienes, and lipid mediators).

[0003] Type I allergies, or immediate-type hypersensitivity reactions, are the result of interactions between allergens and immunoglobulin E (IgE) antibodies, which are bound to mast cells. Allergic symptoms (such as allergic asthma, rhinoconjunctivitis, and atopic dermatitis) have a significant impact on human health and quality of life. It is estimated that 300 to 400 million people worldwide are affected by type I allergies, resulting in medical costs, sick leave, and economic losses of €120 billion (Calderon et al., 2012, Clin. Transl. Allergy 2: 20). These symptoms are triggered by environmental aeroallergens (such as pollen or house dust mites). Birch pollen is widely distributed in Europe, North America, Russia, and northern Japan. More than 100 million people suffer from allergies to birch pollen (Moverare et al., 2005, Int. Arch. Allergy Immunol. 136, 33-38). The main allergen in birch pollen is Bet v 1.

[0004] The Bet v 1 allergen belongs to the PR-10 family of plant pathogenicity-associated proteins. Bet v 1 homologs are also found in various plant-derived food allergens (such as Mal d 1 in apples and Cor a 1 in hazelnuts). Patients with birch pollen allergy usually also have co-occurring allergies to other related tree pollens (such as hazel, alder, beech, and oak), and often suffer from secondary pollen-food syndrome, which negatively impacts health-related quality of life (Popescu, 2015, World J. Methodol. 5, 31-50).

[0005] Allergen immunotherapy (AIT) modulates the allergic immune response based on repeated administration of disease-causing allergens. AIT induces allergen-specific immunoglobulin G, particularly IgG4. IgG4 competes with IgE to bind to the allergen, alleviating allergic symptoms by inhibiting the IgE-mediated hypersensitivity mechanism (Shamji et al., 2012, Allergy 67, 217-226). Bet v 1-specific IgG4 produced by AIT competes with IgE for partially identical or significantly overlapping epitopes (Groh et al., 2017, Clin. Exp. Allergy 47, 693-703).

[0006] Recently, antibodies against birch pollen allergens have emerged as a promising new treatment option for birch pollen allergy. For example, Atanasio et al. (J. Allergy Clin. Immunol. 2020; vol. 149, 1, p. 200-211) reported that a combination of mAbs targeting Bet v 1 can prevent birch allergic reactions. According to their study, REGN5713, REGN5714, and REGN5715, Bet v 1-specific mAbs derived from immunized humanized mice, bind to Bet v 1 with high affinity and non-competitively. A cocktail of all three antibodies (REGN5713 / 14 / 15) has been shown to inhibit IgE binding to Bet v 1, suppressing Bet v 1 and birch pollen extract-induced basophil activation ex vivo, and suppressing mast cell degranulation in vivo.

[0007] Due to the high cross-reactivity observed in patients with birch allergy, antibody approaches targeting birch pollen should aim to target as many Bet v 1 homologous proteins (PR-10 proteins) as possible.

[0008] Based on the above, an object of the present invention is to provide an improved antibody against birch pollen allergen, in particular, an antibody that targets the Bet v 1 allergen and exhibits cross-reactivity with multiple Bet v 1 homologous allergens / proteins. Another object of the present invention is to provide a composition comprising at least two distinct potent antibodies, the antibodies which bind to distinct non-overlapping epitopes on Bet v 1, the major birch allergen, and optionally to at least one further Bet v 1 homologous allergen / protein.

[0009] This objective is achieved by the subject matter described below and in the attached claims.

[0010] The present invention will be described in detail below, but it should be understood that the present invention is not limited to the specific methodologies, protocols, and reagents described herein. Furthermore, it should be understood that the terms used herein are not intended to limit the scope of the present invention, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0011] The elements of the present invention are described below. These elements are listed along with specific embodiments, but it should be understood that they can be combined in any way and in any number to create additional embodiments. The various examples and embodiments described should not be construed as limiting the invention to only the explicitly described embodiments. This specification should be understood to support and encompass embodiments that combine the explicitly described embodiments with any number of disclosed elements. Furthermore, any permutation and combination of all elements described in this application should be deemed disclosed by the description of this application unless the context indicates otherwise.

[0012] Throughout this specification and the subsequent claims, unless otherwise specified in the context, the terms “comprise,” and variations such as “comprises” and “comprising,” are understood to mean including specified members, integers, or steps, but not to mean excluding other unspecified members, integers, or steps. The term “consist of” is a specific embodiment of the term “comprise,” excluding other unspecified members, integers, or steps. In the context of this invention, the term “comprise” encompasses the term “consist of.” Therefore, the term “comprising” encompasses “including” as well as “consisting,” for example, “comprising” X may consist of X alone, or it may include something additional, such as X + Y.

[0013] The terms “a,” “an,” “the,” and similar references used in the context describing the present invention (particularly in the context of the claims) shall be construed to cover both singular and plural forms unless otherwise specifically indicated herein or unless the context clearly contradicts this. The descriptions of value ranges herein are intended merely as abbreviations for individually referring to each individual value that falls within the range. Unless otherwise stated herein, each individual value is incorporated herein as if it were individually stated herein. Nothing in this specification should be construed as indicating a non-claim element essential to the practice of the present invention.

[0014] The term "substantially" does not exclude "completely." For example, a composition that "substantially does not contain" Y may not contain Y at all. Where necessary, the term "substantially" may be excluded from the definition of this invention.

[0015] The term "approximately" in relation to the number x means x ± 10%, for example x ± 5%, or x ± 7%, or x ± 10%, or x ± 12%, or x ± 15%, or x ± 20%.

[0016] As used herein, the term “disease” is generally synonymous with and intended to be used interchangeably with the terms “disorder” and “condition” (in medical terms), in that they reflect an abnormal condition of the body or part thereof of a human or animal that impairs normal function, typically manifests as characteristic signs and symptoms, and reduces the duration or quality of life of the human or animal.

[0017] In this specification, references to “treatment” of a subject or patient are intended to include prevention, prophylaxis, attenuation, improvement, and treatment. In this specification, the terms “subject” or “patient” are used interchangeably to mean all mammals, including humans. Examples of subjects include humans, cattle, dogs, cats, horses, goats, sheep, pigs, and rabbits. In some embodiments, the subject or patient is human.

[0018] Dosage is often expressed in relation to body weight. Therefore, a dosage expressed as [g, mg, or other units] / kg (or g, mg, etc.) usually means [g, mg, or other units] per kg of body weight (or g, mg, etc.), even if the term "body weight" is not explicitly stated.

[0019] The term "binding" and similar references typically mean "specifically binding" and do not encompass nonspecific binding. In particular, specific binding of an antibody means that the antibody recognizes the target antigen and binds to that target with higher affinity (or lower antibody concentration, e.g., EC50) than to structurally different antigens and / or antigens with modified or mutated sequences. Thus, "higher" affinity means at least 2, 3, 4, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 500, 750, 1,000, 1,500, 2,000, 5,000, 7,500, 10,000 or more affinity compared to binding to a control antigen. In some cases, antibody binding to a control antigen may not be detected (below the detection threshold), while antibody binding to a specific antigen may be fully detected / determined.

[0020] As used herein, the term “antibody” encompasses a wide range of forms of antibodies, including but not limited to whole antibodies, antibody fragments (such as antigen-binding fragments), human antibodies, chimeric antibodies, humanized antibodies, recombinant antibodies, and genetically modified antibodies (e.g., variant or mutant antibodies), as long as they retain the characteristic properties of the present invention. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a monoclonal antibody. For example, the antibody may be a human monoclonal antibody.

[0021] As mentioned above, the term "antibody" generally includes antibody fragments. Antibody fragments may retain the antigen-binding activity of the antibody. Such fragments are called "antigen-binding fragments." Antigen-binding fragments include, but are not limited to, single-chain antibodies, Fab, Fab', F(ab')2, Fv, or scFv. Antibody fragments can be obtained from antibodies by methods including digestion with enzymes such as pepsin or papain, and / or cleavage of disulfide bonds by chemical reduction. Alternatively, antibody fragments can be obtained by recombination methods, such as cloning and expressing a portion (fragment) of the heavy chain and / or light chain sequence. The present invention also includes single-chain Fv fragments (scFv) derived from the heavy and light chains of the antibodies of the present invention. For example, the present invention includes scFv containing CDRs derived from the antibodies of the present invention. It also includes monomers and dimers of heavy or light chains, single-domain heavy chain antibodies, single-domain light chain antibodies, and single-chain antibodies, such as single-chain Fv in which the heavy and light chain variable domains are linked by a peptide linker. The antibody fragments of the present invention may be incorporated into various structures known to those skilled in the art. Furthermore, the sequences of the present invention may be components of a multispecific molecule in which the sequences of the present invention target the epitope of the present invention and other regions of the molecule bind to other targets. In this specification, including the claims, there are explicit references to antigen-binding fragments, antibody fragments, antibody variants, and / or antibody derivatives, but the term “antibody” is understood to include all categories of antibodies, namely antigen-binding fragments, antibody fragments, antibody variants, and antibody derivatives.

[0022] Human antibodies are well known through state-of-the-art technology (van Dijk, MA, and van de Winkel, JG, Curr. Opin. Chem. Biol. 5 (2001) 368-374). Human antibodies can also be produced in transgenic animals (e.g., mice and chickens) that can produce the entire or a selected repertoire of human antibodies by immunization in the absence of endogenous immunoglobulin production. Introducing a human germline immunoglobulin gene array into such germline mutant mice enables the production of human antibodies during antigen challenge (see, for example, Jakobovits, A., et al., Proc. Natl. Acad. Sci. USA 90 (1993) 2551-2555; Jakobovits, A., et al., Nature 362 (1993) 255-258; Bruggemann, M., et al., Year Immunol. 7 (1993) 3340). Human antibodies can also be produced using phage display libraries (Hoogenboom, HR, and Winter, G., J. Mol. Biol. 227 (1992) 381-388; Marks, JD, et al., J. Mol. Biol. 222 (1991) 581-597). The techniques of Cole et al. and Boerner et al. are also applicable to the preparation of human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); and Boerner, P., et al., J. Immunol. 147 (1991) 86-95). As used herein, the term "human antibody" typically includes sequence variants of human antibodies that do not exist in nature, obtained by introducing one or more mutations into a (naturally occurring) human antibody. Such mutations include one or more mutations in the CDR or framework region, and Fc modifications (e.g., those known in the art for specific functionality).

[0023] As used herein, the term "variable region" (the variable region of the light chain (V L ), the variable region of the heavy chain (V H )) refers to each of the pairs of light and heavy chains that are directly involved in the binding of an antibody to an antigen.

[0024] The antibodies of the present invention can be of any isotype (e.g., IgA, IgG, IgM, i.e., α, γ or μ heavy chains). Preferably, the antibodies are of the IgG or IgA type. Among the IgG isotypes, the antibodies may be of the IgG1, IgG2, IgG3 or IgG4 subclass, preferably IgG1 or IgG4. The antibodies of the present invention may have κ or λ light chains.

[0025] The antibodies according to the present invention may be provided in a purified form. Typically, the antibodies will be present in a composition that is substantially free of other polypeptides, e.g., less than 90% (weight ratio), usually less than 60%, more usually less than 50% of the composition is composed of other polypeptides.

[0026] The antibodies according to the present invention may be immunogenic in human and / or non-human (or heterologous) hosts, e.g., mice. For example, the antibodies may have idiotopes that are immunogenic in non-human hosts but not in human hosts. The antibodies of the present invention for use in humans include those that cannot be easily isolated from hosts such as mice, goats, rabbits, rats, non-primate mammals, etc., and generally cannot be obtained by humanization or xenomouse.

[0027] In this specification, the term “antigen” refers to any structural substance that functions as a target of an adaptive immune response receptor, particularly an antibody, T cell receptor, and / or B cell receptor. “Epitope,” also known as “antigenic determinant,” is a part (or fragment) of an antigen recognized by the immune system, particularly an antibody, T cell receptor, and / or B cell receptor. Thus, one antigen has at least one epitope; i.e., one antigen has one or more epitopes. An antigen may be (i) a peptide, polypeptide, or protein; (ii) a polysaccharide; (iii) a lipid; (iv) a lipoprotein or lipopeptide; (v) a glycolipid; (vi) a nucleic acid; or (vii) a small molecule drug or toxin. Therefore, the antigen may be a peptide, protein, polysaccharide, lipid, a combination thereof (including lipoproteins and glycolipids), nucleic acid (e.g., DNA, siRNA, shRNA, antisense oligonucleotide, decoy DNA, plasmid), or small molecule drug (e.g., cyclosporine A, paclitaxel, doxorubicin, methotrexate, 5-aminolevulinic acid), or any combination thereof. Preferably, the antigen is selected from (i) peptides, polypeptides, or proteins, (ii) polysaccharides, (iii) lipids, (iv) lipoproteins or lipopeptides, and (v) glycolipids, and more preferably, the antigen is a peptide, polypeptide, or protein.

[0028] As used herein, the term "mutation" refers to a change in a nucleic acid sequence and / or an amino acid sequence as compared to a reference sequence, such as a corresponding genomic sequence. Mutations as compared to a genomic sequence include, for example, (naturally occurring) somatic mutations, spontaneous mutations, mutations induced (e.g., by enzymes, chemicals, radiation, etc.), or mutations obtained by site-directed mutagenesis (a molecular biology technique that results in specific and intentional changes in a nucleic acid sequence and / or an amino acid sequence). Thus, the terms "mutate" or "mutation" are understood to include, for example, physically causing a mutation in a nucleic acid sequence or an amino acid sequence. Mutations include substitutions, deletions, insertions of one or more nucleotides or amino acids, and inversions of multiple consecutive nucleotides or amino acids. To effect a mutation in an amino acid sequence, a mutation may be introduced into the nucleotide sequence encoding the amino acid sequence and a (recombinant) mutant polypeptide may be expressed. Mutations may be achieved, for example, by site-directed mutagenesis to modify the codon of a nucleic acid molecule encoding an amino acid to a codon encoding a different amino acid, or, for example, by designing the synthesis of a nucleic acid molecule containing a nucleotide sequence encoding a polypeptide variant without the need to mutate one or more nucleotides of the nucleic acid molecule encoding the polypeptide, by synthesizing sequence variants.

[0029] As used herein (i.e., throughout this specification), the term “sequence variant” refers to any change compared to a reference sequence. The term “sequence variant” includes nucleotide sequence variants and amino acid sequence variants. Preferably, the reference sequence is one of the sequences listed in the “Sequence and Sequence Number List” (Sequence Number List), i.e., SEQ ID NOs. 1 to SEQ ID NOs. In particular, a sequence variant shares sequence identity with its reference sequence of at least 70% or at least 75%, preferably at least 80% or at least 85%, more preferably at least 90% or at least 93%, even more preferably at least 95% or at least 96%, still more preferably at least 97% or at least 98%, and especially preferably at least 99% (over the entire length of the sequence). In some embodiments, the sequence variant shares at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity. Thus, a higher percentage of identity for the sequence variant is preferable. For example, a sequence variant having at least 84% sequence identity with the reference sequence is preferable to a sequence variant having at least 75% but less than 84% sequence identity with the reference sequence. In some embodiments, the sequence variant maintains the (biological) function of the reference sequence. For example, sequence variants relating to the antibody of the present invention preferably maintain specific binding to Bet v 1 and / or Bet v 1 homologous proteins (such as Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, Mal d 1, Jugr 5, etc.).

[0030] Sequence identity may be calculated as described below. Typically, the sequence variant may retain certain functions of the reference sequence. In some embodiments, the amino acid sequence variant has a modified sequence in which one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) in the reference sequence are deleted or substituted, or one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) are inserted or added to the sequence of the reference amino acid sequence. As a result of the modification, the amino acid sequence variant has an amino acid sequence that is at least 70% or at least 75%, preferably at least 80% or at least 85%, more preferably at least 90% or at least 93%, even more preferably at least 95% or at least 96%, still more preferably at least 97% or at least 98%, and particularly preferably at least 99% identical to the reference sequence. For example, a variant sequence that is at least 90% identical to the reference sequence will not have more than 10 modifications per 100 amino acids, i.e., any combination of deletions, insertions, or substitutions. Of course, the same applies to nucleic acid sequences.

[0031] The "identity percentage" of a sequence variant is typically determined relative to a reference sequence. It is usually calculated with respect to the entire length of the reference sequence (i.e., the sequence described in this application). Identity percentages as referred herein may be determined by methods known in the art, such as BLAST using default parameters specified by the NCBI (National Center for Biotechnology Information; http: / / www.ncbi.nlm.nih.gov / ). [Blosum 62 matrix; gap open penalty = 11 and gap extension penalty = 1].

[0032] Generally, non-conservative amino acid substitutions are also possible, but substitutions are preferably conserved amino acid substitutions, in which the substituted amino acid has structural or chemical properties similar to the corresponding amino acid in the reference sequence. As an example, conservative amino acid substitutions include: substituting one aliphatic or hydrophobic amino acid (such as alanine, valine, leucine, and isoleucine) with another aliphatic or hydrophobic amino acid; substituting one hydroxyl-containing amino acid (such as serine and threonine) with another hydroxyl-containing amino acid; substituting one acidic residue (such as glutamic acid or aspartic acid) with another acidic residue; replacing one amide-containing residue (such as asparagine and glutamine) with another amide-containing residue; replacing one aromatic residue (such as phenylalanine and tyrosine) with another aromatic residue; replacing one basic residue (such as lysine, arginine, and histidine) with another basic residue; and substituting one minor amino acid (such as alanine, serine, threonine, cysteine, and glycine) with another minor amino acid.

[0033] As used herein, "Bet v 1 allergen or Bet v 1 protein" refers to, for example, Bet v 1.0101, Bet v 1.0102, Bet v 1.0103, Bet v 1.0104, Bet v 1.0105, Bet v 1.0106, Bet v 1.0107, Bet v 1.0108, Bet v 1.0109, Bet v 1.0110, Bet v 1.0111, Bet v 1.0112, Bet v 1.0113, Bet v 1.0114, Bet v 1.0115, Bet v 1.0115, Bet v 1.0116, Bet v 1.0117, Bet v 1.0118, Bet v 1.0119, Bet It includes all known isoallergens and variants of Bet v 1, including v 1.0201, Bet v 1.0202, Bet v 1.0203, Bet v 1.0204, Bet v 1.0205, Bet v 1.0206, Bet v 1.0207, and Bet v 1.0301, as well as all known variants of Bet v 1 (e.g., Bet v 1 N160A).

[0034] Allergens or proteins of the Bet v 1 family, which belong to class 10 of the pathogenicity-associated protein (PR10) family,

[0035] [Table 1]

[0036] It includes "Bet v 1 homologous allergens." An example of a "Bet v 1 homologous allergen" is, for example,

[0037] [Table 2] JPEG2026522294000003.jpg175169

[0038] These are some examples. The allergen information on the listed websites is incorporated herein by reference.

[0039] Furthermore, "Bet v 1 homologous allergens" also include all known isoallergens and variants of the allergens listed above. For example, these isoallergens and variants include Aln g 1.0101, Car b 1.1A, Car b 1.2, Car b 1.0101~Car b 1.0113, Car b 1.0201, Car b 1.301, Car b 1.302, Cas s 1.0101, Cor a 1.0101, Cor a 1.0102, Cor a 1.0103, Cor a 1.0104, Cor a 1.0201, Cor a 1.0301, Cor a 1.0401, Cor a 1.0402, Cor a 1.0403, Cor a 1.0404, and Fag. s 1.0101, Ost c 1.0101, Que a 1.0101, Que a 1.0201, Que a 1.0301, Que a 1.0401, Mal d 1.0101~Mal d 1.0109, Mal d 1.0201~Mal d 1.0208, Mal d 1.0301~Mal d 1.0304, Mal d 1.0401~Mal d 1.0403, Jug r 5.0101, Pru p 1.01, and Pru p 1.02.

[0040] Therefore, in this specification, “birch pollen-related allergy” encompasses any allergy triggered by any of the Bet v 1 homologous allergens or isoallergens listed above.

[0041] Throughout this specification, several documents are referenced. Each document referenced herein (including all patents, patent applications, scientific publications, manufacturer specifications, instructions, etc.) is incorporated herein by reference, whether as stated above or below. Nothing in this specification shall be construed as admitting that the present invention has no prior rights to such disclosures or other claims by prior art.

[0042] It should be understood that the present invention is not limited to the specific methodologies, protocols, and reagents described herein, as these may vary. Furthermore, it should be understood that the terms used herein are solely for the purpose of describing specific embodiments and are not intended to limit the scope of the present invention, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0043] [Antibodies and their antigen-binding fragments] In a first embodiment, the present invention provides an isolated antibody or antigen-binding fragment thereof that (specifically) binds to Bet v 1 (Betula berlucosa allergen 1). Bet v 1 is the major birch pollen allergen recognized by serum IgE in more than 95% of patients with birch pollen hypersensitivity. Bet v 1 is a protein of approximately 17 kDa. Several Bet v 1 isoforms have been described (Bet v 1.0101, Bet v 1.0102, Bet v 1.0103, etc., as described above). Accordingly, the antibody or antigen-binding fragment of the present invention binds, in particular, to polypeptides or proteins having the amino acid sequence described in SEQ ID NO: 52 as defined above, and / or to variants of polypeptides or proteins having the amino acid sequence described in SEQ ID NO: 52 as defined above.

[0044] Preferably, the (isolated) antibody or antigen-binding fragment that (specifically) binds to Bet v 1 further (specifically) binds to a Bet v 1 homologous allergen. The Bet v 1 homologous allergen may be another tree pollen allergen. The Bet v 1 homologous tree pollen allergen may be selected from the group consisting of Cor a 1 (Corylus averana allergen 1), Aln g 1 (Alnus glutinosa allergen 1), Fag s 1 (Fagus silvacea allergen 1), Que a 1 (Quercus alba allergen 1), Cas s 1 (Castanea sativa allergen 1), Ost c 1 (Ostria carpinifolia allergen 1), and / or Car b 1 (Carpinus betulus allergen 1). Therefore, preferably, the antibody or antigen-binding fragment of the present invention (i) binds (specifically) to Bet v 1 and (ii) also binds (specifically) to at least one Bet v 1 homologous protein (Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, etc.). Therefore, the antibody or antigen-binding fragment of the present invention particularly binds to polypeptides or proteins having the amino acid sequence described in SEQ ID NO: 52 as defined above, and / or variants of polypeptides or proteins having the amino acid sequence described in SEQ ID NO: 52 as defined above. Furthermore, preferably, it also binds to polypeptides or proteins having the amino acid sequence described in any one of SEQ ID NOs. 53 to 64, and / or variants of polypeptides or proteins having the amino acid sequence described in any one of SEQ ID NOs. 53 to 64.

[0045] Furthermore, the (isolated) antibody or antigen-binding fragment that (specifically) binds to Bet v 1 may further (specifically) bind to a Bet v 1 homologous food-related allergen. The food-related allergen may be selected from any of the Bet v 1 homologous allergens listed above, preferably Mal d 1 (Mars domestica allergen 1), Cor a 1 (Corylus averana allergen 1), Ju r 5 (Yuglans regia allergen 5), and

[0046] [Table 3]

[0047] Selected from.

[0048] The above-mentioned tree pollen allergens, particularly Bet v 1, Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, and / or Car b 1, as well as the above-mentioned food-related allergens (Mal d 1, Cor a 1, Jugr 5, Pru p 1, etc.), may be naturally derived, recombinantly expressed, or synthetic peptides.

[0049] Standard methods for evaluating the binding of an antibody or its antigen-binding fragment according to the present invention are known to those skilled in the art, and include, for example, ELISA (enzyme-linked immunosorbent assay). This allows the relative affinity of antibody binding to be determined by the antibody concentration (EC) required to achieve a maximum binding of 50% in a saturated state. 50 This may be determined by measuring ). Specific examples of ELISAs that may be used to evaluate antibody binding are described in the Examples section of this specification.

[0050] Generally, an antibody or its antigen-binding fragment according to the present invention may contain (at least) three complementarity-determining regions (CDRs) on the heavy chain and (at least) three CDRs on the light chain. Generally, complementarity-determining regions (CDRs) are hypervariable domains located in the heavy chain variable domain and the light chain variable domain, respectively. Typically, the CDRs of the antibody's heavy chain and the linked light chain together form an antigen receptor. Usually, the three CDRs (CDR1, CDR2, CDR3) are arranged discontinuously within the variable domain. Since an antigen receptor typically consists of two variable domains (on two different polypeptide chains (i.e., heavy chain and light chain): heavy chain variable region (VH) and light chain variable region (VL)), each antigen receptor typically has six CDRs (heavy chain: CDRH1, CDRH2, and CDRH3; light chain: CDRL1, CDRL2, and CDRL3). For example, a classical IgG antibody molecule usually has two antigen receptors and therefore contains 12 CDRs. CDRs on the heavy and / or light chains may be separated by framework regions, which are less "variable" regions within the variable domain than the CDRs. For example, a variable region (or each variable region) consists of four framework regions separated by three CDRs.

[0051] The heavy and light chain sequences of exemplary antibodies of the present invention, containing three different CDRs on the heavy chain and three different CDRs on the light chain, were determined. The CDR amino acid sequences of the heavy chain CDR1 (CDRH1), heavy chain CDR2 (CDRH2), heavy chain CDR3 (CDRH3), light chain CDR1 (CDRL1), light chain CDR2 (CDRL2), and light chain CDR3 (CDRL3) of exemplary antibodies 4E1, 25-3D3, and 17B10 are shown in Table 1 below.

[0052] The CDRs identified by the present invention, in particular the three different CDRs on the heavy chain (CDR1, CDR2, and CDR3) and the three different CDRs on the light chain (CDR1, CDR2, and CDR3), may be grafted onto any variable framework region, in particular any variable human framework region, without impairing their specificity.

[0053] The Human Variable Framework Region for Heavy Chain (VH) can be searched from the website: “https: / / www.imgt.org / genedb / resultPage.action;jsessionid=49EB34C22C79EAC51862C76108963216?gene.id.species=Homo+sapiens&molComponent=IG&geneTypeLike=variable&allele.fcode=functional&cloneName=&locusLike=IGH&mainLocusLike=IGH+locus&cosLocusLike=any&groupLike=any&subgroup=-1&geneLike=&selection=any”, and its contents are incorporated herein by reference. Therefore, the VH strand is found in the genes IGHV1-18, IGHV1-2, IGHV1-24, IGHV1-3, IGHV1-45, IGHV1-46, IGHV1-58, IGHV1-69, IGHV1-69-2, IGHV1-69D, IGHV1-8, IGHV2-26, IGHV2-5, IGHV2-70, IGHV2-70D, IGHV3-11, IGHV3-13, IGHV3-15, IGHV3-20, IGHV3-21, IGHV3-23, IGHV3-23D, IGHV3-30, IGHV3-30-3, IGHV3-30-5, IGHV3-33 , may be selected from the group consisting of amino acid sequences encoded by IGHV3-35, IGHV3-43, IGHV3-43D, IGHV3-48, IGHV3-49, IGHV3-53, IGHV3-62, IGHV3-64, IGHV3-64D, IGHV3-66, IGHV3-7, IGHV3-72, IGHV3-73, IGHV3-74, IGHV3-9, IGHV3-NL1, IGHV4-28, IGHV4-30-1, IGHV4-30-2, IGHV4-30-4, IGHV4-31, IGHV4-34, and IGHV4-38-2.

[0054] Human variable framework regions (VK, κ) of light chains can be searched from the website: “https: / / www.imgt.org / genedb / resultPage.action;jsessionid=49EB34C22C79EAC51862C76108963216?gene.id.species=Homo+sapiens&molComponent=IG&geneTypeLike=variable&allele.fcode=functional&cloneName=&locusLike=IGK&mainLocusLike=IGK+locus&cosLocusLike=any&groupLike=any&subgroup=-1&geneLike=&selection=any”, and its contents are incorporated herein by reference. Therefore, the VK(κ) chain contains genes IGKV1-12, IGKV1-13, IGKV1-16, IGKV1-17, IGKV1-27, IGKV1-33, IGKV1-39, IGKV1-5, IGKV1-6, IGKV1-8, IGK V1-9, IGKV1-NL1, IGKV1D-12, IGKV1D-13, IGKV1D-16, IGKV1D-17, IGKV1D-33, IGKV1D-39, IGKV1D-43, IGKV1D-8, IGKV2-24, IGKV2 The amino acid sequences may be selected from the group consisting of those encoded by -28, IGKV2-29, IGKV2-30, IGKV2-40, IGKV2D-26, IGKV2D-28, IGKV2D-29, IGKV2D-30, IGKV2D-40, IGKV3-11, IGKV3-15, IGKV3-20, IGKV3D-11, IGKV3D-15, IGKV3D-20, IGKV3D-7, IGKV4-1, IGKV5-2, IGKV6-21, and IGKV6D-21.

[0055] The human variable framework domain for light chains (VL, λ) can be searched on the website: The contents of “https: / / www.imgt.org / genedb / resultPage.action;jsessionid=49EB34C22C79EAC51862C76108963216?gene.id.species=Homo+sapiens&molComponent=IG&geneTypeLike=variable&allele.fcode=functional&cloneName=&locusLike=IGL&mainLocusLike=IGL+locus&cosLocusLike=any&groupLike=any&subgroup=-1&geneLike=&selection=any” are incorporated herein by reference. Therefore, the VL(λ) strand is found in the genes IGLV1-36, IGLV1-40, IGLV1-44, IGLV1-47, IGLV1-51, IGLV10-54, IGLV2-11, IGLV2-14, IGLV2-18, IGLV2-23, IGLV2-8, IGLV3-1, IGLV3-10, IGLV3-12, IGLV3-16, IGLV3-19, IGLV3-21, IGL The amino acid sequences may be selected from the group consisting of V3-22, IGLV3-25, IGLV3-27, IGLV3-9, IGLV4-3, IGLV4-60, IGLV4-69, IGLV5-37, IGLV5-39, IGLV5-45, IGLV5-52, IGLV6-57, IGLV7-43, IGLV7-46, IGLV8-61, and IGLV9-49.

[0056] The heavy chain and light chain human variable framework regions may include the respective human HJ (heavy chain) and light chain KJ (κ) or LJ (λ) sequences.

[0057] The HJ sequence can be searched on the website: The contents of “https: / / www.imgt.org / genedb / resultPage.action?gene.id.species=Homo+sapiens&molComponent=IG&geneTypeLike=any&allele.fcode=functional&cloneName=&locusLike=IGH&mainLocusLike=IGH+locus&cosLocusLike=any&groupLike=IGHJ&subgroup=-1&geneLike=&selection=any” are incorporated herein by reference. In particular, the HJ sequence may be selected from amino acid sequences encoded by genes consisting of the groups: IGHJ1, IGHJ2, IGHJ3, IGHJ4, IGHJ5, and IGHJ6.

[0058] The KJ sequence can be searched on the website: The contents of “https: / / www.imgt.org / genedb / resultPage.action?gene.id.species=Homo+sapiens&molComponent=IG&geneTypeLike=any&allele.fcode=functional&cloneName=&locusLike=IGK&mainLocusLike=IGK+locus&cosLocusLike=any&groupLike=IGKJ&subgroup=-1&geneLike=&selection=any” are incorporated herein by reference. In particular, the KJ sequence may be selected from amino acid sequences encoded by genes comprising the groups IGKJ1, IGKJ2, IGKJ3, IGKJ4, and IGKJ5.

[0059] The LJ sequence can be searched from the website: The contents of “https: / / www.imgt.org / genedb / resultPage.action?gene.id.species=Homo+sapiens&molComponent=IG&geneTypeLike=any&allele.fcode=functional&cloneName=&locusLike=IGL&mainLocusLike=IGL+locus&cosLocusLike=any&groupLike=IGLJ&subgroup=-1&geneLike=&selection=any” are incorporated herein by reference. In particular, the LJ sequence may be selected from amino acid sequences encoded by genes comprising the groups: IGLJ1, IGLJ2, IGLJ3, IGLJ6, and IGLJ7.

[0060] The heavy chain CDR sequences of the present invention (CDRH1, CDRH2, and CDRH3) may be composed of a combination of a human heavy chain variable framework sequence defined by a human VH sequence described herein and a human HJ sequence described herein. Similarly, the light chain CDR sequences of the present invention (CDRL1, CDRL2, and CDRL3) may be composed of a combination of a human light chain variable framework sequence defined by a human VL or VK sequence described herein and a human LJ or KJ sequence described herein.

[0061] (i) A combination of a human VH sequence (including the CDRH1, CDRH2, and CDRH3 sequences described in the present invention, respectively) and a human VL sequence (including the CDRL1, CDRL2, and CDRL3 sequences of the present invention, respectively), or (ii) a combination of a human VH sequence (including the CDRH1, CDRH2, and CDRH3 sequences of the present invention, respectively) and a human VK sequence (including the CDRL1, CDRL2, and CDRL3 sequences of the present invention, respectively) may thus characterize the variable framework region and its binding region of the antibody according to the present invention. The variable framework region may also be characterized by (i) a combination of a human VH sequence and a human HJ sequence with a human VL sequence and a human LJ sequence, or (ii) a combination of a human VH sequence and a human HJ sequence with a human VK sequence and a human KJ sequence, thereby forming the variable framework region and its binding region of the antibody according to the present invention.

[0062] The CDR sequence of the present invention is inserted into a region of a variable framework sequence that presents the CDR sequence.

[0063] The numbering of residues in the variable region was performed according to the IMGT numbering system (IMGT: http: / / www.imgt.org / ; cf. Lefranc, M.-P. et al. (2009) Nucleic Acids Res. 37, D1006-D1012). The Kabat CDR definition was applied to define the CDR region (Tai Te Wu, Elvin A. Kabat; An analysis of the sequences of the variable regions of Bence Jones proteins and myeloma light chains and their implications for antibody complementarity. J Exp Med 1 August 1970; 132 (2): 211-250; George Johnson, Tai Te Wu, Kabat Database and its applications: 30 years after the first variability plot, Nucleic Acids Research, Volume 28, Issue 1, 1 January 2000, Pages 214-218).

[0064] [Table 4]

[0065] Furthermore, the amino acid sequences of the variable regions of the heavy chain (VH) and light chain (VL) of exemplary antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2 are shown in Table 2 below.

[0066] [Table 5]

[0067] Preferably, the antibody or antigen-binding fragment of the present invention comprises a combination of the six CDR sequences of the exemplary antibody shown in Table 1 (optionally, the VH and VL sequences of the exemplary antibody shown in Table 2), or a variant thereof, as defined herein.

[0068] In some embodiments, the antibody or antigen-binding fragment described in the present invention includes CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6. Preferably, the antibody or antigen-binding fragment includes: The heavy chain CDR1 sequence described in Sequence ID No. 1; The heavy chain CDR2 sequence described in Sequence ID No. 2; The heavy chain CDR3 sequence described in Sequence ID No. 3; Light chain CDR1 sequence as described in Sequence ID No. 4; The light chain CDR2 sequence described in Sequence ID No. 5; and, The light chain CDR3 sequence described in Sequence ID No. 6.

[0069] As shown in the attached examples, such antibodies (e.g., 4E1) specifically bind to Bet v 1 and also specifically bind to or show affinity for multiple Bet v 1 homologous proteins (including tree pollen allergens (Cor a 1, Aln g 1, Fag s 1, Que a 1, Ost c 1, etc.) and food allergens (Mal d 1, Jugr 5, etc.)).

[0070] In particular, as shown in the epitope mapping experiments described later, antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs: 1-6 (e.g., antibody 4E1) specifically bind to epitopes containing residues 46(E), 54(I), 56(K), 69(K), and 88(E) of Bet v 1.

[0071] In some embodiments, such an antibody of the present invention, or its antigen-binding fragment, has an amino acid composition having identity with SEQ ID NO: 7 of 70% or more (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). It may also include a heavy chain variable region (VH) including a column, and a light chain variable region (VL) containing an amino acid sequence having 70% or more identity with SEQ ID NO: 8 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). This preferably preserves the CDR sequences defined above (the heavy chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 1, 2, and 3, respectively; and the light chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 4, 5, and 6, respectively).

[0072] In some embodiments, the antibody, or its antigen-binding fragment, includes a heavy chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 7, and a light chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 8.

[0073] In some embodiments, the antibody or its antigen-binding fragment includes CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14. Preferably, the antibody or its antigen-binding fragment includes: The heavy chain CDR1 sequence described in Sequence ID No. 9; The heavy chain CDR2 sequence described in Sequence ID No. 10; The heavy chain CDR3 sequence described in Sequence ID No. 11; Light chain CDR1 sequence as described in Sequence ID No. 12; The light chain CDR2 sequence described in Sequence ID No. 13; and, The light chain CDR3 sequence described in Sequence ID No. 14.

[0074] As shown in the attached examples, such antibodies (e.g., 25-3D3) specifically bind to Bet v 1 and also specifically bind to or show affinity for multiple Bet v 1 homologous proteins (including Cor a 1, Aln g 1, Fag s1, Cas s1, Ost c 1, Car b1, etc.).

[0075] In particular, as shown in the epitope mapping experiments described later, antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 9-14 (e.g., antibody 25-3D3) specifically bind to epitopes containing residues 33(K), 154(A), 155(H), and 157(D) of Bet v 1.

[0076] In some embodiments, such an antibody of the present invention, or its antigen-binding fragment, has an amino acid sequence having identity with SEQ ID NO: 15 of 70% or more (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). It may also include a heavy chain variable region (VH) containing the above, and a light chain variable region (VL) containing an amino acid sequence having 70% or more identity with SEQ ID NO: 16 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). This preferably preserves the CDR sequences defined above (the heavy chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 9, 10, and 11, respectively; and the light chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 12, 13, and 14, respectively).

[0077] In some embodiments, the antibody, or its antigen-binding fragment, includes a heavy chain variable region comprising the amino acid sequence described in SEQ ID NO: 15, or said amino acid sequence, and a light chain variable region comprising the amino acid sequence described in SEQ ID NO: 16, or said amino acid sequence.

[0078] In some embodiments, the antibody or its antigen-binding fragment includes CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22. Preferably, the antibody or its antigen-binding fragment includes: The heavy chain CDR1 sequence described in Sequence ID No. 17; The heavy chain CDR2 sequence described in Sequence ID No. 18; The heavy chain CDR3 sequence described in Sequence ID No. 19; Light chain CDR1 sequence as described in Sequence ID No. 20; The light chain CDR2 sequence described in Sequence ID No. 21; and, The light chain CDR3 sequence described in Sequence ID No. 22.

[0079] As shown in the attached examples, such antibodies (e.g., 17B10) specifically bind to Bet v 1 and also specifically bind to or show affinity for multiple Bet v 1 homologous proteins (including Cor a 1, Aln g 1, Cas s 1, Ost c 1, Car b 1, etc.).

[0080] In particular, as shown in the epitope mapping experiments described later, antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 17-22 (e.g., antibody 17B10) specifically bind to epitopes containing residues 7(E), 9(E), and 131(A) of Bet v 1.

[0081] In some embodiments, such an antibody of the present invention, or its antigen-binding fragment, has an amino acid sequence having identity with SEQ ID NO: 23 of 70% or more (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). It may also include a heavy chain variable region (VH) containing the above, and a light chain variable region (VL) containing an amino acid sequence having 70% or more identity with SEQ ID NO: 24 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). This preferably preserves the CDR sequences defined above (the heavy chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 17, 18, and 19, respectively; and the light chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 20, 21, and 22, respectively).

[0082] In some embodiments, the antibody, or its antigen-binding fragment, includes a heavy chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 23, and a light chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 24.

[0083] In some embodiments, the antibody or its antigen-binding fragment includes CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71. Preferably, the antibody or its antigen-binding fragment includes: The heavy chain CDR1 sequence described in Sequence ID No. 66; The heavy chain CDR2 sequence described in Sequence ID No. 67; The heavy chain CDR3 sequence described in Sequence ID No. 68; Light chain CDR1 sequence as described in Sequence ID No. 69; The light chain CDR2 sequence described in Sequence ID No. 70; and, The light chain CDR3 sequence described in Sequence ID No. 71.

[0084] As shown in the attached examples, such antibodies (e.g., 6C9) specifically bind to Bet v 1 and also specifically bind to or exhibit affinity for multiple Bet v 1 homologous proteins (including Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Carb b 1, Ost c 1, and Jur r 5, etc.).

[0085] In particular, as shown in the epitope mapping experiments described later, antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 66-71 (e.g., antibody 6C9) specifically bind to epitopes containing residues 9(E), 11(T), 13(V), 106(V), 107(A), 110(D), and 116(K) of Bet v 1.

[0086] In some embodiments, such an antibody of the present invention, or its antigen-binding fragment, has an amino acid sequence having identity with SEQ ID NO: 72 by 70% or more (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). It may also include a heavy chain variable region (VH) containing the above, and a light chain variable region (VL) containing an amino acid sequence having 70% or more identity with SEQ ID NO: 73 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). This preferably preserves the CDR sequences defined above (the heavy chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 66, 67, and 68, respectively; and the light chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 69, 70, and 71, respectively).

[0087] In some embodiments, the antibody, or its antigen-binding fragment, includes a heavy chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 72, and a light chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 73.

[0088] In some embodiments, the antibody or its antigen-binding fragment includes CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO: 79. Preferably, the antibody or its antigen-binding fragment includes: The heavy chain CDR1 sequence described in Sequence ID No. 74; The heavy chain CDR2 sequence described in Sequence ID No. 75; The heavy chain CDR3 sequence described in Sequence ID No. 76; Light chain CDR1 sequence as described in Sequence ID No. 77; The light chain CDR2 sequence described in Sequence ID No. 78; and, The light chain CDR3 sequence described in Sequence ID No. 79.

[0089] As shown in the attached examples, such antibodies (e.g., 3F1) specifically bind to Bet v 1 and also specifically bind to or exhibit affinity for multiple Bet v 1 homologous proteins (including Cor a 1, Fag s 1, Que a 1, Cas s 1, and Jur r 5, etc.).

[0090] In particular, as shown in the epitope mapping experiments described later, the 6C9 clonotype antibody specifically binds to epitopes containing residues 9(E), 11(T), 13(V), 106(V), 107(A), 110(D), and 116(K) of Bet v 1. Antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 74-79 (e.g., antibody 3F1) belong to the same clonal type as 6C9 (as shown by the cross-competition experiment below) and also belong to the same complementarity group. Therefore, it is presumed that antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 74-79 (e.g., antibody 3F1) also specifically bind to the same epitope, namely the epitope containing residues 9(E), 11(T), 13(V), 106(V), 107(A), 110(D), and 116(K) of Bet v 1.

[0091] In some embodiments, such an antibody of the present invention, or its antigen-binding fragment, has an amino acid sequence having identity with SEQ ID NO: 80 of 70% or more (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). It may also include a heavy chain variable region (VH) containing the above, and a light chain variable region (VL) containing an amino acid sequence having 70% or more identity with SEQ ID NO: 81 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). This preferably preserves the CDR sequences defined above (the heavy chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 74, 75, and 76, respectively; and the light chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 77, 78, and 79, respectively).

[0092] In some embodiments, the antibody, or its antigen-binding fragment, includes a heavy chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 80, and a light chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 81.

[0093] In some embodiments, the antibody, or its antigen-binding fragment, includes CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87. Preferably, the antibody, or its antigen-binding fragment, includes: The heavy chain CDR1 sequence described in Sequence ID No. 82; The heavy chain CDR2 sequence described in Sequence ID No. 83; The heavy chain CDR3 sequence described in Sequence ID No. 84; Light chain CDR1 sequence as described in Sequence ID No. 85; The light chain CDR2 sequence described in Sequence ID No. 86; and, The light chain CDR3 sequence described in Sequence ID No. 87.

[0094] As shown in the attached examples, such antibodies (e.g., 13A2) specifically bind to Bet v 1 and also specifically bind to or show affinity for multiple Bet v 1 homologous proteins (including Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Carb b 1, Jur r 5, etc.).

[0095] In particular, as shown in the epitope mapping experiments described later, the 6C9 clone type antibody specifically binds to the epitope containing residues 9(E), 11(T), 13(V), 106(V), 107(A), 110(D), and 116(K) of Bet v 1. Antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 82-87 (e.g., antibody 13A2) belong to the same clone type as 6C9 (as shown in the cross-competition experiments below) and also belong to the same complementarity group. Therefore, it is presumed that antibodies containing the heavy and light chain CDR sequences described in SEQ ID NOs. 82-87 (e.g., antibody 13A2) also specifically bind to the same epitope, namely the epitope containing residues 9(E), 11(T), 13(V), 106(V), 107(A), 110(D), and 116(K) of Bet v 1.

[0096] In some embodiments, such an antibody of the present invention, or its antigen-binding fragment, has an amino acid sequence having 70% or more identity with SEQ ID NO: 88 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). It may also include a heavy chain variable region (VH) containing the above, and a light chain variable region (VL) containing an amino acid sequence having 70% or more identity with SEQ ID NO: 89 (for example, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%). This preferably preserves the CDR sequences defined above (the heavy chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 82, 83, and 84, respectively; and the light chain CDR1, CDR2, and CDR3 sequences described in SEQ ID NOs. 85, 86, and 87, respectively).

[0097] In some embodiments, the antibody, or its antigen-binding fragment, includes a heavy chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 88, and a light chain variable region comprising or consisting of the amino acid sequence described in SEQ ID NO: 89.

[0098] In general, the antibodies of the present invention, or their antigen-binding fragments, may reduce, inhibit, or neutralize allergen-mediated biological activity. In particular, the antibodies may reduce or inhibit the binding of IgE antibodies to birch pollen allergen, especially Bet v 1 as described herein, and optionally to Bet v 1 homologous allergens (other tree pollen allergens and / or food allergens). Thus, the antibodies or their binding fragments according to the present invention may reduce or inhibit the activation of mast cells or basophils, and therefore reduce or prevent the release of mediators (e.g., histamine, lipid mediators, leukotrienes). As a result, the antibodies described herein may inhibit allergic symptoms that would normally occur in a patient after contact with an allergen (e.g., contact with the eyes, nose, or mouth, or ingestion of food). Therefore, the antibodies described herein may reduce, inhibit, or neutralize allergen-mediated biological activity.

[0099] In some embodiments, the CDR, or variable region, of an antibody or its antigen-binding fragment is of human origin or derived from a human CDR or variable region sequence. Exemplary antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2 (wild-type) are human antibodies isolated from human patients. "Human-derived" CDRs or VH / VL sequences include modified human antibody sequences in which mutations have been introduced into the original human CDR or VH / VL sequence. For example, a human-derived CDR may differ from a fully human (wild-type) CDR sequence in that it contains up to five mutations, i.e., one, two, three, four, or five, preferably up to four, more preferably up to three. For example, a human-derived VH or VL sequence may differ from a fully human (wild-type) VH or VL sequence in that it contains up to 10 mutations (e.g., in the framework region), i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations, preferably up to 7, more preferably up to 5.

[0100] In some embodiments, the antibody, or its antigen-binding fragment, is a human antibody. In some embodiments, the antibody, or its antigen-binding fragment, is a monoclonal antibody. For example, the antibody, or its antigen-binding fragment, may be a human monoclonal antibody.

[0101] Human antibodies have advantages over non-human antibodies. This is because non-human antibodies, including chimeric and humanized antibodies, can induce adverse immune responses, causing symptoms such as nausea, diarrhea, and flu-like symptoms. In more severe cases, these side effects can even be fatal. Non-human antibody segments often trigger human immune responses (anti-drug antibodies (ADAs)), causing undesirable side effects and mitigating the effects of non-human antibodies in humans. In contrast, antibodies derived from humans have a higher safety profile because they have demonstrated tolerability in the human body, combined with the typical superior affinity maturation of the human immune system. In this specification, the term “human antibody” includes not only antibodies naturally present in humans but also sequence variants in which specific amino acid residues (not the entire antibody segment) are mutated. In contrast to non-human and humanized antibodies, which typically include entire non-human antibody segments (such as the entire set of CDR sequences), variants in human antibody sequences usually consist only of selective / specific mutations within selected antibody segments (e.g., within the CDR or framework region and / or constant region; for example, to modify the antibody's affinity, functionality, half-life, etc.).

[0102] For example, the human antibody according to the present invention may contain only a limited number of mutations per CDR (e.g., 6 or fewer per CDR, preferably 5 or fewer, more preferably 4 or fewer, even more preferably 3 or fewer, even more preferably 2 or fewer, and particularly preferably only a single mutation) compared to the sequences shown in Table 1. If there are more than two mutations, they do not have to occur consecutively (so as not to form a non-human sequence segment). The same applies to the framework region (or the entire VH / VL sequence) and the constant region. The latter may carry certain modifications known in the art, as described below herein, for example, to modify the functionality of the antibody (related to Fc).

[0103] Preferably, the antibody is an IgG or IgA antibody. IgG and IgA typically compete with IgE for binding sites on allergens, interfering with allergen recognition by IgE bound to Fes receptors on mast cells and basophil surfaces. This may include direct competition by binding to the same epitope or competition due to steric hindrance. Furthermore, IgG antibodies bound to allergens can cause crosslinking of Fcs and inhibitory FcyRIIB receptors, resulting in reduced effector cell activity. Thus, IgG and IgA antibodies or their conjugated fragments according to the present invention can be used for the effective prevention or treatment of allergies. In some embodiments, the variable region or CDR of the antibody as defined herein is derived from a (human) IgE antibody and grafted onto a scaffold of an IgG or IgA antibody. Preferably, the scaffold is of human IgG or IgA. Therefore, the variable region, a portion thereof, or CDR may be human, may be grafted into an antibody framework, which may preferably be human-derived, and may be a different antibody type such as IgG or IgA instead of IgE. Typically, the human-derived portion of the variable region grafted into the antibody framework includes a CDR. Among IgG, IgG1 and IgG4 are preferred.

[0104] Therefore, the antibody or antigen-binding fragment according to the present invention may include an Fc substructure. The Fc substructure may be derived from human sources, such as human IgA or IgG, such as IgG1, IgG2, IgG3, and / or IgG4, such as human IgG4.

[0105] As used herein, the term “Fc substructure” refers to a sequence derived from a portion of the immunoglobulin heavy chain, beginning at the hinge region immediately upstream of the papain cleavage site (e.g., residue 216 in native IgG, with the first residue of the heavy chain constant region being 114) and ending at the C-terminus of the immunoglobulin heavy chain. Therefore, an Fc substructure may be a complete Fc substructure or a part thereof (e.g., a domain). A complete Fc substructure includes at least the hinge domain, the CH2 domain, and the CH3 domain (e.g., EU amino acid positions 216–446). Additional lysine residues (K) may be present at the extreme C-terminus of the Fc substructure, but are often cleaved by mature antibodies.

[0106] In some embodiments, within the context of the present invention, an Fc substructure comprises at least one of a hinge domain (e.g., upper, middle, and / or lower hinge region), a CH2 domain, a CH3 domain, or a variant, part, or fragment thereof. The Fc substructure may comprise at least a hinge domain, a CH2 domain, or a CH3 domain. The Fc substructure may also comprise a complete Fc substructure. The Fc substructure may also comprise one or more amino acid insertions, deletions, or substitutions compared to a naturally occurring Fc substructure. For example, at least one of the hinge domain, CH2 domain, or CH3 domain (or a part thereof) may be deleted.

[0107] Those skilled in the art will understand that an Fc substructure may be modified to retain at least one desirable function conferred by the naturally occurring Fc substructure, even if its amino acid sequence differs from that of the complete Fc substructure of a naturally occurring immunoglobulin molecule. Such functions include Fc receptor (FcR) binding, antibody half-life regulation, ADCC function, protein A binding, protein G binding, and complement binding. The parts of the naturally occurring Fc substructure that perform and / or are essential for such functions are well known to those skilled in the art. In some embodiments, the antibody according to the present invention contains a (complete) Fc substructure / Fc region in which interaction / binding with the Fc receptor is not impaired.

[0108] Generally, antibody binding to Fc receptors can be determined by various methods known to those skilled in the art, such as ELISA (Hessell AJ, Hangartner L, Hunter M, Havenith CEG, Beurskens FJ, Bakker JM, Lanigan CMS, Landucci G, Forthal DN, Parren PWHI, et al.: Fc receptor but not complement binding is important in antibody protection against HIV. Nature 2007, 449: 101-104; Grevys A, Bern M, Foss S, Bratlie DB, Moen A, Gunnarsen KS, Aase A, Michaelsen TE, Sandlie I, Andersen JT: Fc Engineering of Human IgG1 for Altered Binding to the Neonatal Fc Receptor Affects Fc Effector Functions. 2015, 194:5497-5508), or flow cytometry (Perez LG, Costa MR, Todd CA, Haynes BF, Montefiori DC: Utilization of immunoglobulin G Fc receptors by human immunodeficiency virus type 1: a specific role for antibodies against the membrane-proximal external region of gp41. J Virol 2009, 83:7397-7410; Piccoli L, Campo I, Fregni CS, Rodriguez BMF, Minola A, Sallusto F, Luisetti M, Corti D, Lanzavecchia A: Neutralization and clearance of GM-CSF by autoantibodies in pulmonary alveolar proteinosis.It may also be evaluated by sources such as Nat Commun 2015, 6:1-9.

[0109] In some embodiments, the antibody or antigen-binding fragment thereof according to the present invention includes an Fc region. As used herein, the term “Fc region” refers to a portion of an immunoglobulin formed by two or more Fc substructures of an antibody heavy chain. For example, an Fc region may be a monomer or a “single-chain” Fc region (i.e., an scFc region). A single-chain Fc region consists of Fc substructures linked within a single polypeptide chain (e.g., encoded in a single continuous nucleic acid sequence). An exemplary scFc region is disclosed in WO 2008 / 143954 A2. An Fc region may be a dimer. “Dimerated Fc region” or “dcFc” refers to a dimer formed by two distinct Fc substructures of immunoglobulin heavy chains. A dimerated Fc region may be a homodimer of two identical Fc substructures (e.g., Fc regions of naturally occurring immunoglobulins) or a heterodimer of two non-identical Fc substructures.

[0110] In some embodiments, the Fc substructure or Fc region includes, or consists of, an amino acid sequence derived from a human immunoglobulin sequence (e.g., an Fc region or Fc substructure derived from a human IgG molecule). However, the Fc substructure or Fc region may also include one or more amino acids derived from other mammalian species. For example, a primate Fc substructure or primate binding site may be included in the antibody or antigen-binding fragment. Alternatively, one or more mouse amino acids may be present in the Fc substructure or Fc region.

[0111] The Fc substructures of the Fc region may be of the same or different classes and / or subclasses. For example, the Fc substructures may be derived from immunoglobulins of the IgG1, IgG2, IgG3, or IgG4 subclasses (e.g., human immunoglobulins).

[0112] Therefore, the antibody of the present invention may be any isotype (e.g., IgA, IgG, IgM, i.e., α, γ, or μ heavy chain). Preferably, the antibody may be of the IgA or IgG type. Among the IgG isotypes, the antibody may be of the IgG1, IgG2, IgG3, or IgG4 subclass, preferably IgG1 or IgG4, and more preferably IgG4. An exemplary sequence of the IgG4 constant region, which may be useful in the antibodies described herein, is provided in SEQ ID NO: 49. Therefore, the antibody of the present invention may contain the amino acid sequence described in SEQ ID NO: 49, or a variant of that sequence described herein. The human IgG4 constant region sequence of Sequence ID No. 49 contains the stabilizing hinge mutation S228P (S. Angal, DJ King, MW Bodmer, A. Turner, ADG Lawson, G. Roberts, B. Pedley, JR Adair, A single amino acid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody, Molecular Immunology, Volume 30, Issue 1, 1993, Pages 105-108, ISSN 0161-5890, https: / / doi.org / 10.1016 / 0161-5890(93)90432-B). The antibodies of the present invention may have a κ or λ light chain. Exemplary sequences of κ and λ light chain constant regions that may be useful in the antibodies described herein are provided in Sequence ID No. 50 (κLC) and Sequence ID No. 51 (λLC). Therefore, the antibody of the present invention may contain the amino acid sequence of SEQ ID NO: 50 or 51, or a variant of that sequence as described herein.

[0113] As described above, the present invention encompasses antigen-binding fragments. The antigen-binding fragment may or may not include an Fc substructure, particularly a portion of the complete Fc region. In some embodiments, the antibody or its antigen-binding fragment is selected from Fab, Fab', F(ab')2, Fv, or scFv. For example, F(ab')2 (which may be obtained by pepsin cleavage or recombinant expression) and Fab' (which may be obtained from F(ab')2 or by recombinant expression) typically include a hinge region.

[0114] In some embodiments, the antibody, or antigen-binding fragment, may be a single-chain antibody (or fragment). The single-chain antibody (or fragment) may encode a complete set of six CDRs, i.e., comprising three heavy-chain CDRs and three light-chain CDRs. More specifically, the single-chain antibody (or fragment) may also include a heavy-chain variable region (VH) and a light-chain variable region (VL), for example, the VH and VL sequences described above.

[0115] Variant antibodies are also included in the scope of the present invention. Therefore, the sequence variants described in this specification are also included in the scope of the present invention. Such variants include native variants that arise from in vivo somatic mutations during the course of an immune response, or native variants that are generated in vitro during the culture of immortalized B cell clones. Alternatively, variants may arise from degeneracy of the genetic code, or from errors in transcription or translation.

[0116] The antibody or antigen-binding fragment of the present invention may be provided in a purified form. Typically, the antibody or antigen-binding fragment will be present in a composition that is substantially free of other polypeptides (for example, a composition in which less than 90% (by weight), usually less than 60%, and more typically less than 50%, of the composition consists of other polypeptides).

[0117] The antibodies of the present invention may be immunogenic in non-human (or heterologous) hosts, such as mice. In particular, antibodies may have an idiotope that is immunogenic in non-human hosts but not immunogenic in human hosts. In particular, antibodies of the present invention used in humans include those that cannot be easily isolated from hosts (such as mice, goats, rabbits, rats, and non-primate mammals) and cannot generally be obtained by humanization or xenomouse.

[0118] The antibodies of the present invention also include hybrid antibody molecules comprising six CDRs derived from the antibody of the present invention as defined above, and one or more CDRs derived from another antibody against the antigen. For example, the antibody may be multispecific. In other embodiments, the antibody, or its antigen-binding fragment, may be monospecific.

[0119] Therefore, the antibodies or antigen-binding fragments described herein may be multispecific antibodies or multispecific antigen-binding fragments.

[0120] As used herein, the term “multispecificity” refers to the ability to bind to at least two different epitopes, for example, on different antigens or on the same antigen. Conventional monospecific IgG antibodies typically have two identical epitope-binding sites (paratopes) and therefore can only bind to identical epitopes (and not to different epitopes). In contrast, multispecific antibodies have at least two different types of paratopes (antigen-binding sites) and therefore can bind to at least two different epitopes. As used herein, “paratope” refers to the antigen-binding site (or epitope-binding site) of an antibody. Furthermore, a single “specificity” may refer to one, two, three, or more identical paratopes in a single antibody (the actual number of paratopes in a single antibody molecule is called the “valency”). For example, a single native IgG antibody is monospecific and bivalent because it has two identical paratopes. Therefore, a multispecific antibody contains at least two (different) paratopes. Therefore, the term "multispecific antibody" refers to an antibody that has more than one paratope and the ability to bind to two or more different epitopes. The term "multispecific antibody" specifically includes bispecific antibodies, but typically also includes proteins that bind to three or more different epitopes, such as antibody scaffolds, i.e., antibodies that have three or more different paratopes.

[0121] In particular, the multispecific antibody or multispecific antigen-binding fragment of the present invention may contain two or more paratopes, where one or more paratopes may be identical, so that all paratopes of the antibody belong to at least two different types of paratopes, and therefore the antibody has at least two specificities. For example, the multispecific antibody or antigen-binding fragment of the present invention may contain four paratopes, in which case each pair of paratopes is identical (i.e., has the same specificity), and therefore the antibody or fragment is bispecific and tetravalent (two identical paratopes for each of the two specificities). Thus, "one specificity" refers in particular to one or more paratopes exhibiting the same specificity (which typically means that one or more such paratopes are identical), and therefore "two specificities" may be realized by two, three, four, five, six or more paratopes, as long as only two specificities are referred to. In some embodiments, a multispecific antibody contains one single paratope for each (at least two) specificity, i.e., a multispecific antibody contains at least two paratopes in total. For example, a bispecific antibody may contain one single paratope for each of the two specificities, i.e., the antibody may contain two paratopes in total. In other embodiments, the antibody contains two (identical) paratopes for one or more specificities.

[0122] Preferably, the multispecific antibody or multispecific antigen-binding fragment is bispecific or triplicate. As used herein, terms such as “bispecific” and “triplicate” refer to the number of different epitopes to which the antibody can bind. For example, a “bispecific” antibody has exactly two distinct specificities (two distinct antigen-binding sites, each of which may independently occur once or more times, for example, twice). For example, a “triplicate” antibody has exactly three distinct specificities (three distinct antigen-binding sites, each of which may independently occur once or more times, for example, twice).

[0123] Various multispecific antibody formats and methods for obtaining such multispecific antibodies are known in the art. For example, building blocks for bispecific and trispecific antibodies are described in Xiufeng Wu, Stephen J. Demarest, Building blocks for bispecific and trispecific antibodies, Methods, Volume 154, 2019, Pages 3-9, ISSN 1046-2023, https: / / doi.org / 10.1016 / j.ymeth.2018.08.010 (incorporated herein by reference). Methods for obtaining multispecific antibodies and further multispecific antibody formats are described in Amaral M, Hoelper S, Lange C, Jung J, Sjuts H, Weil S, Fischer M, Radoevic K, Rao E. Engineered Technologies and Bioanalysis of multispecific antibody formats. J Appl Bioanal 6(1), 26-51 (2020) (incorporated herein by reference).

[0124] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment of the present invention is a bispecific antibody or bispecific antigen-binding fragment. A bispecific antibody contains (exactly) two specificities. In the context of the present invention, a bispecific antibody may be any bispecific antibody format known in the art, for example, as described in Spiess C., Zhai Q. and Carter PJ (2015) Molecular Immunology 67: 95-106. For example, a bispecific antibody may be a whole antibody, such as a whole IgG-like molecule, or a fragment of that antibody that is not a whole antibody but retains antibody properties. These may also be small recombinant formats, for example, tandem single-strand variable fragment molecules (taFvs), diabodies (Dbs), single-strand diabodies (scDbs), and various other derivatives thereof (for example, as described in Byrne H. et al. (2013) Trends Biotech, 31 (11): 621-632, where various bispecific antibody formats are shown in Figure 2). In some embodiments, the bispecific antibody may be an IgG(H)-scFv fusion, as described in Coloma, M., Morrison, S. Design and production of novel tetravalent bispecific antibodies. Nat Biotechnol 15, 159-163 (1997). https: / / doi.org / 10.1038 / nbt0297-159.

[0125] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment according to the present invention is a triplicate antibody or triplicate antigen-binding fragment. A triplicate antibody has (exactly) three specificities. In the context of the present invention, a triplicate antibody may be any triplicate antibody format known in the art. In some embodiments, the triplicate antibody may be a heterodimer of IgG(H)-scFv having the same Fab domain and using knob-into-hole CH3, as described in Ridgway JB, Presta LG, Carter P. 'Knobs-into-holes' engineering of antibody CH3 domains for heavy chain heterodimerization. Protein Eng. 1996 Jul;9(7):617-21. doi: 10.1093 / protein / 9.7.617.

[0126] Typically, a multispecific antibody or multispecific antigen-binding fragment is at least bivalent; that is, it has at least two paratopes. Preferably, the multispecific antibody or multispecific antigen-binding fragment is bivalent, trivalent, tetravalent, or hexavalent. More preferably, the multispecific antibody or multispecific antigen-binding fragment is tetravalent. Even more preferably, the multispecific antibody or multispecific antigen-binding fragment is tetravalent and bispecific or triplicate.

[0127] Preferably, the multispecific antibody or multispecific antigen-binding fragment binds (specifically) to a distinct non-overlapping epitope of Bet v 1, and optionally, to a distinct non-overlapping epitope of at least one Bet v 1 homologous protein. Therefore, the antigen-binding site (paratope) of the multispecific antibody or multispecific antigen-binding fragment preferably targets the same antigen, i.e., a different non-overlapping epitope on Bet v 1, and optionally, on a different non-overlapping epitope on a Bet v 1 homologous protein. For this purpose, the antigen-binding site (paratope) of the multispecific antibody or multispecific antigen-binding fragment may be derived from antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, respectively, as disclosed herein, and these antibodies preferably bind to a distinct non-overlapping epitope of Bet v 1. For example, as described above, methods for obtaining multispecific antibodies using the binding site of a monospecific antibody are known to those skilled in the art.

[0128] Therefore, a multispecific antibody or multispecific antigen-binding fragment may contain at least two of the following: (i) comprising CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6; in particular comprising CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6 antigen binding site; (ii) comprising CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14; in particular comprising CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14, antigen binding site; (iii) comprising CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22; in particular comprising CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22 antigen binding site; (iv) comprising CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71; in particular comprising CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO: 71 antigen binding site; (v) A CDRH1 having at least 70% identity with SEQ ID NO: 74, a CDRH2 having at least 70% identity with SEQ ID NO: 75, a CDRH3 having at least 70% identity with SEQ ID NO: 76, a CDRL1 having at least 70% identity with SEQ ID NO: 77, a CDRL2 having at least 70% identity with SEQ ID NO: 78, and a CDRL3 having at least 70% identity with SEQ ID NO: 79; in particular, a CDRH1 described in SEQ ID NO: 74, a CDRH2 described in SEQ ID NO: 75, a CDRH3 described in SEQ ID NO: 76, a CDRL1 described in SEQ ID NO: 77, a CDRL2 described in SEQ ID NO: 78, and a CDRL3 described in SEQ ID NO: 79 antigen binding site; (vi) CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87; in particular, CDRH1 as described in SEQ ID NO: 82, CDRH2 as described in SEQ ID NO: 83, CDRH3 as described in SEQ ID NO: 84, CDRL1 as described in SEQ ID NO: 85, CDRL2 as described in SEQ ID NO: 86, and CDRL3 as described in SEQ ID NO: 87 Antigen binding site.

[0129] In a preferred embodiment, the multispecific antibody or multispecific antigen-binding fragment is a trispecific antibody or trispecific antigen-binding fragment comprising three antigen-binding sites selected from (i) to (vi) as defined above, preferably comprising a first antigen-binding site as described in (i), a second antigen-binding site as described in (ii), and a third antigen-binding site selected from any one of (iii) to (vi).

[0130] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment includes at least two of the following: (i) an antigen-binding site comprising VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8, Here, CDRH1 described in SEQ ID NO: 1, CDRH2 described in SEQ ID NO: 2, CDRH3 described in SEQ ID NO: 3, CDRL1 described in SEQ ID NO: 4, CDRL2 described in SEQ ID NO: 5, and CDRL3 described in SEQ ID NO: 6 are preferably retained, and in particular include VH described in SEQ ID NO: 7 and VL described in SEQ ID NO: 8. antigen binding site; (ii) An antigen-binding site comprising VH having at least 70% identity with SEQ ID NO: 15, and VL having at least 70% identity with SEQ ID NO: 16, Here, CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14 are preferably retained, and in particular include VH described in SEQ ID NO: 15 and VL described in SEQ ID NO: 16. antigen binding site; (iii) an antigen-binding site comprising VH having at least 70% identity with SEQ ID NO: 23, and VL having at least 70% identity with SEQ ID NO: 24, Here, CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22 are preferably retained, and in particular include VH described in SEQ ID NO: 23 and VL described in SEQ ID NO: 24. antigen binding site; (iv) An antigen-binding site comprising VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73, Here, CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71 are preferably retained, and in particular include VH described in SEQ ID NO: 72 and VL described in SEQ ID NO: 73. antigen binding site; (v) an antigen-binding site comprising VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81, Here, CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79 are preferably retained, and in particular include VH described in SEQ ID NO: 80 and VL described in SEQ ID NO: 81. antigen binding site; (vi) an antigen-binding site comprising VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89, Here, CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87 are preferably retained, and in particular include VH described in SEQ ID NO: 88 and VL described in SEQ ID NO: 89. Antigen binding site.

[0131] In a preferred embodiment, the multispecific antibody or multispecific antigen-binding fragment is a trispecific antibody or trispecific antigen-binding fragment comprising three antigen-binding sites selected from (i) to (vi) as defined above, preferably comprising a first antigen-binding site as described in (i), a second antigen-binding site as described in (ii), and a third antigen-binding site selected from any one of (iii) to (vi).

[0132] In a preferred embodiment, the multispecific antibody or multispecific antigen-binding fragment includes at least two of the following: (i) Antigen-binding sites of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) Antigen-binding sites of 25-3D3 as described above, comprising (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof; (iii) Antigen-binding sites of 17B10 as described above, including (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof; (iv) Antigen-binding sites of 6C) described above, including (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof; (v) Antigen-binding sites of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) Antigen-binding sites of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0133] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment may be a bispecific antibody or bispecific antigen-binding fragment comprising: (i) an antigen-binding site of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and (ii) Antigen-binding sites of 25-3D3 as described above, comprising (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof.

[0134] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment may be a bispecific antibody or bispecific antigen-binding fragment comprising: (i) an antigen-binding site of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and (iii) Antigen-binding sites of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0135] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment may be a bispecific antibody or bispecific antigen-binding fragment comprising: (ii) Antigen-binding sites of 25-3D3 as described above, including (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof; and (iii) Antigen-binding sites of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0136] In preferred embodiments, the multispecific antibody or multispecific antigen-binding fragment may be a trispecific antibody or trispecific antigen-binding fragment comprising: (i) Antigen-binding sites of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) an antigen-binding site of 25-3D3 described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and a third antigen-binding site selected from the group consisting of (iii) to (vi): (iii) Antigen-binding sites of 17B10 as described above, including (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof; (iv) Antigen-binding sites of 6C9 as described above, comprising (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) VH and VL sequences, or variants thereof; (v) Antigen-binding sites of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) Antigen-binding sites of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0137] In other words, in certain embodiments, the antigen-binding site (paratope) of a multispecific antibody or multispecific antigen-binding fragment may be selected from three different antibodies 4E1, 25-3D3, and 17B10, or their variants, as described above, particularly as shown in Tables 1 and 2. In another particular embodiment, the antigen-binding site (paratope) of a multispecific antibody or multispecific antigen-binding fragment may be selected from three different antibodies 4E1, 25-3D3, and 6C9. In yet another particular embodiment, the antigen-binding site (paratope) of a multispecific antibody or multispecific antigen-binding fragment may be selected from three different antibodies 4E1, 25-3D3, and 3F1. In yet another particular embodiment, the antigen-binding site (paratope) of a multispecific antibody or multispecific antigen-binding fragment may be selected from three different antibodies 4E1, 25-3D3, and 13A2.

[0138] In some embodiments, the multispecific antibody or multispecific antigen-binding fragment is bispecific and contains two distinct antigen-binding sites selected from (i) to (vi). Such a bispecific antibody may be combined at the time of use with a second (monospecific) antibody containing the remaining antigen-binding sites selected from (i) to (vi) (i.e., antigen-binding sites selected from (i) to (vi) that are not present in the (first) bispecific antibody). For example, a bispecific antibody containing two distinct antigen-binding sites selected from (i) to (iii) may be combined with a second antibody so that the combination of the bispecific antibody and the monospecific antibody contains all three antigen-binding sites from (i) to (iii). For this combination, the antibodies may be contained in the same composition or in different compositions (but may be administered in a combined treatment schedule).

[0139] [Nucleic acid] In another embodiment, the present invention also provides nucleic acid molecules comprising polynucleotides that encode the antibodies described above or their antigen-binding fragments.

[0140] In some embodiments, the nucleic acid molecule comprises one or more polynucleotides encoding an exemplary antibody in the present invention (e.g., as described above, particularly shown in Tables 1 and 2) or a variant of its sequence (e.g., having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity).

[0141] Table 3 below shows exemplary nucleic acid sequences encoding the CDR and VH / VL sequences of the exemplary antibodies described herein.

[0142] [Table 6]

[0143] Examples of nucleic acid molecules and / or polynucleotides include recombinant polynucleotides, vectors, oligonucleotides, RNA molecules (such as rRNA, mRNA, miRNA, siRNA, and tRNA), or DNA molecules such as cDNA. The nucleic acid may encode the light and / or heavy chains of an antibody (or single-chain antibody). In other words, the light and heavy chains of an antibody may be encoded by the same nucleic acid molecule (e.g., in the case of a single-chain antibody, or an antibody having separate heavy and light chains in a bisistron configuration, or in an expression cassette containing multiple ribosome entry sites such as IRES). Alternatively, the light and heavy chains of an antibody may be encoded by separate nucleic acid molecules. Similarly, in the case of a multispecific antibody containing two or more immunoglobulin chains, the different chains may be encoded by the same nucleic acid molecule (e.g., in a multicistron configuration, or in an expression cassette containing two or more ribosome entry sites such as IRES). Alternatively, the different immunoglobulin chains of a multispecific antibody may be encoded by separate nucleic acid molecules.

[0144] Due to the redundancy of the genetic code, the present invention also includes variants of nucleic acid sequences encoding the same amino acid sequence. The polynucleotide (or complete nucleic acid molecule) encoding the antibody may be optimized for antibody expression. For example, codon optimization of the nucleotide sequence may be used to improve translation efficiency in the antibody production expression system. Furthermore, the nucleic acid molecule may contain heterologous elements (i.e., elements that do not naturally exist on the same nucleic acid molecule as the coding sequence of the antibody (heavy or light chain)). For example, the nucleic acid molecule may contain heterologous promoters, heterologous enhancers, heterologous UTRs (e.g., for optimal translation / expression), heterologous poly-A-tails, heterologous DNA insulator elements, and so on.

[0145] A nucleic acid molecule is a molecule that contains nucleic acid components. The term nucleic acid molecule usually refers to a DNA or RNA molecule. The term nucleic acid molecule may also be used as a synonym for "polynucleotide." That is, a nucleic acid molecule may consist of polynucleotides that encode antibodies. Alternatively, a nucleic acid molecule may contain additional elements in addition to polynucleotides that encode antibodies. Typically, a nucleic acid molecule is a polymer containing, or composed of, nucleotide monomers covalently bonded to each other by phosphodiester bonds of a sugar / phosphate backbone. The term "nucleic acid molecule" also encompasses modified nucleic acid molecules (such as base-modified, sugar-modified, or backbone-modified ones), DNA, or RNA molecules.

[0146] In general, nucleic acid molecules may be manipulated to insert, delete, or modify specific nucleic acid sequences. Such manipulations include, but are not limited to, changes to introduce restriction sites, changes to modify codon usage, and changes to add or optimize transcriptional and / or translational regulatory sequences. It is also possible to alter nucleic acids to change the encoded amino acids. For example, it may be useful to introduce one or more amino acid substitutions, deletions, and / or insertions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) into the amino acid sequence of an antibody. Such point mutations may alter effector function, antigen-binding affinity, post-translational modification, immunogenicity, etc., introduce amino acids for covalent bonding (e.g., labels), or introduce tags (e.g., for purification purposes). Alternatively, mutations in nucleic acid sequences may be "silent," meaning they are not reflected in the amino acid sequence due to the redundancy of the gene code. Generally, mutations may be introduced at specific sites, or they may be introduced randomly and then selected (e.g., through molecular evolution). For example, one or more nucleic acids encoding either the light or heavy chain of an (exemplary) antibody may be mutated randomly or directionally to introduce different properties into the encoded amino acids. Such changes may result from iterative processes in which the initial change is retained and new changes are introduced at other nucleotide positions. Furthermore, changes achieved in independent processes may be combined.

[0147] In some embodiments, the polynucleotide (i.e., the (complete) nucleic acid molecule) encoding the antibody or its antigen-binding fragment may be codon-optimized. Various tools for codon optimization are well known to those skilled in the art, including, for example, those described in the following publications: Ju Xin Chin, Bevan Kai-Sheng Chung, Dong-Yup Lee, Codon Optimization OnLine (COOL): a web-based multi-objective optimization platform for synthetic gene design, Bioinformatics, Volume 30, Issue 15, 1 August 2014, Pages 2210-2212; or in: Grote A, Hiller K, Scheer M, Munch R, Nortemann B, Hempel DC, Jahn D, JCat: a novel tool to adapt codon usage of a target gene to its potential expression host. Nucleic Acids Res. 2005 Jul 1;33(Web Server issue):W526-31; or, for example, Genscript's OptimumGene® algorithm (described in US2011 / 0091708A1).

[0148] For example, the nucleic acid molecule of the present invention may contain a nucleic acid sequence described in any one of SEQ ID NOs. 25-48 and SEQ ID NOs. 90-113; or a variant of that sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity. Thereafter, the nucleic acid molecule may encode any one of the exemplified antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2 (by combining sequences as shown in Table 3), or variants of those sequences as described herein.

[0149] The present invention also provides a plurality of nucleic acid molecules encoding the antibody or its antigen-binding fragment described herein. Each of the nucleic acid molecules (of the plurality of nucleic acid molecules) contains a polynucleotide encoding the immunoglobulin chain of the antibody or its antigen-binding fragment. Thus, the plurality of nucleic acid molecules together encode the antibody or its antigen-binding fragment (all of its immunoglobulin chain) described herein. In some embodiments, the plurality of nucleic acid molecules encoding the antibody or its antigen-binding fragment described herein may be a combination of first and second nucleic acid molecules, where the first nucleic acid molecule contains a polynucleotide encoding the heavy chain of the antibody or its antigen-binding fragment of the present invention; and the second nucleic acid molecule contains a polynucleotide encoding the corresponding light chain of the same antibody or its same antigen-binding fragment.

[0150] In general, the above description of the (general) characteristics of the nucleic acid molecules of the present invention can be appropriately applied to nucleic acid molecules of multiple nucleic acid molecules. Therefore, one or more polynucleotides encoding the immunoglobulin chain of an antibody or its antigen-binding fragment may be codon-optimized. For example, the multiple may include the nucleic acid sequence described in any one of SEQ ID NOs. 25-48 and SEQ ID NOs. 90-113; or variants of that sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity. As a result, multiple nucleic acid molecules may encode one of the exemplified antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2 (by combining sequences as shown in Table 3), or variants of those sequences as described herein.

[0151] 〔vector〕 The scope of the present invention also includes nucleic acid molecules or vectors containing multiple nucleic acid molecules according to the present invention, such as expression vectors. Typically, vectors contain nucleic acid molecules as described above.

[0152] The present invention also provides a plurality of vectors comprising a plurality of nucleic acid molecules according to the present invention as described above. Thereafter, each vector of the plurality of vectors may contain one or more nucleic acid molecules of the plurality of nucleic acid molecules according to the present invention as described above. In some embodiments, the plurality of vectors may be a combination of a first vector and a second vector, where the first vector comprises the first nucleic acid molecules (with respect to the combination of nucleic acid molecules) as described above, and the second vector comprises the second nucleic acid molecules (with respect to the combination of nucleic acid molecules) as described above.

[0153] Vectors are typically recombinant nucleic acid molecules, i.e., nucleic acid molecules that do not exist in nature. Therefore, vectors may contain heterologous elements (i.e., sequence elements with different origins in nature). For example, a vector may contain multiple cloning sites, heterologous promoters, heterologous enhancers, heterologous selection markers (for identifying cells containing the vector compared to cells without the vector), heterologous origins of replication, heterologous DNA insulating elements, etc. In the context of the present invention, vectors are suitable for incorporating or holding a desired nucleic acid sequence. Such vectors may be storage vectors, expression vectors, cloning vectors, transfer vectors, etc. A storage vector is a vector that enables the convenient storage of nucleic acid molecules. Therefore, a vector may contain sequences corresponding to, for example, a desired antibody (heavy and / or light chain) according to the present invention. An expression vector may be used for the production of expression products of RNA, etc. (e.g., mRNA, or peptides, polypeptides, or proteins). For example, an expression vector may contain sequences necessary for transcription of the vector's sequence extension, such as (heterologous) promoter sequences. A cloning vector is typically a vector containing a cloning site which may be used to incorporate a nucleic acid sequence into the vector. A cloning vector may be, for example, a plasmid vector or a bacteriophage vector. A transfer vector may be a vector suitable for transferring nucleic acid molecules into cells or organisms, such as a viral vector. In the context of this invention, a vector may be, for example, an RNA vector or a DNA vector. For example, a vector in the sense of this application includes a cloning site, a selection marker such as an antibiotic resistance factor, and a sequence suitable for vector proliferation, such as an origin of replication. In the context of this application, a vector may be a plasmid vector.

[0154] As used herein, the term “vector” may refer, for example, to a delivery vector for viral or nonviral delivery of nucleic acids of the present invention; or to a viral or nonviral delivery system. Accordingly, the present invention also provides a delivery vector / system comprising such nucleic acid molecules (or such expression vector). The delivery vector / system may be viral or nonviral. Various examples of viral and nonviral delivery vector / systems are known in the art, including, for example, those described in Nayerossadat N, Maedeh T, Ali PA. Viral and nonviral delivery systems for gene delivery. Adv Biomed Res. 2012;1:27. doi:10.4103 / 2277-9175.98152, which are incorporated herein by reference. Non-limiting examples of viral delivery vectors / systems include retroviral vectors; adenovirus vectors; adeno-associated virus (AAV) vectors, including helper-dependent adenovirus vectors and hybrid adenovirus vectors; herpes simplex virus vectors; lentivirus vectors; poxvirus vectors; and Epstein-Barr virus vectors. Among viral vectors, adenovirus vectors and adeno-associated virus (AAV) vectors are preferred. Non-limiting examples of non-viral delivery vectors / systems include chemical and non-chemical methods. Non-chemical delivery includes physical methods such as electroporation and other methods for transient penetration of cell membranes by mechanical, electrical, ultrasonic, hydrodynamic, or laser-based energy; naked DNA or RNA delivery; gene guns; hydrodynamic delivery; ultrasonic delivery and magnetofection. Chemical non-viral delivery systems include cationic particles, particularly cationic lipids / liposomes, cationic polymers, and lipid / polymer systems. Among non-viral vectors / systems, cationic liposomes are preferred.

[0155] 〔cell〕 In a further embodiment, the present invention also provides (host) cells expressing an antibody or its antigen-binding fragment according to the present invention; and / or (host) cells comprising a vector (or a plurality of vectors) according to the present invention. The (host) cells may be isolated cells that are not part of the body of a human or animal, e.g., a cell line or an artificial cell. These cells may recombinantly, e.g., heterologously express the nucleic acid or vector of the present invention (i.e., these cells / cell types do not naturally express the antibody or antigen-binding fragment).

[0156] Examples of such cells include, but are not limited to, eukaryotic cells such as yeast cells, animal cells, and plant cells. Other examples of such cells include, but are not limited to, prokaryotic cells such as Escherichia coli. In some embodiments, the cells are mammalian cells, such as mammalian cell lines. Examples include human cells, CHO cells, HEK293 cells, PER.C6 cells, NS0 cells, human hepatocytes, myeloma cells, or hybridoma cells.

[0157] Cells may be transfected with vectors according to the present invention, such as expression vectors. The term “transfection” refers to the introduction of nucleic acid molecules, such as DNA or RNA (e.g., mRNA) molecules, into cells (such as eukaryotic or prokaryotic cells). In the context of the present invention, the term “transfection” encompasses any method known to those skilled in the art for introducing nucleic acid molecules into cells, such as mammalian cells. Such methods include, for example, electroporation, lipofection based on cationic lipids and / or liposomes, calcium phosphate precipitation, nanoparticle-based transfection, virus-based transfection, or transfection based on cationic polymers such as DEAE-dextran or polyethyleneimine. In some embodiments, the introduction is nonviral.

[0158] Furthermore, cells of the present invention may be stably or transiently transfected with a vector of the present invention, for example, to express an antibody according to the present invention. In some embodiments, cells are stably transfected with a vector of the present invention encoding an antibody according to the present invention. In other embodiments, cells are transiently transfected with a vector of the present invention encoding an antibody according to the present invention.

[0159] Accordingly, the present invention also provides recombinant host cells that heterologously express the antibody or antigen-binding fragment of the present invention. For example, the cells may be of a different species from the antibody (e.g., CHO cells expressing human antibodies). In some embodiments, the cell type of the cells does not express (such) antibodies in nature. Furthermore, the host cells may confer post-translational modifications (PTMs; e.g., glycosylation) to the antibody that are not present in the native state. Such PTMs may result in functional differences (e.g., reduced immunogenicity). Therefore, the antibody or antigen-binding fragment of the present invention may have post-translational modifications distinct from naturally produced antibodies (e.g., antibodies of the immune response in humans).

[0160] [Antibody production] The antibodies according to the present invention can be produced by any method known in the art. For example, the general methodology for producing monoclonal antibodies using hybridoma technology is well known (Kohler, G. and Milstein, C., 1975; Kozbar et al. 1983).

[0161] Standard molecular biology techniques may be used to prepare the DNA sequences encoding the antibodies or antigen-binding fragments of the present invention. The desired DNA sequences may be synthesized completely or partially, for example, using oligonucleotide synthesis techniques. Site-directed mutagenesis and polymerase chain reaction (PCR) methods may be used as appropriate.

[0162] Any suitable host cell / vector system may be used to express the DNA sequence encoding the antibody molecule of the present invention. Eukaryotic (e.g., mammalian) host cell expression systems may be used for the production of antibody molecules such as complete antibody molecules. Suitable mammalian host cells include, but are not limited to, CHO, HEK293, PER.C6, NS0, myeloma, or hybridoma cells. Prokaryotic (e.g., bacterial) host cell expression systems may also be used for the production of antibody molecules such as complete antibody molecules. Suitable bacterial host cells include, but are not limited to, E. coli cells.

[0163] Therefore, the present invention provides a method for preparing an antibody, antigen-binding fragment, or immunoglobulin chain according to the present invention, the method comprising the following: (i) The process of culturing the host cells as described above; and, (ii) The step of isolating the antibody or immunoglobulin chain from the culture.

[0164] In other words, the present invention also provides a method for producing an antibody molecule according to the present invention, comprising the steps of culturing (heterogeneous) host cells containing a vector encoding the nucleic acid of the present invention under conditions suitable for the expression of a protein from DNA encoding the antibody molecule of the present invention, and isolating the antibody molecule.

[0165] To produce antibodies containing both heavy and light chains, host cells, such as cell lines, may be transfected with two vectors: a first vector encoding a light chain polypeptide and a second vector encoding a heavy chain polypeptide, as described above. Alternatively, a single vector may be used, which contains sequences encoding both light and heavy chain polypeptides (e.g., in a single-chain antibody or bicistronic manner). Similarly, multiple vectors may be used to express multispecific antibodies having two or more immunoglobulin chains.

[0166] Accordingly, the present invention also provides a method for preparing recombinant cells, the method comprising the steps of: (i) providing one or more nucleic acids encoding an antibody of the present invention; (ii) inserting the nucleic acids into an expression vector; and (iii) transfecting the vector into (heterogeneous) host cells to enable the expression of the antibody of interest in the host cells. The nucleic acids in step (i) may, but are not required, be manipulated to introduce restriction sites, alter codon usage, and / or optimize transcriptional and / or translational regulatory sequences.

[0167] Furthermore, the present invention also provides a method for preparing transfected host cells. This method includes the step of transfecting host cells with one or more nucleic acids encoding an antibody of interest. Thus, the procedure of first preparing nucleic acids and then transfecting host cells with them can be carried out in different locations (e.g., different countries), by different people, and at different times.

[0168] Such recombinant cells of the present invention can be used for expression and culture purposes. These cells are particularly useful for antibody expression for large-scale pharmaceutical production. They can also be used as active ingredients in pharmaceutical compositions. Any suitable culture technique can be used, including but not limited to static culture, roller bottle culture, ascites, hollow fiber bioreactor cartridges, modular mini-fermenters, agitated tanks, microcarrier culture, and ceramic core perfusion.

[0169] Transfected host cells are yeast and animal cells, particularly mammalian cells (e.g., CHO cells, NS0 cells, human cells (PER.C6, HEK293, or...). The transfected host cells may be eukaryotic cells, including HKB11 cells, myeloma cells, or human hepatocytes, and plant cells. In some embodiments, the transfected host cells are mammalian cells, such as human cells. In some embodiments, the expression host may glycosylate the antibody of the present invention with a glycan structure that is not immunogenic in itself, particularly in humans. In some embodiments, the transfected host cells may be able to grow in serum-free medium. In further embodiments, the transfected host cells may be able to grow in culture in the absence of animal-derived products. The transfected host cells may be cultured to obtain a cell line.

[0170] The present invention also provides a method for preparing an antibody of interest. The method comprises the steps of culturing or subculturing a transfected host cell population, for example, a stably transfected host cell population, under conditions in which the antibody of interest is expressed, and optionally, purifying the antibody of interest. The transfected host cell population may be prepared by (i) providing a nucleic acid encoding a selected antibody of interest, (ii) inserting the nucleic acid into an expression vector, (iii) transfecting the vector into host cells capable of expressing the antibody of interest, and (iv) culturing or subculturing the transfected host cells containing the inserted nucleic acid for production of the antibody of interest.

[0171] In some embodiments, the antibody according to the present invention may be produced by (i) expressing the nucleic acid sequence according to the present invention in a host cell, for example, by using a vector (or host cell) according to the present invention, and (ii) isolating the expressed antibody product. Furthermore, the method may include (iii) purifying the isolated antibody.

[0172] Therefore, after production, if necessary, the antibodies may be further purified by filtration, centrifugation, and various chromatographic methods (such as HPLC and affinity chromatography). Techniques for purifying antibodies, such as monoclonal antibodies, are known in the art, including techniques for producing pharmaceutical-grade antibodies.

[0173] [Composition] The present invention also provides compositions comprising one or more of the following: (i) The antibody of the present invention or its antigen-binding fragment; (ii) The nucleic acid or multiple nucleic acids of the present invention; (iii) The vector or multiple vectors of the present invention; or, (iv) Cells expressing the antibody according to the present invention, or cells containing the vector according to the present invention.

[0174] The composition may be used for therapeutic or diagnostic purposes. Therefore, the composition may be a pharmaceutical composition or a diagnostic composition. The composition may contain (pharmaceutically acceptable) excipients, diluents, or carriers.

[0175] Accordingly, the present invention also provides pharmaceutical compositions comprising an antibody or its antigen-binding fragment according to the present invention, a nucleic acid or a plurality of nucleic acids according to the present invention, a vector or a plurality of vectors according to the present invention, and / or cells according to the present invention.

[0176] The pharmaceutical composition may optionally include pharmaceutically acceptable carriers, diluents, and / or excipients. While carriers or excipients may facilitate administration, they should not themselves induce the production of antibodies harmful to the individual receiving the composition. They must also not be toxic. Suitable carriers may be large, slowly metabolized polymers such as proteins, polypeptides, liposomes, polysaccharides, polylactic acid, polyglycolic acid, high molecular weight amino acids, amino acid copolymers, and inactive virus particles. In some embodiments, the pharmaceutically acceptable carriers, diluents, and / or excipients in the pharmaceutical composition are not the active ingredients related to birch allergy or related allergy.

[0177] Examples of pharmaceutically acceptable salts that can be used include mineral salts (such as hydrochlorides, hydrobroms, phosphates, and sulfates) or organic salts (such as acetates, propions, malons, and benzoates).

[0178] The pharmaceutically acceptable carrier in the pharmaceutical composition may further include liquids such as water, physiological saline, glycerol, and ethanol. Furthermore, auxiliary substances such as wetting agents, emulsifiers, or pH buffering agents may be present in such compositions. Such carriers allow the pharmaceutical composition to be formulated into tablets, pills, sugar-coated tablets, capsules, liquids, gels, syrups, slurries, suspensions, etc., which can then be administered to the target.

[0179] Pharmaceutical compositions may be prepared in various dosage forms. For example, a composition may be prepared as an injectable liquid solution or suspension. It may also be prepared in a solid form suitable for dissolution or suspension in a liquid vehicle before injection (e.g., lyophilized compositions similar to Synagis® and Herceptin®, which are reconstituted with sterile water containing a preservative). A composition may be prepared for topical administration, for example, as an ointment, cream, or powder. A composition may be prepared for oral administration, for example, as a tablet or capsule, as a spray, or as a (optionally flavored) syrup. A composition may be prepared, for example, as a fine powder or spray for pulmonary administration using an inhaler. A composition may be prepared as a suppository or pessary. A composition may be prepared, for example, as an intravenous infusion for nasal, ear, or eye administration. A composition may also be in kit form, designed so that the combined composition is reconstituted immediately before administration to the subject. For example, lyophilized antibodies may be provided in kit form with sterile water or sterile buffer.

[0180] In some embodiments, the (sole) active ingredient in the composition is an antibody as described herein. In this case, the antibody may be susceptible to degradation in the gastrointestinal tract. Therefore, if the composition is administered via a route using the gastrointestinal tract, the composition may contain a drug that protects the antibody from degradation while releasing the antibody once absorbed from the gastrointestinal tract.

[0181] A thorough discussion of pharmaceutically acceptable carriers can be found in Gennaro (2000) Remington: The Science and Practice of Pharmacy, 20th edition, ISBN: 0683306472.

[0182] The present invention also provides a method for preparing a pharmaceutical composition comprising the following steps: (i) preparing an antibody of the present invention; and (ii) mixing the purified antibody with one or more pharmaceutically acceptable excipients, diluents, or carriers.

[0183] In another embodiment, a method for preparing a pharmaceutical composition includes the step of mixing an antibody with one or more pharmaceutically acceptable carriers, wherein the antibody is a monoclonal antibody.

[0184] The pharmaceutical composition may generally have a pH between 5.5 and 8.5, and in some embodiments, a pH between 6 and 8, for example, about 7. The pH may be maintained by the use of buffering agents. The composition may be sterile and / or pyrogen-free. The composition may be isotonic to humans. In some embodiments, the pharmaceutical composition is supplied in a sealed container.

[0185] The scope of the present invention encompasses compositions that exist in several dosage forms; these forms include, but are not limited to, forms suitable for parenteral administration, such as administration by injection or infusion, such as administration by bolus injection or continuous infusion. When the product is for injection or infusion, the composition may be in the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and may contain formulation agents (such as suspending agents, preservatives, stabilizers, and / or dispersants). Alternatively, the antibody may be in a dry form for reconstitution in a suitable sterile liquid before use.

[0186] A vehicle is typically understood to be a material suitable for storing, transporting, and / or administering pharmaceutically active compounds, particularly compounds such as the antibodies described herein. For example, the vehicle may be a physiologically acceptable liquid suitable for storing, transporting, and / or administering pharmaceutically active compounds, particularly the antibodies described herein. Once formulated, the composition can be administered directly to the subject. In some embodiments, the composition is adapted for administration to mammalian subjects, such as human subjects.

[0187] The pharmaceutical composition may contain an antimicrobial agent, especially if packaged in a multi-dose form. For example, it may contain a cleansing agent such as Tween (polysorbate), such as Tween 80. Cleansing agents are generally present at low levels, for example, less than 0.01%. The composition may also contain a sodium salt (e.g., sodium chloride) to provide a tonic effect. For example, a NaCl concentration of 10 ± 2 mg / ml is typical.

[0188] Furthermore, the pharmaceutical composition may contain, especially when freeze-dried or when it contains materials reconstituted from freeze-dried materials, a sugar alcohol (e.g., mannitol) or disaccharide (e.g., sucrose or trehalose) in an amount of about 15-30 mg / ml (e.g., 25 mg / ml). The pH of the freeze-drying composition may be adjusted to 5-8, or 5.5-7, or around 6.1 before freeze-drying.

[0189] The pharmaceutical composition may be administered by any number of routes, including, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intraperitoneal, intrathecal, intraventricular, transdermal, transcutaneous, topical, subcutaneous, nasal, enteral, sublingual, vaginal, or rectal routes. Optionally, the pharmaceutical composition may be prepared as, for example, tablets or capsules for oral administration, or as, for example, a liquid solution or suspension for topical administration or injection. In some embodiments, the pharmaceutical composition is an injectable preparation. Solid forms suitable for solution or suspension in a liquid vehicle before injection are also included; for example, the pharmaceutical composition may be in lyophilized form.

[0190] For injections, such as intravenous, cutaneous, subcutaneous, or site injections, the active ingredient may be in the form of a parenterally acceptable aqueous solution that is pyrogen-free and has appropriate pH, isotonicity, and stability. Those skilled in the art can prepare a suitable solution using, for example, an isotonic vehicle (such as sodium chloride injection, Ringer's injection, or Ringer's lactate injection). Preservatives, stabilizers, buffers, antioxidants, and / or other additives may be included as needed. Whether administered to an individual is an antibody, peptide, nucleic acid molecule, or other pharmaceutically useful compound, administration is usually carried out in an "effective dose," e.g., a "prophylactically effective dose" or a "therapeutably effective dose" (as applicable), which is sufficient to provide benefit to the individual. Actual dose, administration rate, and administration time depend on the nature and severity of the subject being treated. For injections, the pharmaceutical composition may be supplied, for example, in a pre-filled syringe.

[0191] Pharmaceutical compositions may also be administered orally in any orally acceptable dosage form, including, but not limited to, capsules, tablets, aqueous suspensions, or solutions. For oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. If an aqueous suspension is required for oral administration, the active ingredient, i.e., the antibody as defined above, is used in combination with emulsifiers and suspending agents. Optionally, certain sweeteners, flavorings, or colorings may be added.

[0192] Pharmaceutical compositions may also be administered topically, particularly if the target of treatment includes a contactable area or organ, such as contactable epithelial tissue, where topical administration is easily facilitated. Suitable topical formulations are readily prepared for each of these areas or organs. Pharmaceutical compositions may also be formulated for topical administration in a suitable ointment containing the pharmaceutical composition, particularly the components defined above, suspended or dissolved in one or more carriers. Suitable carriers for topical administration include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying waxes, and water. Alternatively, pharmaceutical compositions may be formulated in a suitable lotion or cream. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0193] The administration treatment may be a single-dose schedule or a multi-dose schedule. In a single-dose administration, for example, daily, weekly, or monthly administration, the amount of antibody in the pharmaceutical composition does not need to exceed 1 g or 500 mg. In some embodiments, in a single-dose administration, the amount of antibody in the pharmaceutical composition does not need to exceed 200 mg or 100 mg. For example, in the case of a single-dose administration, the amount of antibody in the pharmaceutical composition does not need to exceed 50 mg.

[0194] In some embodiments, the composition may contain the antibody of the present invention, which may constitute at least 50% by weight of the total protein in the composition (e.g., 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more). In the composition, the antibody may be in a purified form.

[0195] Instead of delivering antibodies for therapeutic purposes, it is possible to deliver nucleic acids (typically DNA) encoding the target monoclonal antibody to a target, allowing the nucleic acid to be expressed in situ within the target to achieve the desired therapeutic effect. Appropriate gene therapies and nucleic acid delivery vectors are known in the art.

[0196] A pharmaceutical composition typically contains an "effective" amount of one or more antibodies described herein, i.e., an amount sufficient to treat, improve, attenuate, reduce or prevent a desired disease or condition, or to exhibit a detectable therapeutic effect. Therapeutic effects include a reduction or attenuation of the effects of a pathogen or physical symptoms. The exact effective amount for any particular subject depends on the subject's size, weight and health status, the nature and severity of the condition, and the therapeutic agent or combination of therapeutic agents selected for administration. The effective amount for a given situation is determined by routine experimentation and is within the scope of clinician judgment. The effective amount may generally be about 0.005 to about 100 mg / kg, for example, about 0.0075 to about 50 mg / kg, or about 0.01 to about 10 mg / kg. In some embodiments, the effective amount would be about 0.02 to about 5 mg / kg of antibody (e.g., amount of antibody in the pharmaceutical composition) relative to the body weight (e.g., in kg) of the individual being administered.

[0197] Furthermore, the pharmaceutical composition may contain additional active ingredients, which may be additional antibodies or non-antibody components. In other embodiments, the pharmaceutical composition may not contain additional active ingredients (in addition to the antibodies or nucleic acids, vectors, or cells of the present invention as described above).

[0198] Therefore, the pharmaceutical composition may contain one or more additional active ingredients. The antibody of the present invention may be present in the same pharmaceutical composition as the additional active ingredient, or the antibody may be contained in the first pharmaceutical composition and the additional active ingredient may be contained in a second pharmaceutical composition different from the first pharmaceutical composition. Therefore, if multiple additional active ingredients are assumed, each additional active ingredient and antibody may be contained in different pharmaceutical compositions. Such different pharmaceutical compositions may be administered in combination / simultaneously, or may be administered at different times or in different places (e.g., different parts of the body) by any different route of administration.

[0199] Antibodies and additional active ingredients may produce additive therapeutic effects, such as synergistic therapeutic effects. The term "synergistic effect" is used to describe a situation where the combined effect of two or more activators is greater than the sum of the individual effects of each activator. Therefore, when the combined effect of two or more drugs results in "synergistic inhibition" of activity or process, it is intended that the inhibition of activity or process is greater than the sum of the inhibitory effects of each activator. The term "synergistic therapeutic effect" refers to the therapeutic effect observed by a combination of two or more treatments, where the therapeutic effect (measured by several parameters) is greater than the sum of the individual therapeutic effects observed by each individual treatment.

[0200] Preferably, the composition comprises at least two distinct antibodies, or antigen-binding fragments thereof, that bind to distinct non-overlapping epitopes of Bet v1 and optionally to distinct non-overlapping epitopes of the homologous protein of Bet v1. More preferably, the composition comprises at least three distinct antibodies, or antigen-binding fragments thereof, that bind to distinct non-overlapping epitopes of Bet v1 and optionally to distinct non-overlapping epitopes of the homologous protein of Bet v1. As shown in the attached examples, 4E1, 25-3D3, and 17B10 bind to distinct non-overlapping epitopes of Bet v1. Furthermore, cross-competitive ELISA against Bet v1 has shown that antibodies 17B10, 6C9, 3F1, and 13A2 belong to the same complementarity group, and therefore it can be inferred that 17B10, 6C9, 3F1, and 13A2 bind to the same epitope on Bet v1.

[0201] Therefore, the composition may contain at least two antibodies or antigen-binding fragments thereof, and these antibodies or antigen-binding fragments are selected from the group consisting of: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) an antibody or antigen-binding fragment comprising 25-3D3 described above, (a) CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences, and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment described in any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0202] In some embodiments, the composition comprises (exactly) two distinct antibodies, namely: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0203] In some embodiments, the composition comprises (exactly) two distinct antibodies, namely: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0204] In some embodiments, the composition comprises (exactly) two distinct antibodies, namely: (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences of 17B10 as described above, and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0205] Preferably, the composition comprises at least three distinct antibodies or antigen-binding fragments that bind to distinct non-overlapping epitopes of Bet v 1, wherein the antibodies or antigen-binding fragments are selected from the group consisting of: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment as described in any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0206] The present invention also provides a composition comprising at least two distinct antibodies or antigen-binding fragments thereof, wherein the at least two distinct antibodies or antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of a Bet v 1 homologous protein. Herein, the composition comprises at least two distinct antibodies or antigen-binding fragments thereof, and the antibodies or antigen-binding fragments thereof are selected from the group consisting of: (i) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6; (ii) an antibody or antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14; and an antibody or antigen-binding fragment according to any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87.

[0207] Preferably, the composition comprises at least two antibodies or antigen-binding fragments thereof, the antibodies or antigen-binding fragments of which are selected from the group consisting of: (i) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; (ii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 9, CDRH2 as described in SEQ ID NO: 10, CDRH3 as described in SEQ ID NO: 11, CDRL1 as described in SEQ ID NO: 12, CDRL2 as described in SEQ ID NO: 13, and CDRL3 as described in SEQ ID NO: 14; and an antibody or its antigen-binding fragment as described in any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, CDRH3 as described in SEQ ID NO: 19, CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 74, CDRH2 as described in SEQ ID NO: 75, CDRH3 as described in SEQ ID NO: 76, CDRL1 as described in SEQ ID NO: 77, CDRL2 as described in SEQ ID NO: 78, and CDRL3 as described in SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 82, CDRH2 as described in SEQ ID NO: 83, CDRH3 as described in SEQ ID NO: 84, CDRL1 as described in SEQ ID NO: 85, CDRL2 as described in SEQ ID NO: 86, and CDRL3 as described in SEQ ID NO: 87.

[0208] More preferably, the composition comprises at least two antibodies or antigen-binding fragments thereof, wherein the antibodies or antigen-binding fragments are selected from the group consisting of: (i) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; (ii) an antibody or antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 15 and VL having at least 70% identity with SEQ ID NO: 16; and an antibody or antigen-binding fragment according to any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 23, and VL having at least 70% identity with SEQ ID NO: 24; (iv) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; (vi) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89.

[0209] More preferably, the composition comprises at least two antibodies or antigen-binding fragments thereof, wherein the antibodies or antigen-binding fragments are selected from the group consisting of: (i) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 7 and VL as described in SEQ ID NO: 8; (ii) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 15 and VL as described in SEQ ID NO: 16; and an antibody or its antigen-binding fragment as described in any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 23 and VL as described in SEQ ID NO: 24; (iv) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 72 and VL as described in SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 80 and VL as described in SEQ ID NO: 81; (vi) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 88 and VL as described in SEQ ID NO: 89.

[0210] Preferably, the composition comprises at least three antibodies or antigen-binding fragments thereof, the antibodies or antigen-binding fragments of which are selected from the group consisting of: (i) CDRH1 having at least 70% identity with sequence number 1, CDRH2 having at least 70% identity with sequence number 2, CDRH3 having at least 70% identity with sequence number 3, CDRL1 having at least 70% identity with sequence number 4, CDRL2 having at least 70% identity with sequence number 5, and CDRL3 having at least 70% identity with sequence number 6; In particular, CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; In some embodiments, VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; where CDRH1 described in SEQ ID NO: 1, CDRH2 described in SEQ ID NO: 2, CDRH3 described in SEQ ID NO: 3, CDRL1 described in SEQ ID NO: 4, CDRL2 described in SEQ ID NO: 5, and CDRL3 described in SEQ ID NO: 6 are preferably maintained; For example, VH as described in Sequence ID 7, and VL as described in Sequence ID 8, Antibodies or antigen-binding fragments thereof; (ii) CDRH1 having at least 70% identity with sequence number 9, CDRH2 having at least 70% identity with sequence number 10, CDRH3 having at least 70% identity with sequence number 11, CDRL1 having at least 70% identity with sequence number 12, CDRL2 having at least 70% identity with sequence number 13, and CDRL3 having at least 70% identity with sequence number 14; In particular, CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14; In some embodiments, VH has at least 70% identity with SEQ ID NO: 15, and VL has at least 70% identity with SEQ ID NO: 16; where CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14 are preferably maintained; For example, VH as described in Sequence ID No. 15, and VL as described in Sequence ID No. 16, An antibody or its antigen-binding fragment, including (iii) to (vi): (iii) CDRH1 having at least 70% identity with sequence number 17, CDRH2 having at least 70% identity with sequence number 18, CDRH3 having at least 70% identity with sequence number 19, CDRL1 having at least 70% identity with sequence number 20, CDRL2 having at least 70% identity with sequence number 21, and CDRL3 having at least 70% identity with sequence number 22; In particular, CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22; In some embodiments, VH has at least 70% identity with SEQ ID NO: 23, and VL has at least 70% identity with SEQ ID NO: 24; where CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22 are preferably maintained; For example, VH as described in Sequence ID No. 23, and VL as described in Sequence ID No. 24, Antibodies or antigen-binding fragments thereof; (iv) CDRH1 having at least 70% identity with sequence number 66, CDRH2 having at least 70% identity with sequence number 67, CDRH3 having at least 70% identity with sequence number 68, CDRL1 having at least 70% identity with sequence number 69, CDRL2 having at least 70% identity with sequence number 70, and CDRL3 having at least 70% identity with sequence number 71; In particular, CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71; In some embodiments, VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; where CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71 are preferably retained; For example, VH as described in Sequence ID No. 72, and VL as described in Sequence ID No. 73, Antibodies or antigen-binding fragments thereof; (v) CDRH1 having at least 70% identity with sequence number 74, CDRH2 having at least 70% identity with sequence number 75, CDRH3 having at least 70% identity with sequence number 76, CDRL1 having at least 70% identity with sequence number 77, CDRL2 having at least 70% identity with sequence number 78, and CDRL3 having at least 70% identity with sequence number 79; In particular, CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79; In some embodiments, VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; where CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79 are preferably retained; For example, VH as described in Sequence ID No. 80, and VL as described in Sequence ID No. 81, Antibodies or antigen-binding fragments thereof; (vi) CDRH1 having at least 70% identity with sequence number 82, CDRH2 having at least 70% identity with sequence number 83, CDRH3 having at least 70% identity with sequence number 84, CDRL1 having at least 70% identity with sequence number 85, CDRL2 having at least 70% identity with sequence number 86, and CDRL3 having at least 70% identity with sequence number 87; In particular, CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87; In some embodiments, VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89; where CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87 are preferably maintained. For example, VH as described in Sequence ID 88, and VL as described in Sequence ID 89, An antibody or its antigen-binding fragment, including an antibody.

[0211] Preferably, the composition comprises antibodies that bind to three distinct non-overlapping epitopes of Bet v 1. Thus, the composition comprises at least three distinct antibodies or their antigen-binding fragments, preferably comprising the antibody or its antigen-binding fragment as defined above, (i) the antibody or its antigen-binding fragment as defined above, (ii) the antibody or its antigen-binding fragment, and an antibody or its antigen-binding fragment selected from at least one of (iii) to (vi).

[0212] In another preferred embodiment, the composition comprises the antibody or antigen-binding fragment described in (i); the antibody or antigen-binding fragment described in (ii); and an antibody or antigen-binding fragment selected from at least one of (iv) to (vi). As shown in the examples below, the antibodies defined in (iv), (v) and (vi) above, e.g., 6C9, 3F1, and 13A2, belong to the same complementarity group as the antibody defined in (iii) above, e.g., 17B10, and have cross-reactivity to oak antigen and also bind to food-related walnut antigen.

[0213] Therefore, the antibodies included in the composition of the present invention may be selected to target a particularly desired allergen. For example, if targeting oak pollen in addition to its effect against birch pollen, it is advantageous for the composition to include two antibodies that are particularly effective against oak pollen. This can be achieved, for example, by combining the antibody described in (i) above (e.g., 4E1) with an antibody selected from any one of (iv) to (vi) above (e.g., 6C9, 3F1, 13A2).

[0214] As described above, the compositions of the present invention preferably contain antibodies that bind to at least two, preferably three, distinct non-overlapping epitopes of Bet v 1. The antibodies or antigen-binding fragments contained in the composition may be selected from at least two or three different antibodies or variants thereof of 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, as described above, particularly as shown in Tables 1 and 2. Therefore, preferably, the compositions contain two or three (monospecific) antibodies selected from different antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or variants thereof, as described herein and particularly as described above, particularly as shown in Tables 1 and 2. In some embodiments, the compositions contain multispecific antibodies, as described herein.

[0215] The binding of Bet v 1 to three different epitopes may be achieved by a composition comprising: (i) Three distinct monospecific antibodies, or their antigen-binding fragments, that bind to three distinct epitopes of Bet v 1, particularly 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or their variants, as described above, in particular as shown in Tables 1 and 2; (ii) (1) A bispecific antibody or bispecific antigen-binding fragment having two specificities, selected from 4E1, 25-3D3, 17B10, 6C9, 3F1 and 13A2, or their variants, as described above, particularly as shown in Tables 1 and 2, and (2) An additional monospecific antibody conjugating to another epitope on Bet v 1, selected from 4E1, 25-3D3, 17B10, 6C9, 3F1 and 13A2, or their variants, as described above, particularly as shown in Tables 1 and 2; or (iii) As described above, a triplicate antibody or triplicate antigen-binding fragment having three specificities, particularly as shown in Tables 1 and 2, specifically 4E1, 25-3D3, 17B10, 6C9, 3F1 and 13A2, or a selection thereof.

[0216] Alternatively or additionally, the composition may further contain at least one additional agent useful for treating birch pollen allergy and / or related allergies, such as tree pollen allergies (hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy, etc.) and / or related food allergies (apple allergy, hazelnut allergy, and walnut allergy, etc.). The additional agent useful for treating the above allergies may be selected from the group comprising β-adrenergic agonists (e.g., epinephrine), antihistamines, corticosteroids, anti-IgE antibodies, anti-IgE antibody-conjugated fragments, peptide vaccines, and further antibodies capable of binding to birch pollen allergens and / or related (tree pollen and / or food) allergens. In some embodiments, the composition comprises a β-adrenergic agonist such as epinephrine.

[0217] In some embodiments, the composition comprises birch pollen allergen and / or related (tree pollen and / or food) allergens. In some embodiments, the composition comprises an extract comprising birch pollen allergen and / or related (tree pollen and / or food) allergens. The birch pollen allergen may be a birch pollen protein such as Bet v 1. In some embodiments, the composition may be a Bet v 1 homologous protein, such as hazelnut pollen allergen (Cor a 1), alder pollen allergen (Aln g 1), beech pollen allergen (Fag s 1), oak pollen allergen (Que a 1), chestnut pollen allergen (Cas s 1), Japanese hornbeam pollen allergen (Ost c 1), hornbeam pollen allergen (Car b 1), and / or related food allergens, such as apple allergen (Mal d 1), hazelnut allergen (Cor a 1), and / or walnut allergen (Jug r 5), or any other Bet v 1 homologous protein as described above. The birch pollen allergen or related (tree pollen or food) allergen may be naturally derived, recombinantly expressed, or a synthetic peptide. In some embodiments, the anti-Bet v 1 allergen antibody described herein, or its antigen-binding fragment, may be pre-incubated with the birch pollen allergen or related (tree pollen or food) allergen, or administered to the subject as a mixture.

[0218] For example, a combination of birch pollen selected from an extract containing Bet v 1, Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, Mal d 1, and Ju r 5, or a group consisting of mixtures thereof; or the Bet v 1 homologous protein described above; or birch pollen allergens or related (tree pollen or food) allergens as exemplified above, and the antibodies of the present invention enhances the safety of allergen administration (e.g., in desensitization). Furthermore, without being bound by any theory, the inventors assume that the combination of antibodies and their respective allergens (targeted by the antibodies), i.e., the combination of passive (antibody) immunity and active (allergen) immunity, has a synergistic effect. The components, namely the allergen component and the antibody component, may be administered simultaneously, or they may be administered separately at different times, for example, within 15, 30, 60, or 90 minutes.

[0219] The present invention also provides diagnostic compositions comprising antibodies according to the present invention, nucleic acids according to the present invention, vectors according to the present invention, and / or cells according to the present invention. The diagnostic compositions may optionally include appropriate means for detection, such as reagents conventionally used in immunological or nucleic acid-based diagnostic methods.

[0220] The antibodies described herein are suitable, for example, for diagnostic purposes. Therefore, these antibodies may be used in immunoassays, either in liquid phase or conjugated to a solid support. Such immunoassays may be competitive or non-competitive, and may be direct or indirect. Examples of such immunoassays include, but are not limited to, radioimmunoassays (RIA), enzyme-linked immunosorbent assays (ELISA), sandwich immunoassays, immunohistochemistry, flow cytometry, and Western blot assays. For this purpose, the antibodies may be labeled, for example, as described above.

[0221] 〔kit〕 In a further embodiment, the present invention also provides a kit comprising one or more of the following: (i) The antibody or antigen-binding fragment according to the present invention as described above, (ii) The nucleic acid molecule (or multiple nucleic acid molecules) according to the present invention as described above, (iii) The vector (or multiple vectors) according to the present invention as described above, (iv) The cells and / or the cells according to the present invention as described above, (v) The composition according to the present invention as described above.

[0222] Furthermore, the kit may include means for administering the antibody or its antigen-binding fragment according to the present invention, the nucleic acid according to the present invention, the vector according to the present invention, the cell according to the present invention, or the pharmaceutical composition according to the present invention, such as a syringe or container, a leaflet, and / or the co-agent to be administered as described herein. For example, the kit may include a leaflet, for example, an instruction for use. Additionally or alternatively, the kit may include one or more reagents, for example, for use in a suitable diagnostic assay. In some examples, the kit may include a reference agent or control. In some embodiments, the compositions of the present invention may be provided in kit form, for example, designed so that the combined composition is reconstituted immediately before administration to a subject. For example, lyophilized antibodies may be provided in kit form with sterile water or sterile buffer (for example, in a separate container).

[0223] In some embodiments, the kit comprises at least two distinct antibodies (or nucleic acids encoding such antibodies or compositions comprising such antibodies), where the distinct antibodies may be provided in separate containers. Preferably, the kit comprises at least two distinct antibodies or antigen-binding fragments thereof, which antibodies or antigen-binding fragments thereof bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein (or nucleic acids encoding such antibodies or compositions comprising such antibodies). More preferably, the kit comprises at least three distinct antibodies or antigen-binding fragments thereof, which antibodies or antigen-binding fragments thereof bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein (or nucleic acids encoding such antibodies or compositions comprising such antibodies). For this purpose, the antibodies contained in the kit may be selected from the above-mentioned antibodies or variants thereof, for example as shown in Tables 1 and 2. As shown in the appended examples, the antibodies according to the invention may bind to distinct non-overlapping epitopes of Bet v 1.

[0224] Thus, the kit may comprise: (i) at least two distinct antibodies or antigen-binding fragments thereof according to the invention as described above, which bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; (ii) nucleic acid molecules encoding at least two distinct antibodies or antigen-binding fragments thereof according to the invention as described above, which bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; or, (iii) a composition comprising (i) or (ii).

[0225] In the following description, reference is made to kits containing separate antibodies or antigen-binding fragments thereof, although the kits may equally well contain (separate) nucleic acids encoding such antibodies or antigen-binding fragments; or (separate) compositions containing such antibodies or antigen-binding fragments or such nucleic acids.

[0226] In particular, the kit may contain at least two of the following: (i) An antibody or antigen-binding fragment thereof comprising, as described above, for 4E1, (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences, and / or (b) the VH and VL sequences, or variants of those sequences; (ii) An antibody or antigen-binding fragment thereof comprising, as described above, for 25-3D3, (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences, and / or (b) the VH and VL sequences, or variants of those sequences; and an antibody or antigen-binding fragment thereof as described in any one of (iii)-(vi): (iii) An antibody or antigen-binding fragment thereof comprising, as described above, for 17B10, (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences, and / or (b) the VH and VL sequences, or variants of those sequences; (iv) An antibody or antigen-binding fragment thereof comprising, as described above, for 6C9, (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences, and / or (b) the VH and VL sequences, or variants of those sequences; (v) An antibody or antigen-binding fragment thereof comprising, as described above, for 3F1, (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences, and / or (b) the VH and VL sequences, or variants of those sequences; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0227] In some embodiments, the kit may contain (exactly) two distinct antibodies, namely: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0228] In some embodiments, the kit may contain (exactly) two distinct antibodies, namely: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0229] In some embodiments, the kit may contain (exactly) two distinct antibodies, namely: (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0230] Preferably, the kit comprises at least three distinct antibodies or antigen-binding fragments that bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of Bet v 1 homologous proteins; or a nucleic acid encoding such antibodies, or a composition containing such antibodies.

[0231] Therefore, in a preferred embodiment, the kit may contain (exactly) two distinct antibodies, namely: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0232] The present invention also provides a kit comprising at least two distinct antibodies or antigen-binding fragments thereof, wherein the at least two distinct antibodies or antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein. Herein, the composition comprises at least two of the following: (i) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6; (ii) an antibody or antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14; and an antibody or antigen-binding fragment according to any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87.

[0233] Preferably, the kit includes at least two of the following: (i) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; (ii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 9, CDRH2 as described in SEQ ID NO: 10, CDRH3 as described in SEQ ID NO: 11, CDRL1 as described in SEQ ID NO: 12, CDRL2 as described in SEQ ID NO: 13, and CDRL3 as described in SEQ ID NO: 14; and an antibody or its antigen-binding fragment as described in any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, CDRH3 as described in SEQ ID NO: 19, CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 74, CDRH2 as described in SEQ ID NO: 75, CDRH3 as described in SEQ ID NO: 76, CDRL1 as described in SEQ ID NO: 77, CDRL2 as described in SEQ ID NO: 78, and CDRL3 as described in SEQ ID NO: 79; (vi) An antibody or an antigen-binding fragment thereof comprising CDRH1 set forth in SEQ ID NO: 82, CDRH2 set forth in SEQ ID NO: 83, CDRH3 set forth in SEQ ID NO: 84, CDRL1 set forth in SEQ ID NO: 85, CDRL2 set forth in SEQ ID NO: 86, and CDRL3 set forth in SEQ ID NO: 87.

[0234] Preferably, the kit comprises at least two of the following: (i) An antibody or an antigen-binding fragment thereof comprising a VH having at least 70% identity with SEQ ID NO: 7 and a VL having at least 70% identity with SEQ ID NO: 8; (ii) An antibody or an antigen-binding fragment thereof comprising a VH having at least 70% identity with SEQ ID NO: 15 and a VL having at least 70% identity with SEQ ID NO: 16; and an antibody or an antigen-binding fragment thereof as described in any one of (iii) to (vi): (iii) An antibody or an antigen-binding fragment thereof comprising a VH having at least 70% identity with SEQ ID NO: 23 and a VL having at least 70% identity with SEQ ID NO: 24; (iv) An antibody or an antigen-binding fragment thereof comprising a VH having at least 70% identity with SEQ ID NO: 72 and a VL having at least 70% identity with SEQ ID NO: 73; (v) An antibody or an antigen-binding fragment thereof comprising a VH having at least 70% identity with SEQ ID NO: 80 and a VL having at least 70% identity with SEQ ID NO: 81; (vi) An antibody or an antigen-binding fragment thereof comprising a VH having at least 70% identity with SEQ ID NO: 88 and a VL having at least 70% identity with SEQ ID NO: 89.

[0235] More preferably, the kit comprises at least two of the following: (i) An antibody or an antigen-binding fragment thereof comprising the VH set forth in SEQ ID NO: 7 and the VL set forth in SEQ ID NO: 8; (ii) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 15 and VL as described in SEQ ID NO: 16; and an antibody or its antigen-binding fragment as described in any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 23 and VL as described in SEQ ID NO: 24; (iv) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 72 and VL as described in SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 80 and VL as described in SEQ ID NO: 81; (vi) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 88 and VL as described in SEQ ID NO: 89.

[0236] Therefore, the kit preferably includes at least two of the following: (i) CDRH1 having at least 70% identity with sequence number 1, CDRH2 having at least 70% identity with sequence number 2, CDRH3 having at least 70% identity with sequence number 3, CDRL1 having at least 70% identity with sequence number 4, CDRL2 having at least 70% identity with sequence number 5, and CDRL3 having at least 70% identity with sequence number 6; In particular, CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; In some embodiments, VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; where CDRH1 described in SEQ ID NO: 1, CDRH2 described in SEQ ID NO: 2, CDRH3 described in SEQ ID NO: 3, CDRL1 described in SEQ ID NO: 4, CDRL2 described in SEQ ID NO: 5, and CDRL3 described in SEQ ID NO: 6 are preferably maintained; For example, VH as described in Sequence ID 7, and VL as described in Sequence ID 8, Antibodies or antigen-binding fragments thereof; (ii) CDRH1 having at least 70% identity with sequence number 9, CDRH2 having at least 70% identity with sequence number 10, CDRH3 having at least 70% identity with sequence number 11, CDRL1 having at least 70% identity with sequence number 12, CDRL2 having at least 70% identity with sequence number 13, and CDRL3 having at least 70% identity with sequence number 14; In particular, CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14; In some embodiments, VH has at least 70% identity with SEQ ID NO: 15, and VL has at least 70% identity with SEQ ID NO: 16; where CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14 are preferably maintained; For example, VH as described in Sequence ID No. 15, and VL as described in Sequence ID No. 16, An antibody or its antigen-binding fragment, including (iii) to (vi): (iii) CDRH1 having at least 70% identity with sequence number 17, CDRH2 having at least 70% identity with sequence number 18, CDRH3 having at least 70% identity with sequence number 19, CDRL1 having at least 70% identity with sequence number 20, CDRL2 having at least 70% identity with sequence number 21, and CDRL3 having at least 70% identity with sequence number 22; In particular, CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22; In some embodiments, VH has at least 70% identity with SEQ ID NO: 23, and VL has at least 70% identity with SEQ ID NO: 24; where CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22 are preferably maintained; For example, VH as described in Sequence ID No. 23, and VL as described in Sequence ID No. 24, Antibodies or antigen-binding fragments thereof; (iv) CDRH1 having at least 70% identity with sequence number 66, CDRH2 having at least 70% identity with sequence number 67, CDRH3 having at least 70% identity with sequence number 68, CDRL1 having at least 70% identity with sequence number 69, CDRL2 having at least 70% identity with sequence number 70, and CDRL3 having at least 70% identity with sequence number 71; In particular, CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71; In some embodiments, VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; where CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71 are preferably retained; For example, VH as described in Sequence ID No. 72, and VL as described in Sequence ID No. 73, Antibodies or antigen-binding fragments thereof; (v) CDRH1 having at least 70% identity with sequence number 74, CDRH2 having at least 70% identity with sequence number 75, CDRH3 having at least 70% identity with sequence number 76, CDRL1 having at least 70% identity with sequence number 77, CDRL2 having at least 70% identity with sequence number 78, and CDRL3 having at least 70% identity with sequence number 79; In particular, CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79; In some embodiments, VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; where CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79 are preferably retained; For example, VH as described in Sequence ID No. 80, and VL as described in Sequence ID No. 81, Antibodies or antigen-binding fragments thereof; (vi) CDRH1 having at least 70% identity with sequence number 82, CDRH2 having at least 70% identity with sequence number 83, CDRH3 having at least 70% identity with sequence number 84, CDRL1 having at least 70% identity with sequence number 85, CDRL2 having at least 70% identity with sequence number 86, and CDRL3 having at least 70% identity with sequence number 87; In particular, CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87; In some embodiments, VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89; where CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87 are preferably maintained. For example, VH as described in Sequence ID 88, and VL as described in Sequence ID 89, An antibody or its antigen-binding fragment, including an antibody.

[0237] Most preferably, the kit comprises at least three of (i), (ii), and (iii)-(vi) above. In a preferred embodiment, the kit comprises at least three distinct antibodies or antigen-binding fragments thereof, preferably comprising the antibody or antigen-binding fragment thereof as defined above, (i) above, the antibody or antigen-binding fragment thereof as defined above, and at least one of (iii)-(vi) above. In another preferred embodiment, the kit comprises the antibody or antigen-binding fragment thereof as defined above, (i) above, the antibody or antigen-binding fragment thereof as defined above, and at least one of (iv)-(vi) above.

[0238] In particular, the antibodies included in the kit of the present invention may be selected to target a desired allergen. For example, if targeting oak pollen is desired in addition to the effect against birch pollen, it is advantageous for the kit to include two antibodies that are particularly effective against oak pollen. This can be achieved, for example, by combining the antibody described in (i) above (e.g., 4E1) with an antibody selected from any one of (iv) to (vi) above (e.g., 6C9, 3F1, 13A2).

[0239] As described above, the kit of the present invention preferably comprises an antibody that binds to at least two, preferably three, distinct non-overlapping epitopes of Bet v 1, and optionally to a distinct non-overlapping epitope of the Bet v 1 homologous protein. The antibody or antigen-binding fragment contained in the composition may be selected from antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or their variants, as described above, particularly as shown in Tables 1 and 2.

[0240] Therefore, preferably, the kit includes at least two or three (monospecific) antibodies selected from antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or their variants, as described herein and particularly as described above, especially as shown in Tables 1 and 2. In some embodiments, the kit includes multispecific antibodies, as particularly described herein.

[0241] The binding of Bet v 1 to three different epitopes may be achieved by a kit including the following: (i) Three distinct monospecific antibodies, or antigen-binding fragments thereof, that bind to three distinct epitopes of Bet v 1, particularly 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or a selection from their variants, as described above, and optionally bind to distinct non-overlapping epitopes of Bet v 1 homologous proteins; (ii)(1) A bispecific antibody or bispecific antigen-binding fragment having two specificities, selected from 4E1, 25-3D3, 17B10, 6C9, 3F1 and 13A2, or their variants, as described above, particularly as shown in Tables 1 and 2, and (2) An additional monospecific antibody that binds to another epitope on Bet v 1 (optionally on Bet v 1 homologous protein), selected from 4E1, 25-3D3, 17B10, 6C9, 3F1 and 13A2, or their variants, as described above, particularly as shown in Tables 1 and 2; or, (iii) As described above, a triplicate antibody or triplicate antigen-binding fragment having three specificities, particularly as shown in Tables 1 and 2, specifically 4E1, 25-3D3, 17B10, 6C9, 3F1 and 13A2, or a selection thereof.

[0242] Alternatively or additionally, the composition may further contain at least one additional agent useful for treating birch pollen allergy and / or related allergies, such as tree pollen allergies (hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy, etc.) and / or related food allergies (apple allergy, hazelnut allergy, and walnut allergy, etc.). The additional agent useful for treating the above allergies may be selected from the group comprising β-adrenergic agonists (e.g., epinephrine), antihistamines, corticosteroids, anti-IgE antibodies, anti-IgE antibody-conjugated fragments, peptide vaccines, and further antibodies capable of binding to birch pollen allergens and / or related (tree pollen and / or food) allergens. In some embodiments, the composition comprises a β-adrenergic agonist such as epinephrine.

[0243] In some embodiments, the kit includes a birch pollen allergen and / or related allergens. The birch pollen allergen may be a birch pollen protein such as Bet v 1. In some embodiments, the kit may include Bet v 1 homologous tree pollen allergens, such as hazelnut pollen allergen (Cor a 1), alder pollen allergen (Aln g 1), beech pollen allergen (Fag s 1), oak pollen allergen (Que a 1), chestnut pollen allergen (Cas s 1), Japanese hornbeam pollen allergen (Ost c 1), hornbeam pollen allergen (Car b 1), and / or related food allergens, such as apple allergen (Mal d 1), hazelnut allergen (Cor a 1), and walnut allergen (Jug r 5), and / or any other Bet v 1 homologous allergen known in the art or described herein. The birch pollen allergen or related (tree pollen or food) allergen may be of natural origin, recombinantly expressed, or a synthetic peptide. In some embodiments, the antibody or antigen-binding fragment described herein may be provided (in a separate container) with instructions for pre-incubating with the birch pollen allergen and / or related (tree pollen or food) allergen before administering the mixture to the subject. In some embodiments, the antibody or antigen-binding fragment described herein may be provided (in a separate container) with instructions for administering the birch pollen allergen and / or related (tree pollen or food) allergen separately in a combination therapy schedule.

[0244] [Medical procedures and other uses] In a further embodiment, the present invention provides the use of an antibody or its antigen-binding fragment, a nucleic acid molecule (or a plurality of nucleic acid molecules) according to the present invention, a vector (or a plurality of vectors) according to the present invention, a cell according to the present invention, a (pharmaceutical) composition according to the present invention, or a kit according to the present invention as a pharmaceutical.

[0245] In particular, the antibody or antigen-binding fragment thereof according to the present invention, the nucleic acid molecule (or multiple nucleic acid molecules) according to the present invention, the vector (or multiple vectors) according to the present invention, the cell according to the present invention, the (pharmaceutical) composition according to the present invention, or the kit according to the present invention may be used in the prevention and / or treatment of birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy. The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy; or, The antibody, its antigen-binding fragment, the nucleic acid molecule (or multiple nucleic acid molecules), the vector (or multiple vectors), the cells, the (pharmaceutical) composition, or the kit may be used in the prevention and / or treatment of symptoms of birch pollen allergy or related (tree pollen or food) allergy, such as anaphylactic reactions resulting from birch pollen allergy or related (tree pollen or food) allergy.

[0246] Therefore, the present invention also provides a method for treating, improving or alleviating birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy. The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy; or, The present invention provides a method for treating, improving, or alleviating symptoms of birch pollen allergy and / or related allergies, such as anaphylactic reactions caused by birch pollen allergy or related allergies, as described above. The method comprises administering a therapeutically effective dose of an antibody according to the present invention, or its antigen-binding fragment, a nucleic acid molecule (or multiple nucleic acid molecules) according to the present invention, a vector (or multiple vectors) according to the present invention, cells according to the present invention, a (pharmaceutical) composition according to the present invention, or a kit according to the present invention to a target requiring such administration.

[0247] Furthermore, the present invention also provides the use of antibodies or their antigen-binding fragments according to the present invention, nucleic acid molecules (or multiple nucleic acid molecules) according to the present invention, vectors (or multiple vectors) according to the present invention, cells according to the present invention, pharmaceutical compositions according to the present invention, or kits according to the present invention in the manufacture of pharmaceuticals for the prevention, treatment, or reduction of birch pollen allergy and / or related tree pollen allergy and / or related food allergy. The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy; or, The present invention provides, as described herein, the use of an antibody, or its antigen-binding fragment, a nucleic acid molecule (or more nucleic acid molecules), a vector (or more vectors), a cell, a (pharmaceutical) composition, or a kit in the manufacture of a pharmaceutical for the prevention, treatment, or reduction of symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy, such as anaphylactic reactions caused by birch pollen allergy or related (tree pollen and / or food) allergy.

[0248] As used herein, the terms “treat” or “treatment” include therapeutic treatment and prophylactic or preventative measures. Prevention of birch pollen allergy and / or related (tree pollen and / or food) allergy refers in particular to a preventative setting in which the subject has not been diagnosed with birch pollen allergy and / or related (tree pollen and / or food) allergy (no diagnosis was made or the diagnosis was negative), and / or the subject does not exhibit symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy. The prevention of symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy, such as anaphylactic reactions caused by birch pollen allergy and / or related (tree pollen and / or food) allergy, refers in particular to a prophylactic setting when a person has not currently experienced symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy, such as anaphylactic reactions caused by birch pollen allergy and / or related (tree pollen and / or food) allergy, but is likely to experience symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy, such as anaphylactic reactions caused by birch pollen allergy and / or related (tree pollen and / or food) allergy, in the (near) future, due to contact (e.g., via inhalation, ingestion) with an unknown component or a substance known to contain birch pollen allergen and / or related tree pollen allergen and / or related food allergen. In contrast, in a therapeutic setting, the subject is typically diagnosed with and / or related (tree pollen and / or food) allergies as described herein. The terms “treatment” and “therapy” include not only attenuation / reduction of birch pollen allergy (and / or related (tree pollen and / or food) allergies) and / or related symptoms as described herein, but also (complete) cure.

[0249] Generally, the purpose of “treatment” may be to reduce, improve, suppress, prevent, or slow (mitigate or delay) undesirable physiological changes or impairments, such as birch pollen allergy and / or related (tree pollen and / or food) allergy, or symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy, including anaphylactic reactions resulting from birch pollen allergy and / or related (tree pollen and / or food) allergy. Beneficial or desired clinical outcomes of treatment include, but are not limited to, symptom relief, reduction of disease extent, stabilization of disease state (i.e., no exacerbation), delay or slowing of disease progression, improvement or mitigation of disease state, and remission (partial or total), whether detectable or undetectable. “Treatment” may also mean, for example, extending survival compared to the expected survival time without treatment. Patients requiring treatment include not only those who already have a condition or disability, but also those who are prone to developing such a condition or disability, or those for whom the onset or risk of developing such a condition or disability needs to be reduced, delayed, or prevented.

[0250] In some embodiments, the subjects may be human. Such human may be selected from the following groups as described herein: individuals with birch pollen allergy and / or related (tree pollen and / or food) allergy; allergen-sensitized individuals without clinically relevant allergies; individuals with birch pollen allergy and / or related (tree pollen and / or food) allergy who have received immunotherapy; individuals at risk of developing birch pollen allergy and / or related (tree pollen and / or food) allergy; and individuals whose clinical history of birch pollen allergy and / or related (tree pollen and / or food) allergy is unknown.

[0251] Symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy may include, as described herein, one or more of the following: skin rash, itching of the skin, itching or burning in or around the mouth or throat, headache, sneezing, swelling, nausea, diarrhea, or hypersensitivity. Whether a subject has birch pollen allergy and / or related (tree pollen and / or food) allergy may be determined by the correlation of allergic symptoms with contact (e.g., inhalation, ingestion) with the birch pollen allergen or related allergen. In addition or alternatively, allergen skin tests and / or blood tests (for birch pollen allergy IgE) may be performed.

[0252] One method for confirming the effectiveness of a therapeutic treatment involves monitoring disease symptoms after administration of an antibody or composition. The treatment may be a single-dose or multi-dose schedule. In some embodiments, the antibodies, antibody fragments, nucleic acids, vectors, cells, compositions, or kits described herein may be administered to subjects requiring such treatment. Such subjects include, but are not limited to, those at particularly high risk of or susceptible to symptoms of birch pollen allergy and / or related (tree pollen and / or food) allergy, or birch pollen allergy and / or related (tree pollen and / or food) allergy.

[0253] The antibody or its antigen-binding fragment according to the present invention, the nucleic acid molecule (or multiple nucleic acid molecules) according to the present invention, the vector (or multiple vectors) according to the present invention, the cell according to the present invention, the pharmaceutical composition according to the present invention, or the kit according to the present invention may be administered by any route of administration, including but not limited to oral, intravenous, intramuscular, intra-arterial, intraperitoneal, percutaneous, transdermal, topical, subcutaneous, nasal, enteral, sublingual, or rectal routes. In addition, any gene therapy approach may be used, for example, the antibody or its antigen-binding fragment according to the present invention may be administered using, for example, the viral or non-viral vector described above as the nucleic acid or vector encoding the antibody. In some embodiments, the antibody or its antigen-binding fragment according to the present invention, the nucleic acid molecule (or multiple nucleic acid molecules) according to the present invention, the vector (or multiple vectors) according to the present invention, the cell according to the present invention, or the pharmaceutical composition according to the present invention may be administered systemically, for example, by intravenous or subcutaneous administration.

[0254] [Concurrent treatment] In some embodiments, it may be desirable to co-administer or sequentially administer (i) an antibody or antigen-binding fragment according to the present invention, a nucleic acid molecule (or multiple nucleic acid molecules) according to the present invention, a vector (or multiple vectors) according to the present invention, cells according to the present invention, or a pharmaceutical composition according to the present invention, and (ii) a co-agent. In some embodiments, the co-agent may be included in the pharmaceutical composition.

[0255] Preferably, the medical use described above involves administering at least two or three distinct antibodies or antigen-binding fragments that bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; or nucleic acids or vectors encoding such distinct antibodies; or a (combined) administration of a composition containing such distinct antibodies.

[0256] The following description refers to the administration of a separate antibody or its antigen-binding fragment, but it should be understood that the administration may also refer to a (separate) nucleic acid encoding the antibody or antigen-binding fragment; or a (separate) composition containing the antibody or antigen-binding fragment or the nucleic acid.

[0257] Regarding combination therapy, distinct antibodies may be administered in the same composition or in different compositions. Distinct antibodies may be administered via the same route of administration or via separate routes of administration. As used herein, “combined administration” means that distinct antibodies are administered in a combination therapy scheme, i.e., a therapy scheme that includes the administration of distinct antibodies (as opposed to a therapy scheme involving only one antibody followed by a therapy scheme with another (single) antibody). This means that the antibodies may be administered simultaneously (e.g., during the same therapy session) or sequentially on different days.

[0258] Preferably, the above medical use or treatment comprises a (combined) administration of at least two distinct antibodies or their antigen-binding fragments that bind to distinct non-overlapping epitopes of Bet v 1 and / or Bet v 1 homologous protein. More preferably, the above medical use comprises a (combined) administration of at least three distinct antibodies or their antigen-binding fragments that bind to distinct non-overlapping epitopes of Bet v 1 and / or Bet v 1 homologous protein. For this purpose, the antibodies administered in combination may be selected from antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or their variants, as shown above, for example in Tables 1 and 2. As shown in the attached examples, 4E1, 25-3D3, and 17B10 / 6C9 / 3F1 / 13A2 bind to distinct non-overlapping epitopes of Bet v 1.

[0259] Therefore, a medical use or procedure may include at least two (combined) doses: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and / or an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof.

[0260] In some embodiments, the medical use or procedure may involve the administration of two distinct antibodies (in combination), i.e., the two distinct antibodies are: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or sequence variants thereof; and (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or sequence variants thereof.

[0261] In some embodiments, the medical use or procedure may involve the administration of two distinct antibodies (in combination), i.e., the two distinct antibodies are: (i) an antibody or antigen-binding fragment of 4E1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or sequence variants thereof; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof. That is the case.

[0262] In some embodiments, the medical use or procedure may involve the administration of two distinct antibodies (in combination), i.e., the two distinct antibodies are: (ii) an antibody or antigen-binding fragment of 25-3D3 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or antigen-binding fragment of 17B10 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (iv) an antibody or antigen-binding fragment of 6C9 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (v) an antibody or antigen-binding fragment of 3F1 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof; (vi) an antibody or antigen-binding fragment of 13A2 as described above, comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, or variants thereof. That is the case.

[0263] Preferably, the medical use or procedure comprises the administration of a combination of at least three antibodies or antigen-binding fragments, wherein the antibodies or antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein, where the at least three antibodies or antigen-binding fragments are preferably any one selected from (i), (ii), and (iii)-(vi) as defined above.

[0264] The present invention also provides at least three distinct antibodies or antigen-binding fragments for use as pharmaceuticals, preferably for the prevention or treatment of birch pollen allergy or related (tree pollen and / or food) allergy, or symptoms of birch pollen allergy or related (tree pollen and / or food) allergy, each conjugating to a distinct non-overlapping epitope of Bet v 1 and optionally to a distinct non-overlapping epitope of at least one Bet v 1 homologous protein, comprising at least two (combined) doses: (i) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6; (ii) an antibody or antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87.

[0265] Preferably, the medical use / procedure described above includes at least two (combined) doses: (i) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; (ii) an antibody or antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 9, CDRH2 as described in SEQ ID NO: 10, CDRH3 as described in SEQ ID NO: 11, CDRL1 as described in SEQ ID NO: 12, CDRL2 as described in SEQ ID NO: 13, and CDRL3 as described in SEQ ID NO: 14; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, CDRH3 as described in SEQ ID NO: 19, CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 74, CDRH2 as described in SEQ ID NO: 75, CDRH3 as described in SEQ ID NO: 76, CDRL1 as described in SEQ ID NO: 77, CDRL2 as described in SEQ ID NO: 78, and CDRL3 as described in SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 82, CDRH2 as described in SEQ ID NO: 83, CDRH3 as described in SEQ ID NO: 84, CDRL1 as described in SEQ ID NO: 85, CDRL2 as described in SEQ ID NO: 86, and CDRL3 as described in SEQ ID NO: 87.

[0266] More preferably, the medical use / procedure described above includes at least two (combined) doses: (i) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; (ii) an antibody or antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 15 and VL having at least 70% identity with SEQ ID NO: 16; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 23, and VL having at least 70% identity with SEQ ID NO: 24; (iv) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; (vi) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89.

[0267] More preferably, the medical use / procedure described above includes at least two (combined) administrations: (i) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 7 and VL as described in SEQ ID NO: 8; (ii) an antibody or antigen-binding fragment comprising VH as described in SEQ ID NO: 15 and VL as described in SEQ ID NO: 16; and an antibody or antigen-binding fragment selected from any one of (iii) to (vi): (iii) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 23 and VL as described in SEQ ID NO: 24; (iv) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 72 and VL as described in SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 80 and VL as described in SEQ ID NO: 81; (vi) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 88 and VL as described in SEQ ID NO: 89.

[0268] Therefore, in a preferred embodiment, the medical use / treatment described above includes the following (combined) administrations: (i) CDRH1 having at least 70% identity with sequence number 1, CDRH2 having at least 70% identity with sequence number 2, CDRH3 having at least 70% identity with sequence number 3, CDRL1 having at least 70% identity with sequence number 4, CDRL2 having at least 70% identity with sequence number 5, and CDRL3 having at least 70% identity with sequence number 6; In particular, CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; In some embodiments, VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; where CDRH1 described in SEQ ID NO: 1, CDRH2 described in SEQ ID NO: 2, CDRH3 described in SEQ ID NO: 3, CDRL1 described in SEQ ID NO: 4, CDRL2 described in SEQ ID NO: 5, and CDRL3 described in SEQ ID NO: 6 are preferably maintained; For example, VH as described in Sequence ID 7, and VL as described in Sequence ID 8, An antibody or its antigen-binding fragment, including; and (ii) CDRH1 having at least 70% identity with sequence number 9, CDRH2 having at least 70% identity with sequence number 10, CDRH3 having at least 70% identity with sequence number 11, CDRL1 having at least 70% identity with sequence number 12, CDRL2 having at least 70% identity with sequence number 13, and CDRL3 having at least 70% identity with sequence number 14; In particular, CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14; In some embodiments, VH has at least 70% identity with SEQ ID NO: 15, and VL has at least 70% identity with SEQ ID NO: 16; where CDRH1 described in SEQ ID NO: 9, CDRH2 described in SEQ ID NO: 10, CDRH3 described in SEQ ID NO: 11, CDRL1 described in SEQ ID NO: 12, CDRL2 described in SEQ ID NO: 13, and CDRL3 described in SEQ ID NO: 14 are preferably maintained; For example, VH as described in Sequence ID No. 15, and VL as described in Sequence ID No. 16, An antibody or its antigen-binding fragment containing (iii) to (vi); and / or an antibody or its antigen-binding fragment selected from any one of (iii) to (vi): (iii) CDRH1 having at least 70% identity with sequence number 17, CDRH2 having at least 70% identity with sequence number 18, CDRH3 having at least 70% identity with sequence number 19, CDRL1 having at least 70% identity with sequence number 20, CDRL2 having at least 70% identity with sequence number 21, and CDRL3 having at least 70% identity with sequence number 22; In particular, CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22; In some embodiments, VH has at least 70% identity with SEQ ID NO: 23, and VL has at least 70% identity with SEQ ID NO: 24; where CDRH1 described in SEQ ID NO: 17, CDRH2 described in SEQ ID NO: 18, CDRH3 described in SEQ ID NO: 19, CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22 are preferably maintained; For example, VH as described in Sequence ID No. 23, and VL as described in Sequence ID No. 24, Antibodies or antigen-binding fragments thereof; (iv) CDRH1 having at least 70% identity with sequence number 66, CDRH2 having at least 70% identity with sequence number 67, CDRH3 having at least 70% identity with sequence number 68, CDRL1 having at least 70% identity with sequence number 69, CDRL2 having at least 70% identity with sequence number 70, and CDRL3 having at least 70% identity with sequence number 71; In particular, CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71; In some embodiments, VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; where CDRH1 described in SEQ ID NO: 66, CDRH2 described in SEQ ID NO: 67, CDRH3 described in SEQ ID NO: 68, CDRL1 described in SEQ ID NO: 69, CDRL2 described in SEQ ID NO: 70, and CDRL3 described in SEQ ID NO: 71 are preferably retained; For example, VH as described in Sequence ID No. 72, and VL as described in Sequence ID No. 73, Antibodies or antigen-binding fragments thereof; (v) CDRH1 having at least 70% identity with sequence number 74, CDRH2 having at least 70% identity with sequence number 75, CDRH3 having at least 70% identity with sequence number 76, CDRL1 having at least 70% identity with sequence number 77, CDRL2 having at least 70% identity with sequence number 78, and CDRL3 having at least 70% identity with sequence number 79; In particular, CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79; In some embodiments, VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; where CDRH1 described in SEQ ID NO: 74, CDRH2 described in SEQ ID NO: 75, CDRH3 described in SEQ ID NO: 76, CDRL1 described in SEQ ID NO: 77, CDRL2 described in SEQ ID NO: 78, and CDRL3 described in SEQ ID NO: 79 are preferably retained; For example, VH as described in Sequence ID No. 80, and VL as described in Sequence ID No. 81, Antibodies or antigen-binding fragments thereof; (vi) CDRH1 having at least 70% identity with sequence number 82, CDRH2 having at least 70% identity with sequence number 83, CDRH3 having at least 70% identity with sequence number 84, CDRL1 having at least 70% identity with sequence number 85, CDRL2 having at least 70% identity with sequence number 86, and CDRL3 having at least 70% identity with sequence number 87; In particular, CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87; In some embodiments, VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89; where CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87 are preferably maintained. For example, VH as described in Sequence ID 88, and VL as described in Sequence ID 89, An antibody or its antigen-binding fragment, including an antibody.

[0269] Preferably, the medical use or treatment described above comprises an administration of an antibody or an antigen-binding fragment (combined), wherein the antibody or antigen-binding fragment binds to a distinct non-overlapping epitope of Bet v 1 and optionally to three distinct non-overlapping epitopes of at least one Bet v 1 homologous protein. Thus, in a preferred embodiment, the medical use / treatment comprises an administration of three distinct antibodies or antigen-binding fragments (combined), wherein the first antibody or antigen-binding fragment is according to (i), the second antibody or antigen-binding fragment is according to (ii), and the third antibody or antigen-binding fragment is selected from the antibodies or antigen-binding fragments described in any one of (iii) to (vi) above.

[0270] As described above, the medical applications / therapies of the present invention preferably involve the administration of an antibody (combined), wherein the antibody binds to at least two, preferably three, distinct non-overlapping epitopes of Bet v 1 and optionally to at least two, preferably three, distinct non-overlapping epitopes of at least one Bet v 1 homologous protein. The antibody or antigen-binding fragment administered (combined) may be selected from the antibodies 4E1, 25-3D3, 17B10, 6C9, 3F1, and 13A2, or their variants, as described above. Preferably, two or three monospecific antibodies described herein may be administered (combined). In some embodiments, multispecific antibodies, particularly those described herein, may also be administered.

[0271] Alternatively or additionally, at least one additional agent useful for treating birch pollen allergy and / or related (tree pollen and / or food) allergies may be administered (in combination) with the antibodies or antigen-binding fragments described herein. The additional agent useful for treating birch pollen allergy or related (tree pollen and / or food) allergies may be selected from the group comprising β-adrenergic agonists (e.g., epinephrine), antihistamines, corticosteroids, anti-IgE antibodies, anti-IgE antibody-binding fragments, peptide vaccines, and further antibodies capable of binding to birch pollen allergens and / or related (tree pollen and / or food) allergens. In some embodiments, the additional agent is a β-adrenergic agonist such as epinephrine.

[0272] In some embodiments, the birch pollen allergen and / or related (tree pollen and / or food) allergen may be administered (in combination) with the antibody or antigen-binding fragment described herein. The birch pollen allergen and / or related allergen may be Bet v 1 birch pollen allergen and / or related tree pollen allergen and / or related food allergen. The relevant tree pollen allergen is preferably selected from at least one of the following: Cor a 1 hazelnut pollen allergen, Aln g 1 European alder pollen allergen, Fag s 1 beech pollen allergen, Que a 1 oak pollen allergen, Cas s 1 chestnut pollen allergen, Ost c 1 Japanese hornbeam pollen allergen, and Car b 1 hornbeam pollen allergen. The related food allergen is preferably selected from at least one of Mal d 1 apple allergen, Cor a 1 hazelnut allergen, and Jug r 5 walnut allergen. The birch pollen allergen and / or related (tree pollen and / or food) allergen may be of natural origin, recombinantly expressed, or synthetic peptide.

[0273] In some embodiments, the anti-bet v 1 allergen antibody or its antigen-binding fragment described herein may be pre-incubated with birch pollen allergen or related (tree pollen and / or food) allergen, or administered to a subject as a mixture.

[0274] For example, a combination of a birch pollen allergen or related (tree pollen or food) allergen, selected from the group consisting of Bet v 1, Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, Mal d 1, and Jug r 5, or mixtures thereof, with the antibody of the present invention enhances the safety of allergen administration (e.g., in desensitization). Furthermore, without being bound by any theory, the inventors assume that the combination of an antibody and each allergen (targeted by the antibody), i.e., the combination of passive (antibody) immunity and active (allergen) immunity, has a synergistic effect.

[0275] [Further applications] The antibodies and fragments described herein may also be used for the (in-vitro) diagnosis of birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy. The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy.

[0276] The diagnostic method may include contacting an antibody with a sample. Such a sample may be isolated from a subject and may be, for example, an isolated blood sample (whole blood, plasma, or serum). The diagnostic method may also include detection of an antigen / antibody complex, particularly detection after contact between the antibody and the sample. Furthermore, the sample may be tested for whether it contains antibodies that compete with the antibodies described herein, for example, for allergen binding. This is typically performed in vitro, i.e., without contact with the human or animal body. Examples of analytical methods are well known to those skilled in the art, and examples of immunoassays include flow cytometry, dot blot or slot blot, Western blot, ELISA (enzyme-linked immunosorbent assay, e.g., cross-competition), immunohistochemistry, and SDS-PAGE immunocytochemistry following immunoprecipitation. Thus, the diagnosis may be performed in vitro, for example, by using an isolated sample as described above (and the in vitro analytical steps described above).

[0277] The present invention also provides a method for detecting whether a sample contains birch pollen allergens, particularly Bet v 1 or Bet v 1 homologous allergens (such as Cor a 1 hazelnut pollen allergen, Aln g 1 alder pollen allergen, Fag s 1 beech pollen allergen, Que a 1 oak pollen allergen, Cas s 1 chestnut pollen allergen, Ost c 1 Japanese hornbeam pollen allergen, Car b 1 hornbeam pollen allergen, Mal d 1 apple allergen, and Jug r 5 walnut allergen). Such a method may include the following steps: a. The step of contacting a sample with the antibody described herein under conditions acceptable for generating an antibody / antigen complex; and, b. A step to detect the presence of an antibody / antigen complex.

[0278] This may indicate that the presence of detectable antibody / antigen complexes may indicate that the sample contains birch pollen allergens, particularly Bet v 1 and / or Bet v 1 homologous allergens (such as Cor a 1 hazelnut pollen allergen, Aln g 1 alder pollen allergen, Fag s 1 beech pollen allergen, Que a 1 oak pollen allergen, Cas s 1 chestnut pollen allergen, Ost c 1 Japanese hornbeam pollen allergen, Car b 1 hornbeam pollen allergen, Mal d 1 apple allergen, and Jug r 5 walnut allergen).

[0279] In some embodiments, in step (b), the amount of antigen / antibody complex in the test sample may be determined (e.g., measured). This amount may then be compared to a control.

[0280] Therefore, the antibody or antigen-binding fragment of the present invention is, as described above, effective against birch pollen allergens, particularly Bet v 1 and / or Bet v 1 homologous allergens. It may be used in an in-vitro method for detecting the antigen. Similarly, the antibody or antigen-binding fragment of the present invention may be used in an in-vitro method for binding birch pollen allergen, particularly Bet v 1 and / or Bet v 1 homologous allergen, as described above. As described above, the antibody may be brought into contact with an (isolated) sample (i.e., a sample to be tested for the presence of the antigen) in order to detect birch pollen allergen, particularly Bet v 1 and / or Bet v 1 homologous allergen. The antibody and its antigen (as described above, birch pollen allergen, particularly Bet v 1 and / or Bet v 1 homologous allergen) specifically bind to each other, forming an antibody / antigen complex, which can be readily detected by methods known in the art.

[0281] Such detection methods may be used to test (e.g., generate / manufacture) samples such as food, cosmetics, or drug samples. Therefore, the antibodies, antibody fragments, or variants described in the present invention may also be used in non-therapeutic / non-diagnostic contexts, such as in the development or manufacture of various products.

[0282] Furthermore, this detection method may be used to test whether a vaccine sample (e.g., one intended for desensitization) contains Bet v 1 and / or Bet v 1 homologous allergens, which are major birch pollen allergens. This may be useful in the development and / or manufacture of such vaccines. Accordingly, the present invention also provides the use of the antibodies or antigen-binding fragments described herein for testing immunogenic compositions / vaccines, in particular immunogenic compositions / vaccines containing birch pollen allergens (especially Bet v 1 and / or Bet v 1 homologous allergens). For this purpose, the antibodies may be contacted with the immunogenic composition / vaccine, for example, to test for antibody / antigen complexes. Accordingly, the present invention also provides a method for testing an immunogenic composition (vaccine) based on birch pollen allergens (especially Bet v 1 and / or Bet v 1 homologous allergens), where the immunogenic composition / vaccine is contacted with an antibody or its antigen-binding fragment, and optionally, the presence of an antibody / antigen complex is determined. Furthermore, the present invention also encompasses the use of the antibodies or antigen-binding fragments of the present invention for monitoring the quality of immunogenic compositions / vaccines based on birch pollen allergens (particularly Bet v 1 and / or Bet v 1 homologous allergens) by checking whether the immunogenic composition / vaccine contains a desired antigen (e.g., Bet v 1, Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, Mal d 1, and Jug d 5, etc.).

[0283] [Brief explanation of the drawing] The attached drawings will be briefly described below. The drawings are intended to illustrate the present invention in more detail. However, these drawings are not intended to limit the subject matter of the present invention in any way.

[0284] [Figure 1] Figure 1 shows the BLI sensorograms for the binding of MY010 antibodies 4E1, 25-3D3, and 17B10 to Bet v 1 in Example 1. mAb1 was immobilized as shown in the figure, and Bet v 1, mAb2, and mAb3 were injected sequentially as shown in the figure.

[0285] [Figure 2] Figures 2A to 2G show the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared to the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). These affinities were measured by competitive ELISA. Figure 2G is a heat map summarizing the affinities of the antibodies of the present invention and the prior art antibodies.

[0286] [Figure 3] Figures 3A to 3C show the affinity of the MY010 antibodies 25-3D3, 17B10, and 4E1 of the present invention to Bet v 1 and Bet v 1 homologous allergens (hazelnut Cor a 1, alder Aln g 1, beech Fag s 1, oak Que a 1, chestnut Cas s 1, Japanese hornbeam Ost c 1, and hornbeam Car b 1) (Figure 3A) in Example 3, compared to the affinity of the prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (Figure 3B). These affinities were measured by conjugation ELISA. Figure 3C is a heat map summarizing the affinities of the antibodies of the present invention and the prior art antibodies.

[0287] [Figure 4] Figures 4A to 4G show the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jug r 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). These affinities were measured by soluble competitive ELISA. Figure 4G is a heat map summarizing the affinities of the antibodies of the present invention and the comparative antibodies.

[0288] [Figure 5] Figures 5A and 5B show the suppression of mast cell reactivity by "cocktails" containing one (4E1), two (4E1+25-3D3; 4E1+17B10), or three (4E1+17B10+25-3D3) MY010 antibodies of the present invention in Example 5, compared to "cocktails" containing prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (REGN-3). Bone marrow-derived mast cells isolated from transgenic mice expressing human FcERIa were sensitized with a plasma pool derived from birch pollen allergy patients (n=6). Mast cell activation was quantified as the percentage of CD107a-positive cells. Titrated antibody cocktails (80 nM to 0 nM) were pre-incubated with fixed-concentration extracts. The cells were sensitized in a plasma pool overnight at 37°C, and then stimulated with pre-incubated extracts at 37°C for 35 minutes.

[0289] [Figure 6] Figures 6A and 6B show the suppression of mast cell reactivity by the "MY010 cocktail" of the present invention, which contains three antibodies of the present invention (4E1+17B10+25-3D3), in Example 6, compared to the "REGN cocktail" containing prior art antibodies REGN-5713, REGN-5714, and REGN-5715. The titrated antigen / extract was pre-incubated with a fixed concentration (80 nM) of antibody.

[0290] [Figure 7] Figure 7 shows the prevention of basophil degranulation in whole blood samples from patients induced by birch extract (10 ng / ml Bet v 1) using the "MY010 cocktail" (4E1+17B10+25-3D3) of the present invention in Example 7, compared to the prior art REGN cocktail (REGN-5713+REGN-5714+REGN-5715). These preventions were quantified by the percentage of CD63 expression. The bar graph shows basophil activation as the percentage of CD63+ cells (mean ± sd, n=2). White bars: no antibody added. Data from one representative patient. Statistics were performed by one-way Anova and subsequent Dunnett tests comparing each row to 10 ng / ml birch extract.

[0291] [Figure 8] Figure 8 shows the effects of the "MY010 cocktail" on birch-related food allergens (apple Mal d 1, hazelnut Cor a 1, walnut Jugr 5, and peach extract) in Example 8. These effects were measured by a mast cell activation test.

[0292] [Figure 9] Figure 9 shows the competition among the MY010 antibody complementation groups against Bet v 1 in Example 9. This competition was measured by cross-competition ELISA.

[0293] [Figure 10] Figure 10 shows the BLI sensorograms for the binding of MY010 antibodies 4E1, 25-3D3, and 3F1 to Bet v 1 in Example 10. mAb1 was immobilized as shown in the figure, and Bet v 1, mAb2, and mAb3 were injected sequentially in the order shown in the figure.

[0294] [Figure 11] Figures 11A to 11D show the affinity of the MY010 antibodies of the present invention, 6C9 (Figure 11A), 3F1 (Figure 11B), and 13A2 (Figure 11C), to birch extract and birch-related extracts (hazelnut pollen extract, European alder extract, American beech extract, European beech extract, and oak extract) in Example 11. These affinities were measured by soluble competitive ELISA. Figure 11D is a heat map summarizing the affinity of the antibody of the present invention, MY010-17B10.

[0295] [Figure 12] Figures 12A and 12B show the affinity of the MY010 antibodies 6C9, 3F1, and 13A2 of the present invention to Bet v 1 and Bet v 1 homologous allergens (Alder Aln g 1, Hazel Cor a 1.0101 and Cor a 1.0201, Hornbeam recombinant Car b 1 1A (rec Car b 1 1A) and recombinant Car b 1 (rec Car b 1) isoform 2, Japanese ash Ost c 1, Oak Que a 1.0201 and Que a 1.0301, Beech Fag s 1, and Chestnut Cas s 1) (Figure 12A) in Example 12. These affinities were measured by conjugated ELISA. Figure 12B is a heat map summarizing the affinities of the antibodies of the present invention and prior art antibodies.

[0296] [Figure 13] Figures 13A to 13C show the affinity of the MY010 antibodies 6C9 (Figure 13A), 3F1 (Figure 13B), and 13A2 (Figure 13C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1.0108, hazelnut Cor a 1.0401, and walnut Jug r 5.0101) in Example 13. These affinities were measured by soluble competitive ELISA.

[0297] [Figure 14] Figures 14A and 14B show the suppression of mast cell reactivity by different antibody "cocktails" as shown in Example 14. These inhibitory effects were measured by the mast cell activation test (MAT). Figure 14A: Plasma pool from 6 US patients stimulated with birch extract, alder extract, hazel pollen extract, beech extract, and oak extract; Figure 14B: Plasma pool from US, UK, and Swiss patients stimulated with oak extract.

[0298] [Figure 15] Figures 15A to 15D show the effect of the MY010-3 cocktail containing the antibody of the present invention, as shown, in a passive cutaneous anaphylaxis (PCA) model in Example 15, compared to the prior art REGN-3 cocktail. Figure 15A: Experimental setup; Figure 15B: Mice challenged intravenously with birch pollen extract (0.1 μg); Figure 15C: Mice challenged intravenously with hazelnut pollen extract (1 μg); Figure 15D: Mice challenged intravenously with oak pollen extract (1 μg).

[0299] [Figure 16] Figures 16A and 16B show the binding sites of MY010 and the prior art REGN antibody to Bet v 1, as identified by epitope mapping in Example 16. Figure 16A: Scanning mutation introduced into surface-exposed residues of Bet v 1. Figure 16B: Schematic diagram of the Bet v 1 epitope targeted by MY010 and REGN antibodies.

[0300] [Examples] The following are specific examples illustrating various embodiments and aspects of the present invention. However, the present invention is not limited in scope by the specific embodiments described herein. The following constructions and examples are given to enable those skilled in the art to better understand and implement the present invention. However, the present invention is not limited in scope by the exemplary embodiments, and these embodiments are intended only as examples of a single aspect of the present invention, and functionally equivalent methods are within the scope of the present invention. In fact, in addition to those described herein, various modifications of the present invention will be readily apparent to those skilled in the art from the foregoing description, the accompanying figures, and the following examples. All such modifications fall within the scope of the appended claims.

[0301] [Example 1: Antibodies 4E1, 25-3D3, and 17B10 bind to distinct, non-overlapping epitopes on Bet v 1] (1. Antibody cloning) Human lymphocytes were collected from the peripheral blood of allergy patients who voluntarily participated in the study and used as starting material for cloning fully human antibodies. Antibodies specific to major birch pollen-related allergens were isolated by molecular cloning of immunoglobulin genes obtained from single-cell sorted cells derived from short-term oligoclonal cultures of activated memory B cells producing antibodies of interest. Molecular cloning of human antibodies specific to birch pollen-related allergens was performed according to Huang J, Doria-Rose NA, Longo NS, Laub L, Lin CL, Turk E, Kang BH, Migueles SA, Bailer RT, Mascola JR, Connors M. Isolation of human monoclonal antibodies from peripheral blood B cells. Nat Protoc. 2013 Oct;8(10): 1907-15. doi: 10.1038 / nprot.2013.n117, as described in detail in WO 2018 / 234383 A1.

[0302] The CDR and VH / VL sequences (sequence numbers) of the fully human antibodies 4E1, 25-3D3, and 17B10 are shown in Table 4 below.

[0303] [Table 7]

[0304] (2. Antibody expression and purification) Efficient and scalable antibody expression was achieved using suspension culture HEK293 cells, with a method capable of handling expression scales ranging from 20 to 1000 mL. HEK293 cells were transfected with DNA encoding both the light and heavy chains of human IgG4, and polyethyleneimine (PEI Max, Polysciences, USA). To obtain optimal IgG production, transfected cells were supplemented with tryptone N1 (OrganoTechni, France) 24 hours after transfection. After a 5-7 day expression period, the supernatant containing recombinant antibody was collected.

[0305] The purification process used a HiTrap Protein A HP column (Cytiva,US). Protein A, derived from Staphylococcus, exhibits strong affinity for the Fc region of IgG. Binding of protein A to IgG was achieved under neutral pH conditions, after which the antibody was eluted using an acidic pH solution. The purified antibody was dialyzed in PBS, concentrated, and stored at -80°C for long-term storage.

[0306] (3. Biolayer Interferometry) To determine whether antibodies 4E1, 25-3D3, and 17B10 can simultaneously bind to epitopes on Bet v 1, biolayer interferometry (BLI; Forte Bio Octet RED96e; running buffer: 1xPBS) was performed. IgG4 from the mAb cocktail was used. First, mAbs1 (5 μg / ml) was immobilized on the surface of a human Fc-binding BLI sensor, and then blocked with human Fc fragment (50 μg / ml). Next, natural Bet v 1 (1.8 μg / ml) was added, and mAbs2 and mAbs3 (15 μg / ml each) were sequentially injected as shown in the table in Figure 1.

[0307] The sensorogram shown in Figure 1 strongly suggests that all IgGs can simultaneously bind to a single Bet v 1 molecule. Therefore, the BLI data confirmed that all tested IgG antibodies, 4E1, 25-3D3, and 17B10, can simultaneously and non-competitively bind to a single Bet v 1 molecule. Thus, these data indicate that the 4E1, 25-3D3, and 17B10 antibodies bind to distinct, non-overlapping epitopes on Bet v 1.

[0308] [Example 2: Antibodies 4E1, 25-3D3, and 17B10 bind to PR-10 allergen extract with higher affinity and exhibit broader cross-reactivity compared to conventional antibodies REGN-5713, REGN-5714, and REGN-5715.] (1. Preparation of comparative antibodies REGN-5713, REGN-5714, and REGN-5715) The amino acid and DNA sequences of REGN-5713, REGN-5714, and REGN-5715 are derived from patent US10793624B2. Each V region was cloned into an expression vector containing either the coding sequence for human IgG4 (S228P), the coding sequence for VH, or the coding sequence for the human κ constant region or VL sequence. Recombinant antibodies were transiently produced in HEK293 suspension cells and purified by a protein A affinity column as described in Examples 1 and 2. Antibody expression and purification were performed. Epitope specificity of the REGN antibody was confirmed using alanine scanning mutagenesis of Bet v 1 (data not shown).

[0309] (2. Soluble competitive ELISA) The cross-reactivity of MY010 antibodies 4E1, 25-3D3, and 17B10, as well as prior art antibodies REGN-5713, REGN-5714, and REGN-5715, with PR-10 related extracts (birch, hazel, alder, beech, and oak) was evaluated by soluble competitive ELISA.

[0310] Therefore, after coating the plates with streptavidin (1 μg / ml) overnight at 4°C or for 1 hour at 37°C, biotinylated natBet v 1 (1.4 nM) was captured overnight at 4°C. Titrated human MY010-mAbs (4E1, 25-3D3, 17B10) (80 nM~0 nM) and REGN antibodies (REGN-5713, REGN-5714, and REGN-5715) were pre-incubated overnight at 4°C or for 1 hour at room temperature with dilution buffer (no extract = 0 nM) or each extract (14 nM), respectively. Pre-mixed MY010-mAbs / extract and REGN-mAbs / extract were incubated on ELISA plates at room temperature for 15 minutes. The binding of MY010 antibodies (4E1, 25-3D3, 17B10) and REGN antibodies (REGN-5713, REGN-5714, REGN-5715) to immobilized natBet v 1 was quantified using anti-hu-IgG-HRP.

[0311] The results shown in Figures 2A (mAb 4E1), 2B (mAb 25-3D3), and 2C (mAb 17B10) explain that MY010 antibodies 4E1, 25-3D3, and 17B10 bind with high affinity to PR-10 allergen extracts of birch, hazel, alder, beech, and oak, and therefore exhibit broad cross-reactivity to a wide range of tree pollen allergens.

[0312] On the other hand, as shown in Figures 2D to 2F, the prior art mAbs REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F) exhibit low cross-reactivity with birch-related PR-10 allergen extracts, particularly hazelnut, alder, beech, and oak extracts. Therefore, Figures 2A to 2F explain that MY010 antibodies 4E1, 25-3D3, and 17B10 show superior cross-reactivity with multiple birch-related tree pollen extracts compared to the prior art antibodies REGN-5713, REGN-5714, and REGN-5715.

[0313] The results are summarized in the heatmap in Figure 2G, where ΔLogEC50 is plotted: ΔLogEC 50 =LogEC 50 (14nM)-LogEC 50 (0nM).

[0314] [Example 3: Antibodies 4E1, 25-3D3, and 17B10 show high cross-reactivity to recombinant PR-10 proteins derived from different tree pollens.] In this experiment, multiple Bet v 1-related (PR-10) allergens were synthesized and used to define the affinity of MY010 antibodies (4E1, 25-3D3, and 17B10) and REGN antibodies (REGN-5713, REGN-5714, REGN-5715) for birch-related recombinant proteins. These birch-related recombinant proteins are: These are Bet v 1.0101 (birch), Cor a 1.0101 and Cor a 1.0201 (hazel), Aln g 1.0101 (alder), Fag s 1.0101 (beech), Que a 1.0201 (oak), Cas s 1.0101 (chestnut), Ost c 1.0101 (Japanese hornbeam), Car b1.1A and Car b1.2 (hornbeam).

[0315] (1. Expression of recombinant PR-10 protein) PR-10 allergen was synthesized and cloned into the pET21 vector (Twist Biosciences). This vector was used to transform the BL21-derived Rosetta (DE3) strain (Novagen). After inoculating the transformed Rosetta colonies into LB medium, the culture was incubated overnight at 37°C to prepare a stationary phase culture. The following morning, a large-scale LB expression culture was prepared using the stationary phase culture, and this expression culture was incubated with shaking at 37°C until the OD600nm reached 0.7. Subsequently, it was induced with 0.5 mM IPTG and incubated at 37°C for 4 hours. The culture was then centrifuged, and the pellet was frozen at -80°C.

[0316] For the purification of the PR-10 allergen, bacterial pellets were lysed in BugBuster® HT Protein Extraction Reagent (Sigma) supplemented with an EDTA-free protease inhibitor cocktail (Roche) for 1 hour at room temperature. The cell lysate was centrifuged (spinned) to remove debris, and the supernatant was collected and filtered. The histidine-tagged PR-10 protein was purified using TALON® Superflow® resin (Cytiva) in binding buffer (50 mM sodium phosphate, 300 mM NaCl, pH 7.4) according to the manufacturer's recommendations. The protein was then dialyzed overnight in 1x PBS at 4°C.

[0317] (2. Cross-reactivity to PR-10 related proteins by direct binding ELISA) ELISA plates were coated with anti-human FcIgG antibody overnight at 4°C, and then MY010-mAbs (4E1, 25-3D3, 17B10) and REGN-mAbs (REGN-5713, REGN-5714, REGN-5715) were captured at room temperature for 1 hour (2 μg / mL). Titrated histidine-tagged recombinant proteins (3 μg / mL~0) were incubated at room temperature for 1 hour, and binding was evaluated by measurement using anti-histidine-tagged HRP.

[0318] The binding affinity (EC50) to the tested PR-10 allergens is shown in Table 5 below.

[0319] [Table 8]

[0320] Figure 3A shows the binding affinities of 4E1, 25-3D3, and 17B10 to various allergens, and Figure 3B shows the binding affinities of REGN-5713, REGN-5714, and REGN-5715. The results are summarized in the heatmap in Figure 3C, where EC50 (ng / ml) is plotted.

[0321] Therefore, Figures 3A to 3C and Table 5 illustrate that the MY010 antibodies 4E1, 25-3D3, and 17B10 exhibit superior cross-reactivity against multiple recombinant PR-10 proteins derived from different tree pollens compared to the prior art antibodies REGN-5713, REGN-5714, and REGN-5715.

[0322] [Example 4: Antibodies 4E1, 25-3D3, and 17B10 show high cross-reactivity to food-related PR-10 allergens.] (1. Cross-reactivity to food-related PR-10 allergens by direct binding ELISA) The cross-reactivity of MY010 antibodies 4E1, 25-3D3, and 17B10, as well as prior art antibodies REGN-5713, REGN-5714, and REGN-5715, against PR-10-related food allergens (apple Mal d 1.0108, hazelnut Cor a 1.0401, and walnut Jug r 5.0101) was evaluated by soluble competitive ELISA.

[0323] Therefore, after coating the plates with streptavidin (1 μg / ml) overnight at 4°C or for 1 hour at 37°C, biotinylated na tBet v 1 (1.1 nM) was captured overnight at 4°C. Titrated human MY010-mAbs (0-80 nM) and REGN-mAbs were pre-incubated overnight at 4°C or for 1 hour at room temperature with dilution buffer or each PR-10-related dietary protein (Mal d 1, Cor a 1, Ju r 5; 14 nm and 140 nM), respectively. The pre-mixed mAbs / recombinant (rec.) proteins were incubated on ELISA plates at room temperature for 15 minutes. Binding of MY010 antibody to immobilized nat Bet v 1 was quantified using anti-Hu-IgG-HRP. Sources of PR-10 protein: nat Bet v 1 (Indoor Biotechnologies), recombinant Mal d 1.0108 (rec Mal d 1.0108) (Biomay), recombinant Cor a 1.0401 (rec Cor a 1.0401) (Indoor Biotechnologies), recombinant Jug r 5.0101 (rec Jug r 5.0101) (Indoor Biotechnologies).

[0324] The results shown in Figure 4A explain that MY010 antibody 4E1 binds to the food-related PR-10 allergens of apple (Mal d 1), hazelnut (Cor a 1), and walnut (Jur r 5), and therefore exhibits high cross-reactivity to major food-related PR-10 allergens. In addition, MY010 antibody 25-3D3 shows high affinity for hazelnut Cor a 1 (Figure 4B).

[0325] On the other hand, as shown in FIGS. 4D to 4F, among the prior art antibodies REGN-5713, REGN-5714, and REGN-5715, only REGN-5715 shows cross-reactivity with food-related PR-10 allergens (hazelnut Cor a 1). Therefore, the cross-reactivity with food-related PR-10 allergens, particularly apple Mal d 1, hazelnut Cor a 1, and walnut Jug r 5, is lower compared to the MY010 antibodies (4E1 and 25-3D3) of the present invention.

[0326] The results are summarized in the heat map of FIG. 4G, where ΔLogEC50 is plotted: ΔLogEC 50 =LogEC 50 (14 nM) - LogEC 50 (0 nM).

[0327] Therefore, FIGS. 4A to 4G illustrate that the MY010 antibodies 4E1 and 25-3D3 exhibit superior cross-reactivity with multiple food-related PR-10 proteins compared to the prior art antibodies REGN-5713, REGN-5714, and REGN-5715.

[0328] [Example 5: Suppression of reactivity of mast cells by different MY010 antibody cocktails] Next, "cocktails" containing one type (4E1), two types (4E1 + 25-3D3; 4E1 + 17B10), or three types (4E1 + 17B10 + 25-3D3) of MY010 antibodies were examined to determine whether they could suppress allergen-mediated activation of bone marrow-derived mast cells isolated from transgenic mice.

[0329] (1. Comparison by mast cell activation test of antibody effects) Bone marrow-derived mast cells isolated from transgenic mice expressing human FcERIa were sensitized with a plasma pool derived from patients with birch pollen allergy. Mast cell activation was quantified as the percentage of CD107a-positive cells. After sensitizing the cells in the plasma pool overnight at 37°C, they were stimulated for 35 minutes at 37°C with appropriate allergens alone or with allergens pre-incubated with antibodies.

[0330] The titrated antibody (starting at 320 nM) was pre-incubated with the fixed concentration extract. The fixed concentrations of the allergens were as follows: The concentrations are as follows: birch extract: 7.41 ng / mL, hazelnut pollen extract: 22 ng / mL, alder extract: 66.67 ng / mL, oak extract: 13.3 ng / mL, beech (Beach) extract: 13.3 ng / mL, chestnut: 10.2 ng / mL, Japanese hornbeam: 200 ng / mL, and hornbeam: 67 ng / mL.

[0331] The results shown in Figures 5A and 5B demonstrate that "cocktails" containing one, two, or three types of MY010 antibodies effectively suppressed mast cell reactivity in all birch-related extracts tested. Generally, cocktails containing two types of MY010 antibodies (4E1+25-3D3 or 4E1+17B10) performed better than cocktails containing only one type of MY010 antibody (4E1), while cocktails containing three types of antibodies (4E1+17B10+25-3D3) achieved the best cross-reactivity. The results are summarized in Table 6 below.

[0332] [Table 9]

[0333] [Example 6: The MY010 cocktail is effective with all birch-related extracts tested.] Next, the ability of the MY010 cocktail, containing three antibodies (4E1+17B10+25-3D3), to suppress allergen-mediated activation of bone marrow-derived mast cells isolated from transgenic mice was compared with that of a conventional technology cocktail containing three conventional technology antibodies (REGN13-15).

[0334] (1. Comparison of antibody effects using mast cell activation tests) Bone marrow-derived mast cells isolated from transgenic mice expressing human FcERIa were sensitized with a plasma pool derived from patients with birch pollen allergy. Mast cell activation was quantified as the percentage of CD107a-positive cells. After sensitizing the cells in the plasma pool overnight at 37°C, they were stimulated for 35 minutes at 37°C with appropriate allergens alone or with allergens pre-incubated with antibodies.

[0335] The titrated allergen / extract was pre-incubated with a fixed concentration (80 nM) antibody.

[0336] The results shown in Figures 6A and 6B demonstrate that the MY010 cocktail containing the antibody 4E1+17B10+25-3D3 of the present invention is effective against all birch-related extracts / allergens tested and is superior to the prior art REGN cocktail.

[0337] [Example 7: A cocktail of antibodies 4E1, 17B10, and 25-3D3 inhibits allergen-mediated activation of basophils derived from allergy patients.] Next, we investigated whether a cocktail of antibodies 4E1, 17B10, and 25-3D3 could inhibit allergen-mediated activation of basophils derived from allergy patients.

[0338] For this purpose, a whole blood basophil activation assay was performed. Briefly, heparinized whole blood from donors with birch pollen allergy was stimulated with birch extract at 37°C for 30 minutes, either untreated or in the absence or presence of a cocktail of antibodies 4E1, 17B10, and 25-3D3 (80 nM) (pre-incubation of birch extract / antibody was performed at room temperature for at least 1 hour). Cells were stained with CCR3-PE and CD63-FITC before erythrocyte lysis. Basophils were gated as SSClow, CCR3high lymphocytes, and activation was quantified using FACS Calibur (Becton Dickinson AG, Allschwil, Switzerland) as the percentage of CD63+, CCR3high cells. Flow cytometry data were analyzed using FlowJo software (TreeStar, Ashland, Ore).

[0339] The results are shown in Figure 7. The bar graph shows basophil activation as the percentage of CD63+ cells (mean ± sd, n=2). White bars: no antibody added. Data from one representative patient. Statistical analysis was performed using the One-way Anova test, comparing each row to 10 ng / ml birch extract, followed by Dunnett's test.

[0340] In summary, these data indicate that pre-incubation of birch extract with the MY010 antibody cocktail reduces basophil degranulation, and that the MY010 cocktail (4E1, 17B10, and 25-3D3) is superior to conventional REGN cocktails (REGN-5713, REGN-5714, REGN-5715) in inhibiting allergen-mediated activation of basophils from allergy patients.

[0341] [Example 8: A cocktail of antibodies 4E1, 17B10, and 25-3D3 is effective against birch-related food allergens.] The effects of the MY010 antibody cocktail (4E1, 17B10, and 25-3D3) on birch-related food allergens were verified by mast cell activation testing.

[0342] Bone marrow-derived mast cells isolated from transgenic mice expressing human FcERIa were sensitized with a plasma pool derived from patients with birch pollen allergy. Mast cell activation was quantified as the percentage of CD107a-positive cells or as mediator release (sLts, Buehlmann CAST ELISA). Cells were sensitized overnight in a plasma pool at 37°C, and then stimulated for 35 minutes at 37°C with appropriate allergens alone or with allergens pre-incubated with antibodies.

[0343] Titrated allergens / extracts (apple, hazelnut, walnut, peach) were pre-incubated with a fixed concentration (240 nM) of antibody.

[0344] The results shown in Figure 8 demonstrate that cocktails containing MY010 antibodies 4E1, 17B10, and 25-3D3 effectively suppressed mast cell reactivity with all birch-related food allergens / extracts tested. In each case, cocktails containing MY010 antibodies (4E1, 17B10, and 25-3D3) were superior to cocktails containing REGN antibodies (REGN-5713, REGN-5714, and REGN-5715).

[0345] [Example 9: Antibodies 6C9, 3F1, and 13A2 belong to the same clonal type, compete with each other, and also compete with antibody 17B10.] Antibodies 6C9, 3F1, and 13A2 were cloned and expressed as described in Example 1 (see 1. Antibody Cloning; and 2. Antibody Expression and Purification).

[0346] The CDR and VH / VL sequences (sequence numbers) of the fully human antibodies 6C9, 3F1, and 13A2 are shown in Table 7 below.

[0347] [Table 10]

[0348] To investigate whether antibodies 6C9, 3F1, and 13A2 compete with antibodies 4E1, 25-3D3, and 17B10 (MY010 antibody), cross-competitive ELISA assays were performed against Bet v 1. For this purpose, plates were coated with MY010-IgG antibody (all 0.75 ug / mL) overnight at 4°C. MY010 Fab (160 nM) was pre-incubated with histidine-tagged recombinant Bet v 1 (recBet v 1-his-tag) (0.05 ug / ml = 2.38 nM) at room temperature for at least 1 hour. Subsequently, the pre-mixed antibody-allergen was incubated on ELISA plates (2 hours, room temperature). Measurements were performed using anti-histidine-HRP. The table in Figure 9 plots the relative inhibition rate (%) at the highest antibody concentration. The inhibition rate (%) is given by 100 - (OD450Ab / OD450 w / o Ab * 100). Here, OD450Ab is the OD450 value at each point on the titration curve, and OD450 w / o Ab is the average value of the OD450 values ​​obtained without competing antibodies. The relative inhibition rate was calculated by normalizing based on the maximum inhibition value obtained when the coated antibody competed with itself. The results are summarized in Figure 9.

[0349] Figure 9 shows that antibodies MY010-3F1, MY010-6C9, and MY010-13A2 compete with each other and with MY010-17B10. Based on this and the sequence similarity of the VH and VL regions, it can be concluded that these antibodies belong to the same clone type. Furthermore, this clone type belongs to the same complementarity group as MY010-17B10. On the other hand, they do not compete with antibodies from other complementarity groups (25-3D3 and 4E1).

[0350] [Example 10: Antibodies 4E1, 25-3D3, and 3F1 bind to distinct, non-overlapping epitopes on Bet v 1] To investigate whether antibodies 4E1, 25-3D3, and 3F1, representative of the 6C9 clone group, can simultaneously bind to epitopes on Bet v 1, biolayer interferometry (BLI; Forte Bio Octet RED96e; running buffer: 1xPBS) was performed. IgG4 from the mAbs cocktail was used. First, mAbs1 (5 μg / ml) was immobilized on the surface of a human Fc-binding BLI sensor, and then blocked with human Fc fragment (50 μg / ml). Next, natural Bet v 1 (1.8 μg / ml) was added, and mAbs2 and mAbs3 (15 μg / ml each) were sequentially injected as shown in the table in Figure 10.

[0351] The sensorogram shown in Figure 10 strongly suggests that all IgGs can simultaneously bind to a single Bet v 1 molecule. Therefore, BLI data confirmed that all tested IgG antibodies (4E1, 25-3D3, and 3F1, representing the 6C9 clonal group) bound simultaneously and non-competitively to a single Bet v 1 molecule. Thus, these data indicate that the 4E1, 25-3D3, and 3F1 antibodies bind to distinct, non-overlapping epitopes on Bet v 1.

[0352] [Example 11: Antibodies 6C9, 3F1, and 13A2 bind to PR-10 allergen extract with high affinity.] The cross-reactivity of MY010 antibodies 6C9, 3F1, and 13A2, as well as MY010-17B10, which belong to the same complementarity group, with PR-10-related extracts (birch, hazel, alder, beech, and oak) was evaluated by soluble competitive ELISA.

[0353] Therefore, after coating the plates with streptavidin (1 μg / ml) overnight at 4°C or for 1 hour at 37°C, biotinylated natBet v 1 (1.1 nM) was captured overnight at 4°C. Titrated human MY010-mAbs (6C9, 3F1, 13A2, 17B10) (80 nM to 0 nM) were pre-incubated overnight at 4°C or for 1 hour at room temperature with dilution buffer (no extract = 0 nM) or extract (14 nM). The pre-mixed MY010-mAbs / extract was incubated on an ELISA plate at room temperature for 15 minutes. Detection was performed using anti-hu-IgG-HRP.

[0354] The results shown in Figures 11A (mAb 6C9), 11B (mAb 3F1), and 11C (mAb 13A2) explain that MY010 antibodies 6C9, 3F1, and 13A2 bind with high affinity to PR-10 allergen extracts from birch, hazel, beech, and oak, and therefore exhibit broad cross-reactivity to a wide range of tree pollen allergens.

[0355] On the other hand, as shown in Figures 2D to 2F, the prior art mAbs REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F) exhibit low cross-reactivity with birch-related PR-10 allergen extracts, particularly hazelnut, beech, and oak extracts. Therefore, Figures 11A to 11C explain that MY010 antibodies 6C9, 3F1, and 13A2 show superior cross-reactivity with multiple birch-related tree pollen extracts compared to the prior art antibodies REGN-5713, REGN-5714, and REGN-5715.

[0356] Furthermore, a comparison with Figure 2C shows that the 6C9, 3F1, and 13A2 antibodies exhibit superior cross-reactivity with oak compared to antibody 17B10, which belongs to the same complementarity group.

[0357] The results are summarized in the heatmap in Figure 11D, where ΔLogEC50 is plotted: ΔLogEC 50 =LogEC50 (14nM)-LogEC 50 (0nM).

[0358] [Example 12: Antibodies 6C9, 3F1, and 13A2 show high cross-reactivity with birch-associated recombinant PR-10 protein.] In this experiment, the affinity of 6C9 cloned MY010 antibodies (6C9, 3F1, and 13A2) to birch-related recombinant proteins was defined by direct-conjugation ELISA using several Bet v 1-related (PR-10) allergens synthesized according to Example 3. The birch-related recombinant proteins in question are: Bet v 1.0101 (birch); Cor a 1.0101 and Cor a 1.0201 (hazel); Aln g 1.0101 (alder); Fag s 1.0101 (beech); Que a 1.0201 and Que a 1.0301 (oak); Cas s 1.0101 (chestnut); Car b 1.1A and Car b 1.2 (hornbeam); and Ost c 1.0101 (Japanese hornbeam).

[0359] ELISA plates were coated with anti-human FcIgG antibody overnight at 4°C, and then MY010-mAbs (6C9, 3F1, and 13A2) were captured at room temperature for 1 hour (2 μg / mL). Titrated histidine-tagged recombinant proteins (3 μg / mL~0) were incubated at room temperature for 1 hour, and binding was evaluated by measurement using anti-histidine-tagged fusion HRP.

[0360] The binding affinity (EC50) to the tested PR-10 allergens is shown in Table 8 below.

[0361] [Table 11]

[0362] Figure 12A shows the binding affinities of 6C9, 3F1, and 13A2 to various allergens. The results are summarized in the heatmap in Figure 12B, with EC50 (ng / ml) plotted.

[0363] Therefore, Figures 12A and 12B, and Table 8, explain that MY010 antibodies 6C9, 3F1, and 13A2 exhibit superior cross-reactivity to multiple recombinant PR-10 proteins derived from different tree pollens compared to the prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (see Figures 3A and 3C). Furthermore, MY010 antibodies 6C9, 3F1, and 13A2 have higher binding affinity compared to antibody MY010-17B10, which belongs to the same complementarity group (see Figures 3A and 12B). Of MY010 antibodies 6C9, 3F1, and 13A2, MY010-6C9 is the antibody with the highest cross-reactivity.

[0364] [Example 13: Antibodies 6C9, 3F1, and 13A2 show cross-reactivity to food-related walnut allergens.] (1. Cross-reactivity to food-related PR-10 allergens by direct binding ELISA) The cross-reactivity of MY010 antibodies 6C9, 3F1, and 13A2 to PR-10-related food allergens (such as apple Mal d 1.0108, hazelnut Cor a 1.0401, and walnut Jug r 5.0101) was evaluated by soluble competitive ELISA.

[0365] Therefore, after coating the plates with streptavidin (1 μg / ml) overnight at 4°C or for 1 hour at 37°C, biotinylated na tBet v 1 (1.4 nM) was captured overnight at 4°C. Titrated human MY010-mAbs (0-80 nM) were pre-incubated overnight at 4°C or for 1 hour at room temperature with dilution buffer or each recombinant PR-10-related dietary protein (14 nm and 140 nM). The pre-mixed mAbs / recombinant (rec.) proteins were incubated on ELISA plates at room temperature for 15 minutes. Binding of MY010 antibody to immobilized nat Bet v 1 was quantified using anti-Hu-IgG-HRP.

[0366] As shown in Figures 13A-13C, MY010 antibodies 6C9, 3F1, and 13A2 have affinity for the food-related walnut allergen Jur r 5 and therefore exhibit superior cross-reactivity compared to antibody 17B10 (see Figure 4C), which belongs to the same complementarity group, and also compared to prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (see Figures 4D-4F).

[0367] [Example 14: Suppression of mast cell reactivity by different MY010 antibody cocktails] Next, we investigated whether different "cocktails" containing three different MY010 antibodies (17B10+4E1+25-3D3;6C9+4E1+25-3D3;3F1+4E1+25-3D3) could suppress allergen-mediated activation of bone marrow-derived mast cells isolated from transgenic mice.

[0368] (1. Comparison of antibody effects using mast cell activation tests) Bone marrow-derived mast cells isolated from transgenic mice expressing human FcERIa were sensitized with plasma pools from patients with birch pollen allergy. These patients were from the United States (USn) (n=6 patients), the United Kingdom (n=5 patients), and Switzerland (n=10 patients). Mast cell activation was quantified as the percentage of CD107a-positive cells or mediator release (sLts, Buehlmann CAST ELISA). Cells were sensitized overnight in plasma pools at 37°C, and then stimulated for 35 minutes at 37°C with appropriate allergens alone or with allergens pre-incubated with antibodies. Titrated allergens / extracts (birch, alder, hazel, oak, beech, hornbeam, and hornbeam) were pre-incubated with fixed concentrations of antibodies.

[0369] The results shown in Figure 14A demonstrate that cocktails containing three types of MY010 antibodies effectively suppress the reactivity of mast cells in all birch-related extracts tested. In particular, cocktails containing MY010 antibody 6C9 or 3F1 (in addition to MY010 antibodies 4E1 and 25-3D3) show comparable efficacy to cocktails containing MY010 antibody 17B10 (in addition to MY010 antibodies 4E1 and 25-3D3), which belongs to the same complementarity group as 6C9 and 3F1. Compared to prior art REGN antibody cocktails, the MY010 cocktails of the present invention are more effective in suppressing all birch-related allergens tested.

[0370] Furthermore, the results shown in Figure 14B demonstrate that the MY010 antibody cocktail of the present invention exhibits superior efficacy against oak compared to the prior art REGN antibody cocktail. In particular, antibody cocktails containing antibody 6C9 or 3F1, respectively, are more effective in suppressing the reactivity of mast cells in oak extract compared to cocktails containing 17B10, which belongs to the same complementarity group. These experiments support the results of Example 11 shown in Figures 11A and 11B.

[0371] [Example 15: Effects of the MY010-3 cocktail on birch, hazelnut, and oak pollen extracts in a mouse model of passive cutaneous anaphylaxis (PCA)] To investigate the effects of the MY010 cocktail, an in vivo model of passive cutaneous anaphylaxis was established. First, the MY010 cocktail, containing antibodies 17B10, 25-3D3, and 4E1, was titrated against birch extract. This allowed huFceRI transgenic mice (IgE / FceRI humanized mouse strain, Genoway) to be subcutaneously administered different doses (1, 0.1, and 0.01 mg / kg) of the MY010 antibody cocktail, or 1 mg / kg of isotype control. Three days later, the ears of the transgenic mice were intradermally (id) sensitized by injection of human plasma pool. 20 μl of plasma pool from a birch allergy patient was injected into the right ear, and plasma pool from a healthy individual (internal experimental control) was used for the left ear. After 24 hours, the animals were challenged by iv injection (0.1 μg) of birch extract with Evans blue dye (0.5%). Thirty minutes after the challenge, mice were sacrificed and their ears were collected to measure ear thickness. The ears were then placed in formamide solution for dye extraction and quantification. Local allergic reactions in the sensitized ears were monitored by the amount of Evans blue staining exudate. The amount of exuded Evans blue was measured by light absorption at 620 nm and normalized by the weight of the collected ear tissue (Evans blue ng / mg ear tissue).

[0372] In the second experimental set, the effects of the MY010 cocktail and the REGN cocktail on different PR-10 related extracts were compared. As previously described, huFcERI transgenic mice were subcutaneously administered 0.1 or 1 mg / kg of the MY010 cocktail, Regeneron cocktail, or isotype control. Three days later, the mice were intradermally (id) sensitized using plasma from a healthy donor (left ear) or a patient with birch allergy. Twenty-four hours after intradermally (id) sensitization, the mice were ivally challenged with birch pollen-related extracts (0.1 μg birch extract, 1 μg alder extract, 1 μg hazelnut pollen extract, 1 μg oak extract) + Evans blue dye (0.5%). Thirty minutes after the challenge, the mice were sacrificed and the ears were collected to measure ear thickness. The dyes were quantified as previously described. A schematic PCA model is shown in Figure 15A.

[0373] The results shown in Figure 15B demonstrate that the MY010-3 cocktail, containing antibodies 17B10, 25-3D3, and 4E1 according to the present invention, and the REGN-3 cocktail, containing prior art antibodies REGN5713, REGN4715, and REGN5716, have equivalent effects on birch pollen extract.

[0374] However, as shown in Figure 15C, the MY010-3 cocktail containing antibodies 17B10, 25-3D3, and 4E1 according to the present invention has superior efficacy against hazelnut pollen extract compared to the conventional REGN cocktail.

[0375] Furthermore, as shown in Figure 15D, the MY010-3 cocktail containing antibodies 17B10, 25-3D3, and 4E1, and the MY010-3_6C9 cocktail containing antibodies 6C9, 25-3D3, and 4E1, exhibit superior efficacy against oak pollen extract compared to the conventional REGN cocktail.

[0376] Therefore, the superiority of the antibody composition of the present invention against birch-related pollen extract is clearly confirmed in the in vivo PCA mouse model.

[0377] [Example 16: Epitope Mapping] (1. Production and purification of the Bet v 1 mutant library) All surface-exposed Bet v 1 residues were mutated according to the following scheme: D / E to A and K; K / R to A and E; A to S and K; Q / N / H / I / L / M / F / S / T / W / Y / V to A.

[0378] The histidine-tagged and Avi-tagged Bet v 1 mutant was biotinylated in vivo in double-transformed BL21DE3 cells by co-expression of the mutant and the biotinylating enzyme BirA. Expression was performed in a 96-well plate format. Expression was initiated by inoculating 600 μl of preculture medium (LB medium supplemented with kanamycin (25 μg / ml), carbenicillin (25 μg / ml), and glucose (0.5%)) taken from glycerol stock or colonies into each well of a 96-deep-well plate and incubating overnight at 37°C. The following day, 1 ml of expression medium (LB medium supplemented with kanamycin (25 μg / ml), carbenicillin (25 μg / ml), IPTG (1 mM), and biotin (100 μg / ml)) was added to each well, and the culture medium was incubated at room temperature for 24 hours. After expression, cells were collected by centrifugation, and the supernatant was discarded. Subsequently, recombinant Bet v 1 mutants were purified via histidine tagging. To lyse the cells, 100 μl of BugBuster reagent (Novagen®) was added to each well and incubated at room temperature for 20 minutes. Then, 50 μl of binding buffer (phosphate (50 mM), NaCl (300 mM), pH 7.4) was added to each well, and the plate was centrifuged at maximum speed for 20 minutes. The supernatant was transferred to a 96-well plate containing magnetic AmMag® Ni beads (GenScript) prepared according to the manufacturer's instructions. After standing at room temperature for 30 minutes, the beads were collected using a magnetic separation rack, and the supernatant was discarded. The beads were washed three times by adding 250 μl of washing buffer (phosphate (50 mM), imidazole (10 mM), NaCl (300 mM), pH 7.4) to each well. After thorough mixing, the beads were collected using a magnetic rack, and the supernatant was discarded. Finally, 65 μl of elution buffer (phosphate (50 mM), imidazole (150 mM), NaCl (300 mM), pH 7.4) was added, and the mixture was incubated at room temperature for 5 minutes, stirring occasionally. The beads were collected using a magnetic rack, and the eluted proteins were transferred to a clean 96-well plate.

[0379] (2. Testing of the Bet v 1 mutant library for epitope mapping)ELISA was performed to investigate the effect of individual residues in the Bet v 1 mutant library on the binding of MY010 antibodies (4E1, 25-3D3, 17B10, "6C9 clone"). Each library mutant was tested against all MY010 cocktail antibodies, and two of the three antibodies were used as controls for binding and mutant integrity. High-binding 96-well plates (Greiner 675061) were coated with streptavidin (Sigma, S4762-5MG, 1 μg / ml in PBS, 30 μl / well) for 1 hour at 37°C. The plates were then washed once with washing buffer (0.05% Tween 20 in PBS, 150 μl / well). The plates were incubated in blocking buffer (2% BSA in PBS, 100 μl / well) for 1 hour at room temperature, and the blocking solution was discarded. Biotinylated Bet v 1 mutants were captured by adding a 1:150 diluted mutant (30 μl / well) in 0.5% BSA in PBS at 37°C for 1 hour. The plates were then washed four times with washing buffer (150 μl / well). Recombinant Fab of MY010 antibody was added at a concentration of 100 ng / ml (30 μl / well) and incubated at room temperature for 2 hours. The plates were then washed four times with washing buffer (150 μl / well). Subsequently, 30 μl / well of mouse anti-human λ light chain HRP (Abcam JDC-12, 1:1000 dilution) or goat anti-human κ light chain HRP (Invitrogen, 1:5000 dilution) was added and incubated at room temperature for 1 hour. The plates were then washed four times with washing buffer (150 μl / well). To induce color development in the assay, 30 μl of TMB substrate (Sigma T2885, diluted 1:20 with 30 mM citrate (pH 4.1)) was added to each well and incubated for exactly 5 minutes. Color development was stopped by adding 15 μl of H2SO4 (AppliChem, A2699, 1M) to each well. Subsequently, the absorbance of the plate at 450 nm was measured without a reference wavelength using a Multiskan FC ELISA reader (Thermo Scientific). For each Bet v 1 mutant, the binding of all three types of Fab of MY010 was analyzed.If a decrease in binding was observed in one of the three Fab variants of the MY010 antibody compared to the others, the mutant residue of this Bet v 1 variant was designated as part of the binding epitope.

[0380] Figure 16A shows the surface-exposed residues of Bet v 1 identified by scanning mutation introduction. The total number of mutants was 92.

[0381] The results of epitope mapping using scanning mutations are summarized in Table 9 below.

[0382] [Table 12] JPEG2026522294000014.jpg5169

[0383] As shown in Table 9, the MY010 antibodies 6C9, 17B10, 4E1, and 25-3D3 bind to non-overlapping epitopes on Bet v 1. Furthermore, it should be noted that the epitopes to which the MY010 antibodies of the present invention bind are distinct from the epitopes to which the prior art REGN antibodies bind.

[0384] The results described above are schematically shown in Figure 16B.

[0385] [Table 13] JPEG2026522294000016.jpg254169JPEG2026522294000017.jpg241169JPEG2026522294000018.jpg255167JPEG2026522294000019.jpg255169 JPEG2026522294000020.jpg255169JPEG2026522294000021.jpg234169JPEG2026522294000022.jpg255163JPEG2026522294000023.jpg147169 [Brief explanation of the drawing]

[0386] [Figure 1] Figure 1 shows the BLI sensorograms for the binding of MY010 antibodies 4E1, 25-3D3, and 17B10 to Bet v 1 in Example 1. [Figure 2A] Figure 2A shows the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared to the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). [Figure 2B] Figure 2B shows the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared to the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). [Figure 2C] Figure 2C shows the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared to the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). [Figure 2D] Figure 2D shows the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared to the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). [Figure 2E]Figure 2E shows the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared to the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). [Figure 2F] Figure 2F shows the affinity of the MY010 antibodies 4E1 (Figure 2A), 25-3D3 (Figure 2B), and 17B10 (Figure 2C) of the present invention to birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, and oak extract) in Example 2, compared with the affinity of the prior art antibodies REGN-5713 (Figure 2D), REGN-5714 (Figure 2E), and REGN-5715 (Figure 2F). [Figure 2G] Figure 2G is a heat map summarizing the affinity between the antibody of the present invention and the prior art antibody. [Figure 3A] Figure 3A shows the affinity of the MY010 antibodies 25-3D3, 17B10, and 4E1 of the present invention to Bet v 1 and Bet v 1 homologous allergens (hazelnut Cor a 1, alder Aln g 1, beech Fag s 1, oak Que a 1, chestnut Cas s 1, Japanese hornbeam Ost c 1, and hornbeam Car b 1) (Figure 3A) in Example 3, compared to the affinity of the prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (Figure 3B). [Figure 3B] Figure 3B shows the affinity of the MY010 antibodies 25-3D3, 17B10, and 4E1 of the present invention for Bet v 1 and Bet v 1 homologous allergens (hazelnut Cor a 1, alder Aln g 1, beech Fag s 1, oak Que a 1, chestnut Cas s 1, Japanese hornbeam Ost c 1, and hornbeam Car b 1) (Figure 3A) in Example 3, compared to the affinity of the prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (Figure 3B). [Figure 3C]Figure 3C shows a heat map summarizing the affinity between the antibody of the present invention and the prior art antibody. [Figure 4A] Figure 4A shows the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). [Figure 4B] Figure 4B shows the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). [Figure 4C] Figure 4C shows the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). [Figure 4D] Figure 4D shows the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). [Figure 4E]Figure 4E shows the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). [Figure 4F] Figure 4F shows the affinity of the MY010 antibodies 4E1 (Figure 4A), 25-3D3 (Figure 4B), and 17B10 (Figure 4C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5) in Example 4, compared to the affinity of the prior art antibodies REGN-5713 (Figure 4D), REGN-5714 (Figure 4E), and REGN-5715 (Figure 4F). [Figure 4G] Figure 4G shows a heat map summarizing the affinity of the antibody of the present invention and the comparative antibody. [Figure 5A] Figure 5A shows the suppression of mast cell reactivity by a "cocktail" containing one (4E1), two (4E1+25-3D3;4E1+17B10), or three (4E1+17B10+25-3D3) types of the MY010 antibody of the present invention in Example 5, compared to a "cocktail" containing the prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (REGN-3). [Figure 5B] Figure 5B shows the suppression of mast cell reactivity by a "cocktail" containing one (4E1), two (4E1+25-3D3; 4E1+17B10), or three (4E1+17B10+25-3D3) types of the MY010 antibody of the present invention in Example 5, compared to a "cocktail" containing the prior art antibodies REGN-5713, REGN-5714, and REGN-5715 (REGN-3). [Figure 6A]Figure 6A shows the suppression of mast cell reactivity by the "MY010 cocktail" of the present invention, which contains three types of antibodies of the present invention (4E1+17B10+25-3D3), in Example 6, compared to the "REGN cocktail" containing prior art antibodies REGN-5713, REGN-5714, and REGN-5715. [Figure 6B] Figure 6B shows the suppression of mast cell reactivity by the "MY010 cocktail" of the present invention, which contains three types of antibodies of the present invention (4E1+17B10+25-3D3), in Example 6, compared to the "REGN cocktail" containing prior art antibodies REGN-5713, REGN-5714, and REGN-5715. [Figure 7] Figure 7 shows the prevention of basophilic degranulation in patient whole blood samples induced by birch extract (10 ng / ml Bet v 1) using the "MY010 cocktail" (4E1+17B10+25-3D3) of the present invention in Example 7, compared to the prior art REGN cocktail (REGN-5713+REGN-5714+REGN-5715). [Figure 8] Figure 8 shows the effect of the "MY010 cocktail" on birch-related food allergens (apple Mal d 1, hazelnut Cor a 1, and walnut Jugr 5, and peach extract) in Example 8. [Figure 9] Figure 9 shows the competition among the MY010 antibody complementation group against Bet v 1 in Example 9. [Figure 10] Figure 10 shows the BLI sensorograms for the binding of MY010 antibodies 4E1, 25-3D3, and 3F1 to Bet v 1 in Example 10. [Figure 11A] Figure 11A shows the affinity of the MY010 antibodies 6C9 (Figure 11A), 3F1 (Figure 11B), and 13A2 (Figure 11C) of the present invention for birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, beech extract, and oak extract) in Example 11. [Figure 11B]Figure 11B shows the affinity of the MY010 antibodies 6C9 (Figure 11A), 3F1 (Figure 11B), and 13A2 (Figure 11C) of the present invention for birch extract and birch-related extracts (hazelnut pollen extract, European alder extract, American beech extract, European beech extract, and oak extract) in Example 11. [Figure 11C] Figure 11C shows the affinity of the MY010 antibodies 6C9 (Figure 11A), 3F1 (Figure 11B), and 13A2 (Figure 11C) of the present invention for birch extract and birch-related extracts (hazelnut pollen extract, alder extract, beech extract, beech extract, and oak extract) in Example 11. [Figure 11D] Figure 11D shows a heat map summarizing the affinity of the antibody MY010-17B10 of the present invention. [Figure 12A] Figure 12A shows the affinity of the MY010 antibodies 6C9, 3F1, and 13A2 of the present invention for Bet v 1 and Bet v 1 homologous allergens (Alder Aln g 1, Hazel Cor a 1.0101 and Cor a 1.0201, Hornbeam recombinant Car b 1 1A (rec Car b 1 1A) and recombinant Car b 1 (rec Car b 1) isoform 2, Japanese bayberry Ost c 1, Oak Que a 1.0201 and Que a 1.0301, Beech Fag s 1, and Chestnut Cas s 1) (Figure 12A) in Example 12. [Figure 12B] Figure 12B is a heat map summarizing the affinity between the antibody of the present invention and the prior art antibody. [Figure 13A] Figure 13A shows the affinity of the MY010 antibodies 6C9 (Figure 13A), 3F1 (Figure 13B), and 13A2 (Figure 13C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1.0108, hazelnut Cor a 1.0401, and walnut Jug r 5.0101) in Example 13. [Figure 13B]Figure 13B shows the affinity of the MY010 antibodies 6C9 (Figure 13A), 3F1 (Figure 13B), and 13A2 (Figure 13C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1.0108, hazelnut Cor a 1.0401, and walnut Jug r 5.0101) in Example 13. [Figure 13C] Figure 13C shows the affinity of the MY010 antibodies 6C9 (Figure 13A), 3F1 (Figure 13B), and 13A2 (Figure 13C) of the present invention for food-related Bet v 1 homologous allergens (apple Mal d 1.0108, hazelnut Cor a 1.0401, and walnut Jug r 5.0101) in Example 13. [Figure 14A] Figure 14A shows the suppression of mast cell reactivity by different antibody "cocktails" as shown in Example 14. [Figure 14B] Figure 14B shows the suppression of mast cell reactivity by different antibody "cocktails" as shown in Example 14. [Figure 15A] Figure 15A shows the effect of the MY010-3 cocktail containing the antibody of the present invention, as shown, in a passive cutaneous anaphylaxis (PCA) model in Example 15, compared to the prior art REGN-3 cocktail. Figure 15A: Experimental setup. [Figure 15B] Figure 15B shows the effect of the MY010-3 cocktail containing the antibody of the present invention, as shown, in a passive cutaneous anaphylaxis (PCA) model in Example 15, compared to the prior art REGN-3 cocktail. Figure 15B: Mice challenged intravenously with birch pollen extract (0.1 μg). [Figure 15C] Figure 15C shows the effect of the MY010-3 cocktail containing the antibody of the present invention, as shown, in a passive cutaneous anaphylaxis (PCA) model in Example 15, compared to the prior art REGN-3 cocktail. Figure 15C: Mice challenged intravenously with hazelnut pollen extract (1 μg). [Figure 15D]Figure 15D shows the effect of the MY010-3 cocktail containing the antibody of the present invention, as shown, in a passive cutaneous anaphylaxis (PCA) model in Example 15, compared to the prior art REGN-3 cocktail. Figure 15D: Mice challenged intravenously with oak pollen extract (1 μg). [Figure 16A] Figure 16A shows the binding sites of MY010 and the prior art REGN antibody to Bet v 1, as identified by epitope mapping in Example 16. Figure 16A: Scanning mutation introduced into surface-exposed residues of Bet v 1. [Figure 16B] Figure 16B shows the binding sites of MY010 and the prior art REGN antibody to Bet v 1, as identified by epitope mapping in Example 16. Figure 16B: Schematic diagram of the Bet v 1 epitope targeted by MY010 and the REGN antibody.

Claims

1. An antibody or its antigen-binding fragment that specifically binds to Bet v 1 (Betula berlucosa allergen 1).

2. The antibody or antigen-binding fragment according to claim 1, wherein the antibody or antigen-binding fragment further specifically binds to a Bet v 1 homologous allergen.

3. The antibody or antigen-binding fragment according to claim 2, wherein the Bet v 1 homologous allergen is a tree pollen allergen.

4. The antibody or antigen-binding fragment according to claim 3, wherein the Bet v1 homologous tree pollen allergen is selected from the group consisting of Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, and Car b 1.

5. The antibody or antigen-binding fragment according to any one of claims 1 to 4, wherein the antibody or antigen-binding fragment further specifically binds to a Bet v 1 homologous food allergen.

6. The antibody or antigen-binding fragment according to claim 5, wherein the Bet v 1 homologous food allergen is selected from the group consisting of Mal d 1, Cor a 1, and Jug r 5.

7. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment comprises CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO:

6.

8. The antibody or antigen-binding fragment according to any one of claims 1 to 7, wherein the antibody or antigen-binding fragment comprises CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO:

6.

9. The antibody or antigen-binding fragment according to any one of claims 1 to 8, wherein the antibody or antigen-binding fragment comprises VH having at least 70% identity with SEQ ID NO: 7 and VL having at least 70% identity with SEQ ID NO:

8.

10. The antibody or antigen-binding fragment according to any one of claims 1 to 9, wherein the antibody or antigen-binding fragment comprises VH as described in SEQ ID NO: 7 and VL as described in SEQ ID NO:

8.

11. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment comprises CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO:

14.

12. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 11, wherein the antibody or antigen-binding fragment comprises CDRH1 as described in SEQ ID NO: 9, CDRH2 as described in SEQ ID NO: 10, CDRH3 as described in SEQ ID NO: 11, CDRL1 as described in SEQ ID NO: 12, CDRL2 as described in SEQ ID NO: 13, and CDRL3 as described in SEQ ID NO:

14.

13. The antibody or antigen-binding fragment according to any one of claims 1 to 6, 11, and 12, wherein the antibody or antigen-binding fragment comprises VH having at least 70% identity with SEQ ID NO: 15 and VL having at least 70% identity with SEQ ID NO:

16.

14. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 11 to 13, wherein the antibody or antigen-binding fragment comprises VH as described in SEQ ID NO: 15 and VL as described in SEQ ID NO:

16.

15. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment comprises CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO:

22.

16. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 15, wherein the antibody or antigen-binding fragment comprises CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, CDRH3 as described in SEQ ID NO: 19, CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO:

22.

17. The antibody or antigen-binding fragment according to any one of claims 1 to 6, 15, and 16, wherein the antibody or antigen-binding fragment comprises VH having at least 70% identity with SEQ ID NO: 23 and VL having at least 70% identity with SEQ ID NO:

24.

18. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 15 to 17, wherein the antibody or antigen-binding fragment comprises VH as described in SEQ ID NO: 23 and VL as described in SEQ ID NO:

24.

19. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment comprises CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO:

71.

20. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 19, wherein the antibody or antigen-binding fragment comprises CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO:

71.

21. The antibody or antigen-binding fragment according to any one of claims 1 to 6, 19, and 20, wherein the antibody or antigen-binding fragment comprises VH having at least 70% identity with SEQ ID NO: 72 and VL having at least 70% identity with SEQ ID NO:

73.

22. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 19 to 21, wherein the antibody or antigen-binding fragment comprises VH as described in SEQ ID NO: 72 and VL as described in SEQ ID NO:

73.

23. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment comprises CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO:

79.

24. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 23, wherein the antibody or antigen-binding fragment comprises CDRH1 as described in SEQ ID NO: 74, CDRH2 as described in SEQ ID NO: 75, CDRH3 as described in SEQ ID NO: 76, CDRL1 as described in SEQ ID NO: 77, CDRL2 as described in SEQ ID NO: 78, and CDRL3 as described in SEQ ID NO:

79.

25. The antibody or antigen-binding fragment according to any one of claims 1 to 6, 23, and 24, wherein the antibody or antigen-binding fragment comprises VH having at least 70% identity with SEQ ID NO: 80 and VL having at least 70% identity with SEQ ID NO:

81.

26. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 23 to 25, wherein the antibody or antigen-binding fragment comprises VH as described in SEQ ID NO: 80 and VL as described in SEQ ID NO:

81.

27. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the antibody or antigen-binding fragment comprises CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO:

87.

28. The antibody or antigen-binding fragment according to any one of claims 1 to 6 and 27, wherein the antibody or antigen-binding fragment comprises CDRH1 as described in SEQ ID NO: 82, CDRH2 as described in SEQ ID NO: 83, CDRH3 as described in SEQ ID NO: 84, CDRL1 as described in SEQ ID NO: 85, CDRL2 as described in SEQ ID NO: 86, and CDRL3 as described in SEQ ID NO:

87.

29. The antibody or antigen-binding fragment according to any one of claims 1 to 6, 27, and 28, wherein the antibody or antigen-binding fragment comprises a VH having at least 70% identity with SEQ ID NO: 88 and a VL having at least 70% identity with SEQ ID NO:

89.

30. The antibody or its antigen-binding fragment according to any one of claims 1 to 6 and 27 to 29, wherein the antibody or its antigen-binding fragment comprises VH as described in SEQ ID NO: 88 and VL as described in SEQ ID NO:

89.

31. The antibody or antigen-binding fragment according to any one of claims 1 to 30, wherein the CDR or variable region of the antibody or antigen-binding fragment is a human CDR or variable region sequence, or is derived from a human CDR or variable region sequence.

32. The antibody or antigen-binding fragment according to any one of claims 1 to 31, wherein the antibody or antigen-binding fragment is a human antibody.

33. The antibody or antigen-binding fragment according to any one of claims 1 to 32, wherein the antibody or antigen-binding fragment is a monoclonal antibody.

34. The antibody according to any one of claims 1 to 33, wherein the antibody comprises an Fc substructure.

35. The antibody according to any one of claims 1 to 34, wherein the antibody is of the IgG type or the IgA type.

36. The antibody according to claim 23, wherein the antibody is of type IgG1 or type IgG4, and preferably type IgG4.

37. The antibody according to any one of claims 1 to 36, wherein the variable region or the CDR is derived from an IgE antibody and is grafted onto a scaffold of an IgG or IgA antibody.

38. The antibody or antigen-binding fragment according to any one of claims 1 to 37, wherein the antibody or antigen-binding fragment thereof has been purified.

39. The antibody or antigen-binding fragment according to any one of claims 1 to 38, wherein the antibody or antigen-binding fragment is a single-chain antibody.

40. The antibody or antigen-binding fragment according to claim 20, wherein the antibody or antigen-binding fragment is scFv.

41. The antibody or antigen-binding fragment according to any one of claims 1 to 19, wherein the antibody or antigen-binding fragment is selected from Fab, Fab', F(ab')2, and Fv.

42. The antibody or antigen-binding fragment according to any one of claims 1 to 41, wherein the antibody or antigen-binding fragment is a multispecific antibody or a multispecific antigen-binding fragment.

43. The antibody or antigen-binding fragment according to claim 42, wherein the multispecific antibody or multispecific antigen-binding fragment binds to a separate non-overlapping epitope of Bet v 1 and optionally binds to a separate non-overlapping epitope of at least one Bet v 1 homologous allergen.

44. The antibody or antigen-binding fragment according to claim 43, wherein the multispecific antibody or multispecific antigen-binding fragment comprises at least two of the following: (i) an antigen-binding site comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, as defined in any one of claims 7 to 10; (ii) an antigen-binding site comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, as defined in any one of claims 11 to 14; and an antigen-binding site selected from any one of (iii) to (vi): (iii) an antigen-binding site comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, as defined in any one of claims 15 to 18; (iv) an antigen-binding site comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, as defined in any one of claims 19 to 22; (v) an antigen-binding site comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, as defined in any one of claims 23 to 26; (vi) an antigen-binding site comprising (a) the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 sequences and / or (b) the VH and VL sequences, as defined in any one of claims 27 to 30.

45. The CDRH1, CDRH2, and CDRH3 sequences correspond to the genes IGHV1-18, IGHV1-2, IGHV1-24, IGHV1-3, IGHV1-45, IGHV1-46, IGHV1-58, IGHV1-69, IGHV1-69-2, IGHV1-69D, IGHV1-8, and IGHV2-26. , IGHV2-5, IGHV2-70, IGHV2-70D, IGHV3-11, IGHV3-13, IGHV3-15, IGHV3-20, IGH V3-21, IGHV3-23, IGHV3-23D, IGHV3-30, IGHV3-30-3, IGHV3-30-5, IGHV3-33, IGH It is comprised of a human variable heavy chain framework region (VH chain) selected from the group consisting of amino acid sequences encoded by V3-35, IGHV3-43, IGHV3-43D, IGHV3-48, IGHV3-49, IGHV3-53, IGHV3-62, IGHV3-64, IGHV3-64D, IGHV3-66, IGHV3-7, IGHV3-72, IGHV3-73, IGHV3-74, IGHV3-9, IGHV3-NL1, IGHV4-28, IGHV4-30-1, IGHV4-30-2, IGHV4-30-4, IGHV4-31, IGHV4-34 and IGHV4-38-2. The antibody or antigen-binding fragment according to any one of claims 7, 8, 11, 12, 15, 16, 19, 20, 23, 24, 27, or 28.

46. The CDRL1, CDRL2, and CDRL3 sequences are found in the genes IGLV1-36, IGLV1-40, IGLV1-44, IGLV1-47, IGLV1-51, IGLV10-54, IGLV2-11, IGLV2-14, IGLV2-18, IGLV2-23, IGLV2-8, IGLV3-1, IGLV3-10, IGLV3-12, IGLV3-16, IGLV3-19, IGLV3-21, IGLV3-22, I It is contained by a human variable light chain framework region (VL chain) selected from the group consisting of amino acid sequences encoded by GLV3-25, IGLV3-27, IGLV3-9, IGLV4-3, IGLV4-60, IGLV4-69, IGLV5-37, IGLV5-39, IGLV5-45, IGLV5-52, IGLV6-57, IGLV7-43, IGLV7-46, IGLV8-61 and IGLV9-49. The antibody or antigen-binding fragment according to any one of claims 7, 8, 11, 12, 15, 16, 19, 20, 23, 24, 27, 28, or 45.

47. The CDRL1, CDRL2, and CDRL3 sequences correspond to the genes IGKV1-12, IGKV1-13, IGKV1-16, IGKV1-17, IGKV1-27, IGKV1-33, IGKV1-39, IGKV1-5, IGKV1-6, IGKV1-8, IGKV1-9, IGKV1-NL1, IGKV1D-12, IGKV1D-13, IGKV1D-16, IGKV1D-17, IGKV1D-33, IGKV1D-39, IGKV1D-43, IGKV1D-8, IGKV2-24, IGKV2-28, IGK It is comprised of a human variable light chain framework region VK chain selected from the group consisting of amino acid sequences encoded by V2-29, IGKV2-30, IGKV2-40, IGKV2D-26, IGKV2D-28, IGKV2D-29, IGKV2D-30, IGKV2D-40, IGKV3-11, IGKV3-15, IGKV3-20, IGKV3D-11, IGKV3D-15, IGKV3D-20, IGKV3D-7, IGKV4-1, IGKV5-2, IGKV6-21 and IGKV6D-21. The antibody or antigen-binding fragment according to any one of claims 7, 8, 11, 12, 15, 16, 19, 20, 23, 24, 27, 28, or 45.

48. An antibody or antigen-binding fragment according to any one of claims 1 to 47, for use as a pharmaceutical product.

49. An antibody or its antigen-binding fragment used in accordance with claim 48 for the prevention or treatment of birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy.

50. A nucleic acid molecule comprising a polynucleotide encoding an antibody or an antigen-binding fragment according to any one of claims 1 to 47.

51. Nucleic acid sequences shown in any one of sequence numbers 25-48 and 90-111; or, A variant of that sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, The nucleic acid molecule according to claim 50, comprising:

52. A plurality of nucleic acid molecules encoding an antibody or its antigen-binding fragment according to any one of claims 1 to 49, Each of the nucleic acid molecules comprises a polynucleotide encoding the immunoglobulin chain of the antibody or its antigen-binding fragment. Multiple nucleic acid molecules.

53. Nucleic acid sequences shown in any one of sequence numbers 25-48 and 90-111; or, A variant of that sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, A combination of nucleic acid molecules according to claim 52, including the above.

54. A vector comprising a nucleic acid molecule according to claim 50 or 51, or a plurality of nucleic acid molecules according to claim 52 or 53.

55. A plurality of vectors comprising a plurality of nucleic acid molecules according to claim 52 or 53.

56. Expressing an antibody or antigen-binding fragment according to any one of claims 1 to 49, A vector according to claim 54 or a plurality of vectors according to claim 55, host cell.

57. A method for preparing an antibody or its antigen-binding fragment, or its immunoglobulin chain, according to any one of claims 1 to 49, (i) the step of culturing the host cells according to claim 56; and, (ii) A step of isolating the antibody or its immunoglobulin chain from the culture, A method that includes this.

58. A composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 49, a nucleic acid or a plurality of nucleic acids according to any one of claims 50 to 53, a vector or a plurality of vectors according to claim 54 or 55, or a cell according to claim 56.

59. The composition according to claim 58, further comprising a pharmaceutically acceptable excipient, diluent, or carrier.

60. The composition comprises at least two distinct antibodies or their antigen-binding fragments. The composition according to claim 58 or 59, wherein the antibody or its antigen-binding fragment binds to a separate non-overlapping epitope of Bet v 1 and optionally to a separate non-overlapping epitope of at least one Bet v 1 homologous protein.

61. The composition comprises at least two antibodies or their antigen-binding fragments, The antibody or its antigen-binding fragment (i) The antibody or antigen-binding fragment according to any one of claims 7 to 10; (ii) the antibody or antigen-binding fragment according to any one of claims 11 to 14; and the antibody or antigen-binding fragment according to any one of claims (iii) to (vi): (iii) The antibody or antigen-binding fragment according to any one of claims 15 to 18; (iv) The antibody or antigen-binding fragment according to any one of claims 19 to 22; (v) The antibody or antigen-binding fragment according to any one of claims 23 to 26; (vi) The antibody or antigen-binding fragment according to any one of claims 27 to 30, A composition according to claim 60, selected from the group consisting of the following.

62. It comprises at least three distinct antibodies or their antigen-binding fragments, preferably, A combination of the antibody or antigen-binding fragment described in (i) of claim 61, the antibody or antigen-binding fragment described in (ii), and the antibody or antigen-binding fragment described in at least one of (iii) to (vi), The composition according to claim 60 or 61.

63. A composition comprising at least two distinct antibodies or their antigen-binding fragments, The two distinct antibodies or their antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1, and optionally bind to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein. The composition is (i) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6; (ii) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14; and an antibody or its antigen-binding fragment according to any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO: 79; (vi) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87, A composition comprising at least two antibodies or antigen-binding fragments selected from the group consisting of the following.

64. The composition comprises at least two antibodies or their antigen-binding fragments, The antibody or its antigen-binding fragment (i) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; (ii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 9, CDRH2 as described in SEQ ID NO: 10, CDRH3 as described in SEQ ID NO: 11, CDRL1 as described in SEQ ID NO: 12, CDRL2 as described in SEQ ID NO: 13, and CDRL3 as described in SEQ ID NO: 14; and an antibody or its antigen-binding fragment as described in any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, CDRH3 as described in SEQ ID NO: 19, CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 74, CDRH2 as described in SEQ ID NO: 75, CDRH3 as described in SEQ ID NO: 76, CDRL1 as described in SEQ ID NO: 77, CDRL2 as described in SEQ ID NO: 78, and CDRL3 as described in SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87 A composition according to claim 63, selected from the group consisting of the following.

65. The composition comprises at least two antibodies or their antigen-binding fragments, The antibody or its antigen-binding fragment (i) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; (ii) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 15 and VL having at least 70% identity with SEQ ID NO: 16; and the antibody according to any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 23, and VL having at least 70% identity with SEQ ID NO: 24; (iv) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; (vi) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89, A composition according to claim 63 or 64, selected from the group consisting of the following.

66. The composition comprises at least two antibodies or their antigen-binding fragments, The antibody or its antigen-binding fragment (i) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 7 and VL as described in SEQ ID NO: 8; (ii) an antibody or antigen-binding fragment comprising VH as described in SEQ ID NO: 15 and VL as described in SEQ ID NO: 16; and an antibody or antigen-binding fragment as described in any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 23 and VL as described in SEQ ID NO: 24; (iv) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 72 and VL as described in SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 80 and VL as described in SEQ ID NO: 81; (vi) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 88 and VL as described in SEQ ID NO: 89; A composition according to any one of claims 63 to 65, selected from the group consisting of the following.

67. It comprises at least three distinct antibodies or their antigen-binding fragments, preferably, (i) The antibody or antigen-binding fragment described in (ii) The antibody or antigen-binding fragment described in (ii) The antibody or antigen-binding fragment described in at least one of (iii) to (vi) The composition according to any one of claims 63 to 66.

68. The composition according to any one of claims 58 to 67, further comprising at least one additional agent useful for treating birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy.

69. The composition according to claim 68, wherein the additional agent useful for treating birch pollen allergy and / or related allergies is selected from the group consisting of β-adrenergic agonists, epinephrine, antihistamines, corticosteroids, anti-IgE antibodies, anti-IgE antibody-conjugated fragments, peptide vaccines, and further antibodies capable of binding to birch pollen allergens.

70. The composition further comprises birch pollen allergen and / or Bet v 1 homologous allergen, preferably, The allergen is selected from the group consisting of Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, Mal d 1, Jug r 5, and Pru p 1, and / or The composition comprises an extract containing birch pollen allergen and / or Bet v 1 homologous allergen, preferably, The allergen is selected from the group consisting of Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, Mal d 1, Jug r 5, and Pru p 1. The composition according to any one of claims 58 to 69.

71. (i) The antibody or antigen-binding fragment thereof according to any one of claims 1 to 49; (ii) The nucleic acid molecule according to any one of claims 50 to 53; (iii) The vector according to claim 54 or 55; (iv) The cells according to claim 56; and / or, (v) The composition according to any one of claims 59 to 70, A kit containing one or more of the following.

72. The aforementioned kit, (i) at least two antibodies or antigen-binding fragments according to any one of claims 1 to 49, wherein the antibodies or antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; (ii) A nucleic acid molecule encoding at least two antibodies or antigen-binding fragments according to any one of claims 1 to 49, wherein the antibodies or antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; or, Compositions including (iii)(i) or (ii), The kit according to claim 71, including the following:

73. Three antibodies or antigen-binding fragments according to any one of claims 1 to 49, which bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; or, nucleic acid encoding the antibody or a composition containing the antibody, The kit according to claim 72, including the following:

74. A kit comprising at least two antibodies or antigen-binding fragments thereof, The three antibody or antigen-binding fragments bind to distinct non-overlapping epitopes of Bet v 1, and optionally bind to at least one distinct non-overlapping epitope of a Bet v 1 homologous protein. The aforementioned kit, (i) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 1, CDRH2 having at least 70% identity with SEQ ID NO: 2, CDRH3 having at least 70% identity with SEQ ID NO: 3, CDRL1 having at least 70% identity with SEQ ID NO: 4, CDRL2 having at least 70% identity with SEQ ID NO: 5, and CDRL3 having at least 70% identity with SEQ ID NO: 6; (ii) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 9, CDRH2 having at least 70% identity with SEQ ID NO: 10, CDRH3 having at least 70% identity with SEQ ID NO: 11, CDRL1 having at least 70% identity with SEQ ID NO: 12, CDRL2 having at least 70% identity with SEQ ID NO: 13, and CDRL3 having at least 70% identity with SEQ ID NO: 14; and an antibody or its antigen-binding fragment according to any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 17, CDRH2 having at least 70% identity with SEQ ID NO: 18, CDRH3 having at least 70% identity with SEQ ID NO: 19, CDRL1 having at least 70% identity with SEQ ID NO: 20, CDRL2 having at least 70% identity with SEQ ID NO: 21, and CDRL3 having at least 70% identity with SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 66, CDRH2 having at least 70% identity with SEQ ID NO: 67, CDRH3 having at least 70% identity with SEQ ID NO: 68, CDRL1 having at least 70% identity with SEQ ID NO: 69, CDRL2 having at least 70% identity with SEQ ID NO: 70, and CDRL3 having at least 70% identity with SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 74, CDRH2 having at least 70% identity with SEQ ID NO: 75, CDRH3 having at least 70% identity with SEQ ID NO: 76, CDRL1 having at least 70% identity with SEQ ID NO: 77, CDRL2 having at least 70% identity with SEQ ID NO: 78, and CDRL3 having at least 70% identity with SEQ ID NO: 79; (vi) an antibody or its antigen-binding fragment comprising CDRH1 having at least 70% identity with SEQ ID NO: 82, CDRH2 having at least 70% identity with SEQ ID NO: 83, CDRH3 having at least 70% identity with SEQ ID NO: 84, CDRL1 having at least 70% identity with SEQ ID NO: 85, CDRL2 having at least 70% identity with SEQ ID NO: 86, and CDRL3 having at least 70% identity with SEQ ID NO: 87, A kit comprising at least two antibodies or antigen-binding fragments selected from the group consisting of the following.

75. The kit comprises at least two antibodies or antigen-binding fragments thereof. The antibody or its antigen-binding fragment (i) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 1, CDRH2 as described in SEQ ID NO: 2, CDRH3 as described in SEQ ID NO: 3, CDRL1 as described in SEQ ID NO: 4, CDRL2 as described in SEQ ID NO: 5, and CDRL3 as described in SEQ ID NO: 6; (ii) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 9, CDRH2 as described in SEQ ID NO: 10, CDRH3 as described in SEQ ID NO: 11, CDRL1 as described in SEQ ID NO: 12, CDRL2 as described in SEQ ID NO: 13, and CDRL3 as described in SEQ ID NO: 14; and an antibody or its antigen-binding fragment as described in any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, CDRH3 as described in SEQ ID NO: 19, CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO: 22; (iv) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 66, CDRH2 as described in SEQ ID NO: 67, CDRH3 as described in SEQ ID NO: 68, CDRL1 as described in SEQ ID NO: 69, CDRL2 as described in SEQ ID NO: 70, and CDRL3 as described in SEQ ID NO: 71; (v) an antibody or its antigen-binding fragment comprising CDRH1 as described in SEQ ID NO: 74, CDRH2 as described in SEQ ID NO: 75, CDRH3 as described in SEQ ID NO: 76, CDRL1 as described in SEQ ID NO: 77, CDRL2 as described in SEQ ID NO: 78, and CDRL3 as described in SEQ ID NO: 79; (vi) An antibody or its antigen-binding fragment comprising CDRH1 described in SEQ ID NO: 82, CDRH2 described in SEQ ID NO: 83, CDRH3 described in SEQ ID NO: 84, CDRL1 described in SEQ ID NO: 85, CDRL2 described in SEQ ID NO: 86, and CDRL3 described in SEQ ID NO: 87 A kit according to claim 74, selected from the group consisting of the following.

76. The kit comprises at least two antibodies or antigen-binding fragments thereof. The antibody or its antigen-binding fragment (i) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 7, and VL having at least 70% identity with SEQ ID NO: 8; (ii) an antibody or antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 15 and VL having at least 70% identity with SEQ ID NO: 16; and an antibody or antigen-binding fragment according to any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 23, and VL having at least 70% identity with SEQ ID NO: 24; (iv) An antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 72, and VL having at least 70% identity with SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 80, and VL having at least 70% identity with SEQ ID NO: 81; (vi) an antibody or its antigen-binding fragment comprising VH having at least 70% identity with SEQ ID NO: 88, and VL having at least 70% identity with SEQ ID NO: 89, A kit according to claim 74 or 75, selected from the group consisting of the following.

77. The kit comprises at least two antibodies or antigen-binding fragments thereof. The antibody or its antigen-binding fragment (i) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 7 and VL as described in SEQ ID NO: 8; (ii) an antibody or antigen-binding fragment comprising VH as described in SEQ ID NO: 15 and VL as described in SEQ ID NO: 16; and an antibody or antigen-binding fragment as described in any one of (iii) to (vi): (iii) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 23 and VL as described in SEQ ID NO: 24; (iv) An antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 72 and VL as described in SEQ ID NO: 73; (v) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 80 and VL as described in SEQ ID NO: 81; (vi) an antibody or its antigen-binding fragment comprising VH as described in SEQ ID NO: 88 and VL as described in SEQ ID NO: 89 A kit according to any one of claims 74 to 76, selected from the group consisting of the following.

78. It comprises at least three antibodies or antigen-binding fragments, preferably, (i) The antibody or antigen-binding fragment described in (ii) The antibody or antigen-binding fragment described in (ii) The antibody or antigen-binding fragment described in at least one of (iii) to (vi) The kit according to any one of claims 74 to 77.

79. The kit further comprises at least one additional agent useful for treating birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy. The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy. The kit according to any one of claims 71 to 78.

80. The kit further comprises birch pollen allergen, and / or related tree pollen allergen, and / or related food allergen. The aforementioned related tree pollen allergens are Preferably, at least one of the following is selected: hazelnut pollen allergen, alder pollen allergen, beech pollen allergen, oak pollen allergen, chestnut pollen allergen, Japanese hornbeam pollen allergen, and hornbeam pollen allergen. Preferably, selected from the group consisting of Cor a 1, Aln g 1, Fag s 1, Que a 1, Cas s 1, Ost c 1, Car b 1, The aforementioned related food allergens are Preferably, at least one of apple allergens, hazelnut allergens, and walnut allergens is selected. Preferably selected from Mal d 1, Cor a 1, and Jug r 5, The kit according to any one of claims 71 to 79.

81. An antibody or antigen-binding fragment according to any one of claims 1 to 49, a nucleic acid or a plurality of nucleic acids according to any one of claims 50 to 53, a vector or a plurality of vectors according to claim 54 or 55, a cell according to claim 56, a composition according to any one of claims 58 to 70, or a kit according to any one of claims 71 to 80, for use as a pharmaceutical.

82. An antibody or antigen-binding fragment according to any one of claims 1 to 49, a nucleic acid or plurality of nucleic acids according to any one of claims 50 to 53, a vector or plurality of vectors according to claim 54 or 55, a cell according to claim 56, a composition according to any one of claims 58 to 70, or a kit according to any one of claims 71 to 80, for use in the prevention or treatment of birch pollen allergy and / or related tree pollen allergy and / or related food allergy. The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy. Antibodies or their antigen-binding fragments, nucleic acids or multiple nucleic acids, vectors or multiple vectors, cells, compositions, or kits.

83. At least two or three distinct antibodies or antigen-binding fragments that bind to distinct non-overlapping epitopes of Bet v 1 and optionally to distinct non-overlapping epitopes of at least one Bet v 1 homologous protein; or, nucleic acids or vectors encoding the aforementioned separate antibodies; or, A composition comprising the aforementioned separate antibody, It is administered. An antibody or its antigen-binding fragment, a nucleic acid or a plurality of nucleic acids, a vector or a plurality of vectors, a cell, or a composition used in accordance with claim 82.

84. An antibody or its antigen-binding fragment, the nucleic acid or a plurality of nucleic acids, the vector or a plurality of vectors, the cell, or the composition used in accordance with claim 82 or 83, The administration of the antibody or its antigen-binding fragment, the nucleic acid or multiple nucleic acids, the vector or multiple vectors, the cells, or the composition is used in combination with the administration of Bet v 1 birch pollen allergen and / or related tree pollen allergen and / or related food allergen. The aforementioned related tree pollen allergen is preferably selected from at least one of Cor a 1 hazelnut pollen allergen, Aln g 1 alder pollen allergen, Fag s 1 beech pollen allergen, Que a 1 oak pollen allergen, Cas s 1 chestnut pollen allergen, Ost c 1 Japanese hornbeam pollen allergen, and Car b 1 hornbeam pollen allergen. The aforementioned related food allergen is preferably selected from at least one of Mal d 1 apple allergen, Cor a 1 hazelnut allergen, and Jug r 5 walnut allergen. Antibodies or their antigen-binding fragments, nucleic acids or multiple nucleic acids, vectors or multiple vectors, cells, or compositions.

85. An antibody or its antigen-binding fragment, or a composition, used in accordance with claim 84, wherein the antibody or its antigen-binding fragment, or the composition, is administered before or during treatment for desensitization with birch pollen allergen, and / or related tree pollen allergen, and / or related food allergen.

86. The use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 49, a composition according to any one of claims 58 to 70, or a kit according to any one of claims 71 to 80 in the (in-vitro) diagnosis of birch pollen allergy and / or related tree pollen allergy and / or related food allergy, The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy. use.

87. A method for detecting birch pollen allergens and / or related tree pollen allergens and / or related food allergens, comprising the use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 49, a composition according to any one of claims 58 to 70, or a kit according to any one of claims 71 to 80, The aforementioned related tree pollen allergy is preferably selected from at least one of the following: Cor a 1 hazelnut pollen allergen, Aln g 1 alder pollen allergen, Fag s 1 beech pollen allergen, Que a 1 oak pollen allergen, Cas s 1 chestnut pollen allergen, Ost c 1 Japanese hornbeam pollen allergen, and Car b 1 hornbeam pollen allergen. The aforementioned related food allergen is preferably selected from at least one of Mal d 1 apple allergen, Cor a 1 hazelnut allergen, and Jug r 5 walnut allergen. use.

88. The use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 49, a nucleic acid or plurality of nucleic acids according to any one of claims 50 to 53, a vector or plurality of vectors according to claim 54 or 55, a cell according to claim 56, a composition according to any one of claims 58 to 70, or a kit according to any one of claims 71 to 80 in the manufacture of a pharmaceutical for the prevention, treatment or reduction of birch pollen allergy and / or related tree pollen allergy and / or related food allergy, The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy. use.

89. A method for treating, improving or reducing birch pollen allergy, and / or related tree pollen allergy, and / or related food allergy, or a method for reducing the risk of allergic or anaphylactic reactions, The aforementioned related tree pollen allergy is preferably selected from at least one of hazelnut pollen allergy, alder pollen allergy, beech pollen allergy, oak pollen allergy, chestnut pollen allergy, Japanese hornbeam pollen allergy, and hornbeam pollen allergy. The aforementioned related food allergy is preferably selected from at least one of apple allergy, hazelnut allergy, and walnut allergy. The method comprises administering to a target subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 49, a nucleic acid or multiple nucleic acids according to any one of claims 50 to 53, a vector or multiple vectors according to claim 54 or 55, a cell according to claim 56, a composition according to any one of claims 58 to 70, or a kit according to any one of claims 71 to 80. method.

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