Method for isolating allergen-specific antibodies from humans and use thereof

Abstract

In one aspect, a method of generating a human monoclonal antibody that specifically binds an allergen is provided. In some embodiments, the monoclonal antibody is generated from a sequence identified from an isolated single B cell of a human subject allergic to the allergen.

Description

Background Art

[0001] Allergies are a growing health concern worldwide, characterized by a misdirected adaptive immune response against harmless proteins. With food allergies in particular, individuals must diligently avoid allergen exposure, which can lead to potentially fatal anaphylactic reactions. Allergies are incurable, and while desensitization therapies, such as immunotherapy, have shown some clinical benefit, there remains a need for rapid and effective interventions that can improve the quality of life for individuals with allergies. SUMMARY OF THE INVENTION

[0003] In one aspect, the present disclosure provides a method for generating a human monoclonal antibody that specifically binds to an allergen. In some embodiments, the method comprises:

[0004] (a) isolating single B cells from a sample of a human subject, wherein the subject is allergic to the allergen;

[0005] (b) generating cDNA from the single B cell of step (a), wherein the cDNA sequence comprises a first sequence encoding all or part of an immunoglobulin heavy chain and a second sequence encoding all or part of an immunoglobulin light chain;

[0006] (c) determining the sequence of the cDNA from step (b);

[0007] (d) analyzing the sequences determined in step (c) to identify single B cells comprising a first sequence comprising an IgE constant region or an IgG4 constant region;

[0008] (e) identifying, from the single B cell of step (d), (i) a heavy chain variable region sequence in the immunoglobulin heavy chain comprising an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain co-expressed with the immunoglobulin heavy chain in the same single B cell;

[0009] (f) expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (e); and

[0010] (g) identifying one or more antibodies from step (f) that specifically bind to the allergen.

[0011] In some embodiments, step (a) comprises sorting cells in the sample by fluorescence activated cell sorting (FACS). In some embodiments, step (a) comprises selecting single B cells expressing cell surface IgE and / or cell surface IgG4. In some embodiments, step (a) comprises isolating antibody-secreting B cells and / or memory B cells.

[0012] In some embodiments, the method comprises isolating single B cells selected for expressing cell surface IgE and identifying sequences encoding immunoglobulin heavy chains that include IgE constant regions. In some embodiments, step (a) comprises contacting cells from the sample with anti-human CD19 antibodies and anti-human IgE antibodies and selecting cells expressing CD19. + In some embodiments, the method comprises isolating single B cells that express a B cell marker and are negative for non-IgE isotypes. In some embodiments, step (a) comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgM antibodies, anti-human IgG antibodies, anti-human IgA antibodies, and anti-human IgD antibodies and selecting CD19 + IgM - IgG - IgA - IgD - B cells.

[0013] In some embodiments, the method comprises isolating single B cells selected for expressing cell surface IgG4 and identifying sequences encoding immunoglobulin heavy chains comprising IgG4 constant regions. In some embodiments, step (a) comprises contacting cells from the sample with anti-human CD19 antibodies and anti-human IgG4 antibodies and selecting cells expressing CD19. + IgG4 B cells. In some embodiments, the method comprises isolating single B cells that express a B cell marker and are negative for non-IgG4 isotypes. In some embodiments, step (a) comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgM antibodies, anti-human IgE antibodies, anti-human IgA antibodies, anti-human IgD antibodies, anti-human IgG1 antibodies, anti-human IgG2 antibodies, and anti-human IgG3 antibodies and selecting CD19 + IgM - IgE - IgA - IgD - IgG1 - IgG2 - IgG3 - B cells.

[0014] In some embodiments, step (b) comprises reverse transcribing cDNA from RNA from the 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 the single B cell. In some embodiments, the method comprises amplifying the transcriptome of the single B cell.

[0015] In some embodiments, step (f) comprises expressing the heavy chain variable region sequence and the light chain variable region sequence from step (e) in a host cell and purifying the antibody. In some embodiments, step (f) comprises expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (e) and an IgG4 constant region or an IgG1 constant region.

[0016] 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.

[0017] In some embodiments, the sample comprises peripheral blood. In some embodiments, the sample comprises tissue (e.g., tonsil tissue).

[0018] 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 milk allergens, egg allergens, nut allergens, fish allergens, shellfish allergens, legume allergens, legume allergens, seed allergens, and wheat allergens. In some embodiments, the food allergen is a peanut allergen. In some embodiments, the food allergen is a stone fruit 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.

[0019] In another aspect, monoclonal antibodies produced according to the methods disclosed herein are provided.

[0020] In another aspect, a pharmaceutical composition comprising a monoclonal antibody produced according to the methods disclosed herein is provided. In some embodiments, the pharmaceutical composition comprises a plurality of monoclonal antibodies, wherein each monoclonal antibody is produced according to the methods disclosed herein and 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).

[0021] In another aspect, a monoclonal antibody or antigen binding portion thereof that specifically binds to an allergen is provided. In some embodiments, the antibody comprises:

[0022] (a) a heavy chain variable region sequence derived from an immunoglobulin heavy chain from a single IgE- or IgG4-producing B cell of a human subject allergic to the allergen;

[0023] (b) heavy chain IgG constant region sequence;

[0024] (c) a light chain variable region sequence derived from an immunoglobulin light chain from a single IgE-producing or IgG4-producing B cell from a human subject; and

[0025] (d) a light chain constant region sequence that is of the same class as the immunoglobulin light chain of (c).

[0026] In some embodiments, the antibody, or antigen-binding portion thereof, binds to the antigen with a binding affinity (K) of less than 1 nM. D ) specifically binds to the allergen. In some embodiments, the antibody, or antigen-binding portion thereof, has a binding affinity (K) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. D ) specifically binds to the allergen. In some embodiments, the antibody binds to the allergen with a binding affinity (K) of 1 nM to 250 nM. D ) binds to the allergen. 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 milk allergens, egg allergens, nut allergens, fish allergens, shellfish allergens, legume allergens, legume allergens, seed allergens, and wheat allergens. In some embodiments, the food allergen is a peanut allergen or a stone fruit 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 cross-reacts with two different antigens. In some embodiments, the antibody cross-reacts with a first antigen of a first allergen type and a second antigen of a second allergen type different from the first allergen type. In some embodiments, the antibody cross-reacts with a peanut allergen and a stone fruit allergen. In some embodiments, the antibody cross-reacts with two or more stone fruit antigens.

[0027] In some embodiments, the antibody comprises a heavy chain variable region sequence and a light chain variable region sequence derived from an IgE-producing human B cell or an IgG4-producing human B cell.

[0028] In another aspect, a monoclonal antibody or antigen binding portion thereof that specifically binds to peanut allergen is provided. In some embodiments, the antibody binds to peanut allergen with a binding affinity (K) of less than 1 nM. D ) binds to the peanut allergen. In some embodiments, the antibody, or antigen-binding portion thereof, binds to the peanut allergen with a binding affinity (K) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. D ) specifically binds to peanut allergen. In some embodiments, the antibody binds to peanut allergen with a binding affinity (K) of 1 nM to 250 nM.D ) combined with the peanut allergen.

[0029] In some embodiments, the antibody, or antigen binding portion thereof, specifically binds to peanut allergens Ara h 2, Ara h 3, or Ara h 1. In some embodiments, the antibody, or antigen binding portion thereof, binds to peanut allergens Ara h 2, Ara h 3, or Ara h 1 with a K of less than 100 pM. D Specifically binds Ara h 2. 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 h 2 and Ara h 3. In some embodiments, the antibody, or antigen binding portion thereof, cross-reacts with a K of less than 1 nM. D Specific binding to Ara h 2 with a K of less than 1 μM D Specifically binds to Ara h 3. In some embodiments, the antibody, or antigen-binding portion thereof, binds to an epitope comprising the amino acid motif DPYSPS (SEQ ID NO: 704). Furthermore, additional peanut-specific antibodies were discovered in these experiments. Antibodies PA12P3E09 and PA12P3E11 bind to peanut extract and have little binding to the naturally occurring peanut allergen Ara h 2, while antibodies PA12P1D02, PA12P1G11, PA13P1H03, PA12P3C01, and PA12P3E04 bind strongly to both peanut extract and the naturally occurring peanut allergen Ara h 2. In some embodiments, the peanut-specific antibodies bind to peanut extract but do not bind to the naturally occurring peanut allergen Ara h 2.

[0030] In some embodiments, the antibody comprises:

[0031] a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199, 341, 348, 409, 459 or 593;

[0032] a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349, 410, 460, 539 or 594;

[0033] a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201, 350, 411, 461, 540 or 595;

[0034] a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343, 352, 413, 463, 542, or 597;

[0035] 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

[0036] A light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354, 414, 464, 543 or 599.

[0037] In some embodiments, the antibody comprises heavy and light chain CDR sequences contained in 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. In some embodiments, the antibody comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequence:

[0038] (a) SEQ ID NOs: 2, 3, 4, 6, 7 and 8, respectively; or

[0039] (b) SEQ ID NOs: 10, 11, 12, 14, 15 and 8, respectively; or

[0040] (c) SEQ ID NOs: 17, 18, 19, 21, 22 and 23, respectively; or

[0041] (d) SEQ ID NOs: 25, 26, 27, 29, 30 and 31, respectively; or

[0042] (e) SEQ ID NOs: 33, 34, 35, 37, 38 and 39, respectively; or

[0043] (f) SEQ ID NOs: 41, 34, 35, 37, 38 and 39, respectively; or

[0044] (g) SEQ ID NOs: 47, 48, 49, 51, 52 and 53, respectively; or

[0045] (h) SEQ ID NOs: 47, 48, 55, 51, 52 and 53, respectively; or

[0046] (i) SEQ ID NOs: 58, 59, 60, 62, 30 and 63, respectively; or

[0047] (j) SEQ ID NOs: 58, 59, 60, 6, 7 and 8, respectively; or

[0048] (k) SEQ ID NOs: 2, 3, 4, 62, 30 and 63, respectively; or

[0049] (1) SEQ ID NOs: 58, 3, 4, 6, 7 and 8, respectively; or

[0050] (m) SEQ ID NOs: 2, 59, 4, 6, 7 and 8, respectively; or

[0051] (n) SEQ ID NOs: 2, 3, 60, 6, 7 and 8, respectively; or

[0052] (o) SEQ ID NOs: 129, 130, 131, 133, 126 and 134, respectively; or

[0053] (p) SEQ ID NOs: 341, 342, 343, 345, 78 and 346, respectively; or

[0054] (q) SEQ ID NOs: 348, 349, 350, 352, 353 and 354, respectively; or

[0055] (r) SEQ ID NOs: 199, 200, 201, 203, 149 and 204, respectively; or

[0056] (s) are SEQ ID NOs: 409, 410, 411, 413, 86 and 414, respectively; or

[0057] (t) SEQ ID NOs: 459, 460, 461, 463, 196 and 464, respectively; or

[0058] (u) SEQ ID NOs: 113, 539, 540, 542, 196 and 543, respectively; or

[0059] (v) SEQ ID NOs: 593, 594, 595, 597, 598 and 599, respectively.

[0060] 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.

[0061] In some embodiments, the antibody comprises:

[0062] (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

[0063] (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

[0064] (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

[0065] (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

[0066] (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

[0067] (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

[0068] (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

[0069] (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

[0070] (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

[0071] (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

[0072] (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

[0073] (1) 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

[0074] (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

[0075] (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

[0076] (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

[0077] (p) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 128; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 132; or

[0078] (q) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 340; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 344; or

[0079] (r) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 347; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 351; or

[0080] (s) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 406; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 407; or

[0081] (t) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 408; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 412; or

[0082] (u) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 458; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 462; or

[0083] (v) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 538; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 541; or

[0084] (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.

[0085] In yet another aspect, a monoclonal antibody or antigen binding portion thereof that specifically binds to a stone fruit allergen is provided. In some embodiments, the antibody binds to a stone fruit allergen with a binding affinity (K) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. D ) binds to the stone fruit allergen. In some embodiments, the antibody binds to the stone fruit allergen with a binding affinity (K) of less than 1 nM. D ) binds to the stone fruit allergen. In some embodiments, the antibody binds to the stone fruit allergen with a binding affinity (K) of 1 nM to 250 nM. D ) in combination with the stone fruit allergen. In some embodiments, the stone fruit allergen is cashew, pistachio, pecan, walnut, hazelnut and / or macadamia nut.

[0086] In some embodiments, the antibody comprises:

[0087] 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;

[0088] 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;

[0089] 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;

[0090] a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623, 669 or 696;

[0091] 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

[0092] A light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624, 671 or 697.

[0093] In some embodiments, the antibody comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequence:

[0094] (a) SEQ ID NOs: 692, 693, 694, 696, 94 and 697, respectively; or

[0095] (b) SEQ ID NOs: 318, 319, 320, 322, 323 and 324, respectively; or

[0096] (c) SEQ ID NOs: 227, 200, 228, 230, 149 and 231, respectively; or

[0097] (d) SEQ ID NOs: 113, 539, 540, 542, 196 and 543, respectively; or

[0098] (e) SEQ ID NOs: 311, 312, 313, 315, 94 and 316, respectively; or

[0099] (f) SEQ ID NOs: 665, 666, 667, 669, 670 and 671, respectively; or

[0100] (g) SEQ ID NOs: 466, 200, 467, 469, 149 and 470, respectively; or

[0101] (h) SEQ ID NOs: 167, 168, 169, 171, 172 and 173, respectively; or

[0102] (i) SEQ ID NOs: 621, 176, 177, 623, 180 and 624, respectively; or

[0103] (j) SEQ ID NOs: 175, 176, 177, 179, 180 and 181, respectively; or

[0104] (k) are SEQ ID NOs: 438, 439, 440, 442, 30 and 443, respectively.

[0105] 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.

[0106] In some embodiments, the antibody comprises:

[0107] (a) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 691; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 695; or

[0108] (b) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 317; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 321; or

[0109] (c) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 226; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 229; or

[0110] (d) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 538; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 541; or

[0111] (e) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 310; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 314; or

[0112] (f) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 664; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 668; or

[0113] (g) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 465; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 468; or

[0114] (h) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 166; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 170; or

[0115] (i) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 620; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 622; or

[0116] (j) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 174; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 178; or

[0117] (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.

[0118] In 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 a milk allergen with a binding affinity (K) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. D ) binds to the milk allergen. In some embodiments, the antibody binds to the milk allergen with a binding affinity (K) of less than 1 nM. D ) binds to the milk allergen. In some embodiments, the antibody binds to the milk allergen with a binding affinity (K) of 1 nM to 250 nM. D ) combined with the milk allergen.

[0119] In some embodiments, the antibody comprises:

[0120] a heavy chain CDR1 comprising the amino acid sequence of any one 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;

[0121] 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;

[0122] 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;

[0123] a light chain CDR1 comprising the amino acid sequence of any one 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;

[0124] 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

[0125] 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.

[0126] In some embodiments, the antibody comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequence:

[0127] (a) SEQ ID NOs: 800, 801, 802, 804, 110 and 805, respectively; or

[0128] (b) SEQ ID NOs: 121, 826, 827, 829, 830 and 831, respectively; or

[0129] (c) SEQ ID NOs: 833, 826, 827, 835, 149 and 836, respectively; or

[0130] (d) SEQ ID NOs: 853, 854, 855, 857, 662 and 858, respectively; or

[0131] (e) SEQ ID NOs: 860, 861, 862, 864, 865 and 866, respectively; or

[0132] (f) SEQ ID NOs: 868, 378, 869, 871, 682 and 782, respectively.

[0133] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any 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 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.

[0134] In some embodiments, the antibody comprises:

[0135] (a) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 799; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 803; or

[0136] (b) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 825; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 828; or

[0137] (c) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 832; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 834; or

[0138] (d) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 852; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 856; or

[0139] (e) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 859; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 863; or

[0140] (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.

[0141] In some embodiments, the antibody is an antibody that competes with a monoclonal antibody as disclosed herein for binding to an allergen (eg, for binding to a food allergen, such as a peanut allergen, a stone fruit allergen, or a milk allergen).

[0142] 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 with a binding affinity (K) of less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. D ) binds to the fungal allergen (e.g., Aspergillus allergen). In some embodiments, the antibody binds with a binding affinity (K) of less than 1 nM. D ) binds to the fungal allergen (e.g., Aspergillus allergen). In some embodiments, the antibody binds to the fungal allergen with a binding affinity (K) of 1 nM to 250 nM. D ) combined with the fungal allergen.

[0143] 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 f 1).

[0144] In some embodiments, the antibody comprises:

[0145] a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 710, 718, 726, 734 or 742;

[0146] a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 711, 719, 727, 735 or 743;

[0147] a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 712, 720, 728, 736 or 744;

[0148] a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 714, 722, 730, 738 or 746;

[0149] a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 715, 723, 731, 739 or 747; and

[0150] A light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 716, 724, 732, 740 or 748.

[0151] In some embodiments, the antibody comprises heavy and light chain CDR sequences contained in 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. In some embodiments, the antibody comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequences:

[0152] (a) SEQ ID NOs: 710, 711, 712, 714, 715 and 716, respectively; or

[0153] (b) SEQ ID NOs: 718, 719, 720, 722, 723 and 724, respectively; or

[0154] (c) SEQ ID NOs: 726, 7272, 728, 730, 731 and 732, respectively; or

[0155] (d) SEQ ID NOs: 734, 735, 736, 738, 739 and 740, respectively; or

[0156] (e) SEQ ID NOs: 742, 743, 744, 746, 747 and 748, respectively.

[0157] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to any 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 of SEQ ID NOs: 713, 721, 729, 737, or 745.

[0158] In some embodiments, the antibody comprises:

[0159] (a) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 709; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 713; or

[0160] (b) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 717; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 721; or

[0161] (c) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 725; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 729; or

[0162] (d) a heavy chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 733; and a light chain variable region comprising an amino acid sequence that has at least 90% sequence identity to SEQ ID NO: 737; or

[0163] (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.

[0164] In some embodiments, the antibody is an antibody that competes with a monoclonal antibody as disclosed herein for binding to a fungal allergen (eg, for binding to an Aspergillus allergen).

[0165] In another aspect, a pharmaceutical composition is provided comprising a monoclonal antibody or antigen-binding portion as disclosed herein. In some embodiments, the pharmaceutical composition comprises a plurality of monoclonal antibodies as 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 in different types or classes of allergens).

[0166] In another aspect, an antibody-drug conjugate is provided, comprising a monoclonal antibody, or an antigen-binding portion thereof, as disclosed herein. In some embodiments, the antibody-drug conjugate comprises a monoclonal antibody, or an antigen-binding portion thereof, that specifically binds to a fungal allergen as disclosed herein and further comprises an antifungal agent. In some embodiments, the antifungal agent is amphotericin B.

[0167] In yet another aspect, the isolated polynucleotide comprises a nucleotide sequence encoding a monoclonal antibody as disclosed herein. Also provided herein are vectors and host cells comprising a polynucleotide as disclosed herein.

[0168] In another aspect, methods of treatment are provided. In some embodiments, the method of treatment is a method of alleviating one or more allergic symptoms in a subject. In some embodiments, the method of treatment is a method of alleviating one or more allergic symptoms in a subject with a peanut allergy. In some embodiments, the method of treatment is a method of alleviating one or more allergic symptoms in a subject with a stone fruit allergy. In some embodiments, the method of treatment is a method of alleviating one or more allergic symptoms in a subject with a fungal allergy. In some embodiments, the method of treatment is a method of alleviating one or more allergic symptoms in a subject with a milk allergy. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a monoclonal antibody or pharmaceutical composition as disclosed herein.

[0169] In another aspect, a kit is provided. In some embodiments, the kit comprises a monoclonal antibody or pharmaceutical composition as disclosed herein. In some embodiments, the kit is used to practice a method of treatment as disclosed herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0170] Figure 1 . Flowchart of an exemplary workflow of a method for generating allergen-specific monoclonal antibodies.

[0171] 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, gray dots) B cell subsets, which are identified by the expression of established transcription factors and marker genes (right; non-striped for naive / memory B cells and striped for plasmablast (PB) B cells). (B) B cell isotypes, black dots for IGHE and gray dots for other isotypes. (C) Number of cells belonging to each subtype in (A) by isotype (non-striped for naive / memory B cells and striped for plasmablast (PB) B cells). (DG) Analysis of clonal families (CFs). (D) Distribution of cell numbers per CF. (E) Fraction of cells belonging to each isotype in multi-member CFs. (F) Isotype, B cell subtype, mutation frequency, and patient of origin for each cell in multi-member CFs. CFs mentioned in the text are labeled. (G) Heavy chain (right) and light chain (left) CDR3 sequences and similarity heatmaps of CF in (F).

[0172] Figures 3A-3G.Characterization of 89 IgE antibodies from single B cells. (A) Phylogenetic description of antibody heavy chains arranged by IGHV gene (background pattern), patient origin (node ​​pattern), and mutation frequency (node ​​size). (B) Heat map indicating the number of cells with a given heavy and light chain CDR3 length. (C) Heavy chain and light chain mutation frequency per cell. (D) Silent (S) and substitution (R) mutations by region in heavy and light chains. (E) Differential gene expression between IgE PB and other isotype PB. Positive log-fold change indicates genes enriched in IgE PB. (F) Histogram of heavy chain constant region coverage of naive / memory B cells (top) and PB (bottom) by isotype, with loci oriented in a 5' to 3' direction. The average normalized read depth and 95% confidence interval are indicated by the solid line and shaded area, respectively, for the number of cells labeled (n). Membrane exons are the two 3'-most exons of each isotype. (G) Summary of (F), but depicting the proportion of cells with any membrane exon coverage for each isotype.

[0173] Figures 4A-4C Convergence of high-affinity, cross-reactive IgE antibodies in two unrelated individuals (PA12 & PA13). Antibody patterns were conserved between panels. (A) Frequency of silent (S) and substitution (R) mutations by region. (B) Percentile frequency of mutations compared to all class-switched antibodies. (C) Dissociation constants (K) of the major allergen peanut proteins Ara h2 and Ara h3 against six aggregated antibodies and eight variants of PA13P1H08. D Abbreviated antibody variant names are represented as "heavy-light" using the following abbreviations: N = native, R = reverted, FWR = framework region. The "r" prefix indicates that only that region has been reverted.

[0174] Figures 5A-5E Germline transcriptome reveals class switch initiation in individual B cells. (A) Identity of Cε germline transcript splice donors and the number of cells expressing each by isoform. (B) Proportion of cells expressing εGLT by isoform. (C) Example from a single PA11, where identification of phase variants within an IgE constant region exon enabled subsequent validation of biallelic GLT expression in other B cells from the same patient. (D) Global germline transcriptome heatmap representing the proportion of cells expressing a given isoform of a given GLT. Top: GLT isoform expression frequency relative to all GLT isoforms; self-isoform GLT expression is excluded. (E) Histogram of the number of non-self GLT isoforms expressed in each cell.

[0175] Figure 6Study Overview. Plasma was extracted from fresh blood to measure circulating IgE levels, and the cell fraction was enriched for B cells prior to FACS. Single cells were sorted into individual wells of a 96-well plate and processed using scRNA-seq to generate sequencing reads, which 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 the recombinantly expressed antibodies.

[0176] Figures 7A-7C Plasma IgE levels. (A) Allergen specificity and concentration of allergen components (hazelnut, peanut). (B) Total IgE concentration. (C) Positive correlation between total plasma IgE levels and the number of confirmed IgE+ B cells. Each dot represents an individual.

[0177] 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 identity within the final IgE gate determined by heavy chain transcript assembly. ND = Not Determined. (C) For reference, putative basophils (CD19-IgE+) show higher IgE surface expression than IgE+ B cells.

[0178] Figures 9A-9G Overview and quality control of single-cell RNA-seq data. (A) Cells were sequenced in five libraries to a depth of ~1–2 million reads per cell. (B) Histogram of genes per cell. Cells expressing fewer than 950 genes were discarded. (C) Interpretation curves depict the number of genes detected as a function of sequencing depth for eight randomly selected cells from each B cell subtype. The solid line and shaded area represent the mean and 95% confidence interval of gene counts, respectively. (D) Distribution of read mappings for retained cells. Most reads were uniquely mapped (Ensembl reference annotation), while multiply mapped reads primarily belonged to RNA18S5 repeats on chr21 and unplaced backbones. (E) Read mapping across gene bodies shows minimal 3' or 5' bias. (F) Histogram of V gene assembly length by strand. (G) PCA of the top 500 most variable genes before (top) and after (bottom) batch correction.

[0179] Figures 10A-10D Ancillary data supporting B cell subtype classification. Compared to the naive / memory B cell subset (non-striped), PB (striped) has stronger FACS forward and side scatter (A), more cDNA after SmartSeq2 pre-amplification (B), and higher gene expression for antibody light and heavy chains (C). (D) The most differentially expressed genes for each subset.

[0180] Figures 11A-11K Comparison of B cells between isotypes. (AI) Number of cells harboring a given V and J gene by 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 by isotype. (K) Relative utilization of lambda and kappa light chains by isotype.

[0181] Figures 12A-12H . Characterization of antibody specificity and affinity. (A) Semiquantitative indirect ELISA of fusion antibodies, controls, PA13P1H08 variants, and antibodies from other clonal families. A human IgG isotype control (abcam #ab206195) was used as a negative control, while positive controls included an anti-Ara h mouse monoclonal antibody purchased from Indoor Biotechnologies. (BG) Kinetic characterization of antibodies binding to Ara h 2 (BD) and Ara h 3 (EG) using biolayer interferometry. The 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 extracts, the natural peanut allergen Ara h 2, and bovine serum albumin (BSA) (columns). The isotype of each antibody is shown on the left. Higher values ​​indicate stronger binding. OD = optical density. The values ​​depicted represent the values ​​after subtracting the human IgG isotype control. Only any tested antibodies with an OD value above 0.25 are displayed.

[0182] Figures 13A-13E Heavy and light chain sequences used to construct PA13P1H08 antibody variants. (A) Derivation of the original heavy chain CDR3 and surrounding amino acids. (B) Native and reverted heavy chain sequences, except for regions of the heavy chain that have been reverted to the presumed original rearranged sequence. Labels prefixed with "r" simply indicate that the region has been reverted. FWR = framework. (C) Derivation of the original light chain CDR3 and surrounding amino acids. (D) Native and reverted light chain sequences. (E) Light chain sequence taken from another antibody, PA12P4H03, which we confirmed by ELISA does not bind to any peanut allergens.

[0183] Figure 14 GLT splicing for all isoforms. Note that for clarity, only the first three constant region exons are shown for each isoform.

[0184] Figure 15IGV overlay histogram of the ghE constant region locus and splice junctions showing eGLT in single murine B cells stimulated with IL-4, LPS, and BAFF (Wu et al., 2017). Arrows indicate the ghE GLT splice donor.

[0185] Figure 16 Pairwise CDR3 sequence identity of heavy chain CDR3 sequences from clones PA12P3F10, PA12P3D08, PA12P1C07, PA13P1E10, PA13P3G09, and PA13P1H08 (SEQ ID NOs: 19, 35, 42, 12, 27, 4) and three heavy chain CDR3 sequences from multiple patients in a separate peanut allergy immune repertoire sequencing study (62). Each sequence from this separate study had at least 70% identity with one or more sequences from this study. All sequences shared the IGHV3-30 and IGHJ6 gene segments, and the CDR3 was 17 amino acids in length.

[0186] Figure 17A Microarray scanning confirmed the absence of background interactions between the secondary goat anti-human IgG and control mouse monoclonal anti-HA antibodies and the antigen-derived peptides. The control antibodies produced the expected HA control spot pattern (white dots) for the constructed peptide microarrays.

[0187] Figure 17B and 17C Microarray epitope mapping of PA13P1H08 to Ara h 2 ("Ah2") and Ara h 3 ("Ah3") peptides. (B) Microarray scan showing antibody binding to the antigenic peptide (light gray) and the expected HA control spot pattern (white dots). (C) Microarray fluorescence intensity by antigenic peptide. Ara h 2 motifs with high intensity are annotated.

[0188] Figure 18 Plasma IgE levels in milk-allergic subject PA01 and Aspergillus study subject 10033201 to common food allergens, as well as Aspergillus fumigatus and Aspergillus niger. NP = Not Performed. Total IgE for PA01 and 10033201 was 353 kU / L and 3528 kU / L, respectively. The assay was performed by the CLIA-licensed Johns Hopkins University Dermatology, Allergy, and Clinical Immunology Reference Laboratory using the ImmunoCAP system.

[0189] Figure 19Indirect ELISA showing binding of recombinant monoclonal antibodies (columns) to antigens (rows). Antigens include extracts of Aspergillus fumigatus, Aspergillus niger, and Aspergillus nidulans, as well as purified recombinant allergen Aspergillus fumigatus 1 (rAsp f 1). Bovine serum albumin (BSA) served as a negative antigen control. A monoclonal antibody against Asp f 1 ("anti-Asp f 1") served as a positive control against this allergen. Higher values ​​indicate stronger binding. OD = optical density. hIgG = human IgG isotype control.

[0190] Figure 20 Indirect ELISA showing binding of recombinant monoclonal antibodies from subject PA01 (rows) to allergen extracts, the natural peanut allergen Ara h 2, and BSA (columns). The isotype of each antibody is indicated on the left. Higher values ​​indicate stronger binding. OD = optical density. Depicted values ​​represent values ​​after subtraction of a human IgG isotype control.

[0191] Table 1

[0192] Table 1 includes the protein and nucleic acid sequences discussed herein. Polypeptide sequences are provided using standard one-letter codes. One of ordinary skill in the art provided with amino acid sequences will understand that amino acid sequences can be encoded by a set of determined nucleotide sequences, so that readers and inventors have a nucleotide sequence encoding each amino acid sequence. The nucleic acid sequence encoding the polynucleotide can 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 can be a sequence that is codon-optimized for expression in human cells or specific cell types, eukaryotic cells, bacterial cells, etc. Codon optimization can be performed to improve the host's ability to produce a polypeptide encoded by the nucleic acid, which replaces one codon with another codon that encodes the same amino acid and has a higher frequency of occurrence in a specific host cell (see, e.g., Mauro, BioDrugs 32(1):69-81, 2018 and Kato, Int J Mol Sci. 20(4), 2019).

[0193] 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 having a heavy chain variable region and a light chain variable region is described, the heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1, and the light chain variable region comprising an amino acid sequence having 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 method, the identity of two nucleotide or polypeptide sequences or subsequences is calculated as the percentage of positions that are identical or equivalent after the sequences have been aligned, introducing gaps if necessary to achieve the maximum percentage of 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 various sources on EMBnet).

[0194] It is contemplated that, in certain embodiments, the methods or compositions described herein differ from the polypeptide sequences provided herein (e.g., in Table 1) by one or more amino acid substitutions. In some embodiments, the sequences will have at least 90% sequence identity (or other degrees of sequence identity disclosed below) to a sequence or combination of sequences described herein. In one embodiment, the polypeptide sequence differs from a reference sequence (e.g., in Table 1) by one amino acid substitution. In one embodiment, the polypeptide sequence differs from a reference sequence (e.g., in Table 1) by two amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by two amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by three amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by four amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by five amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by six amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by seven amino acid substitutions. In one embodiment, the polypeptide sequence differs from a reference sequence by eight amino acid substitutions. In one embodiment, the difference between the polypeptide sequence and the reference sequence is nine amino acid substitutions. In one embodiment, the difference between the polypeptide sequence and the reference sequence is ten amino acid substitutions. In certain embodiments, the difference between the polypeptide sequence and the reference sequence is 1 to 10 amino acid substitutions, sometimes 1 to 5 amino acid substitutions. In some cases, amino acid substitutions that do not change the basic properties (e.g., binding specificity) relative to the reference sequence are selected. In some cases, amino acid substitutions that change binding affinity but do not change binding specificity are selected. In some cases, substitutions are selected to change properties as 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 identified in the art as replacing one amino acid with another amino acid having similar properties (e.g., polarity, charge, hydrophobicity, and aromaticity). Conservative amino acid substitutions can also be made based on the side chain characteristics of the amino acid (e.g., containing sulfur, hydroxyl, or amide). Non-limiting examples of conservative amino acid substitutions are listed below.

[0195] Detailed Description of the Invention

[0197] I. Introduction

[0198] In one aspect, the present disclosure provides human allergen-specific monoclonal antibodies and methods for generating human allergen-specific monoclonal antibodies from human B cells expressing a single IgE or IgG4. 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 hives to potentially fatal anaphylaxis, are caused by degranulation of mast cells and basophils, which is triggered by recognition of allergenic food proteins by surface-bound IgE antibodies. Despite its central role in immunity and allergic diseases, human IgE antibodies remain poorly characterized due to their scarcity. Fitzsimmons et al., Front Immunol., 2014, 5:61. Similarly, knowledge regarding the IgG4 isotype is lacking, but interest in it is growing due to its potential role in mediating reduced clinical allergen reactivity associated with immunotherapy and early allergen exposure through antigen blocking. Tordesillas et al., Immunity, 2017, 47:32–50.

[0199] The present disclosure provides therapeutic methods for treating a human subject having an allergy or allergic reaction or allergic reaction symptoms in a human subject using one or more allergen-specific monoclonal antibodies, or antigen-binding portions thereof, as disclosed herein. In some embodiments, and 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.

[0200] As described herein, the method of the present disclosure can be used to generate a library of IgE or IgG4 single B cells of genotype confirmation from a sample of a human subject with an allergy to a target antigen, and the IgE or IgG4 single B cells are candidates for producing antibodies with high affinity to the allergen of interest. As described in the Examples section below, it has been found that analyzing the cDNA sequence of the constant region of the immunoglobulin heavy chain to identify the isotype of a single B cell avoids the problem of isotype mischaracterization, which is known to occur when cells are sorted and the B cell isotype is determined based on cell surface markers (e.g., typically performed in FACS cell surface staining). This problem of known isotype mischaracterization is particularly common for 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 methods of the present disclosure generate a library of single B cells that are more likely to produce antibodies with high affinity to allergens. Furthermore, it has been found that the antibodies generated according to the methods disclosed herein are the highest affinity natural human antibodies discovered to date and exhibit cross-reactivity with different antigens.

[0201] II. Definitions

[0202] The terms used herein are for the purpose of describing specific embodiments only and are not intended to be limiting, as the scope of the present invention will be limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art to which the present invention belongs. In this specification and the claims that follow, a number of terms will be mentioned, and unless it is apparent that there is an opposite intention, these terms should be defined as having the following meanings. In some cases, for clarity and / or ease of reference, terms with commonly understood meanings are defined herein, and the inclusion of such definitions herein should not be interpreted as representing substantial differences between the definitions of terms generally understood in the art.

[0203] All numerical designations, such as pH values, temperatures, times, concentrations, and molecular weights, including ranges, are approximate and vary as appropriate by increments of 0.1 or 1.0 (+) or (-). It should be understood, although not always explicitly stated, that all numerical designations are preceded by the term "about."

[0204] 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.

[0205] The term "comprising" is intended to indicate that compounds, compositions, and methods include the recited elements, but not to exclude other elements. When used to define compounds, compositions, and methods, "consisting essentially of" shall mean excluding other elements that would materially affect the basic and novel characteristics of the claimed invention. "Consisting of" shall mean excluding any element, step, or ingredient not specified in the claims. Embodiments defined by each of these transition terms are within the scope of the present invention.

[0206] The term "allergen" refers to a substance that provokes an immune response in a subject, which results in an allergic reaction in the subject.

[0207] As used herein, the term "antibody" refers to a polypeptide encoded by an immunoglobulin gene or a functional fragment thereof that specifically binds to 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 κ, λ, α, γ, δ, ε, and μ constant region genes, as well as various immunoglobulin variable region genes. Light chains are classified as κ or λ. Heavy chains are classified as γ, μ, α, δ, or ε, which in turn define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively.

[0208] An exemplary immunoglobulin (antibody) structural unit comprises two pairs of identical 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 to 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.

[0209] The term "variable region" refers to the domain in an antibody heavy or light chain that confers antigen-binding specificity to the antibody. Typically, an antibody variable region comprises four conserved "framework" regions interspersed with three highly variable "complementarity determining regions."

[0210] 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 antigenic epitopes. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3, and are numbered sequentially from the N-terminus and are also typically identified by the chain in which a particular CDR is located. Thus, a VH CDR3 is located in the variable region of the heavy chain of the antibody in which it is found, while a VL CDR1 is the CDR1 of the variable region of the light chain of the antibody in which it is found.

[0211] As noted, the portion of the variable region not included in the CDRs is called the framework. The "framework regions" of different light or heavy chains are relatively conserved within a species. The framework regions of an antibody, i.e., the combined framework regions of the light and heavy chains, serve to position and arrange the CDRs in three-dimensional space. Framework sequences can be obtained from public DNA databases or published references containing 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.

[0212] 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 have been precisely defined by Kabat et al., Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1983, 1987, and others. See, e.g., 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 Contacts.

[0213] 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 specific binding to an antigen (e.g., an allergen, such as Ara h 2 or Ara h 3). 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 Fv (scFv), disulfide-linked Fv (dsFv), complementarity determining regions (CDRs), VL (light chain variable region), VH (heavy chain variable region), nanobodies, and any combination of those or any other functional portions 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 fragments of the antibodies of the invention include: (a) VH-CH1; (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 OpinBiotechnol., (6):653-8; and Ubah et al., 2016, “Phage Display Derived IgNAR V RegionBinding Domains for Therapeutic Development" Curr Pharm Des. 22(43):6519-6526, each of which is incorporated herein by reference.

[0214] The term "epitope" refers to a region or zone of an antigen to which an antibody specifically binds, i.e., a region or zone in physical contact with the antibody, and can comprise several amino acids or portions of several amino acids, for example, 5 or 6 or more, for example 20 or more amino acids, or portions of those amino acids. In some cases, the epitope comprises, for example, a non-protein component from a carbohydrate, nucleic acid, or lipid. In some cases, the epitope is a three-dimensional portion. Thus, for example, where the target is a protein, the epitope can be composed of continuous amino acids or of amino acids from different portions (e.g., discontinuous epitopes) of the protein that become close by protein folding.

[0215] "Monoclonal antibody" refers to an antibody produced by a single clone of cells or a single cell line, which consists of or consists essentially of antibody molecules that are identical in their primary amino acid sequence. In some embodiments, a monoclonal antibody preparation comprises a population of antibodies that are identical, except for mutations that arise 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.

[0216] "Human antibody" refers to antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present disclosure may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed 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) have been grafted onto human framework sequences.

[0217] The term "specifically binds" means that a molecule (e.g., an antibody or antibody fragment) binds to a target in a sample with greater affinity, avidity, more readily, and / or for a longer 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, such as Ara h 2 or Ara h 3) is an antibody or antigen-binding portion that binds to the target with an affinity that is at least 2 times greater than its affinity for a non-target compound, such as at least 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 25 times, 50 times, or more. For example, in some embodiments, an antibody that specifically binds to an allergen target (such as Ara h 2 or Ara h 3) will typically bind to the allergen target with an affinity that is at least 2 times greater than its affinity for a non-allergen target. One of ordinary skill in the art will understand that an antibody that specifically or preferentially binds to a first target may also specifically or preferentially bind to a second target or may not specifically or preferentially bind to a second target.

[0218] As used herein, the term "binding affinity" refers to the strength of a non-covalent interaction between two molecules, such as an antibody (or antigen-binding fragment thereof) and an antigen. Thus, for example, the term may refer to a 1:1 interaction between an antibody (or antigen-binding fragment thereof) and an antigen, unless otherwise indicated or clear from the context. Binding affinity can be measured by measuring the equilibrium dissociation constant (K D ) is quantified, and the equilibrium dissociation constant is the dissociation rate constant (k d ,time -1 ) divided by the association rate constant (k a ,time -1 M -1 K can be determined by measuring the kinetics of complex formation and dissociation. D , for example using surface plasmon resonance (SPR) methods such as Biacore TM system; kinetic exclusion tests, e.g. and BioLayer interferometry (e.g. using Octet platform). As used herein, "binding affinity" includes not only formal binding affinity, such as formal affinity reflecting a 1:1 interaction between an antibody (or an antigen-binding fragment thereof) and an antigen, but also apparent affinity, which can reflect strong binding, for which K is calculated. D .

[0219] As used herein, the term "cross-reactivity" 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 Arah 2) may exhibit cross-reactivity with a second allergen target (e.g., a second peanut allergen, such as Arah h 3).

[0220] The term "isolated" as used with reference to a nucleic acid or protein (e.g., an antibody) means that the nucleic acid or protein is substantially free of other cellular components with which it is associated in its native state. Preferably, it is 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) has a purity of at least 85%, a purity of at least 90%, a purity of at least 95%, or a purity of at least 99%.

[0221] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The term applies to amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. As used herein, the term encompasses amino acid chains of any length, including full-length proteins, in which the amino acid residues are linked by covalent peptide bonds.

[0222] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are later modified, such as hydroxyproline, γ-carboxyglutamate, 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, carboxyl, amino, and R groups, such as homoserine, norleucine, methionine sulfoxide, and methioninemethyl sulfonium. 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" refers to compounds that have a structure that is different from the general chemical structure of an amino acid but function in a manner similar to naturally occurring amino acids.

[0223] 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. Likewise, nucleotides may be referred to by their commonly accepted single-letter codes.

[0224] 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 also refers to deoxyribonucleotides, ribonucleotides, and naturally occurring variants, and can also refer to synthetic and / or non-naturally occurring nucleic acids (i.e., including nucleic acid analogs or modified backbone residues or linkages), for example, but not limited to: phosphorothioate, phosphoramidate, methylphosphonate, chiral-methylphosphonate, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNA), etc. Unless otherwise indicated, a specific nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences as well as the sequences explicitly indicated. 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)).

[0225] As used herein, the term "sample" refers to a biological sample obtained from a human or non-human mammalian subject. In some embodiments, the sample includes blood, blood components (blood fraction) or blood products (e.g., serum, plasma, platelets, red blood cells, peripheral blood mononuclear cells, etc.); sputum or saliva; feces, urine, other biological fluids (e.g., lymph, saliva, prostatic fluid, gastric juice, intestinal fluid, renal fluid, lung fluid, cerebrospinal fluid, etc.), tissue (e.g., kidney, lung, liver, heart, brain, neural tissue, thyroid, eye, skeletal muscle, cartilage or bone tissue), cultured cells (e.g., primary culture, explant, transformed cell or stem cell) or biopsy sample.

[0226] The terms "subject" and "patient" 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 mammals. In one embodiment, the subject or patient is a human.

[0227] The term "treat" refers to any indicator of successful treatment or alleviation of an injury, disease, or condition, including any objective or subjective parameter, such as ablation; remission; lessening of symptoms or making the injury, disease, or condition more tolerable to the subject; slowing the rate of deterioration or decline; making the final point of decline less debilitating; and / or improving the subject's physical or mental health. Treatment or alleviation of symptoms can be based on objective or subjective parameters. The effects of treatment can be compared to that of an individual or a large group of individuals who have not received treatment, or to the same patient at different times before or during treatment.

[0228] The term "pharmaceutical composition" refers to a composition suitable for administration to a subject. Typically, a pharmaceutical composition is sterile and preferably does not contain contaminants that can cause an undesirable response in a subject. A pharmaceutical composition can be designed to be administered to a subject in need thereof via a variety of different routes of administration, including oral, intravenous, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, inhalation, and the like.

[0229] The term "pharmaceutically acceptable excipient" refers to an inactive drug ingredient that is biologically or pharmacologically compatible with humans or animals, such as, but not limited to, a buffer, a carrier, or a preservative.

[0230] As used herein, a "therapeutic amount" or "therapeutically effective amount" of an agent (e.g., a monoclonal antibody disclosed herein) refers to an amount that treats, alleviates, abates, relieves, improves patient survival, increases survival time or survival rate, reduces symptoms, makes an injury, disease, or condition (e.g., an allergy) more tolerable, slows the rate of degeneration or decline, or improves the patient's physical or mental health. For example, for a given parameter, a therapeutically effective amount will show an increase or decrease in therapeutic effect of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or 100%. Therapeutic efficacy can also be expressed as a "fold" increase or decrease. For example, a therapeutically effective amount can have an effect of at least 1.2-fold, 1.5-fold, 2-fold, 5-fold, or more relative to a control.

[0231] The term "administering" or "dosing" refers to a method of delivering an agent, compound, or composition to the 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 optionally used with the agents and methods described herein include, for example, those discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current edition; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, PA.

[0232] III. Methods for Producing Allergen-Specific Antibodies

[0233] In one aspect, a method for generating allergen-specific monoclonal antibodies from a human sample is provided. In some embodiments, the method comprises:

[0234] (a) isolating single B cells from a sample of a human subject, wherein the subject is allergic to the allergen;

[0235] (b) generating cDNA from the single B cell of step (a), wherein the cDNA sequence comprises a first sequence encoding all or part of an immunoglobulin heavy chain and a second sequence encoding all or part of an immunoglobulin light chain;

[0236] (c) determining the sequence of the cDNA from step (b);

[0237] (d) analyzing the sequences determined in step (c) to identify single B cells comprising a first sequence comprising an IgE constant region or an IgG4 constant region;

[0238] (e) identifying, from the single B cell of step (d), (i) a heavy chain variable region sequence in the immunoglobulin heavy chain comprising an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain co-expressed with the immunoglobulin heavy chain in the same single B cell;

[0239] (f) expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (e); and

[0240] (g) identifying one or more antibodies from step (f) that specifically bind to the allergen.

[0241] Subject population and sample

[0242] 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 a leukapheresis product. In some embodiments, the sample comprises peripheral blood mononuclear cells (PBMCs). In some embodiments, the sample comprises tissue from a human subject, for example, 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.

[0243] In some embodiments, the allergen-specific antibodies are generated from a human subject with an allergy to the allergen. In some embodiments, the human subject with the allergy is an adult. In some embodiments, the human subject with the allergy is a teenager. 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, for example, an allergy to 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, 10, or more allergens. In some embodiments, the human subject has an allergy to two or more different types of antigens (allergens) within a class of allergens, e.g., 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 (e.g., a stone fruit, 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 to 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 non-food allergens, or the subject has an allergy to one or more fungal allergens but not to non-fungal allergens).

[0244] 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 stone fruit allergen, a fish allergen, a shellfish allergen, a legume allergen, a legume allergen, a seed allergen, or a wheat allergen. In some embodiments, the food allergen is a peanut allergen.

[0245] 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, such as Aspergillus fumigatus, Aspergillus niger, or Aspergillus nidulans). In some embodiments, the allergen is a pollen allergen (e.g., tree pollen, grass pollen, or weed 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 bite.

[0246] In some embodiments, the method of generating allergen-specific monoclonal antibodies does not comprise immunizing the human subject with the allergen or exposing the human subject to the allergen prior to obtaining the sample from the subject.

[0247] B cell isolation and screening

[0248] In some embodiments, a single B cell is isolated from a sample from a subject with an allergy to an allergen. In some embodiments, the single B cell is separated into separate compartments, such as separate wells of a multiwell plate, encapsulated into droplets, or dispersed into microwells. In some embodiments, at least 10, 50, 100, 500, 1,000, 5,000, or 10,000 B cells or more are isolated from the sample and separated into separate compartments.

[0249] In some embodiments, the separation step includes sorting 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 the cells are delivered to wells, such as 96-well or 384-well tissue culture or PCR plates. Methods for separating and purifying cell populations by FACS are described in the art. See, for example, Basu et al., J Vis Exp, 2010, 41: 1546, doi: 10.3791 / 1546.

[0250] In some embodiments, the droplet microfluidics platform can be used to distribute 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 unique to the droplet (e.g., a unique molecular identifier barcode nucleotide sequence), thereby enabling downstream processing steps of sequences from multiple B cells to be performed in a single reaction vessel. Methods for encapsulating single cells in droplets are 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.

[0251] In some embodiments, cells are dispersed into microwells designed to capture single cells. Methods for single cell microwell capture are described in the art. See, for example, Han et al., Cell, 2018, 172:5, doi:10.1016 / j.cell.2018.02.001.

[0252] In some embodiments, cells from a sample are screened for the presence, absence, or expression level of one or more markers, and single B cells are isolated based on the presence or expression level of one or more B cell markers (e.g., one, two, three, four, five, six, seven, eight, or more markers). In some embodiments, cells are screened for one or more cell surface B cell markers (e.g., but not limited to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79). In some embodiments, a cell is determined to be a B cell if it is positive for one or more B cell markers, e.g., positive for one or more of CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79. In some embodiments, a single CD19 is isolated. + B cells.

[0253] In some embodiments, cells from the sample are screened for the presence, absence, or expression level of one or more immunoglobulin isotypes (e.g., but not limited to IgE, IgG, IgM, IgA, or IgD, or a subclass thereof). In some embodiments, isolated single B cells are selected for expression of cell surface IgE and / or for expression of cell surface IgG4. In some embodiments, isolated single B cells are not selected for expression of one or more immunoglobulin isotypes (e.g., not selected for expression of cell surface IgE and / or expression of cell surface IgG4).

[0254] IgE selection

[0255] In some embodiments, the method includes separating and selecting single B cells for expressing cell surface IgE. In some embodiments, the separation step includes contacting the cells of the sample with anti-human IgE antibodies and selecting cells expressing IgE on the cell surface. In some embodiments, the separation step includes contacting the cells of the sample with antibodies to B cell markers (e.g., antibodies to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and anti-human IgE antibodies, and selecting cells expressing B cell markers and expressing IgE on the cell surface. In some embodiments, the separation step includes contacting the cells of the sample with anti-human CD19 antibodies and anti-human IgE antibodies, and selecting cells expressing CD19. + IgE B cells.

[0256] In some embodiments, the separation step comprises contacting the cells of the sample with an antibody to a B cell marker (e.g., an antibody to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), an anti-human IgE antibody, and an antibody to 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 subclasses 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 separation step comprises contacting the cells of the sample with an anti-human CD19 antibody; an anti-human IgE antibody; and one or more of an anti-human IgG antibody, an anti-human IgM antibody, an anti-human IgA antibody, or an anti-human IgD antibody, and selecting cells that express CD19 and are negative for cell surface expression of IgG, IgM, IgA, or IgD. + IgE B cells.

[0257] In some embodiments, the isolating step comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgE antibodies, anti-human IgM antibodies, and anti-human IgG antibodies, and selecting cells expressing CD19. + IgM - IgG - In some embodiments, the isolation step comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgE antibodies, anti-human IgM antibodies, anti-human IgG antibodies, anti-human IgA antibodies, and anti-human IgD antibodies, and selecting cells expressing CD19 + IgM - IgG - IgA - IgD - IgE B cells.

[0258] In some embodiments, the separation step comprises contacting the cells of the sample with an antibody to a B cell marker (e.g., an antibody to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and an antibody to 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 the B cell marker and do not express detectable levels of the non-IgE isotype. In some embodiments, the separation step comprises contacting the 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 the B cell marker and do not express detectable levels of the non-IgE isotype. + IgM - IgG - IgA - IgD - B cells.

[0259] In some embodiments, the method includes separating B cells expressing IgE, which are B cells (e.g., plasmablasts or plasma cells) that secrete antibodies. In some embodiments, the method includes separating B cells expressing IgE, which are memory B cells. In some embodiments, the method includes separating B cells expressing IgE that secrete antibodies and expressing IgE memory B cells.

[0260] IgG4 selection

[0261] In some embodiments, the method includes separating and selecting single B cells for expressing cell surface IgG4. In some embodiments, the separation step includes contacting the cells of the sample with anti-human IgG4 antibodies and selecting cells expressing IgG4 on the cell surface. In some embodiments, the separation step includes contacting the cells of the sample with antibodies to B cell markers (e.g., antibodies to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and anti-human IgG4 antibodies, and selecting cells expressing B cell markers and expressing IgG4 on the cell surface. In some embodiments, the separation step includes contacting the cells of the sample with anti-human CD19 antibodies and anti-human IgG4 antibodies, and selecting cells expressing CD19. + IgG4 B cells.

[0262] In some embodiments, the separation step comprises contacting cells from the sample with an antibody to a B cell marker (e.g., an antibody to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), an anti-human IgG4 antibody, and an antibody to 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 the 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 separation 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 the 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 - IgG4 B cells.

[0263] In some embodiments, the isolating step comprises contacting the cells of the sample with an antibody to a B cell marker (e.g., an antibody to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), an anti-human IgG4 antibody, and an antibody to 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 subclasses 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 the cells of the sample with an anti-human CD19 antibody; an anti-human IgG4 antibody; and one or more anti-human IgE antibody, anti-human IgM antibody, anti-human IgA antibody, or anti-human IgD antibody, and selecting cells that are negative for IgE, IgM, IgA, or IgD cell surface expression of CD19. + IgG4-expressing B cells.

[0264] In some embodiments, the isolating step comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgG4 antibodies, anti-human IgM antibodies, and anti-human IgG antibodies, and selecting cells expressing CD19. + IgM - IgE - In some embodiments, the isolation step comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgG4 antibodies, anti-human IgM antibodies, anti-human IgE antibodies, anti-human IgA antibodies, and anti-human IgD antibodies, and selecting cells expressing CD19 + IgM- IgE - IgA - IgD - IgG4 cells.

[0265] In some embodiments, the isolating step comprises contacting the cells of the sample with an antibody to a B cell marker (e.g., an antibody to CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) and an antibody to 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 a non-IgG4 isotype thereof, and selecting cells that express the B cell marker and do not express detectable levels of the non-IgG or non-IgG4 isotype. In some embodiments, the isolating step comprises contacting the 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 CD19 + IgM - IgE - IgA - IgD - In some embodiments, the isolation step comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgM antibodies, anti-human IgE antibodies, anti-human IgA antibodies, anti-human IgD antibodies, anti-human IgG1 antibodies, anti-human IgG2 antibodies, and anti-human IgG3 antibodies, and selecting CD19 + IgM - IgE - IgA - IgD - IgG1 - IgG2 - IgG3 - B cells.

[0266] In some embodiments, the method comprises isolating B cells that express IgG4, which are antibody-secreting B cells. In some embodiments, the method comprises isolating B cells that express IgG4, which are memory B cells. In some embodiments, the method comprises isolating B cells that express IgG4, which are antibody-secreting B cells, and memory B cells that express IgG4.

[0267] Generate and sequence cDNA

[0268] In some embodiments, cDNA is generated from a single B cell isolated from a sample (e.g., a single B cell that has been screened for expression of an immunoglobulin isotype (e.g., IgE or IgG4), or a single B cell that has never been screened for expression of an immunoglobulin isotype). In some embodiments, a cDNA library is prepared from a single B cell. In some embodiments, for each cDNA generated for a single B cell, the cDNA sequence includes a sequence encoding an immunoglobulin heavy chain and a sequence encoding an immunoglobulin light chain.

[0269] In some embodiments, cDNA is generated by reverse transcribing a cDNA sequence (e.g., total RNA or mRNA) from the RNA of a single B cell and amplifying the cDNA sequence. In order to generate cDNA, in some embodiments, a single B cell is cracked and a cDNA sequence is reverse transcribed from the 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.

[0270] In some embodiments, the method includes amplifying the transcriptome of a single B cell. For example, in some embodiments, the method includes reverse transcription of RNA (e.g., polyadenylated mRNA) to synthesize cDNA, and then amplifying the cDNA, for example, by PCR. An exemplary method for reverse transcription of polyadenylated mRNA and amplification of the transcriptome of a single cell is described in Darmanis et al., Cell Reports, 2017, 21: 1399-1410, and Picelli et al., Nature Protocols, 9, 2014, 171-181.

[0271] In some embodiments, the method includes amplifying immunoglobulin heavy chain and light chain sequences from a single B cell. For example, in some embodiments, the method includes reverse transcription RNA (e.g., total RNA) to synthesize cDNA, and then using primers for immunoglobulin heavy chain variable region and constant region, for example, by PCR to amplify cDNA. In some embodiments, the method includes reverse transcription RNA using immunoglobulin-specific primers (e.g., constant region-specific primers) to synthesize cDNA including immunoglobulin sequences, and then using primers for immunoglobulin heavy chain variable region and constant region to amplify cDNA. An exemplary method for amplifying immunoglobulin heavy chain and light chain sequences from a single cell is described in Tiller et al., J. Immunol. Methods, 2008, 329: 112-124.

[0272] In some embodiments, after generating cDNA, the method includes determining the sequence of cDNA. In some embodiments, the cDNA is subjected to sequencing. In some embodiments, the method includes sequencing the transcriptome of a single B cell. In some embodiments, the method includes sequencing the target gene (e.g., immunoglobulin gene, e.g., immunoglobulin heavy chain variable region and constant region and immunoglobulin light chain variable region and constant region).

[0273] Sequencing methods, including methods for high-throughput sequencing, are known in the art. For example, such sequencing technologies include, but are not limited to, pyrosequencing, sequencing-by-ligation, single-molecule sequencing, sequencing-by-synthesis (SBS), massively parallel cloning, massively parallel single-molecule SBS, massively parallel single-molecule real-time, massively parallel single-molecule real-time nanopore technology, and the like. Morozova and Marra provide a review of some of these technologies in Genomics, 92:255 (2008), which is incorporated herein by reference in its entirety.

[0274] In some embodiments, sequencing includes high-throughput sequencing.In high-throughput sequencing, the parallel sequencing reaction using multiple templates and multiple primers allows genome or most of genome to be sequenced quickly.High-throughput sequencing methods include the method for template amplification and the method for not using template amplification.The sequencing method utilizing amplification includes the pyrophosphate sequencing commercialized by Roche as 454 technology platforms (such as GS 20 and GS FLX), the clone array formation and sequencing by synthesis (SBS) chemistry commercialized by Illumina utilizing systems such as NextSeq, and the oligonucleotide connection and detection (SOLiD) platform of the support commercialized by Applied Biosystems.Non-amplification method, also referred to as single molecule sequencing, is commercialized by Helicos BioSciences' HeliScope platform, and the platform commercialized by VisiGen, Oxford Nanopore Technologies Ltd., Life Technologies / Ion Torrent and Pacific Biosciences is demonstrated.

[0275] In some embodiments, an Illumina sequencing platform, such as NextSeq, is used. This sequencing technology utilizes clone array formation and sequencing by synthesis to produce sequences on a large scale. In this method, the sequencing template is fixed to the surface of a circulation cell, and then solid phase amplification produces a copy of each template molecule (up to 1,000 identical copies) in a close adjacent manner to form a dense "cluster" of polynucleotide sequences. In order to sequence the cluster, the clusters on the circulation cell surface are sequenced in parallel using fluorescently labeled nucleotides. For each sequencing cycle, a single labeled dNTP bound to a reversible terminator is added to the nucleic acid chain. The sequence of the incorporated nucleotides is determined by detecting the fluorescence after incorporation, and the fluorescent dye is then removed before the next dNTP addition cycle, thereby performing base-by-base sequencing. Typically, the sequence read length is in the range of about 30 nucleotides to more than 150 nucleotides. For target cDNAs with longer lengths, the sequence can be reassembled in a bioinformatics manner based on the overlap between short sequencing reads to determine the sequence of the full-length target cDNA.

[0276] Identification of B cells with IgE or IgG4 isotype

[0277] In some embodiments, after the cDNA sequence of the individual B cells has been determined, the method includes analyzing the sequence of the cDNA to identify individual B cells that express immunoglobulin heavy chains having an IgE constant region of the IgE isotype and / or the IgG4 isotype. As described herein, it has been discovered 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 a B cell population, thereby resulting in a B cell population that is more likely to produce antibodies that specifically bind to an allergen to which the human subject from whom the B cells were derived is allergic.

[0278] In some embodiments, the method includes the sequence of the immunoglobulin heavy chain that identification coding comprises IgE constant region.In some embodiments, the method includes the sequence of the immunoglobulin heavy chain that identification coding comprises IgG4 constant region.In some embodiments, by comparing cDNA sequence and known IgE constant region sequence or known IgG4 constant region sequence to analyze this cDNA sequence.For example, can compare target cDNA sequence (for example, from " test " sequence of B cell) with known IgE or IgG4 constant region sequence (for example, " reference " sequence) by alignment sequence.The sequence alignment method for comparison is known in the art. Optimal alignment of sequences for comparison can be performed, for example, by the local homology alignment algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by 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. Suitable algorithms for determining percent sequence identity and sequence similarity are BLAST and BLAST 2.0 algorithms, which are 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 National Center for Biotechnology Information (NCBI) website.

[0279] To compare a test sequence to an IgE or IgG4 constant region reference sequence, in some embodiments, the reference sequence is a published sequence, such as an IgE or IgG4 constant region sequence, which is publicly available in the ImMunoGeneTics (IMGT) database. See, for example, Camacho et al., BMC Bioinformatics, 2009, 10:421; Lefranc et al., Nucleic Acids Res, 2009, 37:D1006-1012. Table 1 below also describes methods for analyzing a test sequence to identify sequences of immunoglobulin heavy chain constant regions that include IgE isotypes and / or IgG4 isotypes.

[0280] In some embodiments, in addition to analyzing the sequence to identify and select a single B cell comprising an immunoglobulin heavy chain sequence comprising an IgE constant region or an IgG4 constant region, the method further comprises determining the sequence and / or expression level of one or more other genes in the single B cell. For example, in some embodiments, the method comprises determining the sequence and / or expression level of a set of genes that serve as a "signature" for a particular type of B cell.

[0281] 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.

[0282] Antibody expression

[0283] Typically, for a single B cell identified as having a cDNA comprising an IgE or IgG4 constant region sequence, the heavy chain variable region and light chain variable region sequences from the single B cell are candidate antibody sequences specific for the target allergen. Thus, in some embodiments, the method comprises expressing an antibody comprising the heavy chain variable region and light chain variable region sequences from the single B cell, and identifying whether the expressed antibody specifically binds to the target allergen. 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.

[0284] In some embodiments, the heavy chain variable region and light chain variable region sequences from a single B cell are amplified from a single B cell and cloned into an expression vector. In some embodiments, the heavy chain variable region and light chain variable region sequences from a single B cell are synthesized. In some embodiments, the heavy chain variable region sequence and / or the light chain variable region sequence are codon optimized, for example, to increase antibody expression through an expression system. See, for example, Ayyar et al., Methods, 2017, 116: 51-62.

[0285] The heavy chain variable region 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 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 inrecombinant 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 bicistronic 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 vectors for expressing heavy chain variable region sequences and / or light chain variable region sequences as disclosed herein are vectors comprising the constant regions of the desired heavy chain isotype or light chain subclass. For example, the heavy chain variable region sequences as disclosed herein can be cloned into a vector comprising a human IgG (e.g., IgG1, IgG2, IgG3, or IgG4) heavy chain constant region, and the light chain variable region sequences as disclosed herein can be cloned into a vector comprising a human λ or κ light chain constant region.

[0286] In some embodiments, after expressing and purifying an antibody comprising a heavy chain variable region sequence and a light chain variable region sequence from a single B cell as disclosed herein, the method comprises determining whether the antibody specifically binds to the 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 TM (GE Healthcare, Piscataway, NJ)), kinetic exclusion assays (e.g. ), flow cytometry, fluorescence activated cell sorting (FACS), BioLayer interferometry (e.g. Octet TM(FortéBio, Inc., Menlo Park, CA)) and Western blot analysis. SPR technology is reviewed in, for example, Hahnfeld et al. Determination of Kinetic Data Using SPR Biosensors, Molecular Diagnosis of Infectious Diseases (2004). In a typical SPR experiment, a kind of interacting agent (target or targeted agent) is fixed on an SPR active, gold-plated glass slide in a flow cell, and a sample containing another interacting agent is introduced to flow through the entire surface. When light of a given wavelength is irradiated on the surface, the change in the optical reflectivity of gold represents binding and binding kinetics. In some embodiments, kinetic exclusion is used to determine affinity. This technology is described in, for example, Darling et al., Assay and Drug Development Technologies Vol. 2, No. 6 647-657 (2004). In some embodiments, affinity is determined using BioLayer interferometry. 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).

[0287] In some embodiments, the expressed antibodies specifically bind to the allergen with high affinity. In some embodiments, the antibodies have a binding affinity (K) 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 for the allergen. D In some embodiments, the antibody binds to the molecule with a binding affinity (K) of 1 nM to 250 nM. D ) binds to the allergen.

[0288] Nucleic acids, vectors and host cells

[0289] In some embodiments, the allergen-specific monoclonal antibodies described herein are prepared using recombinant methods. Accordingly, in some aspects, the present 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 have been introduced, which are used to replicate the nucleic acid encoding the antibody and / or express the antibody. In some embodiments, the host cell is a eukaryotic cell, such as a human cell, such as HEK-293.

[0290] In some embodiments, the polynucleotide (e.g., an isolated polynucleotide) comprises a nucleotide sequence encoding an antibody or antigen-binding portion thereof as described herein (e.g., as described in Section IV below). In some embodiments, the polynucleotide comprises a nucleotide sequence encoding one or more amino acid sequences (e.g., CDRs, heavy chain variable regions, or light chain variable regions) disclosed in Table 1 below.

[0291] In another aspect, methods of producing an allergen-specific monoclonal antibody as described herein are provided. In some embodiments, the method comprises 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 subsequently recovered from the host cell (or host cell culture medium).

[0292] Suitable vectors containing polynucleotides encoding antibodies of the present disclosure or fragments thereof include cloning vectors and expression vectors. Although the cloning vector of choice may vary depending on the host cell to be used, useful cloning vectors generally have the ability to self-replicate, may have a single target for a specific restriction endonuclease, and / or may carry a gene for a marker that can be used to select 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 suppliers such as BioRad, Stratagene, and Invitrogen.

[0293] An expression vector is typically a replicable polynucleotide construct comprising a nucleic acid of the present disclosure. An expression vector can replicate in a host cell as an episome or as an integral part of chromosomal DNA. Suitable expression vectors include, but are not limited to, plasmids, viral vectors, including adenoviruses, adeno-associated viruses, retroviruses, and any other vectors.

[0294] IV. Monoclonal antibodies that specifically bind to allergens

[0295] In another aspect, allergen-specific monoclonal antibodies and antigen-binding portions thereof generated from a human sample according to the methods disclosed herein are provided. 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 of 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 the monoclonal antibodies 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.

[0296] 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 include 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 peanut allergens and stone fruit allergens. In some embodiments, the antibodies described herein can bind to peanut allergens and milk allergens. In some embodiments, the antibodies described herein can bind to peanut allergens and fungal allergens. In some embodiments, the antibodies described herein can bind to stone fruit allergens and milk allergens. In some embodiments, the antibodies described herein can bind to stone fruit allergens and fungal allergens. In some embodiments, the antibodies described herein can bind to milk allergens and fungal allergens.

[0297] In some embodiments, the monoclonal antibody, or antigen-binding portion thereof, is an allergen-specific antibody comprising a heavy chain variable region sequence and a light chain variable region sequence identified according to a method comprising:

[0298] (a) isolating single B cells from a sample of a human subject, wherein the subject is allergic to the allergen;

[0299] (b) generating cDNA from the single B cell of step (a), wherein the cDNA sequence comprises a first sequence encoding all or part of an immunoglobulin heavy chain and a second sequence encoding all or part of an immunoglobulin light chain;

[0300] (c) determining the sequence of the cDNA from step (b);

[0301] (d) analyzing the sequences determined in step (c) to identify single B cells comprising a first sequence comprising an IgE constant region or an IgG4 constant region;

[0302] (e) identifying, from the single B cell of step (d), (i) a heavy chain variable region sequence in the immunoglobulin heavy chain comprising an IgE constant region or an IgG4 constant region, and (ii) a light chain variable region sequence in an immunoglobulin light chain co-expressed with the immunoglobulin heavy chain in the same single B cell.

[0303] In some embodiments, the heavy chain variable region and the light chain variable region are from a B cell comprising an immunoglobulin comprising an IgE constant region. In some embodiments, the heavy chain variable region and the light chain variable region are from a B cell comprising an immunoglobulin comprising an IgG4 constant region.

[0304] In some embodiments, the monoclonal antibody, or antigen-binding portion thereof, is an antibody specific for an allergen comprising:

[0305] (a) a heavy chain variable region sequence derived from an immunoglobulin heavy chain from a single IgE- or IgG4-producing B cell of a human subject allergic to the allergen;

[0306] (b) heavy chain IgG constant region sequence;

[0307] (c) a light chain variable region sequence derived from an immunoglobulin light chain from a single IgE- or IgG4-producing B cell from a human subject;

[0308] (d) a light chain constant region sequence that is of the same class as the immunoglobulin light chain of (c).

[0309] In some embodiments, the monoclonal antibody comprises a heavy chain variable region sequence and a light chain variable region sequence derived from a human B cell producing IgE. In some embodiments, the monoclonal antibody comprises a heavy chain variable region sequence and a light chain variable region sequence derived from a human B cell producing IgG4.

[0310] Characteristics of allergen-specific monoclonal antibodies

[0311] In some embodiments, the monoclonal antibody 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 specifically binds to a food allergen (e.g., a milk allergen, an egg allergen, a nut allergen, a fish allergen, a shellfish allergen, a legume allergen, a legume allergen, a seed allergen, or a wheat allergen). In some embodiments, the monoclonal antibody specifically binds to a peanut allergen. In some embodiments, the monoclonal antibody specifically binds to a milk allergen. In some embodiments, the monoclonal antibody specifically binds to an egg allergen.

[0312] In some embodiments, the monoclonal antibody binds to a polypeptide with a binding affinity (K) 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. D ) 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). In some embodiments, the antibody binds to an allergen with a binding affinity (K) of 1 nM to 250 nM. D )Binding allergens.

[0313] In some embodiments, the monoclonal antibody exhibits cross-reactivity with at least two different antigens (e.g., allergens), for example, cross-reactivity with 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 soy allergens, at least two legume allergens, at least two seed allergens, or at least two wheat allergens. One of ordinary skill in the art will recognize that many different allergens, such as many plant food allergens, can be classified into a small number of protein families. For example, more than half of all plant food allergens can be classified into one of four structural protein families: the prolamin superfamily, the cupin superfamily, profilins, and proteins related to Bet v-1. Those skilled in the art will also recognize that for a particular type of allergen (e.g., a "peanut" allergen), there may be more than one peptide or protein that is an allergen. 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, a monoclonal antibody exhibits cross-reactivity with two or more different antigens (which are different types or classes of antigens). As a non-limiting example, in some embodiments, a monoclonal antibody exhibits cross-reactivity with an antigen that is a peanut allergen and an antigen that is a tree nut (e.g., stone fruit) allergen.

[0314] In embodiments wherein the monoclonal antibody exhibits cross-reactivity to at least two different antigens (e.g., allergens), in some embodiments, the monoclonal antibody is expressed with a K 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. D In some embodiments, the monoclonal antibody specifically binds to at least one of the allergens. In some embodiments, the monoclonal antibody has a K 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. DSpecifically binds to a first antigen (e.g., a first allergen). In some embodiments, the monoclonal antibody binds to a first antigen with a K of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. D Specifically binds to a second antigen (eg, a second allergen).

[0315] Engineering changes in the variable region

[0316] In some embodiments, the heavy chain variable region and / or light chain variable region of the monoclonal antibody has a sequence identical to that of a heavy chain variable region and / or light chain variable region encoded by a single B cell producing IgE or IgG4 from a human subject with an allergy to an allergen. In some embodiments, the heavy chain variable region and / or light chain variable region of the monoclonal antibody comprises one or more modifications, e.g., amino acid substitutions, deletions, or insertions.

[0317] As described in the Examples section below, the heavy chain variable region sequence and / or the light chain variable region sequence of the antibodies described herein (e.g., peanut allergen-specific monoclonal antibodies, such as clone PA13P1H08, clone PA13P1E10, clone PA12P3F10, clone PA13P3G09, clone PA12P3D08, clone PA12P1C07, clone PA15P1D12, clone PA15P1D05, or a clone PA13P1H08 variant) can be engineered to include one or more changes in the heavy chain variable region sequence and / or the light chain variable region sequence. In some embodiments, the engineered changes improve the binding affinity of the antibody to the allergen. In some embodiments, the engineered changes improve the cross-reactivity of the antibody to a second allergen.

[0318] In some embodiments, the engineered changes are changes in one or more CDRs, for example, amino acid substitutions in heavy chain CDRs and / or light chain CDRs as described herein. In some embodiments, the engineered changes are changes in one or more framework regions, for example, amino acid substitutions in heavy chain framework regions and / or light chain framework regions. In some embodiments, the engineered changes are reversions of heavy chain and / or light chain sequence regions to the original sequence inferred. Methods for determining the original inferred immunoglobulin sequence are described in the art. See, for example, Magnani et al., PLoS Negl Trop Dis, 2017, 11: e0005655, doi: 10.1371 / journal.pntd.0005655.

[0319] In some embodiments, affinity maturation is used to engineer further mutations that enhance the binding affinity of the antibody to the allergen or enhance the cross-reactivity of the antibody to 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.

[0320] Constant region and isotype switching

[0321] In some embodiments, the monoclonal antibody comprises a heavy chain variable region sequence and a light chain variable region sequence derived from an IgE-producing human B cell or an IgG4-producing human B cell, and further comprises 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-producing human B cell or an IgG4-producing human B cell; as a non-limiting example, if the IgE-producing human B cell comprises a kappa light chain, the monoclonal antibody produced comprises a light chain variable region from an IgE-producing B cell and further comprises a kappa light chain constant region.

[0322] In some embodiments, the monoclonal antibody includes a heavy chain variable region sequence and a light chain variable region sequence derived from a human B cell producing IgE or a human B cell producing IgG4, and further includes 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). One of ordinary skill in the art will appreciate that different heavy chain isotypes (IgA, IgD, IgE, IgG, and IgM) have different effector functions mediated by the heavy chain constant region, and for certain uses, it may be desirable to have an antibody with effector functions of a particular isotype (e.g., IgG).

[0323] In some embodiments, monoclonal antibodies include natural (i.e., wild type) human IgG, IgA, IgM or IgD constant regions. In some embodiments, monoclonal antibodies include natural human IgG1 constant region, natural human IgG2 constant region, natural human IgG3 constant region, natural human IgG4 constant region, natural human IgA1 constant region, natural human IgA2 constant region, natural human IgM constant region or natural human IgD constant region. In some embodiments, monoclonal antibodies include a heavy chain constant region comprising one or more modifications. It will be understood by those of ordinary skill in the art that modification, such as amino acid substitution, can be performed at one or more residues regulating effector functions within the heavy chain constant region. In some embodiments, modification reduces effector functions, such as resulting in a reduction in the ability to induce certain biological functions after binding to Fc receptors expressed on effector cells mediating effector functions. In some embodiments, modification (such as amino acid substitution) prevents Fab arm exchange in vivo, which can introduce undesirable effects and reduce the therapeutic efficacy of antibodies. See, e.g., Silva et al., J Biol Chem, 2015, 280:5462-5469.

[0324] In some embodiments, the monoclonal antibody includes a natural (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 includes one or more modifications that regulate effector function. In some embodiments, the monoclonal antibody includes 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 antibody includes a natural (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 includes one, two, three, four, five, six, seven, eight, nine, ten or more modifications (e.g., amino acid substitutions). In some embodiments, the constant region comprises alterations (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid substitutions) that reduce effector function.

[0325] In some embodiments, the monoclonal antibody comprises a CDR sequence, a heavy chain variable region, and / or a light chain variable region of an antibody from an IgE or IgG4 B cell as described herein (e.g., as disclosed in Table 1 below), and further comprises a heavy chain constant region and / or a light chain constant region that is heterologous to an antibody from an IgE or IgG4 B cell from which the CDR sequence and / or variable region sequence is derived. For example, in some embodiments, the monoclonal antibody comprises a CDR sequence and / or a variable region sequence of an antibody from an IgEB cell, and further comprises a heavy chain constant region and a light chain constant region that are heterologous to an antibody from an IgE B cell [e.g., the heavy chain constant region and / or the light chain constant region are wild-type or modified IgG1, IgG2, IgG3, or IgG4 constant regions, or the heavy chain constant region and / or the light chain constant region comprise one or more modifications (e.g., amino acid substitutions) relative to the native constant region of an antibody from an IgE B cell].

[0326] Antibodies that specifically bind peanut and / or stone fruit allergens

[0327] In some embodiments, a monoclonal antibody, or antigen-binding portion thereof, as disclosed herein specifically binds to a peanut allergen and / or a stone fruit allergen. In some embodiments, the monoclonal antibody specifically binds to a peanut allergen. In some embodiments, the monoclonal antibody specifically binds to a peanut allergen that is Ara h 1, Ara h 2, Ara h 3, or Ara h 6.

[0328] In some embodiments, the monoclonal antibody exhibits cross-reactivity to at least two peanut allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity to two or more of the peanut allergens Ara h 1, Ara h 2, Ara h 3, and Ara h 6 ... D Specifically binds to at least one peanut allergen. In some embodiments, the monoclonal antibody binds to at least one peanut allergen with a K of less than 100 nM (e.g., less than 50 nM, less than 10 nM, less than 5 nM, or less than 1 nM). D Specifically binds to the first peanut allergen with a K of less than 1 μM (e.g., less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM) DSpecifically binds to a second peanut allergen. In some embodiments, the monoclonal antibody binds to a second peanut allergen with a K of less than 100 nM (e.g., less than 50 nM, less than 10 nM, less than 5 nM, or less than 1 nM). D Specifically binds Ara h 2 with a K of less than 1 μM (e.g., less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM). D Specifically binds Ara h 1, Ara h 3, or Ara h 6.

[0329] 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).

[0330] In some embodiments, the monoclonal antibody, or antigen-binding portion thereof, specifically binds to a stone fruit allergen. In some embodiments, the stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity with both cashew and pistachio allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity with both pecan and walnut allergens. In some embodiments, the monoclonal antibody exhibits cross-reactivity with two or more of pecan, walnut, hazelnut, and macadamia nut allergens. In some embodiments, the monoclonal antibodies are expressed with a K 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. D Specifically binds to at least one stone fruit allergen.

[0331] In some embodiments, the monoclonal antibody, or antigen binding portion thereof, specifically binds to a peanut allergen and a stone fruit allergen. In some embodiments, the monoclonal antibody specifically binds to a peanut allergen; and one or more (e.g., 1, 2, 3, 4, or more) of a cashew, pistachio, almond, pine nut, pecan, walnut, hazelnut, or macadamia nut allergen.

[0332] Peanut-specific antibody sequences

[0333] In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen (e.g., specifically binds to Ara h 1, Ara h 2, Ara h 3, or Ara h 6) comprises the heavy chain CDRs and / or light chain CDRs disclosed below in Table 1. In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen comprises the following sequences: a light chain variable region sequence and / or a heavy chain variable region sequence 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 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 a heavy chain variable region sequence disclosed in Table 1 and comprising 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 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 a light chain variable region sequence disclosed in Table 1 and comprising CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0334] In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen (e.g., specifically binds to Ara h 1, Ara h 2, Ara h 3, or Ara h 6) comprises a light chain sequence, or a portion thereof, and / or a heavy chain sequence, or a portion thereof, derived from any of the following antibodies described herein: clone PA13P1H08, clone PA13P1E10, clone PA12P3F10, clone PA13P3G09, clone PA12P3D08, clone PA12P1C07, clone PA15P1D12, clone PA15P1D05, a variant of clone PA13P1H08 (e.g., RR variant, RN variant, NR variant, rCDR1-N variant, rCDR2-N variant, rCDR3-N variant or rFWRs-N variant of clone PA13P1H08), 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 regions (VL), and heavy chain variable regions (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 listed in Table 1 below.

[0335] In some embodiments, monoclonal antibodies that specifically bind to peanut allergens include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0336] (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199, 341, 348, 409, 459 or 593;

[0337] (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349, 410, 460, 539 or 594;

[0338] (c) a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201, 350, 411, 461, 540 or 595;

[0339] (d) a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343, 352, 413, 463, 542 or 597;

[0340] (e) 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

[0341] (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354, 414, 464, 543 or 599.

[0342] In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises heavy chain CDRs 1-3 and light chain CDRs 1-3 comprising the following amino acid sequence:

[0343] (a) SEQ ID NOs: 2, 3, 4, 6, 7 and 8, respectively; or

[0344] (b) SEQ ID NOs: 10, 11, 12, 14, 15 and 8, respectively; or

[0345] (c) SEQ ID NOs: 17, 18, 19, 21, 22 and 23, respectively; or

[0346] (d) SEQ ID NOs: 25, 26, 27, 29, 30 and 31, respectively; or

[0347] (e) SEQ ID NOs: 33, 34, 35, 37, 38 and 39, respectively; or

[0348] (f) SEQ ID NOs: 41, 34, 35, 37, 38 and 39, respectively; or

[0349] (g) SEQ ID NOs: 47, 48, 49, 51, 52 and 53, respectively; or

[0350] (h) SEQ ID NOs: 47, 48, 55, 51, 52 and 53, respectively; or

[0351] (i) SEQ ID NOs: 58, 59, 60, 62, 30 and 63, respectively; or

[0352] (j) SEQ ID NOs: 58, 59, 60, 6, 7 and 8, respectively; or

[0353] (k) SEQ ID NOs: 2, 3, 4, 62, 30 and 63, respectively; or

[0354] (1) SEQ ID NOs: 58, 3, 4, 6, 7 and 8, respectively; or

[0355] (m) SEQ ID NOs: 2, 59, 4, 6, 7 and 8, respectively; or

[0356] (n) SEQ ID NOs: 2, 3, 60, 6, 7 and 8, respectively; or

[0357] (o) SEQ ID NOs: 129, 130, 131, 133, 126 and 134, respectively; or

[0358] (p) SEQ ID NOs: 341, 342, 343, 345, 78 and 346, respectively; or

[0359] (q) SEQ ID NOs: 348, 349, 350, 352, 353 and 354, respectively; or

[0360] (r) SEQ ID NOs: 199, 200, 201, 203, 149 and 204, respectively; or

[0361] (s) are SEQ ID NOs: 409, 410, 411, 413, 86 and 414, respectively; or

[0362] (t) SEQ ID NOs: 459, 460, 461, 463, 196 and 464, respectively; or

[0363] (u) SEQ ID NOs: 113, 539, 540, 542, 196 and 543, respectively; or

[0364] (v) SEQ ID NOs: 593, 594, 595, 597, 598 and 599, respectively.

[0365] 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 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, 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: 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 of NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541 or 596.

[0366] 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 and comprising CDR1, CDR2, and CDR3 identical to the CDRs of the SEQ ID NO. 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 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 comprising a CDR1, CDR2, and CDR3 that are identical to the CDRs of the SEQ ID NO. In some embodiments, a monoclonal antibody that specifically binds to a peanut allergen comprises:

[0367] (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 comprising a CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO; and

[0368] (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 comprising CDR1, CDR2 and CDR3 identical to the CDRs of the SEQ ID NO.

[0369] Clone PA13P1H08

[0370] 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.

[0371] In some embodiments, the antibody comprises:

[0372] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 2, 3 and 4, respectively; and

[0373] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 6, 7 and 8, respectively.

[0374] 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.

[0375] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0376] Clone PA13P1E10

[0377] 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.

[0378] In some embodiments, the antibody comprises:

[0379] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 10, 11 and 12, respectively; and

[0380] (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.

[0381] 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.

[0382] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0383] Clone PA12P3F10

[0384] 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.

[0385] In some embodiments, the antibody comprises:

[0386] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18 and 19, respectively; and

[0387] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 21, 22 and 23, respectively.

[0388] 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.

[0389] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0390] Clone PA13P3G09

[0391] 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.

[0392] In some embodiments, the antibody comprises:

[0393] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 25, 26 and 27, respectively; and

[0394] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 29, 30 and 31, respectively.

[0395] 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.

[0396] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0397] Clone PA12P3D08

[0398] 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.

[0399] In some embodiments, the antibody comprises:

[0400] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 33, 34 and 35, respectively; and

[0401] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 37, 38 and 39, respectively.

[0402] 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.

[0403] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0404] Clone PA12P1C07

[0405] 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.

[0406] In some embodiments, the antibody comprises:

[0407] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 41, 34 and 42, respectively; and

[0408] (b) a light chain variable region comprising a light chain CDR1, CDR2, and CDR3 that have 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 that include the amino acid sequences of SEQ ID NOs: 44, 30, and 45, respectively.

[0409] 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.

[0410] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0411] Clone PA15P1D12

[0412] 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.

[0413] In some embodiments, the antibody comprises:

[0414] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 47, 48 and 49, respectively; and

[0415] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 51, 52 and 53, respectively.

[0416] 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.

[0417] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0418] Clone PA15P1D05

[0419] 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.

[0420] In some embodiments, the antibody comprises:

[0421] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 47, 48 and 55, respectively; and

[0422] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 51, 52 and 53, respectively.

[0423] 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.

[0424] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0425] Variant sequence

[0426] In some embodiments, allergen-specific monoclonal antibodies comprise one or more changes (e.g., amino acid substitutions) in one or more CDR, heavy chain, and / or light chain sequences as disclosed herein (e.g., one or more mutations in one or more CDR, heavy chain, and / or light chain sequences of clones PA13P1H08, PA13P1E10, PA12P3F10, PA13P3G09, PA12P3D08, PA12P1C07, PA15P1D12, and 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, variants were generated from clone PA13P1H08 in which one or more regions in the heavy and / or light chain were reverted to the inferred original rearrangement. Antibody sequences comprising the original light or heavy chain sequences were found to bind to the peanut allergen Ara h 2. It was also surprisingly discovered that, in addition to having subnanomolar affinity for the Ara h 2 peanut allergen, variants of clone PA13P1H08 that included the reverted CDR-H2 sequence also exhibited significantly improved cross-reactivity with a second peanut allergen (Ara h 3). Thus, in some embodiments, the mutation is an amino acid substitution that reverts at least a portion of the cloned sequence from its "native" form (i.e., the cloned CDR, heavy chain variable region, or light chain variable region sequence as disclosed in Table 1) to the putative original immunoglobulin sequence.

[0427] In some embodiments, allergen-specific monoclonal antibodies comprise one or more variant sequences of clone PA13P1H08 variants as disclosed herein. In some embodiments, the antibody comprises one of the sequences of clone PA13P1H08 variant "RR," wherein both the heavy and light chain variable regions of clone PA13P1H08 are regressed back to the inferred original rearrangement. In some embodiments, the antibody comprises a regressed heavy chain variable region sequence comprising SEQ ID NO: 57. In some embodiments, the antibody comprises a regressed light chain variable region sequence comprising SEQ ID NO: 61.

[0428] In some embodiments, the antibody comprises one of the sequences of clone PA13P1H08 variant "RN", wherein the heavy chain variable region of clone PA13P1H08 is regressed to the inferred original rearrangement, and the light chain variable region retains the native 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", wherein the heavy chain variable region retains the native sequence of clone PA13P1H08 (i.e., SEQ ID NO: 1), and the light chain variable region is regressed to the inferred original rearrangement of clone PA13P1H08.

[0429] In some embodiments, the antibody comprises one or more reverted CDR sequences, e.g., one or more reverted heavy chain CDR sequences and / or one or more reverted light chain CDR sequences. In some embodiments, the antibody comprises one or more of a reverted CDR-H1 comprising SEQ ID NO: 58, a reverted CDR-H2 comprising SEQ ID NO: 59, or a reverted CDR-H3 comprising SEQ ID NO: 60. In some embodiments, the antibody comprises one or more of a reverted CDR-L1 comprising SEQ ID NO: 62, a reverted CDR-L2 comprising SEQ ID NO: 30, or a reverted CDR-L3 comprising SEQ ID NO: 63. In some embodiments, the antibody comprises one or more reverted framework regions, e.g., a heavy chain variable region comprising a reverted framework region of SEQ ID NO: 67.

[0430] 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.

[0431] 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 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 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 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 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 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 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 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 a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5.

[0432] In some embodiments, the antibody comprises:

[0433] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 2, 59 and 4, respectively; and

[0434] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 6, 7 and 8, respectively.

[0435] Stone fruit-specific antibody sequences

[0436] In some embodiments, the monoclonal antibody that specifically binds to a stone fruit allergen (e.g., specifically binds to a cashew, pistachio, almond, pine nut, pecan, walnut, hazelnut, or macadamia nut allergen) comprises the heavy chain CDRs and / or light chain CDRs disclosed in Table 1 below. In some embodiments, the monoclonal antibody that specifically binds to a stone fruit allergen comprises the following sequences: a light chain variable region sequence and / or a heavy chain variable region sequence disclosed in Table 1 below. In some embodiments, the monoclonal antibody that specifically binds to a stone fruit 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 a heavy chain variable region sequence disclosed in Table 1, and including 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 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 a light chain variable region sequence disclosed in Table 1, and including CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0437] In some embodiments, the monoclonal antibody that specifically binds to a stone fruit allergen comprises a light chain sequence or a portion thereof and / or a heavy chain sequence or a portion thereof derived 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 regions (VL), and heavy chain variable regions (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 listed in Table 1 below.

[0438] In some embodiments, the monoclonal antibodies that specifically bind to stone fruit allergens include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0439] (a) 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;

[0440] (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;

[0441] (c) a heavy chain CDR3 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 169, 177, 228, 313, 320, 440, 467, 540, 667 or 694;

[0442] (d) a light chain CDR1 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623, 669 or 696;

[0443] (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

[0444] (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624, 671 or 697.

[0445] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit allergen comprises a heavy chain variable region comprising an amino acid sequence comprising SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664 or 691, 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) to any of said SEQ ID NOs. 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 SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695, 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) to any of said SEQ ID NOs. In some embodiments, the monoclonal antibody that specifically binds to a peanut allergen comprises a heavy chain variable region comprising an amino acid sequence comprising a sequence of SEQ ID NOs: 166, 174, 226, 310, 317, 437, 465, 538, 620, 664 or 691, 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) to any of said 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 a sequence of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668 or 695, 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) to any of said SEQ ID NOs: Any of the NOs have at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity).

[0446] In some embodiments, the monoclonal antibody that specifically binds to a stone fruit allergen comprises:

[0447] (a) a heavy chain variable region comprising 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 comprising a CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO; and

[0448] (b) a light chain variable region comprising a CDR1, a CDR2, and a CDR3 that are identical to any of SEQ ID NOs: 170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695 and 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) and that comprises CDR1, CDR2, and CDR3 that are identical to the CDRs of that SEQ ID NO.

[0449] In some embodiments, the monoclonal antibody specifically binds pecan, walnut, hazelnut, and / or macadamia nut allergens and includes one or more (e.g., one, two, three, four, five, or all six) of the following:

[0450] (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 113, 227, 311, 318, 665 or 692;

[0451] (b) a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 200, 312, 319, 539, 666 or 693;

[0452] (c) a heavy chain CDR3 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 228, 313, 320, 540, 667 or 694;

[0453] (d) a light chain CDR1 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 230, 315, 322, 542, 669 or 696;

[0454] (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 94, 149, 196, 323 or 670; and

[0455] (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 231, 316, 324, 543, 671 or 697.

[0456] 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 SEQ ID NOs: 226, 310, 317, 538, 664, or 691, 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) to any of said SEQ ID NOs. 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 SEQ ID NOs: 229, 314, 321, 541, 668, or 695, 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) to any of said SEQ ID NOs. In some embodiments, the monoclonal antibody that specifically binds to pecan, walnut, hazelnut and / or macadamia nut allergens 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) to any of SEQ ID NOs: 226, 310, 317, 437, 538, 664 or 691 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) to any of SEQ ID NOs: 229, 314, 321, 541, 668 or 695. Any of the NOs have at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity).

[0457] In some embodiments, the monoclonal antibody specifically binds cashew and / or pistachio allergens and includes one or more (e.g., one, two, three, four, five, or all six) of the following:

[0458] (a) a heavy chain CDR1 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 167, 175, 227, 438, 466 or 621;

[0459] (b) a heavy chain CDR2 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 168, 176, 200 or 439;

[0460] (c) a heavy chain CDR3 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 169, 177, 440 or 467;

[0461] (d) a light chain CDR1 comprising the amino acid sequence of any one of the following: SEQ ID NOs: 171, 179, 442, 469 or 623;

[0462] (e) a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 30, 149, 172 or 180; and

[0463] (f) a light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 173, 181, 443, 470 or 624.

[0464] 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 SEQ ID NOs: 166, 174, 437, 465, or 620, 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) to any of said SEQ ID NOs. 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 SEQ ID NOs: 170, 178, 441, 468, or 622, 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) to any of said SEQ ID NOs. In some embodiments, the monoclonal antibody that specifically binds to cashew and / or pistachio allergens comprises a heavy chain variable region comprising an amino acid sequence comprising a sequence of SEQ ID NOs: 166, 174, 437, 465 or 620, 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) to any of said SEQ ID NOs; and a light chain variable region comprising an amino acid sequence comprising a sequence of SEQ ID NOs: 170, 178, 441, 468 or 622, 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) to any of said SEQ ID NOs. Any of the NOs have at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity).

[0465] In some embodiments, the monoclonal antibody that specifically binds to a stone fruit allergen comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequence:

[0466] (a) SEQ ID NOs: 692, 693, 694, 696, 94 and 697, respectively; or

[0467] (b) SEQ ID NOs: 318, 319, 320, 322, 323 and 324, respectively; or

[0468] (c) SEQ ID NOs: 227, 200, 228, 230, 149 and 231, respectively; or

[0469] (d) SEQ ID NOs: 113, 539, 540, 542, 196 and 543, respectively; or

[0470] (e) SEQ ID NOs: 311, 312, 313, 315, 94 and 316, respectively; or

[0471] (f) SEQ ID NOs: 665, 666, 667, 669, 670 and 671, respectively; or

[0472] (g) SEQ ID NOs: 466, 200, 467, 469, 149 and 470, respectively; or

[0473] (h) SEQ ID NOs: 167, 168, 169, 171, 172 and 173, respectively; or

[0474] (i) SEQ ID NOs: 621, 176, 177, 623, 180 and 624, respectively; or

[0475] (j) SEQ ID NOs: 175, 176, 177, 179, 180 and 181, respectively; or

[0476] (k) are SEQ ID NOs: 438, 439, 440, 442, 30 and 443, respectively.

[0477] Clone PA14P3H08

[0478] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to pecan, walnut, hazelnut, and / or macadamia nut allergens.

[0479] In some embodiments, the antibody comprises:

[0480] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 692, 693 and 694, respectively; and

[0481] (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.

[0482] 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.

[0483] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0484] Clone PA11P1D11

[0485] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to pecan, walnut, hazelnut, and / or macadamia nut allergens.

[0486] In some embodiments, the antibody comprises:

[0487] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 318, 319 and 320, respectively; and

[0488] (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.

[0489] 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.

[0490] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0491] Clone PA11P1G10

[0492] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to pecan, walnut, hazelnut, and / or macadamia nut allergens.

[0493] In some embodiments, the antibody comprises:

[0494] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 227, 200 and 228, respectively; and

[0495] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 230, 149 and 231, respectively.

[0496] 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.

[0497] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0498] Clone PA12P4D02

[0499] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to pecan, walnut, hazelnut, and / or macadamia nut allergens.

[0500] In some embodiments, the antibody comprises:

[0501] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 113, 539 and 540, respectively; and

[0502] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 542, 196 and 543, respectively.

[0503] 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.

[0504] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0505] Clone PA11P1D12

[0506] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to pecan, walnut, hazelnut, and / or macadamia nut allergens.

[0507] In some embodiments, the antibody comprises:

[0508] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 311, 312 and 313, respectively; and

[0509] (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.

[0510] 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.

[0511] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0512] Clone PA11P1F03

[0513] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to pecan, walnut, hazelnut, and / or macadamia nut allergens.

[0514] In some embodiments, the antibody comprises:

[0515] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 665, 666 and 667, respectively; and

[0516] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 669, 670 and 671, respectively.

[0517] 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.

[0518] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0519] Clone PA11P1C04

[0520] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to cashew and / or pistachio allergens.

[0521] In some embodiments, the antibody comprises:

[0522] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 466, 200 and 467, respectively; and

[0523] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 469, 149 and 470, respectively.

[0524] 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.

[0525] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0526] Clone PA11P1G04

[0527] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to cashew and / or pistachio allergens.

[0528] In some embodiments, the antibody comprises:

[0529] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 167, 168 and 169, respectively; and

[0530] (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.

[0531] 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 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 a light chain variable region comprising the amino acid sequence of SEQ ID NO: 170.

[0532] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0533] Clone PA11P1E01

[0534] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to cashew and / or pistachio allergens.

[0535] In some embodiments, the antibody comprises:

[0536] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 621, 176 and 177, respectively; and

[0537] (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.

[0538] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence 620 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 622 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.

[0539] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0540] Clone PA11P1C11

[0541] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to cashew and / or pistachio allergens.

[0542] In some embodiments, the antibody comprises:

[0543] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 175, 176 and 177, respectively; and

[0544] (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.

[0545] 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.

[0546] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0547] Clone PA11P1C03

[0548] In some embodiments, a monoclonal antibody that specifically binds to a stone fruit 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 stone fruit allergens. In some embodiments, the monoclonal antibody binds to cashew and / or pistachio allergens.

[0549] In some embodiments, the antibody comprises:

[0550] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 438, 439 and 440, respectively; and

[0551] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 442, 30 and 443, respectively.

[0552] 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.

[0553] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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).

[0554] Antibodies that specifically bind to milk allergens

[0555] 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 light chain CDRs disclosed in Table 1 below. In some embodiments, the monoclonal antibody that specifically binds to a milk allergen comprises the following sequences: a light chain variable region sequence and / or a heavy chain variable region sequence disclosed in Table 1 below. In some embodiments, the 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 a heavy chain variable region sequence disclosed in Table 1 and comprising 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 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 a light chain variable region sequence disclosed in Table 1 and comprising CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0556] In some embodiments, the monoclonal antibody that specifically binds to a milk allergen comprises a light chain sequence or a portion thereof and / or a heavy chain sequence or a portion thereof, which is derived from 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 ...12, clone PA01P2E10, clone PA01P2C12, clone PA01P2 01P2E05, clone PA01P2D04, clone PA01P2B12, 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 PA01P The amino acid sequences of the CDRs, light chain variable regions (VL) and heavy chain variable regions (VH) of clone PA01P2B10, clone PA01P2D10, clone PA01P2D09, clone PA01P2B05, clone PA01P4C11, clone PA01P3E08, clone PA01P2E06, clone PA01P2E07, clone PA01P2G07, clone PA01P2B09, clone PA01P2C04 and clone PA01P2H08 are listed in Table 1 below.

[0557] In some embodiments, monoclonal antibodies that specifically bind to peanut allergens include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0558] (a) a heavy chain CDR1 comprising the amino acid sequence of any one 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;

[0559] (b) 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;

[0560] (c) 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;

[0561] (d) a light chain CDR1 comprising the amino acid sequence of any one 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;

[0562] (e) 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

[0563] (f) 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.

[0564] In some embodiments, the monoclonal antibody that specifically binds to a milk allergen comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequence:

[0565] (a) SEQ ID NOs: 750, 751, 752, 754, 196 and 755, respectively; or

[0566] (b) SEQ ID NOs: 757, 758, 759, 761, 762 and 763, respectively; or

[0567] (c) SEQ ID NOs: 765, 766, 767, 769, 30 and 770, respectively; or

[0568] (d) SEQ ID NOs: 772, 773, 774, 776, 389 and 777, respectively; or

[0569] (e) SEQ ID NOs: 779, 532, 780, 782, 404 and 783, respectively; or

[0570] (f) SEQ ID NOs: 785, 786, 787, 789, 790 and 791, respectively; or

[0571] (g) SEQ ID NOs: 793, 794, 795, 797, 30 and 798, respectively; or

[0572] (h) SEQ ID NOs: 800, 801, 802, 804, 110 and 805, respectively; or

[0573] (i) SEQ ID NOs: 807, 808, 809, 811, 509 and 812, respectively; or

[0574] (j) SEQ ID NOs: 814, 815, 816, 818, 94 and 819, respectively; or

[0575] (k) SEQ ID NOs: 821, 822, 787, 776, 790 and 824, respectively; or

[0576] (1) SEQ ID NOs: 121, 826, 827, 829, 830 and 831, respectively; or

[0577] (m) SEQ ID NOs: 833, 826, 827, 835, 149 and 836, respectively; or

[0578] (n) SEQ ID NOs: 838, 839, 840, 842, 843 and 844, respectively; or

[0579] (o) SEQ ID NOs: 846, 847, 848, 850, 196 and 851, respectively; or

[0580] (p) SEQ ID NOs: 853, 854, 855, 857, 662 and 858, respectively; or

[0581] (q) SEQ ID NOs: 860, 861, 862, 864, 865 and 866, respectively; or

[0582] (r) SEQ ID NOs: 868, 378, 869, 871, 682 and 872, respectively; or

[0583] (s) SEQ ID NOs: 874, 875, 876, 878, 22 and 879, respectively; or

[0584] (t) SEQ ID NOs: 881, 882, 883, 885, 886 and 887, respectively; or

[0585] (u) SEQ ID NOs: 889, 890, 891, 523, 762 and 893, respectively; or

[0586] (v) SEQ ID NOs: 895, 896, 897, 899, 900 and 901, respectively; or

[0587] (w) SEQ ID NOs: 903, 904, 905, 907, 908 and 909, respectively; or

[0588] (x) SEQ ID NOs: 911, 912, 913, 915, 149 and 916, respectively; or

[0589] (y) SEQ ID NOs: 918, 919, 920, 922, 923 and 924, respectively; or

[0590] (z) are SEQ ID NOs: 926, 927, 928, 930, 931 and 932, respectively.

[0591] In some embodiments, the 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, the 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, the 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, 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: any one of 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 having an amino acid sequence of at least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity).

[0592] 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 and comprising CDR1, CDR2, and CDR3 identical to the CDRs of the SEQ ID NO. 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 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 the SEQ ID NO.

[0593] In some embodiments, the monoclonal antibody that specifically binds to a milk allergen comprises:

[0594] (a) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 749; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 753; or

[0595] (b) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 756; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 760; or

[0596] (c) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 764; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 768; or

[0597] (d) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 771; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 775; or

[0598] (e) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 778; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 781; or

[0599] (f) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 784; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 788; or

[0600] (g) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 792; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 796; or

[0601] (h) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 799; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 803; or

[0602] (i) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 806; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 810; or

[0603] (j) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 813; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 817; or

[0604] (k) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 820; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 823; or

[0605] (l) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO:825; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO:828; or

[0606] (m) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 832; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 834; or

[0607] (n) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 837; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 841; or

[0608] (o) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 845; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 849; or

[0609] (p) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 852; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 856; or

[0610] (q) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 859; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 863; or

[0611] (r) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 867; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 870; or

[0612] (s) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 873; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 877; or

[0613] (t) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 880; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 884; or

[0614] (u) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 888; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 892; or

[0615] (v) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 894; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 898; or

[0616] (w) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 902; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 906; or

[0617] (x) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 910; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 914; or

[0618] (y) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 917; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 921; or

[0619] (z) a heavy chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 925; and a light chain variable region comprising an amino acid sequence that has 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 has the sequence of, SEQ ID NO: 929.

[0620] Clone PA01P2D09

[0621] In some embodiments, the 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.

[0622] In some embodiments, the antibody comprises:

[0623] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 860, 861 and 862, respectively; and

[0624] (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.

[0625] 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.

[0626] 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: 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).

[0627] Clone PA01P2D04

[0628] In some embodiments, the 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.

[0629] In some embodiments, the antibody comprises:

[0630] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 121, 826 and 827, respectively; and

[0631] (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.

[0632] 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.

[0633] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0634] Clone PA01P2B12

[0635] In some embodiments, the 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.

[0636] In some embodiments, the antibody comprises:

[0637] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 833, 826 and 827, respectively; and

[0638] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 835, 149 and 836, respectively.

[0639] 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.

[0640] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0641] Clone PA01P2B05

[0642] In some embodiments, the 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.

[0643] In some embodiments, the antibody comprises:

[0644] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 868, 378 and 869, respectively; and

[0645] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 871, 682 and 872, respectively.

[0646] 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.

[0647] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0648] Clone PA01P2D10

[0649] In some embodiments, the 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.

[0650] In some embodiments, the antibody comprises:

[0651] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 853, 854 and 855, respectively; and

[0652] (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.

[0653] 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.

[0654] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0655] Clone PA01P2E08

[0656] In some embodiments, the 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.

[0657] In some embodiments, the antibody comprises:

[0658] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 800, 801 and 802, respectively; and

[0659] (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.

[0660] 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.

[0661] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0662] Antibodies that specifically bind to fungal allergens

[0663] 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 f 1), e.g., purified recombinant allergen Aspergillus fumigatus 1 (rAsp f 1).

[0664] 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 exhibits cross-reactivity with 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. D Specifically binds to at least one of the fungal allergens.

[0665] Antibody sequence

[0666] In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen (e.g., specifically binds to an Aspergillus allergen) comprises a heavy chain CDR and / or a light chain CDR disclosed in Table 1 below. In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus allergen) comprises the following sequences: a light chain variable region sequence and / or a heavy chain variable region sequence disclosed in Table 1 below. In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen (e.g., an Aspergillus 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 the heavy chain variable region sequence disclosed in Table 1 and comprising CDRs identical to the heavy chain variable region sequence. and a light chain variable region comprising 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 a light chain variable region sequence disclosed in Table 1 and comprising CDR1, CDR2, and CDR3 that are identical to the CDRs of the light chain variable region sequence.

[0667] In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen (e.g., a monoclonal antibody that specifically binds to an Aspergillus allergen) comprises a light chain sequence or a portion thereof and / or a heavy chain sequence or a portion thereof, which is derived 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 region (VL), and heavy chain variable region (VH) of clone 1003320101_D6, clone 1003320105_D6, clone 1003320107_C5, clone 1003320107_F3, and clone 1003320107_F8 are listed in Table 1 below.

[0668] In some embodiments, the monoclonal antibodies that specifically bind to fungal allergens include one or more (e.g., one, two, three, four, five, or all six) of the following:

[0669] (a) a heavy chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 710, 718, 726, 734 or 742;

[0670] a heavy chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 711, 719, 727, 735 or 743;

[0671] a heavy chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 712, 720, 728, 736 or 744;

[0672] a light chain CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 714, 722, 730, 738 or 746;

[0673] a light chain CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 715, 723, 731, 739 or 747; and

[0674] A light chain CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 716, 724, 732, 740 or 748.

[0675] In some embodiments, a monoclonal antibody that specifically binds to a fungal allergen comprises heavy chain CDR1-3 and light chain CDR1-3 comprising the following amino acid sequence:

[0676] (a) SEQ ID NOs: 710, 711, 712, 714, 715 and 716, respectively; or

[0677] (b) SEQ ID NOs: 718, 719, 720, 722, 723 and 724, respectively; or

[0678] (c) SEQ ID NOs: 726, 7272, 728, 730, 731 and 732, respectively; or

[0679] (d) SEQ ID NOs: 734, 735, 736, 738, 739 and 740, respectively; or

[0680] (e) SEQ ID NOs: 742, 743, 744, 746, 747 and 748, respectively.

[0681] In some embodiments, the 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, the monoclonal antibody that specifically binds to a fungal 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: 713, 721, 729, 737, or 745. In some embodiments, the 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.

[0682] 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 comprising CDR1, CDR2, and CDR3 identical to the CDRs of the SEQ ID NO. In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen comprises a light chain variable region comprising 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: 713, 721, 729, 737, or 745 and comprising CDR1, CDR2, and CDR3 identical to the CDRs of the SEQ ID NO. In some embodiments, the monoclonal antibody that specifically binds to a fungal allergen comprises:

[0683] (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

[0684] (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 comprising CDR1, CDR2 and CDR3 identical to the CDRs of the SEQ ID NO.

[0685] Clone 1003320101_D6

[0686] 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.

[0687] In some embodiments, the antibody comprises:

[0688] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 710, 711 and 712, respectively; and

[0689] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 714, 715 and 716, respectively.

[0690] 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.

[0691] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0692] Clone 1003320105_D6

[0693] 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.

[0694] In some embodiments, the antibody comprises:

[0695] (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: 717 and comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 718, 719 and 720, respectively; and

[0696] (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.

[0697] 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.

[0698] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0699] Clone 1003320107_C5

[0700] 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 f 1). In some embodiments, the antibody cross-reactively binds specifically to more than one of Aspergillus fumigatus, Aspergillus niger, Aspergillus nidulans, or a recombinant Aspergillus antigen (e.g., rAsp f 1).

[0701] In some embodiments, the antibody comprises:

[0702] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 726, 727 and 728, respectively; and

[0703] (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.

[0704] 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.

[0705] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0706] Clone 1003320107_F3

[0707] 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).

[0708] In some embodiments, the antibody comprises:

[0709] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 734, 735 and 736, respectively; and

[0710] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 738, 739 and 740, respectively.

[0711] 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.

[0712] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0713] Clone 1003320107_F8

[0714] 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.

[0715] In some embodiments, the antibody comprises:

[0716] (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 comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 7422, 743 and 744, respectively; and

[0717] (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 comprising light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 746, 747 and 748, respectively.

[0718] 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.

[0719] In some embodiments, the antibody is an antibody that competes for binding with an antibody as 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: 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).

[0720] antigen-binding fragment

[0721] In some embodiments, antibodies as 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 producing antigen-binding fragments. Traditionally, these fragments were derived by 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 isolated directly from recombinant host cell culture. Other techniques for producing antigen-binding fragments are known in the art.

[0722] Antibody conjugates

[0723] In some embodiments, the antibody or antigen-binding fragment can be coupled to another molecule, such as polyethylene glycol (PEGylation) or serum albumin, to provide an 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).

[0724] In some embodiments, an antibody-drug conjugate is provided, comprising a monoclonal antibody or antigen-binding fragment as described herein. In some embodiments, a monoclonal antibody or antigen-binding fragment (e.g., an antibody or antigen-binding fragment that specifically binds to a food allergen or a 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 antifungals include, but are not limited to, amphotericin B, azole antifungals (e.g., ketoconazole, fluconazole, itraconazole, posaconazole, or voriconazole), echinocandins (e.g., anidulafungin, caspofungin, or micafungin), and flucytosine. Methods for manufacturing antibody-drug conjugates are described in, for example, Chudasama et al., Nature Chemistry, 2016, 8: 114-119; WO 2013 / 068874; and US 8,535,678.

[0725] Certain antibodies described herein are derived from B cells isolated from human subjects exposed to an allergen. In certain embodiments, the antibodies, antibody compositions and cells of the present invention can be distinguished from naturally occurring antibodies, compositions and cells in one or more aspects. Distinguishing antibodies, compositions and cells can be referred to as "synthetic" or can be identified by the following conditions: the antibody or composition is "not naturally occurring" or is definitely "non-naturally occurring". As used herein, the terms "corresponding antibody" and "corresponding to" describe the relationship between (1) an antibody characterized by six specific CDR sequences and produced by immune cells of the study subjects described in the Examples below and (2) a synthetic antibody comprising the same six CDR sequences.

[0726] synthetic antibodies

[0727] The synthetic antibodies of the present invention may differ structurally from naturally occurring antibodies having the same CDRs. That is, a synthetic antibody identified by a particular CDR may differ structurally from an antibody comprising the same CDR produced by cells from a study subject as described in the Examples below. Possible differences include:

[0728] Variable region sequences that differ from those of corresponding naturally occurring antibodies

[0729] In one approach, the antibody heavy chain comprises the CDRs of a clone described herein (e.g., PA13P1E10), provided that the antibody heavy chain does not comprise a heavy chain variable region sequence associated with a clone described herein. For illustration, 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), provided that the antibody light chain does not comprise a light chain variable region sequence associated with a clone described herein. For illustration, 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 an antibody of the invention have amino acid sequences that differ from the sequences disclosed herein.

[0730] Lambda and kappa light chains

[0731] In some embodiments, the synthetic antibodies comprise a lambda-type light chain. In some embodiments, the synthetic antibodies comprise a kappa-type light chain.

[0732] Isotype

[0733] In some Implementation In some embodiments, the synthetic antibodies with specified CDRs are of an isotype other than the isotype found to be associated with the study subject (from which the B cells with the specified CDRs were derived). In some embodiments, the antibodies disclosed herein are of an isotype other than IgG1. In some embodiments, the antibodies disclosed herein are of an isotype other than IgG2. In some embodiments, the antibodies disclosed herein are of an isotype other than IgG3. In some embodiments, the antibodies disclosed herein are of an isotype other than IgG4. In some embodiments, the antibodies disclosed herein are of an isotype other than IgM. In some embodiments, the antibodies disclosed herein are of an isotype other than IgA.

[0734] Allotypic

[0735] In some embodiments, the synthetic antibodies having designated CDRs are allotypes other than the allotype found to be associated with the study subject from which the B cells having designated CDRs were derived. In some embodiments, the synthetic antibodies of the invention include 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 include any single 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.

[0736] Table 2: Human immunoglobulin allotypes

[0737]

[0738] Note: Letter designations are given in parentheses. From: Jefferis and Marie-Paule Lefranc, 2009, “Human immunoglobulin allotypes: Possible implications for immunogenicity” mAbs 1(4): 332–338, incorporated herein by reference.

[0739] Constant domain variants

[0740] The synthetic antibodies of the present invention may include changes in the heavy chain constant region to alter the properties of the synthetic antibody relative to the corresponding naturally occurring antibody. Exemplary changes include mutations to modulate antibody effector functions (e.g., complement-based effector functions or FcγR-based effector functions), alter half-life, modulate co-engagement of antigen and FcγR, and introduce or remove glycosylation motifs (glyco-engineering). 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, “IgGFc engineering to modulate antibody effector functions” Protein Cell 2018, 9(1): 63–73; Schlothauer, 2016, “Novel human IgG1 and IgG4Fc-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, incorporated herein by reference for all purposes.

[0741] Synthetic antibody compositions

[0742] The synthetic antibody compositions of the present invention may differ from naturally occurring compositions in at least one or more of the following respects: (i) the composition comprises an antibody that is purified, i.e., isolated from tissue or cellular material with which it is associated in the human body, and optionally in a manufacturing excipient or medium; and / or (ii) the antibody composition of the present invention contains a single species of antibody (which is monoclonal), such that all antibodies in the composition have the same structure and specificity; the cell producing the synthetic antibody

[0743] The antibodies described herein can be produced by recombinant expression in human or non-human cells. Cells that produce synthetic antibodies include non-human cells that express heavy chains, light chains, or both heavy and light chains; human cells that are not immune cell heavy chains, light chains, or both heavy and light chains; and human B cells that produce heavy chains or light chains, but not heavy and light chains. Synthetic antibodies of the present invention can 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, optionally in addition to immune cells, and optionally in cells other than cells in the B cell lineage.

[0744] V. How to use

[0745] In another aspect, the present disclosure provides methods for treating a human subject with one or more allergen-specific monoclonal antibodies disclosed herein, or antigen-binding portions thereof. In some embodiments, methods of treating allergies are provided. In some embodiments, methods of alleviating 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 is often referred to as passive immunotherapy. Without being bound by a particular theory, it is hypothesized that the allergen-specific monoclonal antibodies disclosed herein block the binding of allergens to IgE or outcompete endogenous IgE for allergen binding, which in turn prevents or reduces the initiation of the allergic cascade. In some embodiments, without intending to be limited by a particular mechanism, the antibodies of the present invention provide therapeutic benefit by binding to inhibitory receptors on mast cells and / or basophils.

[0746] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of one or more allergen-specific monoclonal antibodies as disclosed herein (e.g., one or more allergen-specific monoclonal antibodies as disclosed in Section IV above). In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising one or more allergen-specific monoclonal antibodies as disclosed herein (e.g., a pharmaceutical composition as disclosed in Section VI below).

[0747] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of an allergen-specific monoclonal antibody (which is a human IgG isotype, e.g., a human IgG4 isotype) or an antigen-binding portion thereof (which comprises at least a portion of a human IgG or IgG4 isotype constant region sequence).

[0748] In some embodiments, the method comprises administering to the subject a therapeutically effective amount of an allergen-specific monoclonal antibody, or an antigen-binding portion thereof. In some embodiments, the method comprises administering to the subject two or more allergen-specific monoclonal antibodies (e.g., contained in a pharmaceutical composition comprising the two or more allergen-specific monoclonal antibodies). 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.

[0749] In some embodiments, the therapeutic antibody is an antibody comprising the CDR sequences, heavy chain variable region and / or light chain variable region as described herein (e.g., as shown in Table 1 below), and further comprising a native or modified IgM, IgD, IgG3, IgG1, IgA1, IgG2, IgG4 or IgA2 heavy chain constant region.

[0750] In some embodiments, the therapeutic antibody is conjugated to a drug, such as those described above in Section IV.

[0751] In some embodiments, the human subject to be treated is an adult. In some embodiments, the human subject is a teenager.

[0752] 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 legume allergen, a legume 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., tree pollen, grass pollen, or weed 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 bite.

[0753] In some embodiments, the human subject to be treated has an allergy to two or more allergens (e.g., to 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, 10, or more allergens. In some embodiments, the human subject has an allergy to two or more different types of antigens (allergens) within 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 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 an allergy to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more allergens in the same class of allergens, but does not have any known allergies to allergens in other classes of allergens. For example, in some embodiments, the human subject has an allergy to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more food allergens, but does not have any known allergies to non-food allergens. In some embodiments, the human subject has an allergy to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more fungal allergens, but does not have any known allergies to non-fungal allergens.

[0754] In some embodiments, the treatment methods disclosed herein reduce one or more symptoms of an allergy in a subject. One of ordinary skill in the art will appreciate that the symptoms associated with an allergic reaction can vary depending on the type of allergen inducing the allergic reaction. Examples of allergic reaction symptoms include, but are not limited to, hives, rash, eczema flare, redness of the skin, itchy mouth, itchy eyes, nausea, vomiting, diarrhea, stomach pain, nasal congestion, runny nose, nasal congestion, sneezing, coughing, tiredness, sore throat, swelling of the lips, tongue, or throat, headache, difficulty swallowing, shortness of breath, wheezing, decreased blood pressure, or a weak pulse. In some embodiments, the treatment methods disclosed herein reduce the severity of one or more symptoms of an allergy. In some embodiments of the treatment methods described herein, the subject's allergic symptoms include one or more of the following: runny nose, rash, redness of the skin, swelling of the skin, itching or tingling in or around the mouth and / or throat, difficulty swallowing, watery eyes, diarrhea, stomach cramps, nausea, vomiting, throat tightness, shortness of breath or wheezing, rapid breathing, and anaphylactic reaction. In some embodiments, the treatment methods disclosed herein reduce the length of time that one or more symptoms of allergy persist.

[0755] 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., peanut allergen), such as, but not limited to, hives, rash, eczema flare, 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, decreased blood pressure, or a weak pulse. In some embodiments, administration of one or more allergen-specific monoclonal antibodies disclosed herein reduces the severity of one or more symptoms and / or reduces the duration of one or more symptoms.

[0756] In some embodiments, the allergen-specific monoclonal antibodies disclosed herein are administered to human subjects in a therapeutically effective amount or dosage. In some embodiments, a daily dosage 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, the dosage may vary depending on several factors, including the chosen route of administration, the formulation of the composition, the patient's response, the severity of the condition, the subject's weight, and the judgment of the prescribing physician. The dosage may be increased or decreased over time, depending on the needs of the individual patient. In some cases, a patient is initially given a low dose, which is then increased to an effective dose that the patient can tolerate. Determination of an effective amount is well within the capabilities of those skilled in the art.

[0757] The route of administration of an antibody or composition comprising an antibody as described herein can be cutaneous or transdermal, inhalation, enteral, intravenous, intramuscular, intraperitoneal, intrathecal, intralesional, intrabronchial, nasal, ocular or ear 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 by injection (e.g., subcutaneous). In some embodiments, the route of administration of an antibody or composition comprising an antibody by any method described herein is subcutaneous, intravenous or intranasal.

[0758] In some embodiments, the administration of a single dose of an antibody as herein described or a composition comprising an antibody can effectively treat allergies or alleviate one or more symptoms of allergies. In some embodiments, multiple doses of an antibody or composition are administered. In some embodiments, at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days or longer, for example, at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks or longer, is administered as a second dose. In some embodiments, approximately every 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weeks, an antibody as herein described or a composition comprising an antibody is administered to a subject. In some embodiments, over an extended period of time, for example, at least 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350 days or longer, an antibody as herein described or a composition comprising an antibody is administered to a subject.

[0759] In some embodiments, in any of the methods described herein, an additional agent is further administered to the subject, 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., contained in the same pharmaceutical composition or contained in separate pharmaceutical compositions that are administered substantially simultaneously (e.g., administered 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 the additional agent. In some embodiments, the additional agent is administered first, followed by the monoclonal antibody.

[0760] In some embodiments, in any method described herein, the method may further include assessing the alleviation of allergic symptoms (e.g., allergic symptoms associated with peanut allergy, stone fruit allergy, milk allergy, or fungal allergy) in the subject. In certain embodiments, the alleviation of allergic symptoms can be measured by a total nasal symptom score (TNSS), which is based on a patient assessment of four symptoms: congestion, itching, and rhinorrhea, rated on a scale of 0 (none) to 3 (severe), and sneezing, rated on a scale of 0 (none) to 3 (5 or more). Each of the four symptoms is assessed using the following criteria: 0 = none, 1 = mild, 2 = moderate, or 3 = severe. The possible scores for the TNSS are 0-12. In other embodiments, the alleviation of allergic symptoms can be measured by a nasal symptom score on a visual analog scale (VAS), which is commonly used to classify allergy burden as mild, moderate, and severe. A VAS nasal symptom score from 0 (no nasal symptoms) to 100 (maximum nasal symptoms) can be used to assess the severity of combined nasal symptoms. In other embodiments, the reduction of allergic symptoms can be measured by peak nasal inspiratory flow (PNIF), which uses a nasal spirometer to measure the patient's nasal airflow (measured in 1 / min). In yet other embodiments, the reduction of allergic symptoms can be measured by allergen skin testing, such as skin prick testing (SPT), which uses the presence and extent of skin reactivity as a marker of sensitization in target organs (e.g., eyes, nose, lungs, intestines, and skin). When the relevant allergen (e.g., peanut allergen, stone fruit allergen, milk allergen, or fungal allergen) is introduced into the skin, the allergic reaction on the skin produces a blister and flare response, which can be quantified, for example, using the diameter of the blister. In yet other embodiments, the reduction of allergic symptoms can be measured by a basophil activation test, which utilizes flow cytometry to quantify the expression of activated markers on the surface of basophils after allergen stimulation. In yet other embodiments, the reduction of allergic symptoms can be measured by an oral food challenge, which involves administering increasing doses of the allergen to the allergic individual under the supervision of a trained allergist or immunologist. Oral food challenges can be performed in an open, single-blind, or double-blind format, with the gold standard being both double-blind and placebo-controlled.

[0761] In yet another aspect, the present disclosure provides diagnostic and detection methods using one or more allergen-specific monoclonal antibodies, or antigen-binding portions thereof, as disclosed herein. In some embodiments, the allergen-specific monoclonal antibodies, or antigen-bind...

Claims

1. A method for identifying human monoclonal antibodies that specifically bind to allergens, the method comprising: include: (a) obtaining single B cells, wherein the obtaining comprises negatively selecting B cells from peripheral blood of a subject allergic to an allergen and selecting single B cells expressing cell surface IgE; (b) generating cDNA from the single B cell of step (a); (c) determining the sequence of the cDNA from step (b) to identify (i) a heavy chain variable region sequence in an immunoglobulin heavy chain that includes an IgE 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; (d) expressing an antibody comprising the heavy chain variable region sequence and the light chain variable region sequence from step (c); and (e) determining whether the antibody from step (d) specifically binds to the allergen, thereby identifying a human monoclonal antibody comprising an IgE constant region and specifically binding to the allergen.

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

3. The method according to claim 1, wherein step (a) comprises contacting cells from the sample with anti-human CD19 antibodies and anti-human IgE antibodies and selecting cells expressing CD19 + IgE B cells.

4. The method according to claim 1, wherein step (a) comprises contacting cells from the sample with anti-human CD19 antibodies, anti-human IgE antibodies, anti-human IgM antibodies and anti-human IgG antibodies and selecting cells expressing CD19 + IgM - IgG - IgE B cells.

5. The method of claim 1, wherein the method comprises isolating single B cells that express a B cell marker and are negative for non-IgE isotypes.

6. The method according to claim 1, wherein step (a) comprises contacting cells from the sample with anti-human CD19 antibody, anti-human IgM antibody, anti-human IgG antibody, anti-human IgA antibody and anti-human IgD antibody and selecting CD19 + IgM - IgG - IgA - IgD - B cells.

7. The method of claim 1, wherein the method comprises isolating a single B cell that expresses a B cell marker and is negative for a non-IgG4 isotype.

8. The method according to claim 7, wherein step (a) comprises contacting cells from the sample with anti-human CD19 antibody, anti-human IgM antibody, anti-human IgE antibody, anti-human IgA antibody, anti-human IgD antibody, anti-human IgG1 antibody, anti-human IgG2 antibody and anti-human IgG3 antibody and selecting CD19 + IgM - IgE - IgA - IgD - IgG1 - IgG2 - IgG3 - B cells.

9. The method of claim 1, wherein the method further comprises replacing the constant region of the antibody identified in step (c) with a wild-type IgG4 constant region or a modified IgG4 constant region.

10. The method of claim 1, wherein the sample comprises blood, sputum, saliva, and tissue biopsy.

11. 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.

12. The method of claim 11, wherein the allergen is a food allergen selected from the group consisting of milk allergens, egg allergens, nut allergens, fish allergens, shellfish allergens, legume allergens, seed allergens and wheat allergens.

13. The method of claim 11, wherein the allergen is a soy allergen.

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