Methods of isolating allergen-specific antibodies from humans and uses thereof

Abstract

To provide a fast and effective intervention that can improve the quality of life for allergic individuals.SOLUTION: Provided is a method of identifying a human monoclonal antibody that specifically binds to an allergen, the method comprising the steps of: (a) obtaining B cells from a human peripheral blood sample from a human subject allergic to an allergen by performing negative selection of B cells, followed by cell selection based on CD19 expression and cell surface IgE expression; (b) generating cDNA from the RNA of the B cells of the step (a); (c) identifying individual B cells expressing IgE antibodies by sequencing the cDNA from the step (b); (d) expressing an antibody comprising a heavy chain variable region sequence and a light chain variable region sequence from the step (c); and (e) identifying an antibody from the step (d) that specifically binds to the allergen, thereby identifying a human monoclonal antibody that specifically binds to the allergen.SELECTED DRAWING: Figure 7

Description

[Background technology]

[0001] Allergies are a growing health concern worldwide and are characterized by adaptive immune responses misdirected against otherwise harmless proteins. In particular, in the case of food allergies, individuals must be careful to avoid allergen exposure and other risks of potentially fatal allergic reactions. Although there is no cure for allergies and desensitization regimens such as immunotherapy have shown some clinical benefit, there is a need for fast-acting and effective interventions that can improve the quality of life of allergic individuals. Summary of the Invention [Means for solving the problem]

[0002] In one aspect, the disclosure provides a method for generating a human monoclonal antibody that specifically binds to an allergen. In some embodiments, the method comprises: (a) isolating a single B cell from a sample from a human subject, the subject being allergic to an allergen; (b) generating cDNA from the single B cell of step (a), the cDNA sequence comprising a first sequence encoding all or a portion of an immunoglobulin heavy chain and a second sequence encoding all or a portion of an immunoglobulin light chain; (c) determining the sequence of the cDNA from step (b); (d) analyzing the sequence determined in step (c) to identify a single B cell comprising a first sequence comprising an IgE constant region or an IgG4 constant region; (e) identifying from the single B cell of step (d) (i) a heavy chain variable region sequence in an immunoglobulin heavy chain that comprises an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain that is co-expressed with the immunoglobulin heavy chain in the same single B cell; (f) expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (e); and (g) identifying one or more antibodies from step (f) that specifically bind to the allergen; Includes.

[0003] In some embodiments, step (a) comprises sorting cells in the sample by fluorescent activated cell sorting (FACS). In some embodiments, step (a) comprises selecting single B cells for expression of cell surface IgE and / or cell surface IgG4. In some embodiments, step (a) comprises isolating antibody-secreting B cells and / or memory B cells.

[0004] In some embodiments, the method comprises isolating a single B cell selected for expression of cell surface IgE and identifying a sequence encoding an immunoglobulin heavy chain comprising an IgE constant region. In some embodiments, step (a) comprises contacting cells from the sample with an anti-human CD19 antibody and an anti-human IgE antibody, and selecting CD19+IgE-expressing B cells. In some embodiments, the method comprises isolating a single B cell that expresses a B cell marker and is negative for a non-IgE isotype. In some embodiments, step (a) comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgG antibody, an anti-human IgA antibody, and an anti-human IgD antibody, and selecting CD19+IgM-IgG-IgA-IgD-B cells.

[0005] In some embodiments, the method comprises isolating a single B cell selected for cell surface IgG4 expression and identifying a sequence encoding an immunoglobulin heavy chain comprising an IgG4 constant region. In some embodiments, step (a) comprises contacting cells from the sample with an anti-human CD19 antibody and an anti-human IgG4 antibody, and selecting CD19+IgG4-expressing B cells. In some embodiments, the method comprises isolating a single B cell that expresses a B cell marker and is negative for a non-IgG4 isotype. In some embodiments, step (a) comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgE antibody, an anti-human IgA antibody, an anti-human IgD antibody, an anti-human IgG1 antibody, an anti-human IgG2 antibody, and an anti-human IgG3 antibody, and selecting CD19+IgM-IgE-IgA-IgD-IgG1-IgG2-IgG3-B cell.

[0006] In some embodiments, step (b) comprises reverse transcribing cDNA from RNA from a single B cell and amplifying the cDNA. In some embodiments, the RNA is mRNA. In some embodiments, the method comprises amplifying immunoglobulin heavy and light chain sequences from a single B cell. In some embodiments, the method comprises amplifying the transcriptome of a single B cell.

[0007] In some embodiments, step (f) comprises expressing the heavy and light chain variable region sequences from step (e) in a host cell and purifying the antibody. In some embodiments, step (f) comprises expressing an antibody comprising the heavy and light chain variable region sequences from step (e) and an IgG4 constant region or an IgG1 constant region.

[0008] In some embodiments, the method further comprises replacing the constant region of the antibody identified in step (g) with a wild-type IgG4 constant region or a modified IgG4 constant region.

[0009] In some embodiments, the sample comprises peripheral blood. In some embodiments, the sample comprises tissue (e.g., tonsil tissue).

[0010] In some embodiments, the allergen is a food allergen, a plant allergen, a fungal allergen, an animal allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the allergen is a food allergen selected from the group consisting of a milk allergen, an egg allergen, a nut allergen, a fish allergen, a shellfish allergen, a soy allergen, a legume allergen, a seed allergen, and a wheat allergen. In some embodiments, the food allergen is a peanut allergen. In some embodiments, the food allergen is a tree nut allergen. In some embodiments, the food allergen is a milk allergen. In some embodiments, the allergen is a fungal allergen. In some embodiments, the fungal allergen is an Aspergillus allergen.

[0011] In another aspect, there is provided a monoclonal antibody produced according to the methods disclosed herein.

[0012] In yet another aspect, a pharmaceutical composition is provided comprising a monoclonal antibody produced according to the methods disclosed herein. In some embodiments, the pharmaceutical composition comprises a plurality of monoclonal antibodies, each produced according to the methods disclosed herein, wherein the monoclonal antibodies recognize different epitopes or specifically bind to different antigens (e.g., different allergens within the same type or class of allergens or among different types or classes of allergens).

[0013] In yet another aspect, a monoclonal antibody or antigen-binding portion thereof that specifically binds to an allergen is provided. In some embodiments, the antibody is (a) a heavy chain variable region sequence derived from an immunoglobulin heavy chain from an IgE- or IgG4-producing single B cell from a human subject allergic to an allergen; (b) heavy chain IgG constant region sequence; (c) a light chain variable region sequence derived from an immunoglobulin light chain from an IgE- or IgG4-producing single B cell from a human subject, and (d) a light chain constant region sequence of the same class as the immunoglobulin light chain of (c) Includes.

[0014] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the allergen with a binding affinity (KD) of less than 1 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the allergen with a binding affinity (KD) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. In some embodiments, the antibody binds to the allergen with a binding affinity (KD) of between 1 nM and 250 nM. In some embodiments, the allergen is a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the allergen is a food allergen selected from the group consisting of a milk allergen, an egg allergen, a nut allergen, a fish allergen, a shellfish allergen, a soybean allergen, a legume allergen, a seed allergen, and a wheat allergen. In some embodiments, the food allergen is a peanut allergen or a tree nut allergen. In some embodiments, the food allergen is a milk allergen. In some embodiments, the allergen is a fungal allergen. In some embodiments, the fungal allergen is an Aspergillus antigen. In some embodiments, the antibody is cross-reactive with two different antigens. In some embodiments, the antibody is cross-reactive with a first antigen of a first type of allergen and a second antigen of a second type of allergen that is different from the first type of allergen. In some embodiments, the antibody is cross-reactive with a peanut allergen and a tree nut allergen. In some embodiments, the antibody is cross-reactive with two or more tree nut antigens.

[0015] In some embodiments, the antibody comprises heavy and light chain variable region sequences derived from an IgE- or IgG4-producing human B cell.

[0016] In yet another aspect, a monoclonal antibody or antigen-binding portion thereof that specifically binds to a peanut allergen is provided. In some embodiments, the antibody binds to a peanut allergen with a binding affinity (KD) of less than 1 nM. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to a peanut allergen with a binding affinity (KD) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. In some embodiments, the antibody binds to a peanut allergen with a binding affinity (KD) of between 1 nM and 250 nM.

[0017] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to peanut allergens Ara h2, Ara h3, or Ara h1. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to Ara h2 with a KD of less than 100 pM. In some embodiments, the antibody or antigen-binding portion thereof cross-reacts with at least two peanut allergens. In some embodiments, the antibody or antigen-binding portion thereof cross-reacts with Ara h2 and Ara h3. In some embodiments, the antibody or antigen-binding portion thereof specifically binds to Ara h2 with a KD of less than 1 nM and specifically binds to Ara h3 with a KD of less than 1 μM. In some embodiments, the antibody or antigen-binding portion thereof binds to an epitope comprising the amino acid motif DPYSPS (SEQ ID NO: 704). Moreover, additional peanut-specific antibodies were discovered during these experiments. Antibodies PA12P3E09 and PA12P3E11 bound to peanut extract and little or no binding to the natural peanut allergen Ara h2, whereas antibodies PA12P1D02, PA12P1G11, PA13P1H03, PA12P3C01, and PA12P3E04 bound strongly to both peanut extract and the natural peanut allergen Ara h2. In some embodiments, the peanut-specific antibodies bind to the peanut extract but not to the natural peanut allergen Ara h2.

[0018] In some embodiments, the antibody comprises: A heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199, 341, 348, 409, 459, or 593; a heavy chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349, 410, 460, 539, or 594; A heavy chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201, 350, 411, 461, 540, or 595; A light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343, 352, 413, 463, 542, or 597; a light chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 7, 15, 22, 30, 38, 52, 78, 86, 126, 149, 196, 345, 353, or 598; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354, 414, 464, 543, or 599. Includes.

[0019] In some embodiments, the antibody comprises heavy and light chain CDR sequences contained within a heavy chain and light chain variable region sequence pair selected from the group consisting of SEQ ID NOs: 1 and 5, 9 and 13, 16 and 20, 24 and 28, 32 and 36, 40 and 43, 46 and 50, 54 and 56, 57 and 61, 57 and 5, 1 and 61, 64 and 5, 65 and 5, 66 and 5, 67 and 5, 128 and 132, 340 and 344, 347 and 351, 406 and 407, 408 and 412, 458 and 462, 538 and 541, and 592 and 596. (a) SEQ ID NOs: 2, 3, 4, 6, 7, and 8, respectively; or (b) SEQ ID NOs: 10, 11, 12, 14, 15, and 8, respectively; or (c) SEQ ID NOs: 17, 18, 19, 21, 22, and 23, respectively; or (d) SEQ ID NOs: 25, 26, 27, 29, 30, and 31, respectively; or (e) SEQ ID NOs: 33, 34, 35, 37, 38, and 39, respectively; or (f) SEQ ID NOs: 41, 34, 35, 37, 38, and 39, respectively; or (g) SEQ ID NOs: 47, 48, 49, 51, 52, and 53, respectively; or (h) SEQ ID NOs: 47, 48, 55, 51, 52, and 53, respectively; or (i) SEQ ID NOs: 58, 59, 60, 62, 30, and 63, respectively; or (j) SEQ ID NOs: 58, 59, 60, 6, 7, and 8, respectively; or (k) SEQ ID NOs: 2, 3, 4, 62, 30, and 63, respectively; or (l) SEQ ID NOs: 58, 3, 4, 6, 7, and 8, respectively; or (m) SEQ ID NOs: 2, 59, 4, 6, 7, and 8, respectively; or (n) SEQ ID NOs: 2, 3, 60, 6, 7, and 8, respectively; or (o) SEQ ID NOs: 129, 130, 131, 133, 126, and 134, respectively; or (p) SEQ ID NOs: 341, 342, 343, 345, 78, and 346, respectively; or (q) SEQ ID NOs: 348, 349, 350, 352, 353, and 354, respectively; or (r) SEQ ID NOs: 199, 200, 201, 203, 149, and 204, respectively; or (s) SEQ ID NOs: 409, 410, 411, 413, 86, and 414, respectively; or (t) SEQ ID NOs: 459, 460, 461, 463, 196, and 464, respectively; or (u) SEQ ID NOs: 113, 539, 540, 542, 196, and 543, respectively; or (v) SEQ ID NOs: 593, 594, 595, 597, 598, and 599, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0020] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596.

[0021] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:1, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:9, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:13; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 16, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 20; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 24, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 28; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 32, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 36; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 40, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 43; or (g) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 46, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 50; or (h) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 54, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 56; or (i) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 57, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 61; or (j) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 57, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 5; or (k) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 61; or (l) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 64, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 5; or (m) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 65, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 5; or (n) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 66, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 5; or (o) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 67, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 5; or (p) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 128, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 132; or (q) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 340, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 344; or (r) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 347, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 351; or (s) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 406, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 407; or (t) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 408, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 412; or (u) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 458, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 462; or (v) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 538, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 541; or (w) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 592, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 596. Includes.

[0022] In yet another aspect, a monoclonal antibody or antigen-binding portion thereof that specifically binds to a tree nut allergen is provided. In some embodiments, the antibody binds to the tree nut allergen with a binding affinity (KD) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. In some embodiments, the antibody binds to the tree nut allergen with a binding affinity (KD) of less than 1 nM. In some embodiments, the antibody binds to the tree nut allergen with a binding affinity (KD) of between 1 nM and 250 nM. In some embodiments, the tree nut allergen is cashew, pistachio, pecan, walnut, hazelnut, and / or macadamia nut.

[0023] In some embodiments, the antibody comprises: A heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 113, 167, 175, 227, 311, 318, 438, 466, 621, 665, or 692; A heavy chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 168, 176, 200, 312, 319, 439, 539, 666, or 693; a heavy chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 169, 177, 228, 313, 320, 440, 467, 540, 667, or 694; A light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623, 669, or 696; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 94, 149, 172, 180, 196, 323, or 670; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624, 671, or 697. Includes.

[0024] In some embodiments, the antibody comprises: (a) SEQ ID NOs: 692, 693, 694, 696, 94, and 697, respectively; or (b) SEQ ID NOs: 318, 319, 320, 322, 323, and 324, respectively; or (c) SEQ ID NOs: 227, 200, 228, 230, 149, and 231, respectively; or (d) SEQ ID NOs: 113, 539, 540, 542, 196, and 543, respectively; or (e) SEQ ID NOs: 311, 312, 313, 315, 94, and 316, respectively; or (f) SEQ ID NOs: 665, 666, 667, 669, 670, and 671, respectively; or (g) SEQ ID NOs: 466, 200, 467, 469, 149, and 470, respectively; or (h) SEQ ID NOs: 167, 168, 169, 171, 172, and 173, respectively; or (i) SEQ ID NOs: 621, 176, 177, 623, 180, and 624, respectively; or (j) SEQ ID NOs: 175, 176, 177, 179, 180, and 181, respectively; or (k) SEQ ID NOs: 438, 439, 440, 442, 30, and 443, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0025] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695.

[0026] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 691, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 695; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 317, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 321; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 226, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 229; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 538, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 541; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 310, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 314; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 664, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 668; or (g) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 465, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 468; or (h) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 166, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 170; or (i) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 620, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 622; or (j) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 174, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 178; or (k) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 437, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 441. Includes.

[0027] In yet another aspect, a monoclonal antibody or antigen-binding portion thereof that specifically binds to a milk allergen is provided. In some embodiments, the antibody binds to the milk allergen with a binding affinity (KD) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. In some embodiments, the antibody binds to the milk allergen with a binding affinity (KD) of less than 1 nM. In some embodiments, the antibody binds to the milk allergen with a binding affinity (KD) of between 1 nM and 250 nM.

[0028] In some embodiments, the antibody comprises: a heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 121, 750, 757, 765, 772, 779, 785, 793, 800, 807, 814, 821, 833, 838, 846, 853, 860, 868, 874, 881, 889, 895, 903, 911, 918, or 926; a heavy chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 378, 532, 751, 758, 766, 773, 786, 794, 801, 808, 815, 822, 826, 839, 847, 854, 861, 875, 882, 890, 896, 904, 912, 919, or 927; a heavy chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 752, 759, 767, 774, 780, 787, 795, 802, 809, 816, 827, 840, 848, 855, 862, 869, 876, 883, 891, 897, 905, 913, 920, or 928; a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 523, 754, 761, 769, 776, 782, 789, 797, 804, 811, 818, 829, 835, 842, 850, 857, 864, 871, 878, 885, 899, 907, 915, 922, or 930; a light chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 22, 30, 94, 110, 149, 186, 196, 389, 404, 509, 662, 682, 762, 790, 830, 843, 865, 886, 900, 908, 923, or 931; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 755, 763, 770, 777, 783, 791, 798, 805, 812, 819, 824, 831, 836, 844, 851, 858, 866, 872, 879, 887, 893, 901, 909, 916, 924, or 932. Includes.

[0029] In some embodiments, the antibody comprises: (a) SEQ ID NOs: 800, 801, 802, 804, 110, and 805, respectively; or (b) SEQ ID NOs: 121, 826, 827, 829, 830, and 831, respectively; or (c) SEQ ID NOs: 833, 826, 827, 835, 149, and 836, respectively; or (d) SEQ ID NOs: 853, 854, 855, 857, 662, and 858, respectively; or (e) SEQ ID NOs: 860, 861, 862, 864, 865, and 866, respectively; or (f) SEQ ID NOs: 868, 378, 869, 871, 682, and 782, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0030] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 749, 756, 764, 771, 778, 784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873, 880, 888, 894, 902, 910, 917, or 925. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or 929.

[0031] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 799, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 803; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 825, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 828; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 832, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 834; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 852, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 856; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 859, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 863; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 867, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 870. Includes.

[0032] In some embodiments, the antibody is an antibody that competes with a monoclonal antibody disclosed herein for binding to an allergen (e.g., for binding to a food allergen, such as a peanut allergen, a nut allergen, or a milk allergen).

[0033] In yet another aspect, a monoclonal antibody or antigen-binding portion thereof that specifically binds to a fungal allergen is provided. In some embodiments, the fungal allergen is an Aspergillus allergen. In some embodiments, the antibody binds to the fungal allergen (e.g., Aspergillus allergen) with a binding affinity (KD) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. In some embodiments, the antibody binds to the fungal allergen (e.g., Aspergillus allergen) with a binding affinity (KD) of less than 1 nM. In some embodiments, the antibody binds to the fungal allergen (e.g., Aspergillus allergen) with a binding affinity (KD) of between 1 nM and 250 nM.

[0034] In some embodiments, the antibody or antigen-binding portion thereof specifically binds to the allergens Aspergillus fumigatus, Aspergillus niger, and / or Aspergillus nidulans. In some embodiments, the antibody specifically binds to the allergen Aspergillus fumigatus 1 (Asp f1).

[0035] In some embodiments, the antibody comprises: A heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 710, 718, 726, 734, or 742; A heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 711, 719, 727, 735, or 743; A heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 712, 720, 728, 736, or 744; a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 714, 722, 730, 738, or 746; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 715, 723, 731, 739, or 747; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 716, 724, 732, 740, or 748. Includes.

[0036] In some embodiments, the antibody comprises heavy and light chain CDR sequences contained within a heavy chain and light chain variable region sequence pair selected from the group consisting of SEQ ID NOs: 709 and 713, 717 and 721, 725 and 729, 733 and 737, and 741 and 745. (a) SEQ ID NOs: 710, 711, 712, 714, 715, and 716, respectively; or (b) SEQ ID NOs: 718, 719, 720, 722, 723, and 724, respectively; or (c) SEQ ID NOs: 726, 7272, 728, 730, 731, and 732, respectively; or (d) SEQ ID NOs: 734, 735, 736, 738, 739, and 740, respectively; or (e) SEQ ID NOs: 742, 743, 744, 746, 747, and 748, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0037] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 709, 717, 725, 733, or 741. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 713, 721, 729, 737, or 745.

[0038] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 709, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 713; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 717, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 721; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 725, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 729; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 733, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 737; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 741, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 745. Includes.

[0039] In some embodiments, the antibody is an antibody that competes with a monoclonal antibody disclosed herein for binding to a fungal allergen (eg, for binding to an Aspergillus allergen).

[0040] In another aspect, a pharmaceutical composition is provided comprising a monoclonal antibody or antigen-binding portion disclosed herein. In some embodiments, the pharmaceutical composition comprises a plurality of monoclonal antibodies disclosed herein, where the monoclonal antibodies recognize different epitopes or specifically bind to different antigens (e.g., different allergens within the same type or class of allergens or in different types or classes of allergens).

[0041] In another aspect, an antibody-drug conjugate is provided that comprises the monoclonal antibody or antigen-binding portion thereof disclosed herein. In some embodiments, the antibody-drug conjugate comprises a monoclonal antibody or antigen-binding portion thereof that specifically binds to a fungal allergen disclosed herein, and further comprises an antifungal agent. In some embodiments, the antifungal agent is amphotericin B.

[0042] In yet another aspect, an isolated polynucleotide comprising a nucleotide sequence encoding the monoclonal antibody disclosed herein. Also provided herein are vectors and host cells comprising the polynucleotides disclosed herein.

[0043] In another aspect, a method of treatment is provided. In some embodiments, the method of treatment is a method of reducing one or more allergy symptoms in a subject. In some embodiments, the method of treatment is a method of reducing one or more allergy symptoms in a subject with a peanut allergy. In some embodiments, the method of treatment is a method of reducing one or more allergy symptoms in a subject with a tree nut allergy. In some embodiments, the method of treatment is a method of reducing one or more allergy symptoms in a subject with a fungal allergy. In some embodiments, the method of treatment is a method of reducing one or more allergy symptoms in a subject with a milk allergy. In some embodiments, the method comprises administering to a subject a therapeutically effective amount of a monoclonal antibody or pharmaceutical composition disclosed herein.

[0044] In another aspect, a kit is provided. In some embodiments, the kit comprises a monoclonal antibody or pharmaceutical composition disclosed herein. In some embodiments, the kit is for use in practicing a therapeutic method disclosed herein. [Brief description of the drawings]

[0045] [Figure 1] Figure 1. Flow diagram of an exemplary workflow for a method of generating allergen-specific monoclonal antibodies.

[0046] [Figure 2-1] Figures 2A-2G. Characterization of single B cells isolated from fresh human peripheral blood. (A) Principal component analysis (left) separates naive / memory (black dots) and plasmablast (PB, grey dots) B cell subsets identified by expression of established transcription factors and marker genes [right; naive / memory B cells unstriped, plasmablast (PB) B cells striped]. (B) Isotypes of B cells, with IGHE as black dots and other isotypes as grey dots. (C) Number of cells per isotype belonging to each subtype in (A) [naive / memory B cells unstriped, plasmablast (PB) B cells striped]. (D-G) Analysis of clonal families (CFs). (D) Distribution of number of cells per CF. (E) Proportion of cells of each isotype belonging to multi-member CFs. (F) Isotype, B cell subtype, mutation frequency, and patient of origin of each cell within a multi-member CF. CFs mentioned in the text are named. (G) Heavy (right) and light (left) chain CDR3 sequences and similarity heatmap for the CF in (F). [Figure 2-2] Same as above [Figure 2-3] Same as above [Figure 2-4] Same as above

[0047] [Figure 3-1]Figures 3A-3G. Characterization of 89 IgE antibodies from single B cells. (A) Phylogenetic depiction of antibody heavy chains organized by IGHV gene (background pattern), patient of origin (node ​​pattern), and mutation frequency (node ​​size). (B) Heatmap showing the number of cells with given heavy and light chain CDR3 lengths. (C) Heavy and light chain mutation frequency for each cell. (D) Silent (S) and replacement (R) mutations per region within heavy and light chains. (E) Differential gene expression between IgE PB and PB of other isotypes. Positive log fold changes indicate genes enriched in IgE PB. (F) Heavy chain constant region coverage histograms by isotype for naive / memory B cells (top) and PB (bottom) with loci oriented in 5' to 3' direction. Mean normalized read depth and 95% confidence intervals for the number of described cells (n) are indicated by solid lines and shaded areas, respectively. Membrane exons are the two largest 3' exons for each isotype. (G) A summary of (F) showing the percentage of cells of each isotype with any degree of membrane exon coverage. [Figure 3-2] Same as above

[0048] [Figure 4] Figures 4A-4C. Convergence of high affinity and cross-reactive IgE antibodies in two unrelated individuals (PA12 and PA13). Antibody patterns are conserved between panels. (A) Frequency of silent (S) and replacement (R) mutations per region. (B) Mutation frequency percentiles compared to all class-switched antibodies. (C) Dissociation constants (KD) for the major allergen peanut proteins Ara h2 and Ara h3 of six convergent antibodies and eight variants of PA13P1H08. The shortened antibody variant names are designated as "heavy-light" using the following abbreviations: N=unmodified, R=reverted, FWR=framework region. The "r" prefix indicates that only that region is reverted.

[0049] [Diagram 5]5A-5E. Germline transcription reveals class switch priming in single B cells. (A) Identification of Cε germline transcript splice donors by isotype along with the number of cells expressing each isotype. (B) Percentage of cells expressing εGLT by isotype. (C) Example from individual PA11 where identification of phased variants within IgE constant region exons allows subsequent validation of biallelic GLT expression in other B cells from the same patient. (D) Global germline transcriptional status heatmap showing the percentage of cells of a given isotype expressing a given GLT. Top: GLT isotype expression frequency for all GLT isotypes, excluding self isotype GLT expression. (E) Histogram of the number of non-self GLT isotypes expressed in each cell.

[0050] [Figure 6] Figure 6. Study overview. Plasma was extracted from fresh blood to measure circulating IgE levels and cell fractions were enriched for B cells prior to FACS. Single cells were sorted into individual wells of 96-well plates and processed using scRNA-seq to generate sequencing reads that were aligned to the genome to calculate gene expression and assess splicing, and assembled to reconstruct heavy and light chain sequences. Specificity and affinity data were generated for recombinantly expressed antibodies.

[0051] [Figure 7] Figures 7A-7C. Plasma IgE levels. (A) Allergen-specific and allergen component (hazelnut, peanut) concentrations. (B) Total IgE concentrations. (C) Positive correlation between total plasma IgE levels and the number of identified IgE+ B cells. Each point is an individual.

[0052] [Figure 8]Figures 8A-8C. FACS gating and analysis. (A) Gating strategy for sorting single B cells. IgE+ B cells are overlaid as dark grey dots. (B) Isotype identification within the final IgE gate determined per heavy chain transcript assembly. ND = not determined. (C) As a reference, putative basophils (CD19-IgE+) display higher IgE surface expression than IgE+ B cells.

[0053] [Figure 9] Figures 9A-9G. Single-cell RNA-seq data overview and quality control. (A) Cells were sequenced to a depth of approximately 1-2 million reads / cell in five libraries. (B) Histogram of genes per cell. Cells expressing less than 950 genes were discarded. (C) Rarefaction curves depicting the number of detected genes as a function of sequencing depth for eight randomly selected cells in each B-cell subtype. Solid and shaded areas correspond to the mean and 95% confidence interval for gene numbers, respectively. (D) Read mapping distribution for retained cells. The majority of uniquely mapped reads (Ensembl reference annotation) and multiply mapped reads belonged mostly to RNA18S5 repeat sequences on chr21 and unplaced scaffolds. (E) Read mapping across gene bodies showed minimal 3' or 5' bias. (F) V gene assembly length histogram per strand. (G) PCA on the top 500 most variable genes before (top) and after (bottom) batch correction.

[0054] [Figure 10] 10A-10D. Supplementary data supporting B cell subtype classification. PB (striped) have greater FACS forward and side scatter (A), more cDNA after SmartSeq2 preamplification (B), and greater gene expression of antibody light and heavy chains (C) compared to naive / memory B cell subsets (unstriped). (D) Top differentially expressed genes for each subset.

[0055] [Figure 11-1]Figures 11A-11K. B cell comparison across isotypes. (A-I) Number of cells with a given V and J gene per isotype and chain. (A) IGHM. (B) IGHD. (C) IGHG3. (D) IGHG1. (E) IGHA1. (F) IGHG2. (G) IGHG4. (H) IGHE. (I) IGHA2. (J) Heavy chain mutation frequency per isotype. (K) Relative utilization of lambda and kappa light chains per isotype. [Figure 11-2] Same as above [Figure 11-3] Same as above

[0056] [Figure 12-1] Figures 12A-12H. Antibody specificity and affinity characterization. (A) Semi-quantitative indirect ELISA of convergent antibodies, control, PA13P1H08 variants, and antibodies from other clone families. Human IgG isotype control (abcam #ab206195) served as a negative control, and positive controls included anti-Ara h mouse monoclonal antibody purchased from Indoor Biotechnologies. (B-G) Kinetic characterization of antibodies binding to Ara h2 (B-D) and Ara h3 (E-G) using biolayer interferometry. Antibodies are named and described herein. (H) Indirect ELISA showing binding of recombinant monoclonal antibodies from subjects PA11, PA12, PA13, PA14, PA15, and PA16 (rows) to allergen extract, natural peanut allergen Ara h2, and bovine serum albumin (BSA) (columns). The isotype of each antibody is indicated on the left. Higher values ​​indicate stronger binding. OD = optical density. The values ​​shown correspond to those after subtraction of the human IgG isotype control. Only tested antibodies with any OD value above 0.25 are shown. [Figure 12-2] Same as above [Figure 12-3] Same as above [Figure 12-4] Same as above [Figure 12-5] Same as above [Figure 12-6] Same as above

[0057] [Figure 13-1] 13A-13E. Heavy and light chain sequences used in constructing the PA13P1H08 antibody variants. (A) Origin of the predicted naive heavy chain CDR3 and surrounding amino acids. (B) Unmodified and reverted heavy chain sequences, as well as sequences in which a region of the heavy chain has been reverted to the predicted naive rearrangement. Designations with an "r" prefix indicate that only that region has been reverted. FWR=framework. (C) Origin of the predicted naive light chain CDR3 and surrounding amino acids. (D) Unmodified and reverted light chain sequences. (E) Light chain sequence from another antibody, namely PA12P4H03, which we confirmed by ELISA does not bind any peanut allergens. [Figure 13-2] Same as above [Figure 13-3] Same as above

[0058] [Figure 14] Figure 14. GLT splicing for all isotypes. Note that for clarity of illustration, only the first three constant region exons are shown for each isotype.

[0059] [Figure 15] Figure 15. IGV coverage histograms and splice junctions for the ighe constant region locus showing εGLT in single mouse B cells stimulated with IL-4, LPS, and BAFF (Wu et al. 2017). The arrow indicates the ighe GLT splice donor.

[0060] [Figure 16]Figure 16. Pairwise CDR3 sequence identity of heavy chain CDR3 sequences (SEQ ID NOs: 19, 35, 42, 12, 27, 4) from clones PA12P3F10, PA12P3D08, PA12P1C07, PA13P1E10, PA13P3G09, and PA13P1H08, as well as three heavy chain CDR3 sequences from multiple patients in a separate peanut allergy immune repertoire sequencing study (62). Each sequence from this separate study shares at least 70% identity with one or more sequences from this study. All sequences share IGHV3-30 and IGHJ6 gene segments and have a 17 amino acid long CDR3.

[0061] [Figure 17-1] Figure 17A. Microarray scan confirming the absence of background interactions of secondary goat anti-human IgG and control mouse monoclonal anti-HA antibodies with antigen-derived peptides. The control antibodies yielded the expected HA control spot pattern (white dots) that constitute the peptide microarray.

[0062] [Figure 17-2] Figures 17B and 17C. Microarray epitope mapping for Ara h2 ("Ah2") and Ara h3 ("Ah3") peptides of PA13P1H08. (B) Microarray scan illustrating antibody binding to antigenic peptide (light grey) and the expected HA control spot pattern (white dots). (C) Microarray fluorescence intensity per antigenic peptide. Ara h2 motifs with high intensity are annotated.

[0063] [Figure 18]Figure 18. Plasma IgE levels to common food allergens and Aspergillus fumigatus and Aspergillus niger in milk-allergic subject PA01 and Aspergillus study subject 10033201. NP = not performed. Total IgE was 353 kU / L and 3528 kU / L for PA01 and 10033201, respectively. Assays were performed by the CLIA-certified Johns Hopkins University Dermatology, Allergy, and Clinical Immunology Reference Laboratory using the ImmunoCAP system.

[0064] [Figure 19] Figure 19. Indirect ELISA showing binding of recombinant monoclonal antibodies (columns) to antigens (rows). Antigens include extracts of Aspergillus fumigatus, Aspergillus niger, and Aspergillus nidulans, and purified recombinant allergen, Aspergillus fumigatus 1 (rAsp f1). Bovine serum albumin (BSA) serves as a negative antigen control. Monoclonal antibodies against Asp f1 ("anti-Asp f1") serve as a positive control for this allergen. Higher values ​​indicate stronger binding. OD = optical density. hIgG = human IgG isotype control.

[0065] [Figure 20] Figure 20. Indirect ELISA showing binding of recombinant monoclonal antibodies from subject PA01 (rows) to allergen extract, natural peanut allergen Ara h2, and BSA (columns). The isotype of each antibody is indicated on the left. Higher values ​​indicate stronger binding. OD = optical density. Values ​​shown correspond to values ​​after subtraction of a human IgG isotype control. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0066] Table 1 Table 1 includes the protein and nucleic acid sequences discussed herein. Polypeptide sequences are provided using standard one-letter code. Those skilled in the art who are provided with the amino acid sequences will understand that the amino acid sequences may be encoded by a defined set of nucleotide sequences, such that the reader and the inventors have a possession of the nucleotide sequence that encodes each amino acid sequence. The nucleic acid sequence encoding the polynucleotide may be a naturally occurring human sequence. In some embodiments, the nucleic acid sequence encoding the polynucleotide is not a naturally occurring human sequence. The nucleic acid sequence encoding the polynucleotide may be a sequence that is codon-optimized for expression in human cells or specific cell types, eukaryotic cells, bacterial cells, or other. Codon optimization, in which a codon is replaced by another codon that codes for the same amino acid and has a higher frequency of occurrence in a particular host cell, can be performed to improve the host's ability to produce the polypeptide encoded by the nucleic acid [see, for example, Mauro, BioDrugs 32(1):69-81, 2018, and Kato, Int J Mol Sci. 20(4), 2019].

[0067] In certain embodiments, it is contemplated that variant sequences can be used in the methods and compositions disclosed herein.For example, in one aspect, an antibody is described that has a heavy chain variable region that comprises an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:1, and a light chain variable region that comprises an amino acid sequence that has at least 90% sequence identity to SEQ ID NO:5.The degree of sequence identity or similarity can be determined using methods known in the art.In one approach, the identity of two nucleotide or polypeptide sequences or subsequences is calculated as the percentage of identical or equal positions after the sequences are aligned and, if necessary, gaps are introduced to achieve the maximum percent sequence identity. Methods of sequence alignment are known in the art and include, but are not limited to, the Needleman-Wunsch global alignment algorithm [Needleman and Wunsch (1970) J. Mol. Biol. 48(3):443-453 (30)], the Smith and Waterman local homology search algorithm [Smith, Temple F. & Waterman, Michael S. (1981) J. Mol. Biol. 147 (1): 195-197.], manual alignment and inspection, or computerized implementations of these algorithms, such as the "needle" program distributed as part of the EMBOSS software package [Rice, P. et al., Trends in Genetics 16(6): 276-277 (31), version 6.3.1 available from EMBnet for a variety of sources].

[0068] In certain embodiments, it is contemplated that the methods or compositions described herein differ from the polypeptide sequences provided herein (e.g., Table 1) by one or more amino acid substitutions. In some embodiments, the sequences may have at least 90% sequence identity (or other degree of sequence identity as disclosed herein below) to the sequences or combinations of sequences described herein. In one embodiment, the polypeptide sequence differs from the reference sequence (e.g., Table 1) by one amino acid substitution. In one embodiment, the polypeptide sequence differs from the reference sequence (e.g., Table 1) by two amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by two amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by three amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by four amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by five amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by six amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by seven amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by eight amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by 9 amino acid substitutions. In one embodiment, the polypeptide sequence differs from the reference sequence by 10 amino acid substitutions. In certain embodiments, the polypeptide sequence differs from the reference sequence by 1-10 amino acid substitutions, and optionally 1-5 amino acid substitutions. In some cases, amino acid substitutions that do not change the basic properties (e.g., binding specificity) compared to the reference sequence are selected. In some cases, amino acid substitutions that change the binding affinity but not the binding specificity are selected. In some cases, the substitutions are selected to change properties known in the art or described herein below (e.g., substitutions that affect effector function or half-life). In some embodiments, the amino acid substitutions are conservative substitutions. Conservative amino acid substitutions are recognized in the art as the substitution of one amino acid with another amino acid that has similar properties, such as polarity, charge, hydrophobicity, and aromaticity.Conservative amino acid substitutions can also be made on the basis of the side chain characteristics of the amino acid, such as containing sulfur, hydroxyl, or amide. Non-limiting examples of conservative amino acid substitutions are described below. [Table 1]

[0069] I. Introduction In one aspect, the present disclosure provides human allergen-specific monoclonal antibodies and methods for generating human allergen-specific monoclonal antibodies from single IgE or IgG4 expressing human B cells. IgE antibodies, the least abundant class of antibodies in humans, are known to cause symptoms of allergic reactions. For example, food allergy symptoms ranging from urticaria to potentially fatal anaphylaxis are due to degranulation of mast cells and basophils induced by recognition of allergic food proteins by surface-bound IgE antibodies. Despite this central role in immune and allergic diseases, human IgE antibodies remain poorly characterized due to their scarcity. Fitzsimmons et al., Front Immunol., 2014, 5:61. Similarly, knowledge surrounding the IgG4 isotype is lacking, but interest is growing due to its potential role in mediating immunotherapy via antigen blockade and the reduced clinical allergen reactivity associated with early allergen exposure. Tordesillas et al., Immunity, 2017, 47:32-50.

[0070] The present disclosure provides therapeutic methods for treating a human subject with an allergy or alleviating one or more allergic symptoms in a human subject using one or more allergen-specific monoclonal antibodies or antigen-binding portions thereof disclosed herein. In some embodiments, without intending to be bound by a particular mechanism, the allergen-specific monoclonal antibodies disclosed herein are used therapeutically as blocking antibodies, which is often referred to as passive immunotherapy.

[0071] As described herein, the method of the present disclosure can be used to generate a pool of genotype-confirmed IgE or IgG4 single B cells that are candidates for producing antibodies with high affinity for the allergen of interest from a sample from a human subject with an allergy to an antigen of interest. As described in the Examples section below, it has been found that analyzing the cDNA sequence of the immunoglobulin heavy chain constant region to identify the isotype of single B cells avoids the problem of incorrect isotype characterization that is known to occur when B cell isotype is determined based on sorting cells by cell surface markers (e.g., typically performed in FACS cell surface staining). This problem of incorrect isotype characterization is known to be particularly prevalent in IgE B cells because the marker CD23 is a "low affinity" IgE receptor that captures IgE on the surface of non-IgE B cells. See Berkowska et al., J Allergy Clin Immunol, 2014, 134:688-697. Thus, the disclosed method generates a pool of single B cells that are much more likely to produce antibodies with high affinity for allergens. Moreover, the antibodies generated according to the disclosed method are among the highest affinity natural human antibodies discovered to date and have been found to exhibit cross-reactivity to different antigens.

[0072] II. Definition The scope of the present invention is limited only by the appended claims, and therefore the terms used herein are merely for describing certain embodiments and are not intended to be limiting.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.In this specification and the following claims, reference is made to some terms that are defined to have the following meanings unless a contrary intention is apparent.In some cases, terms that have a commonly understood meaning are defined herein for clarity of description and / or for ready reference, and the inclusion of such definitions herein should not be interpreted as amounting to a substantial difference to the definition of the term commonly understood in the art.

[0073] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are approximations that are varied (+) or (-) by 0.1 or 1.0 increments, as appropriate. It is to be understood, although not always explicitly stated, that all numerical designations are preceded by the term "about."

[0074] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes a plurality of compounds.

[0075] The term "comprising" is intended to mean that the compounds, compositions, and methods include the recited elements, but do not exclude other elements. "Consisting essentially of," when used to define compounds, compositions, and methods, is intended to mean excluding other elements that may materially affect the basic and novel characteristics of the claimed invention. "Consisting of" is intended to mean excluding any element, step, or ingredient not specified in the claim. Embodiments defined by each of these transition terms are within the scope of the invention.

[0076] The term "allergen" refers to a substance that induces an immune response in a subject, resulting in an allergic reaction by the subject.

[0077] As used herein, the term "antibody" refers to a polypeptide encoded by an immunoglobulin gene or a functional fragment thereof that specifically binds and recognizes an antigen. As used herein, the term "antibody" also includes antibody fragments that retain binding specificity, including but not limited to Fab, F(ab')2, Fv, and scFv. Recognized immunoglobulin genes include kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as a myriad of immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively.

[0078] An exemplary immunoglobulin (antibody) structural unit comprises two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The N-terminus of each chain defines a variable region of about 100-110 or more amino acids primarily responsible for antigen recognition. Thus, the term "variable heavy chain" or "VH" refers to the variable region of an immunoglobulin heavy chain, including Fv, scFv, dsFv, or Fab, and the term "variable light chain" or "VL" refers to the variable region of an immunoglobulin light chain, including Fv, scFv, dsFv, or Fab.

[0079] The term "variable region" refers to the domain in an antibody heavy or light chain that confers specificity to the antibody with respect to binding to an antigen. Typically, antibody variable regions contain four conserved "framework" regions interspersed with three hypervariable "complementarity determining regions".

[0080] The term "complementarity determining region" or "CDR" refers to the three hypervariable regions in each chain that interrupt the four framework regions established by the light and heavy chain variable regions. The CDRs are primarily responsible for binding to an epitope of an antigen. The CDRs of each chain are typically numbered sequentially from the N-terminus and are referred to as CDR1, CDR2, and CDR3, and are typically identified by the chain in which a particular CDR is located. Thus, the VH CDR3 is located in the variable domain of the heavy chain of the antibody in which it is found, and the VL CDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found.

[0081] As noted, the portion of the variable region that is not contained in the CDR is referred to as the framework. The "framework regions" of different light or heavy chains are relatively conserved in species. The framework regions of an antibody, i.e., the combined framework regions of the constituent light and heavy chains, serve to position and align the CDRs in three-dimensional space. Framework sequences can be obtained from public DNA databases or published references that contain germline antibody gene sequences. For example, germline DNA sequences for human heavy and light chain variable region genes can be found in the "VBASE2" germline variable gene sequence database for human and mouse sequences.

[0082] The amino acid sequences of the CDRs and framework regions can be determined using various definitions well known in the art. The positions and lengths of the CDRs are precisely defined by Kabat et al., Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, 1983, 1987, etc. For example, Johnson and Wu, Nucleic Acids Res. 2000 Jan 1; 28(1): 214-218, Johnson et al., Nucleic Acids Res., 29:205-206 (2001), Chothia & Lesk, (1987) J. Mol. Biol. 196, 901-917, Chothia et al. (1989) Nature 342, 877-883, Chothia et al. (1992) J. Mol. Biol. 227, 799-817, Al-Lazikani et al., J. Mol. Biol 1997, 273(4)), and MacCallum et al., J. Mol. Biol., 262:732-745 (1996). See also International Immunogenetics Database (IMGT), AbM, and Observed Antigen Contact.

[0083] The terms "antigen-binding portion" and "antigen-binding fragment" are used interchangeably herein and refer to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., an allergen, such as Ara h2 or Ara h3). Examples of antibody-binding fragments include, but are not limited to, Fab fragments (monovalent fragments consisting of the VL, VH, CL, and CH1 domains), F(ab')2 fragments (bivalent fragments comprising two Fab fragments linked by a disulfide bridge at the hinge region), single chain Fvs (scFvs), disulfide-linked Fvs (dsFvs), complementarity determining regions (CDRs), VL (light chain variable region), VH (heavy chain variable region), nanobodies, and any combinations thereof, or any other functional portion of an immunoglobulin peptide capable of binding to a target antigen. Antibodies and antigen-binding portions thereof include domain-specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g. monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains. Exemplary configurations of variable and constant domains that may be found within the antigen-binding fragment of an antibody of the invention include (a) VH-CHI, (b) VH-CH2, (c) VH-CH3, (d) VH-CH1-CH2, (e) VH-Ch1-Ch2-Ch3, (f) VH-Ch2-Ch3, (g) VH-CL, (h) VL-CH1, (i) VL-Ch2, (X) VL-Ch3, (j) VL-CH1-CH2, (k) VL-CH1-CH2-CH3, (l) VL-CH2-CH3, and (m) VL-CL [see, e.g., FUNDAMENTAL IMMUNOLOGY (Paul ed., 4th ed. 2001), Gruber et al. (1994) J Immunol. 152:5368-5374, McCartney, et al., 1995 Protein Eng. 8:301-314, Shukra et al., 2014, “Production of recombinant antibodies using bacteriophages” Eur J Microbiol Immunol (Bp). 4(2): 91-98, Todorovska, 2001, “Design and application of diabodies, triabodies and tetrabodies for cancer targeting” J Immunol Methods; 248(1-2):47-66, Salvador et al., 2019, “Nanobody: outstanding features for diagnostic and therapeutic applications” Anal Bioanal Chem. 411(9):1703-1713, Gill et al., 2006, “Biopharmaceutical drug discovery using novel protein scaffolds.” Curr Opin Biotechnol., (6):653-8, and Ubah et al., 2016, “Phage Display Derived IgNAR V Region Binding Domains for Therapeutic Development” Curr Pharm Des. 22(43):6519-6526. See also.

[0084] The term "epitope" refers to the area or region of an antigen to which an antibody specifically binds, i.e., the area or region that is in physical contact with the antibody, and may include a few amino acids or a portion of a few amino acids, such as 5 or 6 or more, such as 20 or more amino acids, or a portion of those amino acids. In some cases, epitopes include non-proteinaceous components, such as from carbohydrates, nucleic acids, or lipids. In some cases, epitopes are three-dimensional sites. Thus, for example, if the target is a protein, the epitope may be composed of consecutive amino acids, or amino acids from different parts of the protein that are in close proximity due to protein folding (e.g., discontinuous epitopes).

[0085] "Monoclonal antibody" refers to an antibody that consists or consists essentially of antibody molecules that are produced by a single clone of cells or a single cell line and that are identical in primary amino acid sequence. In some embodiments, a monoclonal antibody preparation comprises a population of antibodies that are identical except for mutations that occur during monoclonal antibody production and that bind to the same epitope of an antigen. Unless otherwise specified or clear from the context, the term "monoclonal antibody" includes synthetic antibodies and antigen-binding fragments thereof.

[0086] "Human antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies of the present disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). The term "human antibody" is not intended to include chimeric or humanized antibodies in which CDR sequences derived from the germline or immune cells of a non-human species (e.g., mouse) are grafted onto human framework sequences.

[0087] The term "specifically binds" refers to a molecule (e.g., an antibody or antibody fragment) that binds to a target in a sample with greater affinity, avidity, more readily, and / or with greater duration than it binds to a non-target compound. In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a target (e.g., an allergen, e.g., Ara h2 or Ara h3) is an antibody or antigen-binding portion thereof that binds to the target with at least 2-fold greater affinity than a non-target compound, e.g., at least 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 25-fold, or 50-fold or more greater affinity. For example, in some embodiments, an antibody that specifically binds to an allergen target, e.g., Ara h2 or Ara h3, can typically bind to the allergen target with at least 2-fold greater affinity than to a non-allergen target. It will be understood by those skilled in the art that an antibody that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target.

[0088] The term "binding affinity" as used herein refers to the strength of non-covalent interactions between two molecules, such as an antibody (or an antigen-binding fragment thereof) and an antigen. Thus, for example, the term may refer to a 1:1 interaction between an antibody (or an antigen-binding fragment thereof) and an antigen, unless otherwise indicated or clear from the context. Binding affinity can be quantified by measuring the equilibrium dissociation constant (KD), which refers to the dissociation rate constant (kd, time-1) divided by the association rate constant (ka, time-1M-1). KD can be determined, for example, by measuring the kinetics of complex formation and dissociation using surface plasmon resonance (SPR) methods, such as the Biacore™ system; binding equilibrium exclusion methods such as KinExA®; and biolayer interferometry (e.g., using the ForteBio® Octet platform). As used herein, "binding affinity" includes not only regular binding affinity, e.g., binding affinity that reflects a 1:1 interaction between an antibody (or antigen-binding fragment thereof) and an antigen, but also apparent affinity from which a KD is calculated, which may reflect strong binding.

[0089] The term "cross-react" as used herein refers to the ability of an antibody to bind to two or more antigens. As a non-limiting example, in some embodiments, an antibody that specifically binds to a first allergen target (e.g., a first peanut allergen, such as Ara h2) can exhibit cross-reactivity with a second allergen target (e.g., a second peanut allergen, such as Ara h3).

[0090] The term "isolated" when used in connection with a nucleic acid or protein (e.g., an antibody) refers to the nucleic acid or protein being essentially free of other cellular components with which it is naturally associated. The nucleic acid or protein is preferably in a homogeneous state. Purity and homogeneity are typically determined using analytical chemistry techniques, such as electrophoresis (e.g., polyacrylamide gel electrophoresis) or chromatography (e.g., high performance liquid chromatography). In some embodiments, the isolated nucleic acid or protein (e.g., an antibody) is at least 85% pure, at least 90% pure, at least 95% pure, or at least 99% pure.

[0091] The terms "polypeptide", "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. These terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of corresponding naturally occurring amino acids, as well as naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. As used herein, these terms encompass any length of amino acid chain, including full-length proteins, in which amino acid residues are linked by covalent peptide bonds.

[0092] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are subsequently modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as naturally occurring amino acids, i.e., a carbon bound to a hydrogen, a carboxyl group, an amino group, and an R group, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids. "Amino acid mimetics" refer to chemical compounds that have a structure that differs from the general chemical structure of amino acids, but function similarly to naturally occurring amino acids.

[0093] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Similarly, nucleotides may be referred to by their commonly accepted single-letter codes.

[0094] As used herein, the terms "nucleic acid" and "polynucleotide" are used interchangeably. The use of the term "polynucleotide" includes oligonucleotides (i.e., short polynucleotides). The term can also refer to deoxyribonucleotides, ribonucleotides, and naturally occurring variants, including, but not limited to, synthetic and / or non-naturally occurring nucleic acids (i.e., nucleic acid analogs or modified backbone residues or linkages), such as phosphorothioates, phosphoramidates, methyl phosphonates, chiral methyl phosphonates, 2-O-methyl ribonucleotides, peptide nucleic acids (PNAs), etc. Unless otherwise indicated, a particular nucleic acid sequence implicitly encompasses not only the sequence explicitly shown, but also conservatively modified variants (e.g., degenerate codon substitutions) and complementary sequences of the particular nucleic acid sequence. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (see, e.g., Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al, J. Biol. Chem. 260:2605-2608 (1985); Rossolini et al, Mol. Cell. Probes 8:91-98 (1994)).

[0095] The term "sample" as used herein refers to a biological sample obtained from a human or non-human mammalian subject. In some embodiments, the sample includes blood, blood fractions, or blood products (e.g., serum, plasma, platelets, red blood cells, peripheral blood mononuclear cells, etc.); sputum or saliva; stool, urine, other biological fluids (e.g., lymphatic fluid, saliva, prostatic fluid, gastric fluid, intestinal fluid, renal fluid, lung fluid, cerebrospinal fluid, etc.), tissues (e.g., kidney, lung, liver, heart, brain, neural tissue, thyroid, eye, skeletal muscle, cartilage, or bone tissue), cultured cells (e.g., primary cultures, explants, transformed cells, or stem cells), or biopsy samples.

[0096] The terms "subject" and "patient", as used interchangeably herein, refer to mammals, including, but not limited to, humans, non-human primates, rodents (e.g., rats, mice, and guinea pigs), rabbits, cows, pigs, horses, and other mammalian species. In one embodiment, the subject or patient is a human.

[0097] The terms "treat", "treating" and "treatment" refer to any indication of success in treating or improving an injury, disease or condition, including any objective or subjective parameter, such as remission; relief; reduction in symptoms or making the injury, disease or condition more tolerable to the subject; slowing down the rate of deterioration or exacerbation; making the end point of deterioration less debilitating; and / or improving the physical or mental health of the subject. The treatment or improvement of symptoms can be based on objective or subjective parameters. The effect of treatment can be compared to an individual or pool of individuals not receiving treatment, or to the same patient at different time points before or during treatment.

[0098] The term "pharmaceutical composition" refers to a composition suitable for administration to a subject. Generally, pharmaceutical compositions are sterile and preferably free of contaminants that may induce an undesirable response in a subject. Pharmaceutical compositions can be designed for administration to a subject in need thereof via a number of different routes of administration, including oral, intravenous, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, inhalation, and the like.

[0099] The term "pharmaceutical acceptable excipient" refers to non-active pharmaceutical ingredients that are biologically or pharmacologically compatible for use in humans or animals, such as, but not limited to, buffers, carriers, or preservatives.

[0100] As used herein, a "therapeutic amount" or "therapeutically effective amount" of an agent (e.g., a monoclonal antibody disclosed herein) is an amount of agent that treats, improves, ameliorates, alleviates, improves patient survival, increases survival time or survival rate, reduces symptoms, makes an injury, disease, or condition (e.g., allergy) more tolerable, slows the rate of deterioration or exacerbation, or improves the patient's physical or mental health. For example, with respect to a given parameter, a therapeutically effective amount may exhibit at least a 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or 100% increase or decrease in therapeutic efficacy. The therapeutic efficacy may also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount may have at least 1.2-fold, 1.5-fold, 2-fold, or 5-fold or more efficacy over control.

[0101] The terms "administer", "administered" or "administering" refer to a method of delivering an agent, compound or composition to a desired site of biological action. These methods include, but are not limited to, topical delivery, parenteral delivery, intravenous delivery, intradermal delivery, intramuscular delivery, rectal delivery, or intraperitoneal delivery. Administration techniques that may be used with the agents and methods described herein include, for example, those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon, and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, PA.

[0102] III. Methods for generating allergen-specific antibodies In one aspect, a method for generating allergen-specific monoclonal antibodies from a human sample is provided. In some embodiments, the method comprises: (a) isolating a single B cell from a sample from a human subject, the subject being allergic to an allergen; (b) generating cDNA from the single B cell of step (a), the cDNA sequence comprising a first sequence encoding all or a portion of an immunoglobulin heavy chain and a second sequence encoding all or a portion of an immunoglobulin light chain; (c) determining the sequence of the cDNA from step (b); (d) analyzing the sequence determined in step (c) to identify a single B cell comprising a first sequence comprising an IgE constant region or an IgG4 constant region; (e) identifying from the single B cell of step (d) (i) a heavy chain variable region sequence in an immunoglobulin heavy chain that comprises an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain that is co-expressed with the immunoglobulin heavy chain in the same single B cell; (f) expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (e); and (g) identifying one or more antibodies from step (f) that specifically bind to the allergen; Includes.

[0103] Study population and samples In some embodiments, the method of generating allergen-specific monoclonal antibodies comprises isolating B cells from a biological sample from a human subject. In some embodiments, the sample comprises whole blood, peripheral blood, or leukoreduction preparations. In some embodiments, the sample comprises peripheral blood mononuclear cells (PBMCs). In some embodiments, the sample comprises tissue from a human subject, such as tonsil tissue, spleen, or bone marrow. Methods for isolating B cells from blood and tissue samples are described in the art. See, for example, Heine et al., Curr Protoc. Immunol., 2011, 94:7.5.1-7.5.14, and Zuccolo et al., BMC Immunol, 2009, 10:30, doi:10.1186 / 1471-2172-10-30.

[0104] In some embodiments, the allergen-specific antibody is generated from a human subject having an allergy to an allergen. In some embodiments, the human subject having an allergy is an adult. In some embodiments, the human subject having an allergy is a minor. In some embodiments, the human subject has an allergy to a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the human subject has an allergy to two or more allergens, such as two or more of a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the human subject has an allergy to 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more allergens. In some embodiments, the human subject has an allergy to two or more different types of antigens (allergens) in a class of allergens, for example, an allergy to two or more different food allergens (e.g., an allergy to two or more different peanut antigens, or an allergy to a peanut allergen and a non-peanut allergen, such as a nut, egg, or milk allergen), or an allergy to two or more different fungal allergens (e.g., an allergy to two or more different species of Aspergillus). In some embodiments, the human subject has an allergy to two or more different classes of allergens (e.g., an allergy to one or more food allergens and one or more plant allergens). In some embodiments, the human subject has an allergy to only one class of allergens (e.g., the subject has an allergy to one or more food allergens but not to a non-food allergen, or the subject has an allergy to one or more fungal allergens but not to a non-fungal allergen).

[0105] In some embodiments, the human subject has an allergy to a food allergen.In some embodiments, the food allergen is a milk allergen, an egg allergen, a nut allergen, a tree nut allergen, a fish allergen, a shellfish allergen, a soybean allergen, a legume allergen, a seed allergen, or a wheat allergen.In some embodiments, the food allergen is a peanut allergen.

[0106] In some embodiments, the human subject has an allergy to a plant allergen or a fungal allergen. In some embodiments, the allergen is a fungal allergen (e.g., Aspergillus, e.g., Asp. fumigatus, Asp. niger, or Asp. nidulans). In some embodiments, the allergen is a pollen allergen (e.g., tree pollen, grass pollen, or grass pollen) or a mold allergen. In some embodiments, the human subject has an allergy to an animal allergen. In some embodiments, the allergen is a dander allergen or an insect sting.

[0107] In some embodiments, the method for producing allergen-specific monoclonal antibodies does not include immunizing a human subject with or exposing a human subject to an allergen prior to obtaining a sample from the subject.

[0108] B cell isolation and screening In some embodiments, single B cells are isolated from a sample from a subject with allergy to an allergen.In some embodiments, single B cells are separated into separate compartments, such as separate wells of a multi-well plate, encapsulated in droplets, or dispersed in microwells.In some embodiments, at least 10, 50, 100, 500, 1,000, 5,000, or 10,000 or more B cells are isolated from a sample and separated into separate compartments.

[0109] In some embodiments, the isolating step comprises sorting the cells in the sample by fluorescence-activated cell sorting (FACS). FACS sorting can be used to sort cells based on cell surface marker expression, cell size, and / or granularity, and deliver the cells individually to wells, such as 96-well or 384-well tissue culture or PCR plates. Methods for isolating and purifying cell populations by FACS have been described in the art. See, for example, Basu et al., J Vis Exp, 2010, 41:1546, doi:10.3791 / 1546.

[0110] In some embodiments, the droplet microfluidic platform can be used to dispense single B cells into separate droplets. In some embodiments, the nucleic acid (e.g., mRNA) of a single cell in a droplet is labeled with a nucleotide sequence that is unique to the droplet, such as a unique molecular identifier barcode nucleotide sequence, thereby allowing downstream processing steps for sequences from multiple B cells to be performed in a single reaction vessel. Methods for encapsulating single cells into droplets have been described in the art. See, for example, Macosko et al., Cell, 2015, 161: 1202-1214; Zhang et al., Scientific Reports, 2017, 7:41192, doi:10.1038 / srep41192.

[0111] In some embodiments, the cells are distributed into microwells designed to capture single cells. Methods for single cell microwell capture are described in the art. See, e.g., Han et al., Cell, 2018, 172:5, doi:10.1016 / j.cell.2018.02.001.

[0112] In some embodiments, cells from the sample are screened for the presence, absence, or level of expression of one or more markers, and single B cells are isolated based on the presence or level of expression of one or more B cell markers (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 or more markers). In some embodiments, cells are screened for the presence, absence, or level of expression of one or more cell surface B cell markers, such as, but not limited to, CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79. In some embodiments, cells are determined to be B cells if they are positive for one or more of the B cell markers, for example, positive for one or more of CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79. In some embodiments, single CD19+ B cells are isolated.

[0113] In some embodiments, cells from the sample are screened for the presence, absence, or level of expression of one or more immunoglobulin isotypes, such as, but not limited to, IgE, IgG, IgM, IgA, or IgD, or subclasses thereof. In some embodiments, the isolated single B cells are selected for expression of cell surface IgE and / or expression of cell surface IgG4. In some embodiments, the single B cells are isolated without selection for expression of one or more immunoglobulin isotypes (e.g., without selection for expression of cell surface IgE and / or expression of cell surface IgG4).

[0114] IgE selection In some embodiments, the method comprises isolating a single B cell selected for expression of cell surface IgE. In some embodiments, the isolating step comprises contacting the cells of the sample with an anti-human IgE antibody and selecting cells expressing IgE on the cell surface. In some embodiments, the isolating step comprises contacting the cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and an anti-human IgE antibody, and selecting cells expressing the B cell marker and expressing IgE on the cell surface. In some embodiments, the isolating step comprises contacting the cells of the sample with an anti-human CD19 antibody and an anti-human IgE antibody, and selecting CD19+IgE-expressing B cells.

[0115] In some embodiments, the isolating step comprises contacting cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), an anti-human IgE antibody, and an antibody against one or more immunoglobulin isotypes (e.g., an anti-human IgG antibody, an anti-human IgM antibody, an anti-human IgA antibody, and / or an anti-human IgD antibody) or an antibody against a subclass thereof, and selecting cells that express the B cell marker, express IgE on the cell surface, and do not express detectable levels of one or more other immunoglobulin isotypes being screened. In some embodiments, the isolating step comprises contacting cells of the sample with one or more of an anti-human CD19 antibody, an anti-human IgE antibody, and an anti-human IgG antibody, an anti-human IgM antibody, an anti-human IgA antibody, or an anti-human IgD antibody, and isolating cells of the sample that are negative for IgG, IgM, IgA, or IgD cell surface expression. + Selecting IgE-expressing B cells.

[0116] In some embodiments, the isolating step comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgE antibody, an anti-human IgM antibody, and an anti-human IgG antibody, and selecting CD19+IgM-IgG-IgE expressing B cells. In some embodiments, the isolating step comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgE antibody, an anti-human IgM antibody, an anti-human IgG antibody, an anti-human IgA antibody, and an anti-human IgD antibody, and selecting CD19+IgM-IgG-IgE expressing B cells. + IgM - IgG - IgA - IgD - Selecting IgE-expressing B cells.

[0117] In some embodiments, the isolating step comprises contacting cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and an antibody against a non-IgE isotype (e.g., an anti-human IgG antibody, an anti-human IgM antibody, an anti-human IgA antibody, and an anti-human IgD antibody) or a subclass thereof, and selecting cells that express a B cell marker and do not express detectable levels of a non-IgE isotype. In some embodiments, the isolating step comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgG antibody, an anti-human IgA antibody, and an anti-human IgD antibody, and selecting cells that express a B cell marker and do not express detectable levels of a non-IgE isotype. + IgM - IgG - IgA - IgD - Selecting B cells.

[0118] In some embodiments, the method includes isolating IgE-expressing B cells that are antibody-secreting B cells (e.g., plasmablasts or plasma cells). In some embodiments, the method includes isolating IgE-expressing B cells that are memory B cells. In some embodiments, the method includes isolating IgE-expressing antibody-secreting B cells and IgE-expressing memory B cells.

[0119] IgG4 selection In some embodiments, the method comprises isolating a single B cell that is selected for expression of cell surface IgG4. In some embodiments, the isolating step comprises contacting the cells of the sample with an anti-human IgG4 antibody and selecting cells that express IgG4 on the cell surface. In some embodiments, the isolating step comprises contacting the cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and an anti-human IgG4 antibody, and selecting cells that express a B cell marker and express IgG4 on the cell surface. In some embodiments, the isolating step comprises contacting the cells of the sample with an anti-human CD19 antibody and an anti-human IgG4 antibody, and selecting cells that express a CD19 cell marker and express IgG4 on the cell surface. + Selecting IgG4-expressing B cells.

[0120] In some embodiments, the isolating step comprises contacting cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), an anti-human IgG4 antibody, and an antibody against one or more IgG subclasses (e.g., an anti-human IgG1 antibody, an anti-human IgG2 antibody, an anti-human IgG3 antibody), and selecting cells that express a B cell marker, express IgG4 on the cell surface, and do not express detectable levels of one or more other IgG subclasses being screened. In some embodiments, the isolating step comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgG1 antibody, an anti-human IgG2 antibody, an anti-human IgG3 antibody, and an anti-human IgG4 antibody, and selecting cells that express a B cell marker, express IgG4 on the cell surface, and do not express detectable levels of one or more other IgG subclasses being screened. + IgG1 - IgG2 - IgG3 - Selecting IgG4-expressing B cells.

[0121] In some embodiments, the isolating step comprises contacting cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), an anti-human IgG4 antibody, and an antibody against one or more immunoglobulin isotypes (e.g., an anti-human IgE antibody, an anti-human IgM antibody, an anti-human IgA antibody, and / or an anti-human IgD antibody) or an antibody against a subclass thereof, and selecting cells that express the B cell marker, express IgG4 on the cell surface, and do not express detectable levels of one or more other immunoglobulin isotypes being screened. In some embodiments, the isolating step comprises contacting cells of the sample with one or more of an anti-human CD19 antibody, an anti-human IgG4 antibody, and an anti-human IgE antibody, an anti-human IgM antibody, an anti-human IgA antibody, or an anti-human IgD antibody, and isolating cells of the sample that are negative for IgE, IgM, IgA, or IgD cell surface expression. + Selecting IgG4-expressing B cells.

[0122] In some embodiments, the isolating step comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgG4 antibody, an anti-human IgM antibody, and an anti-human IgG antibody, and detecting CD19 + IgM - IgE - In some embodiments, the isolating step includes contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgG4 antibody, an anti-human IgM antibody, an anti-human IgE antibody, an anti-human IgA antibody, and an anti-human IgD antibody, and selecting IgG4-expressing B cells. + IgM - IgE - IgA - IgD - Selecting IgG4-expressing B cells.

[0123] In some embodiments, the isolating step comprises contacting cells of the sample with an antibody against a B cell marker (e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and an antibody against a non-IgG isotype (e.g., an anti-human IgE antibody, an anti-human IgM antibody, an anti-human IgA antibody, and an anti-human IgD antibody) or an antibody against a non-IgG4 isotype, and selecting cells that express a B cell marker and do not express detectable levels of non-IgG or non-IgG4 isotypes. In some embodiments, the isolating step comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgE antibody, an anti-human IgA antibody, and an anti-human IgD antibody, and selecting cells that express a B cell marker and do not express detectable levels of non-IgG or non-IgG4 isotypes. + IgM - IgE - IgA - IgD - In some embodiments, the isolating step includes contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgE antibody, an anti-human IgA antibody, an anti-human IgD antibody, an anti-human IgG1 antibody, an anti-human IgG2 antibody, and an anti-human IgG3 antibody, and selecting CD19 B cells. + IgM - IgE - IgA - IgD - IgG1 - IgG2 - IgG3 - Selecting B cells.

[0124] In some embodiments, the method comprises isolating IgG4-expressing B cells that are antibody-secreting B cells. In some embodiments, the method comprises isolating IgG4-expressing B cells that are memory B cells. In some embodiments, the method comprises isolating IgG4-expressing antibody-secreting B cells and IgG4-expressing memory B cells.

[0125] cDNA generation and sequencing In some embodiments, cDNA is generated from a single B cell isolated from a sample (e.g., from a single B cell that has been screened for expression of an immunoglobulin isotype, such as IgE or IgG4, or from a single B cell that has not been screened for expression of an immunoglobulin isotype). In some embodiments, a cDNA library is prepared from the single B cell. In some embodiments, in the cDNA generated for each single B cell, the cDNA sequence comprises a sequence encoding an immunoglobulin heavy chain and a sequence encoding an immunoglobulin light chain.

[0126] In some embodiments, cDNA is generated by reverse transcribing cDNA sequences from RNA (e.g., total RNA or mRNA) from a single B cell and amplifying the cDNA sequences. With respect to generating cDNA, in some embodiments, a single B cell is lysed and a cDNA sequence is reverse transcribed from mRNA present in the cell lysate. In some embodiments, RNA is isolated from a single B cell and cDNA is reverse transcribed from the isolated RNA.

[0127] In some embodiments, the method comprises amplifying the transcriptome of single B cell.For example, in some embodiments, the method comprises reverse transcribing RNA (e.g., polyadenylated mRNA) to synthesize cDNA, and then amplifying the cDNA, for example, by PCR.Exemplary methods for reverse transcribing polyadenylated mRNA and amplifying the transcriptome of single cell are described in Darmanis et al., Cell Reports, 2017, 21:1399-1410, and Picelli et al., Nature Protocols, 9, 2014, 171-181.

[0128] In some embodiments, the method includes amplifying immunoglobulin heavy and light chain sequences from a single B cell. For example, in some embodiments, the method includes reverse transcribing RNA (e.g., total RNA) to synthesize cDNA, and then amplifying the cDNA, for example, by PCR using primers for immunoglobulin heavy chain variable and constant regions. In some embodiments, the method includes reverse transcribing RNA using immunoglobulin-specific primers (e.g., constant region-specific primers) to synthesize cDNA comprising immunoglobulin sequences, and then amplifying the cDNA using primers for immunoglobulin heavy chain variable and constant regions. An exemplary method for amplifying immunoglobulin heavy and light chain sequences from a single cell is described in Tiller et al., J. Immunol. Methods, 2008, 329:112-124.

[0129] After the cDNA is generated, in some embodiments, the method comprises sequencing the cDNA. In some embodiments, the cDNA is subjected to sequencing. In some embodiments, the method comprises sequencing the transcriptome of a single B cell. In some embodiments, the method comprises sequencing a target gene (e.g., an immunoglobulin gene, such as an immunoglobulin heavy chain variable and constant region, and an immunoglobulin light chain variable and constant region).

[0130] Sequencing methods, including methods for high-throughput sequencing, are known in the art.For example, such sequencing techniques include, but are not limited to, pyrosequencing, sequencing by ligation, single molecule sequencing, sequence-by-synthesis (SBS), massively parallel clone, massively parallel single molecule SBS, massively parallel single molecule real-time, massively parallel single molecule real-time nanopore technology, etc.Morozova and Marra provide a review of some such techniques in Genomics, 92: 255 (2008), the entirety of which is incorporated herein by reference.

[0131] In some embodiments, the sequencing comprises high-throughput sequencing. In high-throughput sequencing, parallel sequencing reactions using multiple templates and multiple primers allow rapid sequencing of genomes or large portions of genomes. High-throughput sequencing methods include methods that typically use and do not use template amplification. Sequencing methods that utilize amplification include pyrosequencing, as commercialized by Roche in its 454 technology platform (e.g., GS20 and GS FLX), clonal array formation and sequencing by synthesis (SBS) chemistry, as commercialized by Illumina with systems such as NextSeq, and the Supported Oligonucleotide Ligation and Detection (SOLiD) platform, as commercialized by Applied Biosystems. Non-amplification approaches, also known as single molecule sequencing, are exemplified by the HeliScope platform, as commercialized by Helicos BioSciences, and the platforms commercialized by VisiGen, Oxford Nanopore Technologies Ltd., Life Technologies / Ion Torrent, and Pacific Biosciences, respectively.

[0132] In some embodiments, an Illumina sequencing platform such as NextSeq is used. This sequencing technology utilizes clonal array formation and sequencing by synthesis to generate sequences on a large scale. In this method, sequencing templates are immobilized on a flow cell surface, and then solid-phase amplification creates copies (up to 1,000 identical copies) of each template molecule in close proximity to form dense "clusters" of polynucleotide sequences. For sequencing clusters, fluorescently labeled nucleotides are used to sequence the clusters on the flow cell surface in parallel. For each sequencing cycle, a single labeled reversible terminator-linked dNTP is added to the nucleic acid strand. The sequence of the incorporated nucleotide is determined by detecting the fluorescence after incorporation, and the fluorescent dye is then removed before the addition of the next cycle of dNTPs, resulting in base-by-base sequencing. Typically, sequence read lengths range from about 30 nucleotides to more than 150 nucleotides. If the target cDNA of interest has a longer length, the sequences can be bioinformatically reassembled based on the overlap between the short sequencing reads to determine the sequence of the full-length target cDNA.

[0133] Identification of B cells with IgE or IgG4 isotypes In some embodiments, after the sequence of the cDNA has been determined for the single B cell, the method includes analyzing the sequence of the cDNA to identify a single B cell that expresses an immunoglobulin heavy chain having a constant region of the IgE isotype and / or IgG4 isotype. As described herein, it has been found that determining the isotype of a B cell based on the sequence of the heavy chain transcript rather than FACS immunoglobulin surface staining substantially reduces the number of false positive IgE cells in the B cell population, thus resulting in a population of B cells that are much more likely to produce antibodies that specifically bind to an allergen to which the human subject from whom the B cells originated is allergic.

[0134] In some embodiments, the method includes identifying a sequence encoding an immunoglobulin heavy chain that includes an IgE constant region. In some embodiments, the method includes identifying a sequence encoding an immunoglobulin heavy chain that includes an IgG4 constant region. In some embodiments, the cDNA sequence is analyzed by comparing the sequence to a known IgE constant region sequence or a known IgG4 constant region sequence. For example, a cDNA sequence of interest (e.g., a "test" sequence from a B cell) can be compared to a known IgE or IgG4 constant region sequence (e.g., a "reference" sequence) by aligning the sequence. Methods of aligning sequences for comparison are known in the art. Optimal alignment of sequences for comparison can be performed, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by manual alignment and visual inspection. Preferred algorithms for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms described in Altschul et al. (1990) J. Mol. Biol. 215: 403-410 and Altschul et al. (1977) Nucleic Acids Res. 25: 3389-3402, respectively. Software for performing BLAST analyses is publicly available through the website of the National Center for Biotechnology Information (NCBI).

[0135] With regard to comparing the test sequence to an IgE or IgG4 constant region reference sequence, in some embodiments the reference sequence is a published sequence, such as a publicly available IgE or IgG4 constant region sequence in the ImMunoGeneTics (IMGT) database. See, e.g., Camacho et al., BMC Bioinformatics, 2009, 10:421; Lefranc et al., Nucleic Acids Res, 2009, 37:D1006-1012. Methods for analyzing test sequences to identify sequences that contain immunoglobulin heavy chain constant regions of the IgE and / or IgG4 isotypes are also described in Table 1 below.

[0136] In some embodiments, in addition to analyzing the sequence to identify and select single B cells that contain an immunoglobulin heavy chain sequence that includes an IgE constant region or an IgG4 constant region, the method further comprises determining the sequence and / or level of expression of one or more other genes in the single B cell. For example, in some embodiments, the method comprises determining the sequence and / or level of expression of a set of genes that are a "signature" for a particular type of B cell.

[0137] In some embodiments, for a B cell identified as having an immunoglobulin heavy chain comprising an IgE constant region or an IgG4 constant region, the method further comprises identifying, from the same B cell, a heavy chain variable region sequence expressed by the cell and a light chain variable region sequence expressed by the cell.

[0138] Antibody expression Typically, for a single B cell identified as having a cDNA containing an IgE or IgG4 constant region sequence, the heavy and light chain variable region sequences from the single B cell are candidate antibody sequences with specificity for the allergen of interest. Thus, in some embodiments, the method includes expressing an antibody containing the heavy and light chain variable region sequences from the single B cell, and identifying whether the expressed antibody specifically binds to the allergen of interest. Methods for expressing and purifying recombinant antibodies are described in the art. See, for example, Frenzel et al., Front Immunol., 2013, 4:217, doi:10.3389 / fimmu.2013.00217; Siegemund et al., Methods Mol Biol., 2014, 1131:273-295.

[0139] In some embodiments, the heavy and light chain variable region sequences from a single B cell are amplified from the single B cell and cloned into an expression vector. In some embodiments, the heavy and light chain variable region sequences from a single B cell are synthesized. In some embodiments, the heavy and / or light chain variable region sequences are codon-optimized to increase antibody expression, for example, by an expression system. See, e.g., Ayyar et al., Methods, 2017, 116:51-62.

[0140] The heavy and light chain variable region sequences from a single B cell can be expressed using any number of expression systems, including prokaryotic and eukaryotic expression systems. In some embodiments, the expression system is a mammalian cell expression, such as a hybridoma, or a CHO or HEK293 cell expression system. Many such systems are widely available from commercial suppliers. Cell expression systems are also described in the art. See, for example, Kunert and Reinhart, 2016, "Advances in recombinant antibody manufacturing" Appl Microbiol Biotechnol. 100:3451-61; Jager et al., BMC Proc., 2015, 9:P40, doi:10.1186 / 1753-6561-9-S9-P40, and references cited therein. In some embodiments, the heavy and light chains are expressed using a single vector, for example in a dicistronic expression unit or under the control of different promoters. In other embodiments, the heavy and light chains are expressed using separate vectors. In some embodiments, the expression vector for expressing the heavy and / or light chain variable region sequences disclosed herein is a vector that contains the constant region of the desired heavy chain isotype or light chain subclass. For example, the heavy chain variable region sequence disclosed herein can be cloned into a vector that contains the human IgG (e.g., IgG1, IgG2, IgG3, or IgG4) heavy chain constant region, and the light chain variable region sequence disclosed herein can be cloned into a vector that contains the human lambda or kappa light chain constant region.

[0141] After an antibody comprising the heavy and light chain variable region sequences from a single B cell disclosed herein is expressed and purified, in some embodiments, the method includes determining whether the antibody specifically binds to an allergen. Methods for analyzing binding affinity and binding kinetics are known in the art. See, for example, Ernst et al., Determination of Equilibrium Dissociation Constants, Therapeutic Monoclonal Antibodies (Wiley & Sons ed. 2009). These methods include, but are not limited to, solid-phase binding assays (e.g., ELISA assays), immunoprecipitation, surface plasmon resonance [SPR, e.g., Biacore™ (GE Healthcare, Piscataway, NJ)], equilibrium exclusion assays [e.g., KinExA®], flow cytometry, fluorescence-activated cell sorting (FACS), biolayer interferometry [e.g., Octet™ (ForteBio, Inc., Menlo Park, CA)], and Western blot analysis. SPR techniques are reviewed, for example, in Hahnfeld et al. Determination of Kinetic Data Using SPR Biosensors, Molecular Diagnosis of Infectious Diseases (2004). In a typical SPR experiment, one interactant (target or targeting agent) is immobilized on an SPR-active gold-coated glass slide in a flow cell, and a sample containing the other interactant is introduced to flow over the surface. When light of a given wavelength is directed at the surface, changes in the optical reflectance of the gold indicate binding and binding kinetics. In some embodiments, the equilibrium exclusion method is used to determine affinity. This technique is described, for example, in Darling et al., Assay and Drug Development Technologies Vol. 2, number 6 647-657 (2004).In some embodiments, biolayer interferometry assays are used to determine affinity. This technique is described, for example, in Wilson et al., Biochemistry and Molecular Biology Education, 38:400-407 (2010); Dysinger et al., J. Immunol. Methods, 379:30-41 (2012).

[0142] In some embodiments, the expressed antibody specifically binds to the allergen with high affinity. In some embodiments, the antibody has a binding affinity (KD) for the allergen of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM. In some embodiments, the antibody binds to the allergen with a binding affinity (KD) of between 1 nM and 250 nM.

[0143] Nucleic Acids, Vectors, and Host Cells In some embodiments, the allergen-specific monoclonal antibodies described herein are prepared using recombinant methods. Thus, in some aspects, the invention provides isolated nucleic acids comprising a nucleic acid sequence encoding any of the allergen-specific monoclonal antibodies described herein (e.g., any one or more of the CDRs described herein); vectors comprising such nucleic acids; and host cells into which the nucleic acids are introduced and which are used to replicate the nucleic acids encoding the antibodies and / or to express the antibodies. In some embodiments, the host cells are eukaryotic, e.g., human cells, e.g., HEK-293.

[0144] In some embodiments, a polynucleotide (e.g., an isolated polynucleotide) comprises a nucleotide sequence that encodes an antibody or antigen-binding portion thereof described herein (e.g., described in Section IV, below). In some embodiments, a polynucleotide comprises a nucleotide sequence that encodes one or more amino acid sequences (e.g., a CDR, a heavy chain variable region, or a light chain variable region) disclosed in Table 1, below.

[0145] In a further aspect, methods of making an allergen-specific monoclonal antibody as described herein are provided. In some embodiments, the methods include culturing a host cell as described herein (e.g., a host cell expressing a polynucleotide or vector as described herein) under conditions suitable for expression of the antibody. In some embodiments, the antibody is then removed from the host cell (or host cell culture medium).

[0146] Suitable vectors containing a polynucleotide encoding an antibody or fragment thereof of the present disclosure include cloning vectors and expression vectors. The cloning vector selected may vary depending on the host cell intended for use, but useful cloning vectors generally have the ability to replicate autonomously, may have a single target for a specific restriction endonuclease, and / or may carry a gene as a marker that can be used in selecting clones containing the vector. Examples include plasmids and bacterial viruses, such as pUC18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322, pMB9, ColE1, pCR1, RP4, phage DNA, and shuttle vectors such as pSA3 and pAT28. Cloning vectors are available from commercial vendors such as BioRad, Stratagene, and Invitrogen.

[0147] Expression vector generally refers to a replicable polynucleotide construct that contains the nucleic acid of the present disclosure.Expression vector can replicate in host cell either as episome or integral part of chromosomal DNA.Suitable expression vector includes but is not limited to plasmid, adenovirus, adeno-associated virus, retrovirus, and any other vector.

[0148] IV. Monoclonal antibodies that specifically bind to allergens In another aspect, allergen-specific monoclonal antibodies and antigen-binding portions thereof are provided that are generated from a human sample according to the methods disclosed herein. In some embodiments, the monoclonal antibodies are antibodies generated according to the methods disclosed in Section III above. In some embodiments, the monoclonal antibodies are antibodies generated from a sample from a human subject with an allergy to a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen, and specifically bind to the food allergen, the plant allergen, the fungal allergen, the animal allergen, the dust mite allergen, the drug allergen, the cosmetic allergen, or the latex allergen.

[0149] In some embodiments, the antibodies described herein are full length antibodies, Fab, Fab', F(ab')2, Fab'-SH, Fv, single chain antibodies, or single chain Fv (scFv) antibodies. In some embodiments, the antibodies described herein comprise an IgG4 constant region. In some embodiments, the antibodies described herein are monospecific antibodies. In some embodiments, the antibodies described herein are multispecific antibodies. In particular, the antibodies described herein can be bispecific antibodies that bind to two different allergens. For example, in some embodiments, the antibodies described herein can bind to a peanut allergen and a tree nut allergen. In some embodiments, the antibodies described herein can bind to a peanut allergen and a milk allergen. In some embodiments, the antibodies described herein can bind to a peanut allergen and a fungal allergen. In some embodiments, the antibodies described herein can bind to a tree nut allergen and a milk allergen. In some embodiments, the antibodies described herein can bind to a tree nut allergen and a fungal allergen. In some embodiments, the antibodies described herein are capable of binding to milk allergens and fungal allergens.

[0150] In some embodiments, the monoclonal antibody or antigen-binding portion thereof comprises: (a) isolating a single B cell from a sample from a human subject, the subject being allergic to an allergen; (b) generating cDNA from the single B cell of step (a), the cDNA sequence comprising a first sequence encoding all or a portion of an immunoglobulin heavy chain and a second sequence encoding all or a portion of an immunoglobulin light chain; (c) determining the sequence of the cDNA from step (b); (d) analyzing the sequence determined in step (c) to identify a single B cell comprising a first sequence comprising an IgE constant region or an IgG4 constant region; (e) identifying from the single B cell of step (d) (i) a heavy chain variable region sequence in an immunoglobulin heavy chain that comprises an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain that is co-expressed with the immunoglobulin heavy chain in the same single B cell. The allergen-specific antibody comprises a heavy chain variable region sequence and a light chain variable region sequence identified according to a method comprising:

[0151] In some embodiments, the heavy and light chain variable regions are from a B cell that comprises an immunoglobulin that comprises an IgE constant region. In some embodiments, the heavy and light chain variable regions are from a B cell that comprises an immunoglobulin that comprises an IgG4 constant region.

[0152] In some embodiments, the monoclonal antibody or antigen-binding portion thereof comprises: (a) a heavy chain variable region sequence derived from an immunoglobulin heavy chain from an IgE- or IgG4-producing single B cell from a human subject allergic to an allergen; (b) heavy chain IgG constant region sequence; (c) a light chain variable region sequence derived from an immunoglobulin light chain from an IgE- or IgG4-producing single B cell from a human subject; (d) a light chain constant region sequence of the same class as the immunoglobulin light chain of (c) It is an allergen-specific antibody comprising:

[0153] In some embodiments, the monoclonal antibody comprises heavy and light chain variable region sequences derived from an IgE-producing human B cell. In some embodiments, the monoclonal antibody comprises heavy and light chain variable region sequences derived from an IgG4-producing human B cell.

[0154] Characterization of allergen-specific monoclonal antibodies In some embodiments, the monoclonal antibody is an antibody that specifically binds to a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the monoclonal antibody is an antibody that specifically binds to a food allergen, such as a milk allergen, an egg allergen, a nut allergen, a fish allergen, a shellfish allergen, a soy allergen, a legume allergen, a seed allergen, or a wheat allergen. In some embodiments, the monoclonal antibody is an antibody that specifically binds to a peanut allergen. In some embodiments, the monoclonal antibody is an antibody that specifically binds to a milk allergen. In some embodiments, the monoclonal antibody is an antibody that specifically binds to an egg allergen.

[0155] In some embodiments, the monoclonal antibody specifically binds to an allergen (e.g., a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen) with a binding affinity (KD) of less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM. In some embodiments, the antibody binds to an allergen with a binding affinity (KD) of between 1 nM and 250 nM.

[0156] In some embodiments, the monoclonal antibody exhibits cross-reactivity with at least two different antigens (e.g., allergens), such as at least two food allergens, at least two plant allergens, at least two fungal allergens, at least two animal allergens, at least two dust mite allergens, at least two drug allergens, at least two cosmetic allergens, or at least two latex allergens.In some embodiments, the monoclonal antibody exhibits cross-reactivity with at least two milk allergens, at least two egg allergens, at least two nut allergens, at least two fish allergens, at least two shellfish allergens, at least two soybean allergens, at least two legume allergens, at least two seed allergens, or at least two wheat allergens.It will be understood by those skilled in the art that many different allergens, such as many plant food allergens, can be broadly classified into a small number of protein families. For example, more than half of all plant food allergens can be categorized into one of the following four structural protein families: prolamin superfamily, cupin superfamily, profilin, and Bet v-1 related protein. It will also be understood by those skilled in the art that for a particular type of allergen (e.g., "peanut" allergen), there may be more than one allergenic peptide or protein. As a non-limiting example, there are 12 known peanut allergens. See Mueller et al., Curr Allergy Asthma Rep, 2014, 14:429. In some embodiments, the monoclonal antibody exhibits cross-reactivity with two or more different antigens that are different types or classes of antigens. As a non-limiting example, in some embodiments, the monoclonal antibody exhibits cross-reactivity with an antigen that is a peanut allergen and an antigen that is a nut (e.g., tree nut) allergen.

[0157] In embodiments in which a monoclonal antibody exhibits cross-reactivity with at least two different antigens (e.g., allergens), in some embodiments the monoclonal antibody specifically binds to at least one allergen with a KD of less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM. In some embodiments, the monoclonal antibody specifically binds to a first antigen (e.g., a first allergen) with a KD of less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM. In some embodiments, the monoclonal antibody specifically binds to a second antigen (e.g., a second allergen) with a KD of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM.

[0158] Engineered mutations in variable regions In some embodiments, the heavy and / or light chain variable regions of the monoclonal antibody have sequences identical to those encoded by an IgE- or IgG4-producing single B cell from a human subject with an allergy to an allergen. In some embodiments, the heavy and / or light chain variable regions of the monoclonal antibody contain one or more modifications, such as amino acid substitutions, deletions, or insertions.

[0159] As described in the Examples section below, the heavy and / or light chain variable region sequences of the antibodies described herein (e.g., peanut allergen-specific monoclonal antibodies, e.g., clone PA13P1H08, clone PA13P1E10, clone PA12P3F10, clone PA13P3G09, clone PA12P3D08, clone PA12P1C07, clone PA15P1D12, clone PA15P1D05, or clone PA13P1H08 variants) can be engineered to include one or more mutations in the heavy and / or light chain variable region sequences. In some embodiments, the engineered mutations improve the binding affinity of the antibody for an allergen. In some embodiments, the engineered mutations improve the cross-reactivity of the antibody for a second allergen.

[0160] In some embodiments, the engineered mutation is a mutation in one or more CDRs, such as an amino acid substitution in the heavy and / or light chain CDRs described herein. In some embodiments, the engineered mutation is a mutation in one or more framework regions, such as an amino acid substitution in the heavy and / or light chain framework regions. In some embodiments, the engineered mutation is a reversion of a region of the heavy and / or light chain sequence to a predicted naive sequence. Methods for determining predicted naive immunoglobulin sequences are described in the art. See, for example, Magnani et al., PLoS Negl Trop Dis, 2017, 11:e0005655, doi:10.1371 / journal.pntd.0005655.

[0161] In some embodiments, affinity maturation is used to engineer additional mutations that enhance the binding affinity of the antibody for an allergen or enhance the cross-reactivity of the antibody for a second allergen. Methods for performing affinity maturation are known in the art. See, for example, Renaut et al., Methods Mol Biol, 2012, 907:451-461.

[0162] Constant regions and isotype switching In some embodiments, the monoclonal antibodies comprise heavy and light chain variable region sequences derived from an IgE- or IgG4-producing human B cell, and further comprise a kappa or lambda light chain constant region. In some embodiments, the light chain constant region (kappa or lambda) is from the same type of light chain (i.e., kappa or lambda) as the light chain variable region derived from an IgE- or IgG4-producing human B cell; as a non-limiting example, if the IgE-producing human B cell contains a kappa light chain, then the monoclonal antibodies produced comprise the light chain variable region from an IgE-producing B cell and further comprise a kappa light chain constant region.

[0163] In some embodiments, the monoclonal antibody comprises heavy and light chain variable region sequences derived from an IgE- or IgG4-producing human B cell, and further comprises a heavy chain constant region having an IgG isotype (e.g., IgG4), an IgA isotype (e.g., IgA1), an IgM isotype, an IgD isotype, or derived from an IgG, IgA, IgM, or IgD isotype (e.g., a modified IgG4 constant region). It will be appreciated by those skilled in the art that different heavy chain isotypes (IgA, IgD, IgE, IgG, and IgM) have different effector functions mediated by the heavy chain constant region, and that for certain uses it may be desirable to have an antibody having an effector function of a particular isotype (e.g., IgG).

[0164] In some embodiments, the monoclonal antibody comprises an unmodified (i.e., wild-type) human IgG, IgA, IgM, or IgD constant region. In some embodiments, the monoclonal antibody comprises an unmodified human IgG1 constant region, an unmodified human IgG2 constant region, an unmodified human IgG3 constant region, an unmodified human IgG4 constant region, an unmodified human IgA1 constant region, an unmodified human IgA2 constant region, an unmodified human IgM constant region, or an unmodified human IgD constant region. In some embodiments, the monoclonal antibody comprises a heavy chain constant region that comprises one or more modifications. It will be understood by those skilled in the art that modifications, such as amino acid substitutions, can be made at one or more residues in the heavy chain constant region that modulate effector function. In some embodiments, the modifications reduce effector function, for example, resulting in a reduced ability to induce a certain biological function upon binding to an Fc receptor expressed on an effector cell that mediates the effector function. In some embodiments, the modifications (e.g., amino acid substitutions) prevent Fab arm exchange in vivo, which may introduce undesirable effects and reduce the therapeutic effectiveness of the antibody. See, e.g., Silva et al., J Biol Chem, 2015, 280:5462-5469.

[0165] In some embodiments, the monoclonal antibodies comprise an unmodified (i.e., wild-type) human IgM constant region, a human IgD constant region, a human IgG constant region derived from IgG1, IgG2, IgG3, or IgG4, or a human IgA constant region derived from IgA1 or IgA2, and comprise one or more modifications that modulate effector function. In some embodiments, the monoclonal antibodies comprise a human IgM constant region, a human IgD constant region, a human IgG constant region derived from IgG1, IgG2, IgG3, or IgG4, or a human IgA constant region derived from IgA1 or IgA2. In some embodiments, the monoclonal antibodies comprise an unmodified (i.e., wild-type) human IgM constant region, a human IgD constant region, a human IgG constant region derived from IgG1, IgG2, IgG3, or IgG4, or a human IgA constant region derived from IgA1 or IgA2, and comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more modifications (e.g., amino acid substitutions). In some embodiments, the constant region comprises mutations (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more amino acid substitutions) that reduce effector function.

[0166] In some embodiments, the monoclonal antibody comprises CDR sequences, heavy chain variable region, and / or light chain variable region from an antibody derived from an IgE or IgG4 B cell as described herein (e.g., disclosed in Table 1 below), and further comprises a heavy chain constant region and / or a light chain constant region that is heterologous to the antibody from an IgE or IgG4 B cell from which the CDR sequences and / or variable region sequences are derived. For example, in some embodiments, the monoclonal antibody comprises CDR sequences and / or variable region sequences of an antibody from an IgE B cell, and further comprises a heavy chain constant region and a light chain constant region that is heterologous to the antibody from an IgE B cell (e.g., the heavy chain constant region and / or light chain constant region is a wild-type or modified IgG1, IgG2, IgG3, or IgG4 constant region, or the heavy chain constant region and / or light chain constant region comprises one or more modifications (e.g., amino acid substitutions) compared to the unmodified constant region of an antibody from an IgE B cell).

[0167] Antibodies that specifically bind to peanut and / or tree nut allergens In some embodiments, the monoclonal antibodies or antigen-binding portions thereof disclosed herein specifically bind to a peanut allergen and / or a tree nut allergen. In some embodiments, the monoclonal antibodies specifically bind to a peanut allergen. In some embodiments, the monoclonal antibodies specifically bind to a peanut allergen that is Ara h1, Ara h2, Ara h3, or Ara h6.

[0168] In some embodiments, the monoclonal antibody exhibits cross-reactivity with at least two peanut allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity with two or more of the peanut allergens Ara h1, Ara h2, Ara h3, and Ara h6. In some embodiments, the monoclonal antibody specifically binds to at least one of the peanut allergens with a KD of less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM. In some embodiments, the monoclonal antibody specifically binds to a first peanut allergen with a KD of less than 100 nM (e.g., less than 50 nM, less than 10 nM, less than 5 nM, or less than 1 nM) and to a second peanut allergen with a KD of less than 1 μM (e.g., less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM). In some embodiments, the monoclonal antibody specifically binds to Ara h2 with a KD of less than 100 nM (e.g., less than 50 nM, less than 10 nM, less than 5 nM, or less than 1 nM) and to Ara h1, Ara h3, or Ara h6 with a KD of less than 1 μM (e.g., less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM).

[0169] In some embodiments, the monoclonal antibody recognizes an epitope comprising or consisting of the amino acid motif DPYSPS (SEQ ID NO: 704). In some embodiments, the monoclonal antibody recognizes an epitope comprising or consisting of the amino acid sequence DSYGRDPYSPS (SEQ ID NO: 705), YSPSQDPYSPS (SEQ ID NO: 706), or PDRRDPYSPS (SEQ ID NO: 707).

[0170] In some embodiments, the monoclonal antibody or antigen-binding portion thereof specifically binds to a tree nut allergen. In some embodiments, the tree nut allergen is a cashew, pistachio, almond, pine nut, pecan, walnut, hazelnut, or macadamia nut allergen. In some embodiments, the monoclonal antibody exhibits cross-reactivity with at least two tree nut allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity with both a cashew allergen and a pistachio allergen. In some embodiments, the monoclonal antibody exhibits cross-reactivity with both a pecan allergen and a walnut allergen. In some embodiments, the monoclonal antibody exhibits cross-reactivity with two or more of a pecan, walnut, hazelnut, and macadamia nut allergen. In some embodiments, the monoclonal antibody specifically binds to at least one of the above tree nut allergens with a KD of less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM.

[0171] In some embodiments, the monoclonal antibody or antigen-binding portion thereof specifically binds to a peanut allergen and a tree nut allergen, hi some embodiments, the monoclonal antibody specifically binds to a peanut allergen and one or more (e.g., 1, 2, 3, or more) of a cashew, pistachio, almond, pine nut, pecan, walnut, hazelnut, or macadamia nut allergen.

[0172] Peanut-specific antibody sequences In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen (e.g., specifically binds to Ara h1, Ara h2, Ara h3, or Ara h6) comprises the heavy chain and / or light chain CDRs disclosed below in Table 1. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises the light chain and / or heavy chain variable region sequences disclosed below in Table 1. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain variable region comprising an amino acid sequence that has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a heavy chain variable region sequence disclosed in Table 1 and that contains CDR1, CDR2, and CDR3 that are identical to the CDRs of the heavy chain variable region sequence, and a light chain variable region comprising an amino acid sequence that has at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a light chain variable region sequence disclosed in Table 1 and that contains CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0173] In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen (e.g., that specifically binds Ara h1, Ara h2, Ara h3, or Ara h6) is selected from the group consisting of the following antibodies described herein: clone PA13P1H08, clone PA13P1E10, clone PA12P3F10, clone PA13P3G09, clone PA12P3D08, clone PA12P1C07, clone PA15P1D12, clone PA15P1D05, mutants of clone PA13P1H08 (e.g., RR mutant, RN mutant, NR mutant, r mutant, r variant ... and / or rFWR-N mutant), clone PA12P4D02, clone PA12P3E09, clone PA12P3E11, clone PA12P1D02, clone PA12P1G11, clone PA13P1H03, clone PA12P3C01, or clone PA12P3E04. The amino acid sequences of the CDRs, light chain variable domain (VL), and heavy chain variable domain (VH) of clone PA13P1H08, clone PA13P1E10, clone PA12P3F10, clone PA13P3G09, clone PA12P3D08, clone PA12P1C07, clone PA15P1D12, clone PA15P1D05, clone PA13P1H08 variant, clone PA12P4D02, clone PA12P3E09, clone PA12P3E11, clone PA12P1D02, clone PA12P1G11, clone PA13P1H03, clone PA12P3C01, and clone PA12P3E04 are set forth in Table 1 below.

[0174] In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen is (a) a heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199, 341, 348, 409, 459, or 593; (b) a heavy chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349, 410, 460, 539, or 594; (c) a heavy chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201, 350, 411, 461, 540, or 595; (d) a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343, 352, 413, 463, 542, or 597; (e) a light chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 7, 15, 22, 30, 38, 52, 78, 86, 126, 149, 196, 345, 353, or 598; and (f) a light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354, 414, 464, 543, or 599. The compound may include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0175] In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen is (a) SEQ ID NOs: 2, 3, 4, 6, 7, and 8, respectively; or (b) SEQ ID NOs: 10, 11, 12, 14, 15, and 8, respectively; or (c) SEQ ID NOs: 17, 18, 19, 21, 22, and 23, respectively; or (d) SEQ ID NOs: 25, 26, 27, 29, 30, and 31, respectively; or (e) SEQ ID NOs: 33, 34, 35, 37, 38, and 39, respectively; or (f) SEQ ID NOs: 41, 34, 35, 37, 38, and 39, respectively; or (g) SEQ ID NOs: 47, 48, 49, 51, 52, and 53, respectively; or (h) SEQ ID NOs: 47, 48, 55, 51, 52, and 53, respectively; or (i) SEQ ID NOs: 58, 59, 60, 62, 30, and 63, respectively; or (j) SEQ ID NOs: 58, 59, 60, 6, 7, and 8, respectively; or (k) SEQ ID NOs: 2, 3, 4, 62, 30, and 63, respectively; or (l) SEQ ID NOs: 58, 3, 4, 6, 7, and 8, respectively; or (m) SEQ ID NOs: 2, 59, 4, 6, 7, and 8, respectively; or (n) SEQ ID NOs: 2, 3, 60, 6, 7, and 8, respectively; or (o) SEQ ID NOs: 129, 130, 131, 133, 126, and 134, respectively; or (p) SEQ ID NOs: 341, 342, 343, 345, 78, and 346, respectively; or (q) SEQ ID NOs: 348, 349, 350, 352, 353, and 354, respectively; or (r) SEQ ID NOs: 199, 200, 201, 203, 149, and 204, respectively; or (s) SEQ ID NOs: 409, 410, 411, 413, 86, and 414, respectively; or (t) SEQ ID NOs: 459, 460, 461, 463, 196, and 464, respectively; or (u) SEQ ID NOs: 113, 539, 540, 542, 196, and 543, respectively; or (v) SEQ ID NOs: 593, 594, 595, 597, 598, and 599, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0176] In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592. and a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596.

[0177] In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592, and comprises a heavy chain variable region comprising an amino acid sequence that includes CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a light chain variable region that has at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596 and comprises an amino acid sequence that includes CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO. (a) a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592, and including CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO; and (b) a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596, and including CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO. Includes.

[0178] Clone PA13P1H08 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:2, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:3, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:4, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:6, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:7, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:8.

[0179] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:1, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs:2, 3, and 4, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:5, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs:6, 7, and 8, respectively. Includes.

[0180] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:1, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:5. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:1, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:5.

[0181] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 2, 3, 4, 6, 7, and 8, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5).

[0182] Clone PA13P1E10 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 14, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 15, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 8.

[0183] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 9, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 13, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 14, 15, and 8, respectively. Includes.

[0184] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:9, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:13. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:9, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:13.

[0185] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 10, 11, 12, 14, 15, and 8, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 13).

[0186] Clone PA12P3F10 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 17, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 18, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 19, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 22, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 23.

[0187] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 16, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 20, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 21, 22, and 23, respectively. Includes.

[0188] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 16, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 20. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 16, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:20.

[0189] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, 19, 21, 22, and 23, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 16 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 20).

[0190] Clone PA13P3G09 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:25, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:26, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:27, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:29, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:30, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:31.

[0191] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 24, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 25, 26, and 27, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 28, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 29, 30, and 31, respectively. Includes.

[0192] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:24, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:28. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:24, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:28.

[0193] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 25, 26, 27, 29, 30, and 31, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 24 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 28).

[0194] Clone PA12P3D08 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 33, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 35, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 37, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 38, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 39.

[0195] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 32, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 33, 34, and 35, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 36, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 37, 38, and 39, respectively. Includes.

[0196] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 32, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 36. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 32, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:36.

[0197] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 33, 34, 35, 37, 38, and 39, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 32 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 36).

[0198] Clone PA12P1C07 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 41, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 34, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 42, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 44, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 45.

[0199] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 40, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 41, 34, and 42, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 43, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 44, 30, and 45, respectively. Includes.

[0200] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 40, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 43. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 40, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 43.

[0201] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 41, 34, 42, 44, 30, and 45, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 40 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 43).

[0202] Clone PA15P1D12 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 47, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 48, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 49, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 52, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 53.

[0203] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 46, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 47, 48, and 49, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 50, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 51, 52, and 53, respectively. Includes.

[0204] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 46, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 50. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 46, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:50.

[0205] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 41, 34, 42, 44, 30, and 45, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 46 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 50).

[0206] Clone PA15P1D05 In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 47, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 48, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 55, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 51, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 52, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 53.

[0207] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 54, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 47, 48, and 55, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 56, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 51, 52, and 53, respectively. Includes.

[0208] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 54, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 56. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:56.

[0209] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 47, 48, 55, 51, 52, and 53, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 54 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 56).

[0210] Mutant sequences In some embodiments, the allergen-specific monoclonal antibody comprises one or more mutations (e.g., amino acid substitutions) in one or more CDR, heavy chain, and / or light chain sequences disclosed herein (e.g., one or more mutations in one or more CDR, heavy chain, and / or light chain sequences of clone PA13P1H08, clone PA13P1E10, clone PA12P3F10, clone PA13P3G09, clone PA12P3D08, clone PA12P1C07, clone PA15P1D12, clone PA15P1D05). In some embodiments, one or more substitutions are made in the CDR, heavy chain, or light chain sequences of clone PA13P1H08. As described in the Examples section below, mutants were made from clone PA13P1H08, in which one or more regions in the heavy and / or light chain were reverted to the predicted naive rearrangement. It was found that antibody sequences comprising naive light or heavy chain sequences are capable of binding to the peanut allergen Ara h2. It was also surprisingly found that variants of clone PA13P1H08 comprising reverted CDR-H2 sequences exhibit significantly improved cross-reactivity to a second peanut allergen (Ara h3) in addition to having sub-nanomolar affinity for the Ara h2 peanut allergen. Thus, in some embodiments, the mutations are amino acid substitutions that revert at least a portion of the sequence from the "unmodified" form of the clone (i.e., the CDR, heavy chain variable region, or light chain variable region sequence of the clone disclosed in Table 1) to the predicted naive immunoglobulin sequence.

[0211] In some embodiments, the allergen-specific monoclonal antibody comprises one or more mutant sequences of clone PA13P1H08 mutants disclosed herein. In some embodiments, the antibody comprises one of the sequences of clone PA13P1H08 mutant "RR", in which both the heavy and light chain variable regions of clone PA13P1H08 have been reverted to the predicted naive rearrangement. In some embodiments, the antibody comprises a reverted heavy chain variable region sequence comprising SEQ ID NO:57. In some embodiments, the antibody comprises a reverted light chain variable region sequence comprising SEQ ID NO:61.

[0212] In some embodiments, the antibody comprises one of the sequences of clone PA13P1H08 variant "RN" in which the heavy chain variable region of clone PA13P1H08 has been reverted to the predicted naive rearrangement and the light chain variable region retains the unmodified sequence of clone PA13P1H08 (i.e., SEQ ID NO: 5). In some embodiments, the antibody comprises one of the sequences of clone PA13P1H08 variant "NR" in which the heavy chain variable region retains the unmodified sequence of clone PA13P1H08 (i.e., SEQ ID NO: 1) and the light chain variable region has been reverted to the predicted naive rearrangement of clone PA13P1H08.

[0213] In some embodiments, the antibody comprises one or more restored CDR sequences, e.g., one or more restored heavy chain CDR sequences, and / or one or more restored light chain CDR sequences. In some embodiments, the antibody comprises one or more of restored CDR-H1 comprising SEQ ID NO:58, restored CDR-H2 comprising SEQ ID NO:59, or restored CDR-H3 comprising SEQ ID NO:60. In some embodiments, the antibody comprises one or more of restored CDR-L1 comprising SEQ ID NO:62, restored CDR-L2 comprising SEQ ID NO:30, or restored CDR-L3 comprising SEQ ID NO:63. In some embodiments, the antibody comprises a heavy chain variable region comprising one or more restored framework regions, e.g., the restored framework region of SEQ ID NO:67.

[0214] In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:58, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:59, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:60, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:6 or SEQ ID NO:62, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:7 or SEQ ID NO:30, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:8 or SEQ ID NO:63.

[0215] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:1, SEQ ID NO:57, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:67, and comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:5 or SEQ ID NO:61. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:57, and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:61. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:57, and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:5. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:1 and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:61. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:64 and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:5. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:65 and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:5. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:66 and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:5. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:67 and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:5.

[0216] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 65, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 2, 59, and 4, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO:5, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs:6, 7, and 8, respectively. Includes.

[0217] Nut-specific antibody sequences In some embodiments, a monoclonal antibody that specifically binds to a tree nut allergen (e.g., that specifically binds to a cashew, pistachio, almond, pine nut, pecan, walnut, hazelnut, or macadamia nut allergen) comprises the heavy chain and / or light chain CDRs disclosed below in Table 1. In some embodiments, a monoclonal antibody that specifically binds to a tree nut allergen comprises the light chain and / or heavy chain variable region sequences disclosed below in Table 1. In some embodiments, a monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain variable region comprising an amino acid sequence that has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a heavy chain variable region sequence disclosed in Table 1 and that contains CDR1, CDR2, and CDR3 that are identical to the CDRs of the heavy chain variable region sequence, and a light chain variable region comprising an amino acid sequence that has at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a light chain variable region sequence disclosed in Table 1 and that contains CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0218] In some embodiments, a monoclonal antibody that specifically binds to a nut allergen comprises a light chain sequence or portion thereof and / or a heavy chain sequence or portion thereof from any of the following antibodies described herein: clone PA14P3H08, clone PA11P1D11, clone PA11P1G10, clone PA12P4D02, clone PA11P1D12, clone PA11P1F03, clone PA11P1C04, clone PA11P1G04, clone PA11P1E01, clone PA11P1C11, or clone PA11P1C03. The amino acid sequences of the CDRs, light chain variable domain (VL), and heavy chain variable domain (VH) of clone PA14P3H08, clone PA11P1D11, clone PA11P1G10, clone PA12P4D02, clone PA11P1D12, clone PA11P1F03, clone PA11P1C04, clone PA11P1G04, clone PA11P1E01, clone PA11P1C11, and clone PA11P1C03 are set forth in Table 1 below.

[0219] In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen is (a) a heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 113, 167, 175, 227, 311, 318, 438, 466, 621, 665, or 692; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 168, 176, 200, 312, 319, 439, 539, 666, or 693; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 169, 177, 228, 313, 320, 440, 467, 540, 667, or 694; (d) a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623, 669, or 696; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 94, 149, 172, 180, 196, 323, or 670; and (f) a light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624, 671, or 697. The compound may include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0220] In some embodiments, a monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain variable region comprising an amino acid sequence comprising or having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) of any one of SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises a light chain variable region comprising an amino acid sequence comprising or having at least 75% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) of any one of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises or has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691. and a light chain variable region comprising an amino acid sequence comprising any one of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695, or having at least 75% sequence identity thereto (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto).

[0221] In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen is (a) a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691, and including CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO; and (b) a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695, and including CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO. Includes.

[0222] In some embodiments, the monoclonal antibody specifically binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen; (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 113, 227, 311, 318, 665, or 692; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 200, 312, 319, 539, 666, or 693; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 228, 313, 320, 540, 667, or 694; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 230, 315, 322, 542, 669, or 696; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 94, 149, 196, 323, or 670; and (f) a light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 231, 316, 324, 543, 671, or 697. The compound may include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0223] In some embodiments, a monoclonal antibody that specifically binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen comprises a heavy chain variable region comprising an amino acid sequence comprising or having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) of any one of SEQ ID NOs: 226, 310, 317, 538, 664, or 691. In some embodiments, a monoclonal antibody that specifically binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen comprises a light chain variable region comprising an amino acid sequence comprising or having at least 75% sequence identity thereto (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) of any one of SEQ ID NOs: 229, 314, 321, 541, 668, or 695. In some embodiments, a monoclonal antibody that specifically binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen comprises or shares at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or more) to any one of SEQ ID NOs: 226, 310, 317, 437, 538, 664, or 691. and a light chain variable region comprising an amino acid sequence having at least 75% sequence identity thereto (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) thereto, wherein the heavy chain variable region comprises an amino acid sequence having at least 75% sequence identity thereto (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto).

[0224] In some embodiments, the monoclonal antibody specifically binds to a cashew and / or pistachio allergen; (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 167, 175, 227, 438, 466, or 621; (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 168, 176, 200, or 439; (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 169, 177, 440, or 467; (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 171, 179, 442, 469, or 623; (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 149, 172, or 180; and (f) a light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 173, 181, 443, 470, or 624. The compound may include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0225] In some embodiments, a monoclonal antibody that specifically binds to a cashew and / or pistachio allergen comprises a heavy chain variable region comprising an amino acid sequence comprising any one of SEQ ID NOs: 166, 174, 437, 465, or 620, or having at least 75% sequence identity thereto (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto). In some embodiments, a monoclonal antibody that specifically binds to a cashew and / or pistachio allergen comprises a light chain variable region comprising an amino acid sequence comprising any one of SEQ ID NOs: 170, 178, 441, 468, or 622, or having at least 75% sequence identity thereto (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity thereto). In some embodiments, a monoclonal antibody that specifically binds to a cashew and / or pistachio allergen comprises a heavy chain variable region comprising an amino acid sequence comprising or having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) of any one of SEQ ID NOs: 166, 174, 437, 465, or 620, and a light chain variable region comprising an amino acid sequence comprising or having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) of any one of SEQ ID NOs: 170, 178, 441, 468, or 622.

[0226] In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen is (a) SEQ ID NOs: 692, 693, 694, 696, 94, and 697, respectively; or (b) SEQ ID NOs: 318, 319, 320, 322, 323, and 324, respectively; or (c) SEQ ID NOs: 227, 200, 228, 230, 149, and 231, respectively; or (d) SEQ ID NOs: 113, 539, 540, 542, 196, and 543, respectively; or (e) SEQ ID NOs: 311, 312, 313, 315, 94, and 316, respectively; or (f) SEQ ID NOs: 665, 666, 667, 669, 670, and 671, respectively; or (g) SEQ ID NOs: 466, 200, 467, 469, 149, and 470, respectively; or (h) SEQ ID NOs: 167, 168, 169, 171, 172, and 173, respectively; or (i) SEQ ID NOs: 621, 176, 177, 623, 180, and 624, respectively; or (j) SEQ ID NOs: 175, 176, 177, 179, 180, and 181, respectively; or (k) SEQ ID NOs: 438, 439, 440, 442, 30, and 443, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0227] Clone PA14P3H08 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 692, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 693, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 694, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 696, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 94, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 697. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen.

[0228] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 691, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 692, 693, and 694, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 695, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 696, 94, and 697, respectively. Includes.

[0229] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 691, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 695. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 691, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 695.

[0230] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 692, 693, 694, 696, 95, and 697, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 691 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 695).

[0231] Clone PA11P1D11 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 318, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 319, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 320, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 322, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 323, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 324. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen.

[0232] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 317, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 318, 319, and 320, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 321, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 322, 323, and 324, respectively. Includes.

[0233] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 317, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 321. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 317, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 321.

[0234] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 318, 319, 320, 322, 323, and 324, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 317 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 321).

[0235] Clone PA11P1G10 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 227, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 200, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 228, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 230, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 149, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 231. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen.

[0236] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 226, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 227, 200, and 228, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 229, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 230, 149, and 231, respectively. Includes.

[0237] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 226, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 229. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 226, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 229.

[0238] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 227, 200, 228, 230, 149, and 231, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 226 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 229).

[0239] Clone PA12P4D02 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 539, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 540, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 542, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 196, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 543. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen.

[0240] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 538, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 113, 539, and 540, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 541, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 542, 196, and 543, respectively. Includes.

[0241] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 538, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 541. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 538, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 541.

[0242] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 113, 539, 540, 542, 196, and 543, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 538 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 541).

[0243] Clone PA11P1D12 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 692, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 693, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 694, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 696, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 94, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 697. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen.

[0244] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 310, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 311, 312, and 313, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 314, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 315, 94, and 316, respectively. Includes.

[0245] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 310, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 314. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 310, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 314.

[0246] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 311, 312, 313, 315, 94, and 316, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 310 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 314).

[0247] Clone PA11P1F03 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 665, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 666, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 667, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 669, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 670, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 671. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a pecan, walnut, hazelnut, and / or macadamia nut allergen.

[0248] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 664, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 665, 666, and 667, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 668, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 669, 670, and 671, respectively. Includes.

[0249] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 664, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 668. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 664, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 668.

[0250] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 665, 666, 667, 669, 670, and 671, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 664 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 668).

[0251] Clone PA11P1C04 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 466, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 200, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 467, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 469, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 149, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 470. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a cashew and / or pistachio allergen.

[0252] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 465, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 466, 200, and 467, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 468, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 469, 149, and 470, respectively. Includes.

[0253] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 465, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 468. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 465, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 468.

[0254] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 466, 200, 467, 469, 149, and 470, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 465 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 468).

[0255] Clone PA11P1G04 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 167, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 168, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 169, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 171, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 172, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 173. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a cashew and / or pistachio allergen.

[0256] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 166, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 167, 168, and 169, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 170, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 171, 172, and 173, respectively. Includes.

[0257] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 166, and comprises a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 170. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 166, and comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:170.

[0258] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 167, 168, 169, 171, 172, and 173, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 166 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 170).

[0259] Clone PA11P1E01 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 621, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 176, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 177, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 623, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 180, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 624. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a cashew and / or pistachio allergen.

[0260] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 620, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 621, 176, and 177, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 622, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 623, 180, and 624, respectively. Includes.

[0261] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 620, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 622. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 620, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 622.

[0262] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 621, 176, 177, 623, 180, and 624, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 620 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 622).

[0263] Clone PA11P1C11 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 175, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 176, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 177, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 179, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 180, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 181. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a cashew and / or pistachio allergen.

[0264] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 174, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 175, 176, and 177, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 178, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 179, 180, and 181, respectively. Includes.

[0265] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 174, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 178. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 174, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 178.

[0266] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 175, 176, 177, 179, 180, and 181, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 174 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 178).

[0267] Clone PA11P1C03 In some embodiments, the monoclonal antibody that specifically binds to a tree nut allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 438, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 439, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 440, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 442, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 443. In some embodiments, the monoclonal antibody binds to two or more tree nut allergens. In some embodiments, the monoclonal antibody binds to a cashew and / or pistachio allergen.

[0268] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 437, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 438, 439, and 440, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 441, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 442, 30, and 443, respectively. Includes.

[0269] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 437, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 441. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 437, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 441.

[0270] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of SEQ ID NOs: 438, 439, 440, 442, 30, and 443, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 437 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 441).

[0271] Antibodies that specifically bind to milk allergens In some embodiments, the monoclonal antibody or antigen-binding portion thereof specifically binds to a milk allergen (e.g., a cow's milk allergen). In some embodiments, the monoclonal antibody that specifically binds to a milk allergen comprises the heavy chain CDRs and / or the light chain CDRs disclosed in Table 1 below. In some embodiments, the monoclonal antibody that specifically binds to a milk allergen comprises the light chain variable region sequence and / or the heavy chain variable region sequence disclosed in Table 1 below. In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain variable region comprising an amino acid sequence that has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a heavy chain variable region sequence disclosed in Table 1 and that contains CDR1, CDR2, and CDR3 that are identical to the CDRs of the heavy chain variable region sequence; and a light chain variable region comprising an amino acid sequence that has at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a light chain variable region sequence disclosed in Table 1 and that contains CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0272] In some embodiments, the monoclonal antibody that specifically binds to a milk allergen is any of the following antibodies described herein: clone PA01P2C05, clone PA01P2B03, clone PA01P2A12, clone PA01P2C12, clone PA01P2E10, clone PA01P2C09, clone PA01P2D06, clone PA01P2E08, clone PA01P2A05, clone PA01P2B04, clone PA01P2E05, clone PA01P2D04, clone PA01P2B1 2, comprising a light chain sequence or portion thereof and / or a heavy chain sequence or portion thereof from any of clone PA01P2D11, clone PA01P2B10, clone PA01P2D10, clone PA01P2D09, clone PA01P2B05, clone PA01P4C11, clone PA01P3E08, clone PA01P2E06, clone PA01P2E07, clone PA01P2G07, clone PA01P2B09, clone PA01P2C04, or clone PA01P2H08. Clone PA01P2C05, Clone PA01P2B03, Clone PA01P2A12, Clone PA01P2C12, Clone PA01P2E10, Clone PA01P2C09, Clone PA01P2D06, Clone PA01P2E08, Clone PA01P2A05, Clone PA01P2B04, Clone PA01P2E05, Clone PA01P2D04, Clone PA01P2B12, Clone PA01P2D11, Clone PA01P2B10, The amino acid sequences of the CDRs, light chain variable domain (VL), and heavy chain variable domain (VH) of clone PA01P2D10, clone PA01P2D09, clone PA01P2B05, clone PA01P4C11, clone PA01P3E08, clone PA01P2E06, clone PA01P2E07, clone PA01P2G07, clone PA01P2B09, clone PA01P2C04, and clone PA01P2H08 are set forth in Table 1 below.

[0273] In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen is (a) a heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 121, 750, 757, 765, 772, 779, 785, 793, 800, 807, 814, 821, 833, 838, 846, 853, 860, 868, 874, 881, 889, 895, 903, 911, 918, or 926; (b) a heavy chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 378, 532, 751, 758, 766, 773, 786, 794, 801, 808, 815, 822, 826, 839, 847, 854, 861, 875, 882, 890, 896, 904, 912, 919, or 927; (c) a heavy chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 752, 759, 767, 774, 780, 787, 795, 802, 809, 816, 827, 840, 848, 855, 862, 869, 876, 883, 891, 897, 905, 913, 920, or 928; (d) a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 523, 754, 761, 769, 776, 782, 789, 797, 804, 811, 818, 829, 835, 842, 850, 857, 864, 871, 878, 885, 899, 907, 915, 922, or 930; (e) a light chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 22, 30, 94, 110, 149, 186, 196, 389, 404, 509, 662, 682, 762, 790, 830, 843, 865, 886, 900, 908, 923, or 931; and (f) a light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 755, 763, 770, 777, 783, 791, 798, 805, 812, 819, 824, 831, 836, 844, 851, 858, 866, 872, 879, 887, 893, 901, 909, 916, 924, or 932. The compound may include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0274] In some embodiments, the monoclonal antibody that specifically binds to a milk allergen is (a) SEQ ID NOs: 750, 751, 752, 754, 196, and 755, respectively; or (b) SEQ ID NOs: 757, 758, 759, 761, 762, and 763, respectively; or (c) SEQ ID NOs: 765, 766, 767, 769, 30, and 770, respectively; or (d) SEQ ID NOs: 772, 773, 774, 776, 389, and 777, respectively; or (e) SEQ ID NOs: 779, 532, 780, 782, 404, and 783, respectively; or (f) SEQ ID NOs: 785, 786, 787, 789, 790, and 791, respectively; or (g) SEQ ID NOs: 793, 794, 795, 797, 30, and 798, respectively; or (h) SEQ ID NOs: 800, 801, 802, 804, 110, and 805, respectively; or (i) SEQ ID NOs: 807, 808, 809, 811, 509, and 812, respectively; or (j) SEQ ID NOs: 814, 815, 816, 818, 94, and 819, respectively; or (k) SEQ ID NOs: 821, 822, 787, 776, 790, and 824, respectively; or (l) SEQ ID NOs: 121, 826, 827, 829, 830, and 831, respectively; or (m) SEQ ID NOs: 833, 826, 827, 835, 149, and 836, respectively; or (n) SEQ ID NOs: 838, 839, 840, 842, 843, and 844, respectively; or (o) SEQ ID NOs: 846, 847, 848, 850, 196, and 851, respectively; or (p) SEQ ID NOs: 853, 854, 855, 857, 662, and 858, respectively; or (q) SEQ ID NOs: 860, 861, 862, 864, 865, and 866, respectively; or (r) SEQ ID NOs: 868, 378, 869, 871, 682, and 872, respectively; or (s) SEQ ID NOs: 874, 875, 876, 878, 22, and 879, respectively; or (t) SEQ ID NOs: 881, 882, 883, 885, 886, and 887, respectively; or (u) SEQ ID NOs: 889, 890, 891, 523, 762, and 893, respectively; or (v) SEQ ID NOs: 895, 896, 897, 899, 900, and 901, respectively; or (w) SEQ ID NOs: 903, 904, 905, 907, 908, and 909, respectively; or (x) SEQ ID NOs: 911, 912, 913, 915, 149, and 916, respectively; or (y) SEQ ID NOs: 918, 919, 920, 922, 923, and 924, respectively; or (z) SEQ ID NOs: 926, 927, 928, 930, 931, and 932, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0275] In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 749, 756, 764, 771, 778, 784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873, 880, 888, 894, 902, 910, 917, or 925. In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or 929.In some embodiments, a monoclonal antibody that specifically binds to a milk allergen has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 749, 756, 764, 771, 778, 784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873, 880, 888, 894, 902, 910, 917, or 925. and a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or 929.

[0276] In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain variable region that comprises an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 749, 756, 764, 771, 778, 784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873, 880, 888, 894, 902, 910, 917, or 925 and includes CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO. In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a light chain variable region that comprises an amino acid sequence having at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or 929 and comprising CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO.

[0277] In some embodiments, the monoclonal antibody that specifically binds to a milk allergen is (a) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to, or having the sequence of, SEQ ID NO: 749, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to, or having the sequence of SEQ ID NO: 753; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to or having the sequence of SEQ ID NO: 756, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to or having the sequence of SEQ ID NO: 760; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 764 or having the sequence of SEQ ID NO: 764, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 768 or having the sequence of SEQ ID NO: 768; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 771 or having the sequence of SEQ ID NO: 771, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 775 or having the sequence of SEQ ID NO: 775; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to, or having the sequence of, SEQ ID NO: 778, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to, or having the sequence of SEQ ID NO: 781; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 784 or having the sequence of SEQ ID NO: 784, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 788 or having the sequence of SEQ ID NO: 788; or (g) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 792 or having the sequence of SEQ ID NO: 792, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 796 or having the sequence of SEQ ID NO: 796; or (h) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to or having the sequence of SEQ ID NO: 799, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to or having the sequence of SEQ ID NO: 803; or (i) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to or having the sequence of SEQ ID NO: 806, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to or having the sequence of SEQ ID NO: 810; or (j) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 813 or having the sequence of SEQ ID NO: 813, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 817 or having the sequence of SEQ ID NO: 817; or (k) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 820 or having the sequence of SEQ ID NO: 820, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 823 or having the sequence of SEQ ID NO: 823; or (l) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 825 or having the sequence of SEQ ID NO: 825, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 828 or having the sequence of SEQ ID NO: 828; or (m) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 832 or having the sequence of SEQ ID NO: 832, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 834 or having the sequence of SEQ ID NO: 834; or (n) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 837 or having the sequence of SEQ ID NO: 837, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 841 or having the sequence of SEQ ID NO: 841; or (o) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 845 or having the sequence of SEQ ID NO: 845, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 849 or having the sequence of SEQ ID NO: 849; or (p) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 852 or having the sequence of SEQ ID NO: 852, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 856 or having the sequence of SEQ ID NO: 856; or (q) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to, or having the sequence of, SEQ ID NO: 859, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to, or having the sequence of, SEQ ID NO: 863; or (r) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 867 or having the sequence of SEQ ID NO: 867, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 870 or having the sequence of SEQ ID NO: 870; or (s) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 873 or having the sequence of SEQ ID NO: 873, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 877 or having the sequence of SEQ ID NO: 877; or (t) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 880 or having the sequence of SEQ ID NO: 880, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 884 or having the sequence of SEQ ID NO: 884; or (u) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 888 or having the sequence of SEQ ID NO: 888, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 892 or having the sequence of SEQ ID NO: 892; or (v) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 894 or having the sequence of SEQ ID NO: 894, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 898 or having the sequence of SEQ ID NO: 898; or (w) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 902 or having the sequence of SEQ ID NO: 902, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 906 or having the sequence of SEQ ID NO: 906; or (x) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 910 or having the sequence of SEQ ID NO: 910, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 914 or having the sequence of SEQ ID NO: 914; or (y) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 917 or having the sequence of SEQ ID NO: 917, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 921 or having the sequence of SEQ ID NO: 921; or (z) a heavy chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 925 or having the sequence of SEQ ID NO: 925, and a light chain variable region comprising an amino acid sequence having at least 90% (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 929 or having the sequence of SEQ ID NO: 929. Includes.

[0278] Clone PA01P2D09 In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 860, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 861, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 862, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 864, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 865, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 866.

[0279] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 859, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 860, 861, and 862, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 863, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 864, 865, and 866, respectively. Includes.

[0280] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 859, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 863. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 859, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:863.

[0281] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 860, 861, 862, 864, 865, and 866, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 859 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 863).

[0282] Clone PA01P2D04 In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 121, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 826, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 827, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 829, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 830, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 831.

[0283] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 825, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 121, 826, and 827, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 828, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 829, 830, and 831, respectively. Includes.

[0284] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 825, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 828. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 825, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:828.

[0285] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 121, 826, 827, 829, 830, and 831, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 825 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 828).

[0286] Clone PA01P2B12 In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 833, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 826, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 827, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 835, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 149, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 836.

[0287] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 832, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 833, 826, and 827, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 834, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 835, 149, and 836, respectively. Includes.

[0288] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 832, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 834. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 832, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 834.

[0289] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 833, 826, 827, 835, 149, and 836, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 832 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 834).

[0290] Clone PA01P2B05 In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 868, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 378, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 869, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 871, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 682, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 872.

[0291] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 867, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 868, 378, and 869, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 870, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 871, 682, and 872, respectively. Includes.

[0292] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 867, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 870. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 867, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 870.

[0293] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 868, 378, 869, 871, 682, and 872, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 867 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 870).

[0294] Clone PA01P2D10 In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 853, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 854, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 855, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 857, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 662, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 858.

[0295] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 852, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 853, 854, and 855, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 856, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 857, 662, and 858, respectively. Includes.

[0296] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 852, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 856. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 852, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:856.

[0297] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 853, 854, 855, 857, 662, and 858, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 852 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 856).

[0298] Clone PA01P2E08 In some embodiments, a monoclonal antibody that specifically binds to a milk allergen comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 800, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 801, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 802, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 804, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 110, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 805.

[0299] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 799, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 800, 801, and 802, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 803, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 804, 110, and 805, respectively. Includes.

[0300] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 799, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 803. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 799, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 803.

[0301] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 800, 801, 802, 804, 110, and 805, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 799 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 803).

[0302] Antibodies that specifically bind to fungal allergens In some embodiments, the monoclonal antibody or antigen-binding portion thereof specifically binds to a fungal allergen. In some embodiments, the monoclonal antibody specifically binds to a fungal allergen that is an Aspergillus fumigatus, Aspergillus niger, or Aspergillus nidulans allergen (e.g., an extract of Aspergillus fumigatus, Aspergillus niger, or Aspergillus nidulans). In some embodiments, the fungal allergen is Aspergillus fumigatus 1 (Asp f1), e.g., purified recombinant allergen Aspergillus fumigatus 1 (rAsp f1).

[0303] In some embodiments, the monoclonal antibody exhibits cross-reactivity with at least two fungal allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity with two or more Aspergillus allergens (e.g., two or more species of Aspergillus). In some embodiments, the monoclonal antibody exhibits cross-reactivity with two or more of the fungal allergens Aspergillus fumigatus, Aspergillus niger, and Aspergillus nidulans. In some embodiments, the monoclonal antibody specifically binds to at least one of the fungal allergens with a KD of less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about 10 pM, or less than 1 pM.

[0304] Antibody sequence In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., specifically binds to an Aspergillus allergen) comprises the heavy and / or light chain CDRs disclosed in Table 1 below. In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises the light and / or heavy chain variable region sequences disclosed in Table 1 below. In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises the light and / or heavy chain variable region sequences disclosed in Table 1 below. In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) has at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to the heavy chain variable region sequences disclosed in Table 1, and has CDR1, CDR2, and CDR3 that are identical to the CDRs of the heavy chain variable region sequence. The heavy chain variable region comprises an amino acid sequence that comprises DR3, and a light chain variable region that comprises an amino acid sequence that has at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to a light chain variable region sequence disclosed in Table 1 and that comprises CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0305] In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., that specifically binds to an Aspergillus allergen) comprises a light chain sequence or portion thereof and / or a heavy chain sequence or portion thereof from any of the following antibodies described herein: clone 1003320101_D6, clone 1003320105_D6, clone 1003320107_C5, clone 1003320107_F3, or clone 1003320107_F8. The amino acid sequences of the CDRs, light chain variable domain (VL), and heavy chain variable domain (VH) of clone 1003320101_D6, clone 1003320105_D6, clone 1003320107_C5, clone 1003320107_F3, and clone 1003320107_F8 are set forth in Table 1 below.

[0306] In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen is (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 710, 718, 726, 734, or 742; A heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 711, 719, 727, 735, or 743; A heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 712, 720, 728, 736, or 744; a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 714, 722, 730, 738, or 746; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 715, 723, 731, 739, or 747; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 716, 724, 732, 740, or 748. The compound may include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0307] In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen is (a) SEQ ID NOs: 710, 711, 712, 714, 715, and 716, respectively; or (b) SEQ ID NOs: 718, 719, 720, 722, 723, and 724, respectively; or (c) SEQ ID NOs: 726, 7272, 728, 730, 731, and 732, respectively; or (d) SEQ ID NOs: 734, 735, 736, 738, 739, and 740, respectively; or (e) SEQ ID NOs: 742, 743, 744, 746, 747, and 748, respectively. The heavy chain CDR1 to 3 and the light chain CDR1 to 3 contain the amino acid sequences of:

[0308] In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen comprises a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 709, 717, 725, 733, or 741. In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen comprises a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 713, 721, 729, 737, or 745. In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen comprises a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 709, 717, 725, 733, or 741, and a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 713, 721, 729, 737, or 745.

[0309] In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen comprises a heavy chain variable region that comprises an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 709, 717, 725, 733, or 741 and that comprises CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO. In some embodiments, a monoclonal antibody that specifically binds a fungal allergen comprises a light chain variable region that has at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 713, 721, 729, 737, or 745 and comprises an amino acid sequence that includes CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO. In some embodiments, a monoclonal antibody that specifically binds a fungal allergen comprises (a) a heavy chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 709, 717, 725, 733, or 741, and comprising CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO; and (b) a light chain variable region comprising an amino acid sequence having at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 713, 721, 729, 737, or 745, and including CDR1, CDR2, and CDR3 identical to the CDRs of that SEQ ID NO. Includes.

[0310] Clone 1003320101_D6 In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 710, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 711, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 712, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 714, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 715, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 716. In some embodiments, the antibody specifically binds to the fungal allergen Aspergillus fumigatus.

[0311] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 709, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 710, 711, and 712, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 713, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 714, 715, and 716, respectively. Includes.

[0312] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 709, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 713. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 709, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 713.

[0313] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 710, 711, 712, 714, 715, and 716, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 709 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 713).

[0314] Clone 1003320105_D6 In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 718, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 719, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 720, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 722, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 723, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 724. In some embodiments, the antibody specifically binds to the fungal allergen Aspergillus fumigatus.

[0315] In some embodiments, the antibody comprises: (a) having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 717, and SEQ ID NOs: 718, 719, and 720, respectively. a heavy chain variable region comprising an amino acid sequence comprising heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequence of (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 721, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 722, 723, and 724, respectively. Includes.

[0316] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 717, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 721. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 717, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 721.

[0317] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 718, 719, 720, 722, 723, and 724, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 717 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 721).

[0318] Clone 1003320107_C5 In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 726, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 727, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 728, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 730, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 731, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 732. In some embodiments, the antibody specifically binds to the fungal allergen Aspergillus fumigatus. In some embodiments, the antibody specifically binds to the fungal allergen Aspergillus niger. In some embodiments, the antibody specifically binds to the fungal allergen Aspergillus nidulans. In some embodiments, the antibody specifically binds to a recombinant Aspergillus antigen (e.g., rAsp f1). In some embodiments, the antibody specifically binds to two or more of Aspergillus fumigatus, Aspergillus niger, Aspergillus nidulans, or recombinant Aspergillus antigen (eg, rAsp f1) through cross-reactivity.

[0319] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 725, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 726, 727, and 728, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 729, and comprising light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 730, 731, and 732, respectively. Includes.

[0320] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 725, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 729. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 725, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 729.

[0321] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 726, 727, 728, 730, 731, and 732, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 725 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 729).

[0322] Clone 1003320107_F3 In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 734, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 735, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 736, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 738, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 739, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 740. In some embodiments, the antibody specifically binds to a recombinant Aspergillus antigen (e.g., rAsp f1).

[0323] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 733, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 734, 735, and 736, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 737, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 738, 739, and 740, respectively. Includes.

[0324] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 733, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 737. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 733, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 737.

[0325] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 734, 735, 736, 738, 739, and 740, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 733 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 737).

[0326] Clone 1003320107_F8 In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 742, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 743, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 744, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 746, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 747, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 748. In some embodiments, the antibody specifically binds to the fungal allergen Aspergillus fumigatus.

[0327] In some embodiments, the antibody comprises: (a) a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 741, and including heavy chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 7422, 743, and 744, respectively; and (b) a light chain variable region comprising an amino acid sequence having at least 70% sequence identity (e.g., at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 745, and including light chain CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 746, 747, and 748, respectively. Includes.

[0328] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 741, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to SEQ ID NO: 745. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 741, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 745.

[0329] In some embodiments, the antibody is an antibody that competes for binding with an antibody described herein (e.g., an antibody comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequence of SEQ ID NO: 742, 743, 744, 746, 747, and 748, respectively, or an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 741 and further comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 745).

[0330] antigen binding fragment In some embodiments, the antibodies disclosed herein (e.g., antibodies that bind to food allergens, plant allergens, fungal allergens, animal allergens, dust mite allergens, drug allergens, cosmetic allergens, or latex allergens disclosed in Section IV) are antigen-binding portions (also referred to herein as antigen-binding fragments). Examples of antigen-binding fragments include, but are not limited to, Fab, F(ab')2, Fv, scFv, bivalent scFv, single domain antibodies, or diabodies. Various techniques have been developed for the production of antigen-binding fragments. Traditionally, these fragments were derived via proteolytic digestion of intact antibodies (see, e.g., Morimoto et al., J. Biochem. Biophys. Meth., 24:107-117 (1992), and Brennan et al., Science, 229:81 (1985)). However, these fragments can now be produced directly using recombinant host cells. For example, antigen-binding fragments can be isolated from antibody phage libraries. Alternatively, Fab'-SH fragments can be directly recovered from E. coli cells and chemically coupled to form F(ab')2 fragments (see, e.g., Carter et al., BioTechnology, 10:163-167 (1992)). According to another approach, F(ab')2 fragments can be directly isolated from recombinant host cell culture. Other techniques for the production of antigen-binding fragments are known in the art.

[0331] Antibody conjugates In some embodiments, the antibody or antigen-binding fragment can be conjugated to another molecule, such as polyethylene glycol (PEGylation) or serum albumin, to achieve extended half-life in vivo. Examples of PEGylation of antigen-binding fragments are provided in Knight et al. Platelets 15:409, 2004 (for abciximab), Pedley et al., Br. J. Cancer 70:1126, 1994 (for anti-CEA antibodies), Chapman et al., Nature Biotech. 17:780, 1999, and Humphreys, et al., Protein Eng. Des. 20: 227, 2007).

[0332] In some embodiments, an antibody-drug conjugate is provided that comprises the monoclonal antibody or antigen-binding fragment described herein. In some embodiments, the monoclonal antibody or antigen-binding fragment (e.g., an antibody or antigen-binding fragment that specifically binds to a food allergen or fungal allergen) is covalently linked to a cytotoxic drug. In some embodiments, the antibody or antigen-binding fragment is an antibody that specifically binds to a fungal allergen, and the drug is an antifungal drug. Suitable antifungal drugs include, but are not limited to, amphotericin B, azole antifungals (e.g., ketoconazole, fluconazole, isavuconazole, itraconazole, posaconazole, or voriconazole), echinocandins (e.g., anidulafungin, caspofungin, or micafungin), and flucytosine. Methods for making antibody-drug conjugates are described, for example, in Chudasama et al., Nature Chemistry, 2016, 8:114-119, WO2013 / 068874, and US8,535,678.

[0333] Synthetic antibodies, antibody compositions, and antibody-producing cells Certain antibodies described herein are derived from B cells isolated from human subjects exposed to allergens. In certain embodiments, the antibodies, antibody compositions, and cells of the present invention are distinguishable in one or more respects from naturally occurring antibodies, compositions, and cells. Distinguishable antibodies, compositions, and cells may be referred to as "synthetic" and the antibody or composition may be identified as "non-naturally occurring" by the condition "not naturally occurring" or positively. As used herein, the terms "corresponding antibody" and "corresponding to" describe the relationship between (1) an antibody that is characterized by six specified CDR sequences and is produced by the immune cells of the study described in the following examples, and (2) a synthetic antibody that contains the same six CDR sequences.

[0334] synthetic antibody The synthetic antibodies of the invention may differ in structure from naturally occurring antibodies with the same CDRs. That is, a synthetic antibody identified by a specified CDR may be structurally different from an antibody containing the specified CDR produced by the cells studied in the Examples below. Possible differences include:

[0335] Variable region sequences that differ from the corresponding naturally occurring antibodies In one approach, the antibody heavy chain comprises the CDRs of a clone described herein (e.g., PA13P1E10), but does not comprise the heavy chain variable region sequence associated with said clone described herein. For example, in one embodiment, an antibody comprising the CDRs of clone PA13P1E10 does not have a heavy chain variable region comprising SEQ ID NO:9. In another approach, the antibody light chain comprises the CDRs of a clone described herein (e.g., PA13P1E10), but does not comprise the light chain variable region sequence associated with said clone described herein. For example, in one embodiment, an antibody comprising the CDRs of clone PA13P1E10 does not have a light chain variable region comprising SEQ ID NO:13). In one approach, both the heavy and light chain variable regions of the antibody of the invention have amino acid sequences other than those disclosed herein.

[0336] Lambda and kappa light chains In some embodiments, the synthetic antibody comprises a lambda-type light chain. In some embodiments, the synthetic antibody comprises a kappa-type light chain.

[0337] Isotype

[0338] In some embodiments, the synthetic antibody with the specified CDRs is an isotype other than the isotype found to be associated with the study subject from which the B cell with the specified CDRs was derived. In some embodiments, the antibody disclosed herein is an isotype other than IgG1. In some embodiments, the antibody disclosed herein is an isotype other than IgG2. In some embodiments, the antibody disclosed herein is an isotype other than IgG3. In some embodiments, the antibody disclosed herein is an isotype other than IgG4. In some embodiments, the antibody disclosed herein is an isotype other than IgM. In some embodiments, the antibody disclosed herein is an isotype other than IgA.

[0339] Allotypes In some embodiments, the synthetic antibodies with specified CDRs are of an allotype other than the allotype found to be associated with the study subject from which the B cells with the specified CDRs were derived. In some embodiments, the synthetic antibodies of the invention comprise an allotype selected from those listed in Table 2 below that is different from the allotype of the antibody from the corresponding study subject. In some embodiments, the synthetic antibodies of the invention comprise any individual allotype selected from those listed in Table 2, provided that the allotype is different from the corresponding allotype of the antibody from the study subject. [Table 2]

[0340] Constant domain mutants The synthetic antibodies of the invention may contain mutations in the heavy chain constant region to alter the properties of the synthetic antibody compared to the corresponding naturally occurring antibody. Exemplary changes include mutations that modulate antibody effector functions (e.g., complement-based effector functions or FcγR-based effector functions), alter half-life, modulate antigen-FcγR interaction, or introduce or remove glycosylation motifs (glycoengineering). See Fonseca et al., 2018, "Boosting half-life and effector functions of therapeutic antibodies by Fc-engineering: An interaction-function review" Int J Biol Macromol. 19:306-311; Wang et al., 2018, "IgG Fc engineering to modulate antibody effector functions" Protein Cell 2018, 9(1):63-73; Schlothauer, 2016, "Novel human IgG1 and IgG4 Fc-engineered antibodies with completely abolished immune effector functions," Protein Engineering, Design and Selection 29(10):457-466; Tam et al., 2017, "Functional, Biophysical, and Structural Characterization of Human IgG1 and IgG4 Fc Variants with Ablated Immune Functionality" Antibodies 6, 12, each of which is incorporated by reference herein for all purposes.

[0341] synthetic antibody composition The synthetic antibody compositions of the invention may differ from naturally occurring compositions in at least one or more of the following respects: (i) the compositions comprise antibodies that are purified, i.e., separated from tissue or cellular material with which they are associated in the human body, optionally in manufactured excipients or medium, and / or (ii) the antibody compositions of the invention contain a single antibody species (are monoclonal), such that all antibodies in the composition have the same structure and specificity.

[0342] Synthetic antibody producing cells The antibodies described herein may be produced by recombinant expression in human or non-human cells.Synthetic antibody-producing cells include non-human cells that express heavy chains, light chains, or both heavy and light chains; human cells that are not immune cells that produce heavy chains, light chains, or both heavy and light chains; and human B cells that produce heavy chains or light chains, but not both heavy and light chains.Synthetic antibodies of the present invention may be heterologously expressed in vitro or in vivo in cells other than human B cells, such as non-human cells, and human cells other than B cells, appropriately other than immune cells, and appropriately other than cells in the B cell lineage.

[0343] V. Method of Use In another aspect, the disclosure provides therapeutic methods for treating a human subject with one or more allergen-specific monoclonal antibodies or antigen-binding portions thereof disclosed herein. In some embodiments, methods for treating allergy are provided. In some embodiments, methods for reducing one or more allergic symptoms in a subject are provided. In some embodiments, the allergen-specific monoclonal antibodies disclosed herein are used therapeutically as blocking antibodies, which are often referred to as passive immunotherapy. Without being bound to a particular theory, it is hypothesized that the allergen-specific monoclonal antibodies disclosed herein block allergen binding to IgE or outcompete endogenous IgE for allergen binding, which prevents or reduces the initiation of the allergy cascade. Without intending to be bound to a particular mechanism, in some embodiments, the antibodies of the present invention provide therapeutic benefit by binding to inhibitory receptors on mast cells and / or basophils.

[0344] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of one or more allergen-specific monoclonal antibodies disclosed herein (e.g., one or more allergen-specific monoclonal antibodies disclosed in Section IV above). In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition disclosed in Section VI below) comprising one or more allergen-specific monoclonal antibodies.

[0345] In some embodiments, the methods include administering to the subject a therapeutically effective amount of an allergen-specific monoclonal antibody that is of a human IgG isotype, such as a human IgG4 isotype, or an antigen-binding portion thereof that comprises at least a portion of a human IgG or IgG4 isotype constant region sequence.

[0346] In some embodiments, the method comprises administering a therapeutically effective amount of an allergen-specific monoclonal antibody or antigen-binding portion thereof to the subject. In some embodiments, the method comprises administering two or more allergen-specific monoclonal antibodies (e.g., in a pharmaceutical composition comprising two or more allergen-specific monoclonal antibodies) to the subject. In some embodiments, the method comprises administering two or more antibodies that specifically bind to the same allergen. In some embodiments, the method comprises administering two or more antibodies that specifically bind to different epitopes of the same allergen. In some embodiments, the method comprises administering two or more antibodies that specifically bind to two or more different allergens.

[0347] In some embodiments, a therapeutic antibody is an antibody that comprises the CDR sequences, heavy chain variable region, and / or light chain variable region described herein (e.g., as disclosed in Table 1 below), and further comprises an unmodified or modified IgM, IgD, IgG3, IgG1, IgA1, IgG2, IgG4, or IgA2 heavy chain constant region.

[0348] In some embodiments, the therapeutic antibody is conjugated to a drug, for example, as described in Section IV above.

[0349] In some embodiments, the human subject being treated is an adult. In some embodiments, the human subject is a minor.

[0350] In some embodiments, the human subject to be treated has an allergy to a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the human subject has an allergy to a food allergen. In some embodiments, the food allergen is a milk allergen, an egg allergen, a nut allergen, a fish allergen, a shellfish allergen, a soybean allergen, a bean allergen, a seed allergen, or a wheat allergen. In some embodiments, the food allergen is a peanut allergen. In some embodiments, the food allergen is a milk allergen. In some embodiments, the food allergen is an egg allergen. In some embodiments, the human subject has an allergy to a plant allergen or a fungal allergen (e.g., an Aspergillus allergen). In some embodiments, the allergen is a pollen allergen (e.g., a tree pollen, a grass pollen, or a grass pollen) or a mold allergen. In some embodiments, the human subject has an allergy to an animal allergen, hi some embodiments, the allergen is a dander allergen or an insect sting.

[0351] In some embodiments, the human subject to be treated has an allergy to two or more allergens, such as food allergens, plant allergens, fungal allergens, animal allergens, dust mite allergens, drug allergens, cosmetic allergens, or latex allergens. In some embodiments, the human subject has an allergy to two or more different types of antigens (allergens) in a class of allergens, such as an allergy to two or more different food allergens (e.g., an allergy to two or more different peanut antigens, or an allergy to a peanut allergen and a non-peanut allergen, such as an egg or milk allergen). In some embodiments, the human subject has an allergy to two or more different classes of allergens (e.g., an allergy to one or more food allergens and one or more plant allergens). In some embodiments, the human subject has allergies to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more allergens in the same allergen class, but does not have any known allergies to allergens in other allergen classes. For example, in some embodiments, the human subject has allergies to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more food allergens, but does not have any known allergies to non-food allergens. In some embodiments, the human subject has allergies to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more fungal allergens, but does not have any known allergies to non-fungal allergens.

[0352] In some embodiments, the therapeutic methods disclosed herein reduce one or more symptoms of allergy in a subject. It will be understood by those skilled in the art that symptoms associated with an allergic reaction may vary depending on the type of allergen that induces the allergic reaction. Examples of allergic reaction symptoms include, but are not limited to, hives, skin rash, eczema erythema, redness of the skin, itchy mouth, itchy eyes, nausea, vomiting, diarrhea, stomach pain, nasal congestion, runny nose, stuffy nose, sneezing, coughing, fatigue, sore throat, swelling of the lips, tongue, or throat, headache, trouble swallowing, shortness of breath, wheezing, low blood pressure, or weak pulse. In some embodiments, the therapeutic methods disclosed herein reduce the severity of one or more symptoms of allergy. In some embodiments of the methods of treatment described herein, the allergy symptoms in the subject include one or more of runny nose, skin rash, skin redness, skin swelling, itching or stinging in or around the mouth and / or throat, difficulty swallowing, watery eyes, diarrhea, stomach cramps, nausea, vomiting, tightness in the throat, shortness of breath or wheezing, shortness of breath, and anaphylaxis. In some embodiments, the methods of treatment disclosed herein reduce the length of time of one or more symptoms of allergy.

[0353] In some embodiments, the treatment methods disclosed herein reduce one or more symptoms of an allergic reaction to an allergen, such as a food allergen (e.g., a peanut allergen), including, but not limited to, a rash, skin rash, eczema erythema, redness of the skin, itchy mouth, nausea, vomiting, diarrhea, stomach pain, nasal congestion, runny nose, sneezing, dry cough, swelling of the lips, tongue, or throat, difficulty swallowing, shortness of breath, wheezing, low blood pressure, or weak pulse. In some embodiments, administration of one or more allergen-specific monoclonal antibodies disclosed herein reduces the severity of one or more of the above symptoms and / or reduces the length of time that one or more of the above symptoms last.

[0354] In some embodiments, the allergen-specific monoclonal antibodies disclosed herein are administered to a human subject in a therapeutically effective amount or dose. In some embodiments, a daily dose range of about 0.01 mg / kg to about 500 mg / kg, or about 0.1 mg / kg to about 200 mg / kg, or about 1 mg / kg to about 100 mg / kg, or about 10 mg / kg to about 50 mg / kg may be used. However, dosage may vary depending on several factors, including the selected route of administration, the formulation of the composition, the patient response, the severity of the condition, the weight of the subject, and the judgment of the prescribing physician. Dosage may be increased or decreased over time as required by the individual patient. In certain cases, the patient is initially administered a low dose, which is then increased to an effective dosage that is tolerable to the patient. The determination of an effective amount is well within the capabilities of one of ordinary skill in the art.

[0355] The route of administration of the antibody or composition comprising the antibody described herein may be dermal or transdermal, inhalation, intestinal, intravenous, intramuscular, intraperitoneal, intrathecal, intralesional, intrabronchial, nasal, ocular or otic delivery, oral, rectal, subcutaneous, topical, transmucosal, or any other method known in the art. In some embodiments, the antibody or composition is administered by infusion (e.g., intravenous) or injection (e.g., subcutaneous). In some embodiments, the route of administration of the antibody or composition comprising the antibody in any of the methods described herein is subcutaneous, intravenous, or intranasal.

[0356] In some embodiments, administration of a single dose of the antibody or composition comprising the antibody described herein is effective for treating allergy or alleviating one or more symptoms of allergy. In some embodiments, multiple doses of the antibody or composition are administered. In some embodiments, the second dose is administered at least 1, 2, 3, 4, 5, 6, or 7 days or more, for example, at least 1, 2, 3, 4, 5, 6, 7, or 8 weeks or more after administration of the first dose. In some embodiments, the antibody or composition comprising the antibody described herein is administered to the subject about every 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, the antibody or composition comprising the antibody described herein is administered to the subject for an extended period of time, for example, for at least 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, or 350 days or more.

[0357] In some embodiments, in any of the methods described herein, the subject is further administered an additional agent, such as an antihistamine, epinephrine, a decongestant, a bronchodilator, or a corticosteroid. In some embodiments, the monoclonal antibody and the additional agent are administered substantially simultaneously, i.e., in the same pharmaceutical composition or in separate pharmaceutical compositions that are administered at substantially the same time (e.g., within a few seconds of each other). In some embodiments, the monoclonal antibody and the additional agent are administered separately. In some embodiments, the monoclonal antibody is administered first, followed by administration of the additional agent. In some embodiments, the additional agent is administered first, followed by administration of the monoclonal antibody.

[0358] In some embodiments, in any of the methods described herein, the method can further include a step of evaluating the allergy symptom reduction in the subject (e.g., allergy symptoms associated with peanut allergy, tree nut allergy, milk allergy, or fungal allergy). In some embodiments, the allergy symptom reduction can be measured by a Total Nasal Symptom Score (TNSS) that is generated from the patient's assessment of four symptoms graded on a scale of 0 (none) to 3 (severe) for nasal congestion, itching, and rhinorrhea, and 0 (none) to 3 (5 or more sneezes) for sneezing. Each of the four symptoms is rated using the following scale: 0=none, 1=mild, 2=moderate, or 3=severe. The TNSS has possible scores of 0 to 12. In other embodiments, the allergy symptom reduction can be measured by a visual analog scale (VAS) nasal symptom score, which is often used to classify allergy burden into mild, moderate, and severe. A VAS nasal symptom score ranging from 0 (no nasal symptoms) to 100 (maximal nasal symptoms) can be used to assess the severity of the combined nasal symptoms. In other embodiments, the reduction in allergy symptoms can be measured by peak nasal inspiratory flow (PNIF), which uses a nasal spirometer to measure the nasal airflow (measured as 1 / min) in a patient. In yet other embodiments, the reduction in allergy symptoms can be measured by allergen skin tests, such as skin prick tests (SPT), which use the presence and degree of skin reactivity as a marker for sensitization in target organs such as the eye, nose, lungs, gut, and skin. When a relevant allergen (e.g., peanut allergen, tree nut allergen, milk allergen, or fungal allergen) is introduced to the skin, the allergic reaction in the skin causes a wheal and erythema response, which can be quantified, for example, using the diameter of the wheal. In yet other embodiments, the reduction in allergy symptoms can be measured by a basophil activation test, which utilizes flow cytometry to quantify the expression of a marker of activation on the surface of basophils following allergen stimulation.In yet other embodiments, reduction in allergy symptoms can be measured by oral food challenge, which involves administering increasing doses of allergen to an allergic individual under the supervision of a trained allergist or immunologist. Oral food challenge can be performed according to an open-label, single-blind, or double-blind format, although the gold standard is both double-blind and placebo-controlled.

[0359] In yet another aspect, the disclosure provides diagnostic and detection methods using one or more allergen-specific monoclonal antibodies or antigen-binding portions thereof disclosed herein. In some embodiments, the allergen-specific monoclonal antibodies or antigen-binding portions thereof are used to detect whether a sample from a subject has allergic reactivity to an allergen (e.g., a food allergen such as a peanut allergen, a nut allergen, or a milk allergen), a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen. In some embodiments, the allergen-specific monoclonal antibodies or antigen-binding portions thereof are used to detect whether a sample from a subject has allergic reactivity to a specific epitope of an allergen (e.g., using an antibody known to bind to a specific epitope of an allergen). In some embodiments, the method includes contacting a sample from a subject (e.g., a blood or plasma sample) with an allergen-specific monoclonal antibody or antigen-binding portion thereof disclosed herein.

[0360] VI. Compositions and Kits In another aspect, compositions and kits are provided that comprise one or more allergen-specific monoclonal antibodies or antigen-binding portions thereof generated from human B cells.

[0361] Pharmaceutical Compositions In some embodiments, a pharmaceutical composition is provided that includes one or more allergen-specific monoclonal antibodies or antigen-binding portions thereof. In some embodiments, the pharmaceutical composition includes a monoclonal antibody described herein, e.g., disclosed in Section IV above. In some embodiments, the pharmaceutical composition is for use in a method of reducing one or more allergy symptoms in a subject (e.g., allergy symptoms resulting from allergy to a food allergen, a plant allergen, a fungal allergen, an animal allergen, a dust mite allergen, a drug allergen, a cosmetic allergen, or a latex allergen). In some embodiments, the pharmaceutical composition is for use in a method of reducing one or more allergy symptoms in a subject with a food allergy, e.g., a peanut allergy. In some embodiments, the pharmaceutical composition is for use in a method of reducing one or more allergy symptoms in a subject with an allergy to two or more allergens (e.g., two or more food allergens, e.g., a peanut allergy and a tree nut allergy). In some embodiments, the pharmaceutical composition is for use in a method of reducing one or more allergy symptoms in a subject with a fungal allergy.

[0362] In some embodiments, the pharmaceutical composition comprises two or more monoclonal antibodies or antigen-binding portions thereof described herein (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more antibodies or antigen-binding portions thereof). In some embodiments, the composition comprises two or more monoclonal antibodies that specifically bind to the same allergen. In some embodiments, the composition comprises two or more monoclonal antibodies that specifically bind to different epitopes of the same allergen. In some embodiments, the composition comprises two or more monoclonal antibodies that specifically bind to two or more different allergens. It will also be understood by those skilled in the art that for a particular type or class of allergen, e.g., a type of food allergen, e.g., peanut allergen, there may be more than one substance (e.g., peptide or protein) within that type or class of allergen that induces an allergic response. In some embodiments, the composition comprises two or more monoclonal antibodies that specifically bind to different allergens within a particular type or class of allergens, for example, two or more different peptides or proteins that are allergens of the same type or class (e.g., two or more different proteins that are pea...

Claims

1. 1. A method for producing a human monoclonal antibody that specifically binds to an allergen, comprising the steps of: (a) isolating a single B cell from a sample from a human subject, the subject being allergic to an allergen; (b) generating cDNA from the single B cell of step (a), wherein the cDNA sequence comprises a first sequence encoding all or a portion of an immunoglobulin heavy chain and a second sequence encoding all or a portion of an immunoglobulin light chain; (c) determining the sequence of the cDNA from step (b); (d) analyzing the sequences determined in step (c) to identify a single B cell that comprises a first sequence that comprises an IgE constant region or an IgG4 constant region; (e) identifying from the single B cell of step (d) (i) a heavy chain variable region sequence in an immunoglobulin heavy chain that comprises an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain that is co-expressed with the immunoglobulin heavy chain in the same single B cell; (f) expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (e); and (g) identifying one or more antibodies from step (f) that specifically bind to the allergen; A method comprising:

2. 2. The method of claim 1, wherein step (a) comprises sorting cells in the sample by fluorescence-activated cell sorting (FACS).

3. 2. The method of claim 1, wherein step (a) comprises selecting single B cells for expression of cell surface IgE and / or cell surface IgG4.

4. 2. The method of claim 1, comprising isolating a single B cell that is selected for expression of cell surface IgE and identifying a sequence encoding an immunoglobulin heavy chain that contains an IgE constant region.

5. Step (a) comprises contacting cells from the sample with an anti-human CD19 antibody and an anti-human IgE antibody, and ＋ The method of claim 4, comprising selecting IgE-expressing B cells.

6. Step (a) comprises contacting cells from the sample with an anti-human CD19 antibody, an anti-human IgE antibody, an anti-human IgM antibody, and an anti-human IgG antibody, and detecting CD19 ＋ IgM － IgG － The method of claim 4, comprising selecting IgE-expressing B cells.

7. 2. The method of claim 1, comprising isolating a single B cell that expresses a B cell marker and is negative for a non-IgE isotype.

8. Step (a) contacts cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgG antibody, an anti-human IgA antibody, and an anti-human IgD antibody, and ＋ IgM － IgG － IgA － IgD － The method of claim 7, comprising selecting B cells.

9. 2. The method of claim 1, comprising isolating a single B cell that is selected for expression of cell surface IgG4, and identifying a sequence encoding an immunoglobulin heavy chain that comprises an IgG4 constant region.

10. Step (a) contacting cells from the sample with an anti-human CD19 antibody and an anti-human IgG4 antibody, and ＋ 10. The method of claim 9, comprising selecting IgG4 expressing B cells.

11. 2. The method of claim 1, comprising isolating a single B cell that expresses B cell markers and is negative for a non-IgG4 isotype.

12. Step (a) contacts cells from the sample with an anti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgE antibody, an anti-human IgA antibody, an anti-human IgD antibody, an anti-human IgG1 antibody, an anti-human IgG2 antibody, and an anti-human IgG3 antibody, and ＋ IgM － IgE － IgA － IgD － IgG1 － IgG2 － IgG3 － The method of claim 11, comprising selecting B cells.

13. 2. The method of claim 1, wherein step (a) comprises isolating antibody-secreting B cells and / or memory B cells.

14. 2. The method of claim 1, wherein step (b) comprises reverse transcribing cDNA from RNA from a single B cell and amplifying the cDNA.

15. The method of claim 14, wherein the RNA is mRNA.

16. 15. The method of claim 14, comprising amplifying immunoglobulin heavy and light chain sequences from a single B cell.

17. 15. The method of claim 14, comprising amplifying the transcriptome of a single B cell.

18. 2. The method of claim 1, wherein step (f) comprises expressing the heavy and light chain variable region sequences from step (e) in a host cell and purifying the antibody.

19. 2. The method of claim 1, wherein step (f) comprises expressing an antibody comprising the heavy and light chain variable region sequences from step (e) and an IgG1 constant region.

20. 2. The method of claim 1, further comprising replacing the constant region of the antibody identified in step (g) with a wild-type IgG4 constant region or a modified IgG4 constant region.

21. The method of claim 1 , wherein the sample comprises peripheral blood.

22. 2. The method of claim 1, wherein the allergen is a food allergen, a plant allergen, a fungal allergen, an animal allergen, a drug allergen, a cosmetic allergen, or a latex allergen.

23. 23. The method of claim 22, wherein the allergen is a food allergen selected from the group consisting of a milk allergen, an egg allergen, a nut allergen, a fish allergen, a shellfish allergen, a soy allergen, a legume allergen, a seed allergen, and a wheat allergen.

24. 24. The method of claim 23, wherein the food allergen is a peanut allergen.

25. 24. The method of claim 23, wherein the food allergen is a nut allergen.

26. 24. The method of claim 23, wherein the food allergen is a milk allergen.

27. 23. The method of claim 22, wherein the allergen is a fungal allergen.

28. 28. The method of claim 27, wherein the fungal allergen is an Aspergillus allergen.

29. Binding affinity (K Ｄ ) a monoclonal antibody or antigen-binding portion thereof that specifically binds to a peanut allergen and / or a tree nut allergen, the antibody comprising: a heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199, 341, 348, 409, 459, or 593; a heavy chain CDR2 comprising any one of the amino acid sequences of SEQ ID NOs: 3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349, 410, 460, 539, or 594; a heavy chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201, 350, 411, 461, 540, or 595; a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343, 352, 413, 463, 542, or 597; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 7, 15, 22, 30, 38, 52, 78, 86, 126, 149, 196, 345, 353, or 598; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354, 414, 464, 543, or 599. A monoclonal antibody or an antigen-binding portion thereof comprising:

30. 30. The monoclonal antibody or antigen-binding portion thereof of claim 29, comprising heavy and light chain CDR sequences contained within a heavy chain variable region and light chain variable region sequence pair selected from the group consisting of SEQ ID NOs: 1 and 5, 9 and 13, 16 and 20, 24 and 28, 32 and 36, 40 and 43, 46 and 50, 54 and 56, 57 and 61, 57 and 5, 1 and 61, 64 and 5, 65 and 5, 66 and 5, 67 and 5, 128 and 132, 340 and 344, 347 and 351, 406 and 407, 408 and 412, 458 and 462, 538 and 541, and 592 and 596.

31. (a) SEQ ID NOs: 2, 3, 4, 6, 7, and 8, respectively; or (b) SEQ ID NOs: 10, 11, 12, 14, 15, and 8, respectively; or (c) SEQ ID NOs: 17, 18, 19, 21, 22, and 23, respectively; or (d) SEQ ID NOs: 25, 26, 27, 29, 30, and 31, respectively; or (e) SEQ ID NOs: 33, 34, 35, 37, 38, and 39, respectively; or (f) SEQ ID NOs: 41, 34, 35, 37, 38, and 39, respectively; or (g) SEQ ID NOs: 47, 48, 49, 51, 52, and 53, respectively; or (h) SEQ ID NOs: 47, 48, 55, 51, 52, and 53, respectively; or (i) SEQ ID NOs: 58, 59, 60, 62, 30, and 63, respectively; or (j) SEQ ID NOs: 58, 59, 60, 6, 7, and 8, respectively; or (k) SEQ ID NOs: 2, 3, 4, 62, 30, and 63, respectively; or (l) SEQ ID NOs: 58, 3, 4, 6, 7, and 8, respectively; or (m) SEQ ID NOs: 2, 59, 4, 6, 7, and 8, respectively; or (n) SEQ ID NOs: 2, 3, 60, 6, 7, and 8, respectively; or (o) SEQ ID NOs: 129, 130, 131, 133, 126, and 134, respectively; or (p) SEQ ID NOs: 341, 342, 343, 345, 78, and 346, respectively; or (q) SEQ ID NOs: 348, 349, 350, 352, 353, and 354, respectively; or (r) SEQ ID NOs: 199, 200, 201, 203, 149, and 204, respectively; or (s) SEQ ID NOs: 409, 410, 411, 413, 86, and 414, respectively; or (t) SEQ ID NOs: 459, 460, 461, 463, 196, and 464, respectively; or (u) SEQ ID NOs: 113, 539, 540, 542, 196, and 543, respectively; or (v) SEQ ID NOs: 593, 594, 595, 597, 598, and 599, respectively.

30. The monoclonal antibody or antigen-binding portion thereof according to claim 29, comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of:

32. 30. The monoclonal antibody or antigen-binding portion thereof of claim 29, comprising a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592.

33. 30. The monoclonal antibody or antigen-binding portion thereof of claim 29, comprising a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596.

34. (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:1, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:9, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:13; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 16, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 20; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:24, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:28; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 32, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 36; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 40, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 43; or (g) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 46, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 50; or (h) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:54, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:56; or (i) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 57, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 61; or (j) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:57, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5; or (k) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:1, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:61; or (l) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:64, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5; or (m) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:65, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5; or (n) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 66, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 5; or (o) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:67, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:5; or (p) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 128, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 132; or (q) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 340, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 344; or (r) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 347, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 351; or (s) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 406, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 407; or (t) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 408, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 412; or (u) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 458, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 462; or (v) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 538, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 541; or (w) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:592, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

596.

30. The monoclonal antibody or antigen-binding portion thereof of claim 29, comprising:

35. The antigen-binding portion is Fab, F(ab') ２ 35. The monoclonal antibody or antigen-binding portion thereof of any one of claims 29 to 34, which is an Fv, scFv, a bivalent scFv, a single domain antibody, or a diabody.

36. 36. A pharmaceutical composition comprising the monoclonal antibody or antigen-binding portion thereof of any one of claims 29 to 35 and a pharma- ceutically acceptable carrier.

37. 36. A pharmaceutical composition comprising a plurality of monoclonal antibodies according to any one of claims 29 to 35, wherein the monoclonal antibodies recognise different epitopes or specifically bind to different allergens.

38. Binding affinity (K Ｄ ) a monoclonal antibody or antigen-binding portion thereof that specifically binds to a tree nut allergen, a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 113, 167, 175, 227, 311, 318, 438, 466, 621, 665, or 692; a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 168, 176, 200, 312, 319, 439, 539, 666, or 693; a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 169, 177, 228, 313, 320, 440, 467, 540, 667, or 694; a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623, 669, or 696; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 94, 149, 172, 180, 196, 323, or 670; and A light chain CDR3 comprising any one of the amino acid sequences of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624, 671, or 697. A monoclonal antibody or an antigen-binding portion thereof comprising:

39. 39. The monoclonal antibody or antigen-binding portion thereof of claim 38, comprising heavy and light chain CDR sequences contained within a heavy chain variable region and light chain variable region sequence pair selected from the group consisting of SEQ ID NOs: 166 and 170, 174 and 178, 226 and 229, 310 and 314, 317 and 321, 437 and 441, 465 and 468, 538 and 541, 620 and 622, 664 and 668, and 691 and 695.

40. (a) SEQ ID NOs: 692, 693, 694, 696, 94, and 697, respectively; or (b) SEQ ID NOs: 318, 319, 320, 322, 323, and 324, respectively; or (c) SEQ ID NOs: 227, 200, 228, 230, 149, and 231, respectively; or (d) SEQ ID NOs: 113, 539, 540, 542, 196, and 543, respectively; or (e) SEQ ID NOs: 311, 312, 313, 315, 94, and 316, respectively; or (f) SEQ ID NOs: 665, 666, 667, 669, 670, and 671, respectively; or (g) SEQ ID NOs: 466, 200, 467, 469, 149, and 470, respectively; or (h) SEQ ID NOs: 167, 168, 169, 171, 172, and 173, respectively; or (i) SEQ ID NOs: 621, 176, 177, 623, 180, and 624, respectively; or (j) SEQ ID NOs: 175, 176, 177, 179, 180, and 181, respectively; or (k) SEQ ID NOs: 438, 439, 440, 442, 30, and 443, respectively.

39. The monoclonal antibody or antigen-binding portion thereof according to claim 38, comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of:

41. 39. The monoclonal antibody or antigen-binding portion thereof of claim 38, comprising a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691.

42. 39. The monoclonal antibody or antigen-binding portion thereof of claim 38, comprising a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695.

43. The antibody, (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:691, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:695; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 317, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 321; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 226, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 229; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 538, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 541; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 310, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 314; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:664, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:668; or (g) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 465, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 468; or (h) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 166, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 170; or (i) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 620, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 622; or (j) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 174, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 178; or (k) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 437, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

441.

39. The monoclonal antibody or antigen-binding portion thereof of claim 38, comprising:

44. The antigen-binding portion is Fab, F(ab') ２ 44. The monoclonal antibody or antigen-binding portion thereof of any one of claims 38 to 43, which is an Fv, scFv, a bivalent scFv, a single domain antibody, or a diabody.

45. 45. A pharmaceutical composition comprising the monoclonal antibody or antigen-binding portion thereof of any one of claims 38 to 44 and a pharma- ceutically acceptable carrier.

46. Binding affinity (K Ｄ ) a monoclonal antibody or antigen-binding portion thereof that specifically binds to an Aspergillus allergen, a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 710, 718, 726, 734, or 742; a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 711, 719, 727, 735, or 743; a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 712, 720, 728, 736, or 744; a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 714, 722, 730, 738, or 746; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 715, 723, 731, 739, or 747; and A light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 716, 724, 732, 740, or 748. A monoclonal antibody or an antigen-binding portion thereof comprising:

47. 47. The monoclonal antibody or antigen-binding portion thereof of claim 46, comprising heavy and light chain CDR sequences contained within a heavy chain variable region and light chain variable region sequence pair selected from the group consisting of SEQ ID NOs: 709 and 713, 717 and 721, 725 and 729, 733 and 737, and 741 and 745.

48. (a) SEQ ID NOs: 710, 711, 712, 714, 715, and 716, respectively; or (b) SEQ ID NOs: 718, 719, 720, 722, 723, and 724, respectively; or (c) SEQ ID NOs: 726, 7272, 728, 730, 731, and 732, respectively; or (d) SEQ ID NOs: 734, 735, 736, 738, 739, and 740, respectively; or (e) SEQ ID NOs: 742, 743, 744, 746, 747, and 748, respectively.

47. The monoclonal antibody or antigen-binding portion thereof according to claim 46, comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of:

49. 47. The monoclonal antibody or antigen-binding portion thereof of claim 46, comprising a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 709, 717, 725, 733, or 741.

50. 47. The monoclonal antibody or antigen-binding portion thereof of claim 46, comprising a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 713, 721, 729, 737, or 745.

51. The antibody, (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 709, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 713; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 717, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 721; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 725, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 729; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 733, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 737; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 741, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

745.

47. The monoclonal antibody or antigen-binding portion thereof of claim 46, comprising:

52. The antigen-binding portion is Fab, F(ab') ２ 52. The monoclonal antibody or antigen-binding portion thereof of any one of claims 46 to 51, which is an Fv, scFv, a bivalent scFv, a single domain antibody, or a diabody.

53. 53. A pharmaceutical composition comprising the monoclonal antibody or antigen-binding portion thereof of any one of claims 46 to 52 and a pharma- ceutically acceptable carrier.

54. 53. A pharmaceutical composition comprising a plurality of monoclonal antibodies according to any one of claims 46 to 52, wherein the monoclonal antibodies recognise different epitopes or specifically bind to different allergens.

55. Binding affinity (K Ｄ ) a monoclonal antibody or antigen-binding portion thereof that specifically binds to a milk allergen, a heavy chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 121, 750, 757, 765, 772, 779, 785, 793, 800, 807, 814, 821, 833, 838, 846, 853, 860, 868, 874, 881, 889, 895, 903, 911, 918, or 926; a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 378, 532, 751, 758, 766, 773, 786, 794, 801, 808, 815, 822, 826, 839, 847, 854, 861, 875, 882, 890, 896, 904, 912, 919, or 927; a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 752, 759, 767, 774, 780, 787, 795, 802, 809, 816, 827, 840, 848, 855, 862, 869, 876, 883, 891, 897, 905, 913, 920, or 928; a light chain CDR1 comprising any one of the amino acid sequences of SEQ ID NOs: 523, 754, 761, 769, 776, 782, 789, 797, 804, 811, 818, 829, 835, 842, 850, 857, 864, 871, 878, 885, 899, 907, 915, 922, or 930; a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 22, 30, 94, 110, 149, 186, 196, 389, 404, 509, 662, 682, 762, 790, 830, 843, 865, 886, 900, 908, 923, or 931; and A light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 755, 763, 770, 777, 783, 791, 798, 805, 812, 819, 824, 831, 836, 844, 851, 858, 866, 872, 879, 887, 893, 901, 909, 916, 924, or 932. A monoclonal antibody or an antigen-binding portion thereof comprising:

56. (a) SEQ ID NOs: 800, 801, 802, 804, 110, and 805, respectively; or (b) SEQ ID NOs: 121, 826, 827, 829, 830, and 831, respectively; or (c) SEQ ID NOs: 833, 826, 827, 835, 149, and 836, respectively; or (d) SEQ ID NOs: 853, 854, 855, 857, 662, and 858, respectively; or (e) SEQ ID NOs: 860, 861, 862, 864, 865, and 866, respectively; or (f) SEQ ID NOs: 868, 378, 869, 871, 682, and 782, respectively.

56. The monoclonal antibody or antigen-binding portion thereof according to claim 55, comprising heavy chain CDR1-3 and light chain CDR1-3 comprising the amino acid sequences of:

57. 56. The monoclonal antibody or antigen-binding portion thereof of claim 55, comprising a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 749, 756, 764, 771, 778, 784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873, 880, 888, 894, 902, 910, 917, or 925.

58. 56. The monoclonal antibody or antigen-binding portion thereof of claim 55, comprising a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or 929.

59. The antibody, (a) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 799, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 803; or (b) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 825, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 828; or (c) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 832, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 834; or (d) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 852, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 856; or (e) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 859, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 863; or (f) a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 867, and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

870.

56. The monoclonal antibody or antigen-binding portion thereof of claim 55, comprising:

60. The antigen-binding portion is Fab, F(ab') ２ 60. The monoclonal antibody or antigen-binding portion thereof of any one of claims 55 to 59, which is an Fv, scFv, a bivalent scFv, a single domain antibody, or a diabody.

61. 61. A pharmaceutical composition comprising the monoclonal antibody or antigen-binding portion thereof of any one of claims 55 to 60 and a pharma- ceutically acceptable carrier.

62. 61. An antibody-drug conjugate comprising the monoclonal antibody or antigen-binding portion thereof of any one of claims 29 to 35, 38 to 44, 46 to 52, or 55 to 60.

63. 63. The antibody-drug conjugate of claim 62, comprising the monoclonal antibody or antigen-binding portion thereof of any one of claims 46 to 52, wherein the drug is an antifungal agent.

64. The antibody-drug conjugate of claim 63, wherein the antifungal agent is amphotericin B.

65. 11. An isolated polynucleotide comprising a nucleotide sequence encoding the monoclonal antibody of any one of claims 29 to 35, 38 to 44, 46 to 52, or 55 to 60.

66. A vector comprising the polynucleotide of claim 65.

67. 67. A host cell comprising the polynucleotide of claim 65 or the vector of claim 66.

68. A kit comprising a monoclonal antibody of any one of claims 29 to 35, 38 to 44, 46 to 52, or 55 to 60, a pharmaceutical composition of any one of claims 36, 37, 45, 53, 54, or 61, or an antibody-drug conjugate of any one of claims 62 to 64.

69. A method for alleviating one or more allergic symptoms in a subject, comprising administering to the subject a therapeutically effective amount of a monoclonal antibody or antigen-binding portion thereof described in any one of claims 29 to 35, 38 to 44, 46 to 52, or 55 to 60, a pharmaceutical composition described in any one of claims 36, 37, 45, 53, 54, or 61, or an antibody-drug conjugate described in any one of claims 62 to 64.

70. A method for reducing one or more allergic symptoms in a subject having a peanut allergy, comprising administering to the subject a therapeutically effective amount of a monoclonal antibody or antigen-binding portion thereof described in any one of claims 29 to 35, or a pharmaceutical composition described in claim 36 or 37.

71. A method for reducing one or more allergic symptoms in a subject having a tree nut allergy, comprising administering to the subject a therapeutically effective amount of a monoclonal antibody or antigen-binding portion thereof described in any one of claims 38 to 44, or a pharmaceutical composition described in claim 45.

72. A method for reducing one or more allergic symptoms in a subject with a fungal allergy, comprising administering to a subject a therapeutically effective amount of a monoclonal antibody or antigen-binding portion thereof described in any one of claims 46 to 52, a pharmaceutical composition described in claim 53 or 54, or an antibody-drug conjugate described in any one of claims 62 to 64.

73. A method for reducing one or more allergic symptoms in a subject having a milk allergy, comprising administering to a subject a therapeutically effective amount of a monoclonal antibody or antigen-binding portion thereof described in any one of claims 55 to 60, a pharmaceutical composition described in claim 61, or an antibody-drug conjugate described in any one of claims 62 to 64.

74. 74. The method of any one of claims 69 to 73, wherein the monoclonal antibody or antigen-binding portion thereof is administered subcutaneously, intravenously, or intranasally.

75. 75. The method of any one of claims 69 to 74, wherein the allergic symptoms include one or more of: runny nose, skin wheals, skin redness, skin swelling, itching or stinging in or around the mouth and / or throat, difficulty swallowing, watery eyes, diarrhea, stomach cramps, nausea, vomiting, throat tightness, shortness of breath or wheezing, shortness of breath, and anaphylaxis.

76. 76. The method of any one of claims 69 to 75, wherein the subject is further administered an additional agent.

77. 77. The method of claim 76, wherein the additional agent is an antihistamine, epinephrine, a decongestant, a bronchodilator, or a corticosteroid.

78. 78. The method of claim 76 or 77, wherein the monoclonal antibody or antigen-binding portion thereof and the additional agent are administered substantially simultaneously or separately.

79. 79. The method of any one of claims 69 to 78, further comprising evaluating the alleviation of allergic symptoms in the subject.

80. 80. The method of claim 79, wherein evaluating comprises measuring relief of allergy symptoms by Total Nasal Symptom Score (TNSS), visual analog scale (VAS) nasal symptom score, improvement in peak nasal inspiratory flow (PNIF), allergen skin test, basophil activation test, or oral food challenge.

81. A monoclonal antibody or antigen-binding portion thereof according to any of claims 29 to 35, 38 to 44, 46 to 52, and 55 to 60, or a pharmaceutical composition according to any of claims 36, 37, 45, 54, or 61, or a method according to any of claims 69 to 80, wherein the monoclonal antibody is of the IgM, IgD, IgA, or IgG isotype.

82. 82. The monoclonal antibody, antigen-binding portion, pharmaceutical composition, or method of claim 81, wherein the antibody is an IgG4.

83. 82. The monoclonal antibody, antigen-binding portion, pharmaceutical composition, or method of claim 81, wherein the antibody comprises a modified IgM, IgD, IgG3, IgG1, IgA1, IgG2, IgG4, or IgA2 heavy chain constant region.

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