Compositions and methods for neutralization of antigens

Description

Compositions and Methods for Antigen Neutralization Cross-Reference

[001] This application claims the benefit of U.S. Provisional Application No. 63 / 496,508, filed April 17, 2023, the application for which is incorporated herein by reference. Sequence Listing

[002] This application contains a Sequence Listing that was submitted electronically in XML file format and is incorporated herein by reference in its entirety. The said XML file, created on April 2, 2024, is named 65491-701_601_SL.xml and is 70,130 bytes in size. Foundation

[003] Many people are allergic to animals, plants, or dust mites, especially those with other allergies or asthma. Despite efforts to develop approaches that reduce, minimize, or prevent an allergic response to animal allergens, there remains an unmet need to overcome the limitations inherent in conventional methods at reasonable cost. The present disclosure addresses these needs and offers related advantages. Summary

[004] An allergen-binding protein that binds to or neutralizes an allergen is provided herein, wherein the allergen-binding protein comprises a nanobody, a monobody, a DARPin, or a small peptide less than about 25 kilodaltons (kDa) in molecular weight or less than about 300 amino acids in length.

[005] Also provided here is an allergen-binding protein that binds to or neutralizes an allergen, wherein the allergen-binding protein comprises a nanobody or small peptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group Petition 870250113096, dated 09 / 12 / 2025, p. 9 / 183 2 / 119 which consists of SEQ ID Nos: 1-57 and 67-73.

[006] In some embodiments, the allergen-binding protein is formulated to coat a food or to spray, spray or brush onto an animal or household surface.

[007] In some forms, the food is pet food.

[008] In some forms, the allergen is a pet allergen.

[009] In some embodiments, the allergen-binding protein is formulated to be applied to the food as a coating.

[010] In some embodiments, the allergen-binding protein is formulated to be mixed into the food.

[011] In some embodiments, the allergen-binding protein is at a concentration of about 0.01 milligrams per milliliter (mg / ml) to about 500 mg / ml in solution or after suspension.

[012] In some forms, the allergen induces an allergic reaction in a human being.

[013] In some forms, the allergen comprises an environmental allergen.

[014] In some forms, the allergen comprises an animal allergen.

[015] In some forms, the allergen comprises a pet allergen.

[016] In some forms, the allergen comprises an allergen from a cat, dog, rabbit, mouse or cockroach.

[017] In some forms, the allergen is selected from the group consisting of Fel d 1, Fel d 2, Fel d 3, Fel d 4, Can f 1, Can f 2, Can f 4, Can f 7, Ory C 1, Mus M 1 and Bla G 2.

[018] In some forms, the allergen is Fel d 1.

[019] In some embodiments, the allergen-binding protein comprises a nanobody comprising a sequence Petition 870250113096, dated 09 / 12 / 2025, p. 10 / 183 3 / 119 amino acids with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence selected from the group consisting of SEQ ID Nos: 49-57.

[020] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO: 49.

[021] In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID NO: 49 sequence.

[022] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO: 57.

[023] In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID NO: 57 sequence.

[024] In some forms, the allergen is Can f 1.

[025] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 67-73.

[026] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 67. Petition 870250113096, dated 09 / 12 / 2025, p. 11 / 183 4 / 119

[027] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO: 67.

[028] In some forms, the allergen is Can f 2.

[029] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-23.

[030] In some embodiments, wherein the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 16.

[031] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence consisting of SEQ ID NO: 16.

[032] In some forms, the allergen comprises a dust allergen.

[033] In some embodiments, the dust allergen comprises Der p1 or Der p2.

[034] In some forms, the dust allergen is Der p1.

[035] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 24-34.

[036] In some embodiments, the allergen-binding protein comprises a peptide with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least Petition 870250113096, dated 09 / 12 / 2025, p. 12 / 183 5 / 119 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID Nos: 35-48.

[037] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 27.

[038] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO: 27.

[039] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 28.

[040] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO: 28.

[041] In some forms, the dust allergen is Der p2.

[042] In some embodiments, the allergen-binding protein comprises a nanobody with at least 80%, at least 85%, at least 90% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 18.

[043] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 3.

[044] In some embodiments, the binding protein to Petition 870250113096, dated 09 / 12 / 2025, p. 13 / 183 6 / 119 allergens comprises a nanobody with a sequence consisting of SEQ ID NO: 3.

[045] In some embodiments, the allergen comprises a plant allergen or a plant pollen allergen.

[046] In some embodiments, the plant allergen or pollen allergen is selected from the group consisting of Bet v1, Phl p 5, Phl p 1, Poa p 1, Cyn d 1, Bet v 2, Ole e 1, Amb a 1, Amb a 11 and Art v 1.

[047] In some forms, the allergen comprises a mold allergen.

[048] In some embodiments, the mold allergen is selected from the group consisting of Alt a 1, Asp f 1, Asp f 2, Cla h 8, Pen ch 13 and Pen ch 18.

[049] In some forms, the allergen is a food allergen.

[050] In some embodiments, the food allergen is selected from the group consisting of Pen a 1, Ara h 1, Ara h 3.

[051] In some embodiments, the allergen-binding protein comprises at least one modified amino acid.

[052] In some embodiments, the allergen-binding protein comprises about 1, about 5, about 10, about 15, about 20, about 25 or about 30 modified amino acids.

[053] In some embodiments, the modified amino acid comprises a non-canonical amino acid.

[054] In some embodiments, the non-canonical amino acid is selected from the group consisting of para-benzoylphenylalanine, 3,4-dihydroxyphenylalanine, a tetrazine, a chloroctene, homopropargylglycine, para-proparglyoxyphenylalanine, paraazidophenylalanine, para-isothiocyanate phenylalanine, parabenzoylphenylalanine, para-cyanophenylalanine, paranitrophenylalanine, a halogenated m-tyrosine analog, a halogenated proline analog, a tryptophan analog Petition 870250113096, dated 09 / 12 / 2025, page 14 / 183 7 / 119 halogenated and a halogenated leucine analogue.

[055] In some embodiments, the solubility of the allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600% or more compared to an unmodified allergen-binding protein.

[056] In some embodiments, the binding affinity of the allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to an unmodified allergen-binding protein.

[057] In some embodiments, the thermal stability of the allergen-binding protein is increased by about 10%, 25%, 50%, 75%, 100%, 200%, 300% or more compared to an unmodified allergen-binding protein.

[058] In some embodiments, the allergen-binding protein is a multivalent allergen-binding protein.

[059] In some embodiments, the multivalent allergen-binding protein is a bivalent allergen-binding protein.

[060] In some embodiments, the binding affinity of the multivalent allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to a monovalent allergen-binding protein.

[061] In some embodiments, the allergen-binding protein additionally comprises a carrier.

[062] In some embodiments, the carrier comprises a solvent, a diluent, a dispersion medium or a coating.

[063] In some embodiments, the carrier comprises silica.

[064] In some embodiments, the allergen-binding protein is in dry form.

[065] In some forms, the carrier includes water.

[066] In some embodiments, the binding protein to Petition 870250113096, dated 09 / 12 / 2025, p. 15 / 183 8 / 119 allergens comprises a liquid.

[067] In some embodiments, the allergen-binding protein additionally comprises a preservative.

[068] In some embodiments, the preservative comprises potassium sorbate, EDTA, benzoic acid, phenoxyethanol or maltol.

[069] In some embodiments, the allergen-binding protein additionally comprises a stabilizing or thickening agent.

[070] In some embodiments, the stabilizing or thickening agent comprises dextrin, maltodextrin, glycerol, glucose, sucrose or trehalose.

[071] In some embodiments, the allergen-binding protein additionally comprises an isotonic agent.

[072] In some embodiments, the allergen-binding protein is suspended or resuspended in a solvent.

[073] In some embodiments, the solvent comprises water.

[074] Also provided here is a composition comprising: an allergen-binding protein that binds to or neutralizes an allergen, wherein the allergen-binding protein comprises a nanobody, a monobody, a DARPin or a small peptide less than about 25 kilodaltons in molecular weight or less than about 300 amino acids in length.

[075] Also provided here is a composition comprising: an allergen-binding protein that binds to or neutralizes an allergen, wherein the allergen-binding protein comprises a nanobody or a small peptide less than about 25 kilodaltons in molecular weight or less than about 300 amino acids in length, wherein the allergen-binding protein comprises a sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 1-57 and 67-73. Petition 870250113096, dated 09 / 12 / 2025, p. 16 / 183 9 / 119

[076] In some embodiments, the composition is formulated to coat a food or to spray, spray or brush onto an animal or domestic surface.

[077] In some forms, the food is pet food.

[078] In some forms, the allergen is a pet allergen.

[079] In some embodiments, the composition is formulated to be applied over the food as a coating.

[080] In some forms, the composition is formulated to be mixed with food.

[081] In some embodiments, the allergen-binding protein is at a concentration of about 0.01 milligrams per milliliter (mg / ml) to about 500 mg / ml in solution or after suspension.

[082] In some forms, the allergen induces an allergic reaction in a human being.

[083] In some forms, the allergen comprises an environmental allergen.

[084] In some forms, the allergen comprises an animal allergen.

[085] In some forms, the allergen comprises a pet allergen.

[086] In some forms, the allergen comprises an allergen from a cat, dog, rabbit, mouse or cockroach.

[087] In some forms, the allergen is selected from the group consisting of Fel d 1, Fel d 2, Fel d 3, Fel d 4, Can f 1, Can f 2, Can f 4, Can f 7, Ory C 1, Mus M 1 and Bla G 2.

[088] In some forms, the allergen is Fel d 1.

[089] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% of Petition 870250113096, dated 09 / 12 / 2025, p. 17 / 183 10 / 119 sequence identity with a sequence selected from the group consisting of SEQ ID Nos: 49-57.

[090] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO: 49.

[091] In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID NO: 49 sequence.

[092] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO: 57.

[093] In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID NO: 57 sequence.

[094] In some forms, the allergen is Can f 1.

[095] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 67-73.

[096] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 67.

[097] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting Petition 870250113096, dated 09 / 12 / 2025, p. 18 / 183 11 / 119 in SEQ ID NO: 67.

[098] In some forms, the allergen is Can f 2.

[099] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 9-23.

[100] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 16.

[101] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence consisting of SEQ ID NO: 16.

[102] In some forms, the allergen comprises a dust allergen.

[103] In some embodiments, the dust allergen comprises Der p1 or Der p2.

[104] In some forms, the dust allergen is Der p1.

[105] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 24-34.

[106] In some embodiments, the allergen-binding protein comprises a peptide with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 35-48. Petition 870250113096, dated 09 / 12 / 2025, p. 19 / 183 12 / 119

[107] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 27.

[108] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO: 27.

[109] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 28.

[110] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO: 28.

[111] In some forms, the dust allergen is Der p2.

[112] In some embodiments, the allergen-binding protein comprises a nanobody with at least 80%, at least 85%, at least 90% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 18.

[113] In some embodiments, the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO: 3.

[114] In some embodiments, the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO: 3. Petition 870250113096, dated 09 / 12 / 2025, p. 20 / 183 13 / 119

[115] In some embodiments, the allergen comprises a plant allergen or a plant pollen allergen.

[116] In some embodiments, the plant allergen or pollen allergen is selected from the group consisting of Bet v1, Phl p 5, Phl p 1, Poa p 1, Cyn d 1, Bet v 2, Ole e 1, Amb a 1, Amb a 11 and Art v 1.

[117] In some forms, the allergen comprises a mold allergen.

[118] In some embodiments, the mold allergen is selected from the group consisting of Alt a 1, Asp f 1, Asp f 2, Cla h 8, Pen ch 13 and Pen ch 18.

[119] In some forms, the allergen is a food allergen.

[120] In some embodiments, the food allergen is selected from the group consisting of Pen a 1, Ara h 1, Ara h 3.

[121] In some embodiments, the allergen-binding protein comprises at least one amino acid modification.

[122] In some embodiments, the allergen-binding protein comprises about 1, about 5, about 10, about 15, about 20, about 25 or about 30 amino acid modifications.

[123] In some embodiments, amino acid modification involves the introduction of a non-canonical amino acid.

[124] In some embodiments, the non-canonical amino acid is selected from the group consisting of para-benzoylphenylalanine, 3,4-dihydroxyphenylalanine, a tetrazine, chloroctenes, homopropargylglycine, para-proparglyoxyphenylalanine, paraazidophenylalanine, para-isothiocyanate phenylalanine, parabenzoylphenylalanine, para-cyanophenylalanine, para-nitrophenylalanine, a halogenated m-tyrosine analog, a halogenated proline analog, a halogenated tryptophan analog and a halogenated leucine analog.

[125] In some forms, the solubility of the composition is Petition 870250113096, dated 09 / 12 / 2025, page 21 / 183 14 / 119 increased by approximately 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600% or more compared to an unmodified allergen-binding protein.

[126] In some embodiments, the binding affinity of the allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to an unmodified allergen-binding protein.

[127] In some embodiments, the thermal stability of the allergen-binding protein is increased by about 10%, 25%, 50%, 75%, 100%, 200%, 300% or more compared to an unmodified allergen-binding protein.

[128] In some embodiments, the allergen-binding protein is a multivalent allergen-binding protein.

[129] In some embodiments, the multivalent allergen-binding protein is a bivalent allergen-binding protein.

[130] In some embodiments, the binding affinity of the multivalent allergen-binding protein composition is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to a monovalent allergen-binding protein.

[131] In some forms, the composition additionally includes a carrier.

[132] In some embodiments, the carrier comprises a solvent, a diluent, a dispersion medium or a coating.

[133] In some embodiments, the carrier comprises silica.

[134] In some forms, the composition is in dry form.

[135] In some forms, the carrier includes water.

[136] In some embodiments, the composition comprises a liquid.

[137] In some forms, the composition comprises Petition 870250113096, dated 09 / 12 / 2025, page 22 / 183 15 / 119 plus a preservative.

[138] In some embodiments, the preservative comprises potassium sorbate, EDTA, benzoic acid, phenoxyethanol or maltol.

[139] In some embodiments, the composition additionally comprises a stabilizing or thickening agent.

[140] In some embodiments, the stabilizing or thickening agent comprises dextrin, maltodextrin, glycerol, glucose, sucrose or trehalose.

[141] In some forms, the composition additionally includes an isotonic agent.

[142] In some embodiments, the composition is suspended or resuspended in a solvent.

[143] In some embodiments, the solvent comprises water.

[144] Also provided here is a method comprising: spraying or spraying the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments, or placing the surface in contact with the composition of any of the foregoing embodiments.

[145] In some forms, contact involves coating or brushing.

[146] In some embodiments, the surface comprises the surface of an air filter or humidifier.

[147] In some forms, the surface comprises the allergen.

[148] In some forms, the composition neutralizes the allergen on the surface.

[149] In some forms, the surface comprises food.

[150] In some forms, the food is consumed by an animal that is the source of the antigen.

[151] In some forms, the surface comprises a Petition 870250113096, dated 09 / 12 / 2025, p. 23 / 183 16 / 119 pet accessory (e.g., collar or brush) or an area / product where an animal can sit or walk (e.g., a dog bed).

[152] In some forms, nebulization is carried out by a humidifier or a sprayer.

[153] In some embodiments, about 0.01 milliliter (ml) to about 20 ml of the sprayed composition per square meter of area neutralises the allergenicity of the antigen.

[154] Also provided here is a method of preparing allergen-binding protein of any of the foregoing embodiments or of the composition of any of the foregoing embodiments comprising: harvesting the allergen-binding protein from an engineered microbe or from a secretion of the microbe, wherein the microbe comprises a heterologous nucleic acid encoding the allergen-binding protein.

[155] In some embodiments, the method further comprises incorporating the heterologous nucleic acid encoding the allergen-binding protein into a cell-free protein expression system.

[156] In some forms, the microbe comprises a yeast or bacterium.

[157] In some embodiments, the microbe is a bacterium and comprises E. coli.

[158] In some forms, the microbe is a yeast and comprises Pichia pastoris.

[159] In some embodiments, the allergen-binding protein is purified or concentrated from a secretion by the microbe.

[160] In some forms, purification or concentration includes a filtration step.

[161] In some embodiments, the filtration step involves passing the secretion through a filter of about 0.01 nm, 0.1 nm, 1 nm, 5 nm or more in size. Petition 870250113096, dated 09 / 12 / 2025, p. 24 / 183 17 / 119

[162] In some embodiments, the method does not include a centrifugation step.

[163] Also provided here is a method for treating an allergy in an individual in need, the method comprising administering to the individual the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments.

[164] Also provided here is a method for treating an allergy in an individual in need, the method comprising (i) administering to the individual a therapeutically effective amount of a microbe designed to produce the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments.

[165] In some forms, the microbe is a yeast or bacterium.

[166] In some embodiments, the microbe is a bacterium and comprises E. coli.

[167] In some forms, the microbe is a yeast and comprises Pichia pastoris.

[168] In some forms, the microbe is administered to the individual orally.

[169] In some forms, the microbe produces the allergen-binding protein or composition when ingested.

[170] Also provided here is a method of neutralizing an allergen, the method comprising (i) aerosolizing the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments into a mist and (ii) bringing the mist into contact with a surface comprising an allergen.

[171] In some forms, aerosolization is carried out by an aerosolization machine.

[172] In some forms, the aerosolization machine is used by an individual. Petition 870250113096, dated 09 / 12 / 2025, p. 25 / 183 18 / 119

[173] In some embodiments, the surface comprises a pet accessory or an area / product where an animal can sit or walk.

[174] Also provided here is a method of neutralizing an allergen, the method comprising (i) aerosolizing the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments into a mist and (ii) bringing the mist into contact with a food.

[175] In some forms, the food is pet food.

[176] In some forms, the allergen is a pet allergen.

[177] In some forms, the mist is applied to the food as a coating.

[178] In some forms, the mist is mixed with the food.

[179] Also provided here is a method for treating an allergy in an individual in need, the method comprising aerosolizing the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments into a mist.

[180] In some forms, aerosolization is carried out by an aerosolization machine.

[181] In some forms, an aerosolization machine is used.

[182] In some forms, the aerosolization machine is used by the individual.

[183] ​​In some forms, the method additionally involves the individual inhaling the mist.

[184] In some forms, the individual is a human.

[185] In some forms, the individual is a non-human animal.

[186] A pharmaceutical composition is also provided here. Petition 870250113096, dated 09 / 12 / 2025, page 26 / 183 19 / 119 comprising (i) the allergen-binding protein of any of the foregoing embodiments or the composition of any of the foregoing embodiments and (ii) a pharmaceutically acceptable excipient.

[187] Also provided here is an allergen-binding protein comprising a sequence of either of the SEQ ID NOs: 1-57 or 67-73.

[188] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 3.

[189] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 16.

[190] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 27.

[191] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 28.

[192] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 49.

[193] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 57.

[194] Also provided here is an allergen-binding protein comprising a sequence with SEQ ID NO: 67.

[195] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the detailed description that follows, in which only illustrative embodiments of the present disclosure are shown and described. As will be seen, the present disclosure is susceptible to other and different embodiments, and its various details are susceptible to modifications in several obvious respects, all without departing from the disclosure. Consequently, the drawings and description should be considered illustrative in nature, and not restrictive. Incorporation by Reference

[196] All publications, patents and patent applications Petition 870250113096, dated 09 / 12 / 2025, page 27 / 183 20 / 119 mentioned in this descriptive report are incorporated herein by reference to the same extent as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated by reference. Brief Description of the Drawings

[197] The innovative features of the invention are presented in particular in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by referring to the detailed description below, which presents illustrative embodiments in which the principles of the invention are used, and to the attached drawings (also referred to as Figure and FIG. in this document), of which: Figure 1 shows a schematic illustration of a functional test, according to one or more embodiments of the present disclosure.

[198] Figure 2 represents a Western Blot SDSPAGE image of a recombinant Der p2 protein labeled with His, according to one or more embodiments of the present disclosure.

[199] Figure 3A illustrates the results of a Der p2 yeast display VHH library representing the sequence alignment of 50 randomly selected clones, according to one or more embodiments of the present disclosure.

[200] Figures 3B-3I are binding curves obtained from ELISA assays performed using the identified anti-Der p2 VHH clones, according to one or more embodiments of the present disclosure.

[201] Figure 4 represents a Western Blot SDSPAGE image of a recombinant Can f1 protein labeled with His, according to one or more embodiments of the present disclosure.

[202] Figure 5A illustrates the results of a VHH library displaying Can f1 yeast, representing the sequence alignment of 50 randomly selected clones, according to one or more embodiments of the present disclosure. Petition 870250113096, dated 09 / 12 / 2025, page 28 / 183 21 / 119

[203] Figures 5B-5H are binding curves obtained from ELISA assays performed using the identified VHH anti-Can f1 clones, according to one or more embodiments of the present disclosure.

[204] Figure 6 represents a Western Blot SDSPAGE image of a recombinant Can f2 protein labeled with His, according to one or more embodiments of the present disclosure.

[205] Figure 7A illustrates the results of a VHH library displaying Can f2 yeast representing the sequence alignment of 50 randomly selected clones, according to one or more embodiments of the present disclosure.

[206] Figures 7B-7P are binding curves obtained from ELISA assays performed using the identified VHH anti-Can f2 clones, according to one or more embodiments of the present disclosure.

[207] Figure 8 represents a Western Blot SDSPAGE image of a recombinant Der p1 protein labeled with His, according to one or more embodiments of the present disclosure.

[208] Figures 9A-9K are binding curves obtained from ELISA assays performed using the identified anti-Der p2 VHH clones, according to one or more embodiments of the present disclosure.

[209] Figure 10 illustrates the results of a Der p2 activity assay when Der p2 is incubated with the indicated anti-Der p2 VHH clones, according to one or more embodiments of the present disclosure.

[210] Figures 11A-11H illustrate the results of Der p2 activity assays when Der p2, at varying concentrations, is incubated with the indicated anti-Der p2 VHH clones, according to one or more embodiments of the present disclosure.

[211] Figure 12 illustrates the results of a Der p1 phage display 7 amino acid peptide library, representing the sequence alignment of the resulting peptides, according to one or more embodiments of the present disclosure.

[212] Figure 13 is a bond curve obtained from a test. Petition 870250113096, dated 09 / 12 / 2025, page 29 / 183 22 / 119 ELISA performed using the identified peptide clone 2-D3, according to one or more embodiments of the present disclosure.

[213] Figure 14 represents a Western Blot SDSPAGE image of a recombinant Fel d1 protein labeled with His, according to one or more embodiments of the present disclosure.

[214] Figure 15A illustrates the results of a Fel d1 yeast display VHH library, representing the sequence alignment of 50 randomly selected clones, according to one or more embodiments of the present disclosure.

[215] Figures 15B-15D are binding curves obtained from ELISA assays performed using the identified anti-Fel d1 VHH clones specific for the Fel d1 epitope ENARILKNCVDAKM (SEQ ID NO: 61), according to one or more embodiments of the present disclosure.

[216] Figure 16 is a binding curve obtained from an ELISA assay performed using the identified anti-Fel d1 VHH clone specific for the Fel d1 epitope FAVANGNELLLDLS (SEQ ID NO: 59), according to one or more embodiments of the present disclosure.

[217] Figures 17A-17E are binding curves obtained from ELISA assays performed using the identified anti-Fel d1 VHH clones specific for the Fel d1 epitope AKMTEEDKENALS (SEQ ID NO: 60), according to one or more embodiments of the present disclosure.

[218] Figure 18 is a binding curve obtained from an ELISA assay performed using the VHH anti-Fel d1 clone identified specific for the Fel d1 epitope VAQYKALPVVLENA (SEQ ID NO: 58).

[219] Figures 19A-19I illustrate the results of a thermostability test performed using the identified VHH anti-Fel d1 clones, according to one or more embodiments of the present disclosure.

[220] Figures 20A-20C illustrate size exclusion chromatography (SEC) plots obtained by testing the neutralization capacity of the VHH anti-Fel d1 clone C3, according to a Petition 870250113096, dated 09 / 12 / 2025, page 30 / 183 23 / 119 or more modalities of the present disclosure. Detailed Description

[221] An allergy can involve an abnormal immune system response to exposure to an allergen. The human body’s natural immune system can create antibodies. Antibodies can include Y-shaped proteins that bind to different foreign proteins or chemicals that interact with them. An allergic reaction can occur when the immune system reacts strangely to a foreign substance (allergen) and enters a hyperactive state of defense (TH2 cell immune response, etc.). This defense can sometimes be triggered by immunoglobulin E (IgE) antibodies that mast cells use to detect foreign substances, in this case, the allergen. When people inhale or come into contact with an allergen, the immune system can react, causing an inflammatory response in the nasal passages or lungs. Prolonged exposure to allergens can cause persistent inflammation associated with asthma.

[222] About 7 out of 10 households in the United States have a pet, and approximately 15 to 30 percent of the population is allergic to pets, specifically dogs or cats. Cat and dog allergens can be found in the animal's shed skin cells (dander), saliva, urine, sweat, and fur. Dandelion can cause a problem because it is very small and can remain airborne. Cats can be a major source of indoor inhalant allergens. The global incidence of cat allergies is rapidly increasing and has been considered a major public health problem. Some examples of cat allergens may include Fel d 1 to Fel d 8. More than about 95% of cat allergies may be caused by a secretoglobin, called Fel d 1. Cats can secrete the Fel d1 protein from their salivary and sebaceous glands and spread it throughout their fur during regular tongue baths. Fel d 1 Petition 870250113096, dated 09 / 12 / 2025, p. 31 / 183 24 / 119 can be easily transported through the air and remain in the indoor environment. Canis familiaris allergen 1 (Can f1) and Canis familiaris allergen 2 (Can f2) may be the two allergens present in dog hair or dander extracts. Canine allergens may also be present in dander, saliva, urine, and blood. Allergen levels can vary between breeds, and all breeds can trigger allergies, including even hairless dogs. There are Group I protein allergens (e.g., Der p I and Der f I) and Group II protein allergens (e.g., Der p II and Der f II) in house dust mite allergy.

[223] Several other allergens can also induce allergic reactions. Examples of pet allergens include, but are not limited to, Fel d 1, Fel d 2, Fel d 3, Fel d 4, Can f 1, Can f 2, Can f 4, Can f 7, Ory C 1, Mus M 1, or Bla G 2. Examples of plant or pollen allergens include, but are not limited to, Bet v1, Phl p 5, Phl p 1, Poa p 1, Cyn d 1, Bet v 2, Ole e 1, Amb a 1, Amb a 11, and Art v 1. Examples of mold allergens include, but are not limited to, Alt a 1, Asp f 1, Asp f 2, Cla h 8, Pen ch 13, and Pen ch 18. Examples of food allergens include, but are not limited to, Pen a 1, Ara h 1, and Ara h 3.

[224] The human body can produce antibodies that bind to the allergen and neutralize the allergic reaction. Antibodies can be produced by cells outside the human body and used to neutralize allergens. However, antibody production can be expensive, as it is done with human, mammalian, or avian cells. For example, a conventional approach might involve immunizing eggs / chickens with Fel d 1 and feeding these eggs to cats to reduce production costs. This approach may temporarily neutralize Fel d 1 in the cat's mouth, but the results are not perfect.

[225] Alpacas, llamas, and camels can produce similar immune proteins. Scientists can isolate a small section Petition 870250113096, dated 09 / 12 / 2025, page 32 / 183 25 / 119 of immune proteins, called single-domain antibodies (sdAbs) or nanobodies. The usefulness and advantages of single-domain antibodies or nanobodies may include, but are not limited to, their smaller size, greater number of accessible epitopes, relatively low production costs due to production in bacteria or yeast, and greater robustness compared to their full-length antibodies. These can also be produced from bacteria or yeast at a very low cost. Peptides with short amino acid chains can also be designed to bind to allergens. There are other small protein ligands that can be designed to bind and neutralize allergens, such as monobodies and engineered ankyrin repeat proteins (DARPs). Allergen-Binding Proteins

[226] In certain aspects, the present disclosure provides an allergen-binding protein. In some embodiments, the allergen-binding protein can bind to or neutralize the allergen. In some embodiments, the allergen-binding protein comprises an antibody fragment. The antibody fragment can be monoclonal. The antibody fragment can be polyclonal. In some embodiments, the allergen-binding protein comprises a nanobody, a monobody, a DARPin, or a small peptide. In some embodiments, the allergen-binding protein can be synthetic. In some embodiments, the allergen-binding protein can be engineered. In some embodiments, the allergen-binding protein can be recombinant.

[227] In some embodiments, the present disclosure provides an allergen-binding protein that binds to or neutralizes the allergen. In some embodiments, the allergen-binding protein comprises a nanobody, a monobody, a DARPin, or a small peptide less than about 25 kilodaltons (kDa) in size. Petition 870250113096, dated 09 / 12 / 2025, p. 33 / 183 26 / 119 molecular weight or less of about 300 amino acids in length. In some embodiments, the allergen-binding protein may be formulated to coat a food or to spray, mist, or brush onto the surface of an animal or home. In some embodiments, the food is a pet food. In some embodiments, the allergen is a pet allergen. In some embodiments, the allergen-binding protein is formulated to be applied to the food as a coating. In some embodiments, the allergen-binding protein is formulated to be mixed into the food.

[228] In some embodiments, the allergen-binding protein may be a nanobody. In some embodiments, the allergen-binding protein may be a monobody. In some embodiments, the allergen-binding protein may be a DARPin. In some embodiments, DARPins are small, single-domain proteins of about 14 kDa that can be selected to bind to any target protein with high affinity and specificity. In some embodiments, the allergen-binding protein may be a small peptide.

[229] In some embodiments, the allergen-binding protein described herein may be a nanobody. In some embodiments, the allergen-binding protein may be a single-domain antibody. The utility and advantages of single-domain antibodies (sdAbs) may include, but are not limited to, their smaller size, greater number of accessible epitopes, relatively low production costs, or greater robustness, compared to their full-length antibodies.

[230] In some embodiments, the allergen-binding protein comprises a nanobody or peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% of Petition 870250113096, dated 09 / 12 / 2025, page 34 / 183 27 / 119 sequence identity with a sequence selected from the group consisting of SEQ IDs 1-57 and 67-73. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 1. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 2. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 3.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 4. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 5. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 6.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% of... Petition 870250113096, dated 09 / 12 / 2025, page 35 / 183 28 / 119 sequence identity with a sequence of SEQ ID NO: 7. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 8. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 9. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 10.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 11. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 12. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 13.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 14. In some embodiments, a. Petition 870250113096, dated 09 / 12 / 2025, p. 36 / 183 29 / 119 Allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 15. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 16. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 17.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 18. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 19. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 20.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 21. In some embodiments, the allergen-binding protein comprises a nanobody with at least... Petition 870250113096, dated 09 / 12 / 2025, p. 37 / 183 30 / 119 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 22. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 23. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO: 24.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 25. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 26. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 27.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 28. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100% sequence identity. Petition 870250113096, dated 09 / 12 / 2025, page 38 / 183 31 / 119 less than 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 29. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 30. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 31. In some embodiments, the allergen-binding protein Allergens comprise a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 32.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 33. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 34. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 49.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with. Petition 870250113096, dated 09 / 12 / 2025, page 39 / 183 32 / 119 a sequence of SEQ ID NO: 50. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 51. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 52. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO: 53.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 54. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 55. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 56.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 57. In some embodiments, the allergen-binding protein... Petition 870250113096, dated 09 / 12 / 2025, p. 40 / 183 33 / 119 comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 67. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 68. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 69.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 70. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 71. In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 72.In some embodiments, the allergen-binding protein comprises a nanobody with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence of SEQ ID NO: 73.

[231] In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence Petition 870250113096, dated 09 / 12 / 2025, p. 41 / 183 34 / 119 NO: 1. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 2. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 3. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 4. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 5. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 6. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 7. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 8. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO: 9.In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 10. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 11. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 12. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 13. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 14. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 15. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 16. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 17. 17.In some forms, the protein binds to allergens. Petition 870250113096, dated 09 / 12 / 2025, p. 42 / 183 35 / 119 comprises a nanobody with a SEQ ID sequence number: 18. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 19. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 20. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 21. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 22. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 23. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 24. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 25. No. 25.In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 26. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 27. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 28. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 29. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 30. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 31. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number 32. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence number: 33.In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 34. Petition 870250113096, dated 09 / 12 / 2025, p. 43 / 183 36 / 119 In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 49. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 50. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 51. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 52. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 53. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 54. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 55. In some embodiments, the allergen-binding protein comprises a nanobody with a sequence of SEQ ID NO: 56.In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 57. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 67. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 68. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 69. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 70. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 71. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 72. In some embodiments, the allergen-binding protein comprises a nanobody with a SEQ ID sequence of 71. 73.

[232] In some embodiments, the binding protein to Petition 870250113096, dated 09 / 12 / 2025, p. 44 / 183 37 / 119 allergens comprise a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 35. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 36. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 37.In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 38. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 39. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 40.In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 41. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% of... Petition 870250113096, dated 09 / 12 / 2025, page 45 / 183 38 / 119 sequence identity with SEQ ID NO: 42. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 43. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 44. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with SEQ ID NO: 45.In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 46. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 47. In some embodiments, the allergen-binding protein comprises a peptide with at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with SEQ ID NO: 48.

[233] In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID sequence NO: 35. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID sequence NO: 36. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID sequence NO: 37. In Petition 870250113096, dated 09 / 12 / 2025, page 46 / 183 39 / 119 In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 38 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 39 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 40 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 41 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 42 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 43 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: 44 sequence. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID NO: sequence. 45.In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID sequence of 46. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID sequence of 47. In some embodiments, the allergen-binding protein comprises a peptide with a SEQ ID sequence of 48.

[234] In some embodiments, the allergen-binding protein described herein may be a humanized antibody fragment, a nanobody, a variant or a derivative thereof, which may, for example, be formulated for administration to a human being. In some embodiments, the humanized antibody may be a chimeric humanized antibody or a fully human antibody, for example, comprising an amino acid sequence of or similar to an amino acid sequence of a human antibody and a non-human amino acid sequence. For example, a portion of the heavy and / or light chain of a chimeric humanized antibody may be identical or similar to a sequence Petition 870250113096, dated 09 / 12 / 2025, p. 47 / 183 40 / 119 corresponding to a human antibody, while the remainder of the chain(s) may be non-human, for example, identical or similar to a corresponding sequence in an antibody derived from another species or belonging to another class or subclass of antibodies. The non-human sequence may be humanized to reduce the likelihood of immunogenicity while preserving target specificity, for example, by incorporating human DNA into the genetic sequence of the antibody-producing genes in the non-human animal. The humanized antibody may be a fully human antibody, for example, containing an amino acid sequence that is an amino acid sequence of a human antibody.

[235] In some embodiments, the allergen-binding protein described herein comprises a signal peptidase or a peptide protein fusion. Signal peptides can result in increased expression and / or secretion of proteins by a cell. Signal peptidases can cleave a signal peptide from the antibody or its antigen-binding fragment, for example, during a secretion process, generating a mature antibody that does not comprise the signal peptide sequence.

[236] The constant regions of the allergen-binding protein described here may mediate various effector functions and may be minimally involved in antigen binding.

[237] The allergen-binding protein described herein comprises constant regions that are selected or modified to provide suitable protein characteristics, for example, characteristics suitable for treating a disease or condition as disclosed herein.

[238] Variable regions (V) can mediate antigen binding and define the specificity of a particular antibody for an antigen.

[239] Within the hypervariable regions, there are amino acid residues that mainly determine the Petition 870250113096, dated 09 / 12 / 2025, page 48 / 183 41 / 119 antibody binding specificity. The sequences comprising these residues are known as complementarity-determining regions (CDRs). An antigen-binding site of an allergen-binding protein may be composed of several variable loops that confer specificity to the allergen-binding protein.

[240] In some embodiments, the allergen-binding protein described herein comprises variants or derivatives thereof. For example, a non-human animal, bacterium, yeast, or plant may be genetically modified to produce protein variants or derivatives. In some embodiments, an allergen-binding protein may be a single-domain antibody (sdAb), for example, a VHH heavy chain antibody (HCAb), nanobody, monobody, DARPin, small peptide, or scFV.

[241] In other embodiments, the allergen-binding protein described herein may be a binding fragment thereof. In some cases, the allergen-binding protein described herein may be a humanized antibody or a binding fragment thereof, a chimeric antibody or a binding fragment thereof, a monoclonal antibody or a binding fragment thereof, a multispecific antibody or a binding fragment thereof, a bispecific antibody or a binding fragment thereof, or a single-domain antibody (e.g., nanobody®). In some cases, the allergen-binding protein described here may be monovalent Fab', divalent Fab2, F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv)2, diabody, minibody, nanobody, triabody, tetrabody, disulfide-stabilized Fv protein (dsFv), single-domain antibody (sdAb), NAR Ig, camelid antibody or a binding fragment thereof, or a chemically modified derivative thereof.

[242] In some embodiments, the allergen-binding protein described herein may be a multispecific antibody. In some cases, the multispecific protein comprises two or Petition 870250113096, dated 09 / 12 / 2025, page 49 / 183 42 / 119 plus target-binding fractions, in which each of the two or more target-binding fractions binds specifically to one antigen, and the two or more antigens are different. In some cases, the multispecific antibody comprises target-binding fractions that bind specifically to three or more different antigens, four or more different antigens, or five or more different antigens. In some embodiments, the antibody may be a bispecific antibody.In some cases, the bispecific antibody or binding fragment includes, but is not limited to, Knobs-into-Holes (KiH), Asymmetric Reengineering Technology-Immunoglobulin (ART-Ig), Triomab quadroma, bispecific monoclonal antibody (BiMAb, BsmAb, BsAb, bsMab, BS-Mab or Bi-MAb), FcΔAdp, XmAb, Azymetric, Bispecific T-Cell Receptor-Based Antibody Engagement (BEAT), Bispecific T-Cell Engager (BiTE), Biclonics, Fab-scFv-Fc, Fab Two-in-One / Dual-Action (DAF), FinomAb, scFv-Fc(Fab)-fusion, Dock-aNd-Lock (DNL), Adaptir (formerly SCORPION), Tandem diAbody (TandAb), Dual-affinity-ReTargeting (DART) or nanobody. In some embodiments, the bispecific antibody may be a trifunctional antibody or a bispecific miniantibody. In some cases, the bispecific antibody may be a trifunctional antibody. The trifunctional antibody may be a full-length monoclonal antibody comprising binding sites for two different antigens.

[243] In some embodiments, the allergen-binding protein described herein comprises one or more mutations to stabilize the protein and / or increase the half-life.

[244] In some embodiments, the allergen-binding protein described herein comprises a humanized antibody or its binding fragment, or a chimeric antibody or its binding fragment. In some embodiments, the protein comprises a multispecific antibody or its binding fragment. Petition 870250113096, dated 09 / 12 / 2025, p. 50 / 183 43 / 119 binding. In some embodiments, the antibody comprises a bispecific antibody or its binding fragment. In some embodiments, the antibody may be an IgG-scFv, nanobody, BiTE, diabody, DART, TandAb, scDiabody, scDiabody-CH3, triple body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, FabscFv-Fc KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab')2, F(ab')2-scFv2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, diabodyFc, tandem scFv-Fc, or intrabody. In some cases, the antibody is a monovalent Fab', divalent Fab2, F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv)2, diabody, minibody, nanobody, triabody, tetrabody, disulfide-stabilized Fv protein (dsFv), single-domain antibody (e.g., a nanobody), Ig NAR, camelid antibody or binding fragment thereof, or a chemically modified derivative thereof.

[245] In some embodiments, the allergen-binding protein described herein (e.g., single-domain antibody) may bind to an epitope expressed by the target cell associated with the disease or condition described herein. In some embodiments, the antibody or its antigen-binding fragment may bind to an epitope associated with the microenvironment described herein.

[246] In some embodiments, the exogenous protein can function as an agonist or an antagonist, wherein upon binding to any of the epitopes described herein, the binding of the antibody or single-domain antibody induces an agonist or antagonist effect.

[247] In some embodiments, the antigen-binding protein may be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 nm in length. In some embodiments, the antigen-binding protein may be at most 1, at most 2, at most 3, at most 4, at most 5, at most 6, at most 7, at Petition 870250113096, dated 09 / 12 / 2025, p. 51 / 183 44 / 119 maximum 8, maximum 9, or maximum 10 nm in length. In some embodiments, the antigen-binding protein may be at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 nm long. In some embodiments, the antigen-binding protein may be between two of the values ​​described above, for example, between about 1 and about 10, about 2 and about 9, about 3 and about 8, about 4 and about 7, or about 5 and about 6 nm in length. In some embodiments, the antigen-binding protein may be between two of the values ​​described above, for example, between 1 and 10, 2 and 9, 3 and 8, 4 and 7, or 5 and 6 nm in length. In some embodiments, the antigen-binding protein can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nm in length.

[248] In some embodiments, the antigen-binding protein may have a molecular weight of about 9, 11, 13, 15, 17, 19, 21, 25, or 30 kDa. In some embodiments, the antigen-binding protein may have a molecular weight of at most 9, 11, 13, 15, 17, 19, 21, 25, or 30 kDa. In some embodiments, the antigen-binding protein may have a molecular weight of at least 9, 11, 13, 15, 17, 19, 21, 25, or 30 kDa. In some embodiments, the antigen-binding protein may have a molecular weight between two of the values ​​described above, for example, between approximately 9 and approximately 30, approximately 11 and approximately 25, approximately 13 and approximately 21, approximately 15 and approximately 19, or approximately 17 and approximately 30 kDa.In some forms, the antigen-binding protein can have a molecular weight of 9, 11, 13, 15, 17, 19, 21, 25, or 30 kDa.

[249] In some embodiments, the antigen-binding protein comprises about 5, about 10, about 20, about Petition 870250113096, dated 09 / 12 / 2025, p. 52 / 183 45 / 119 of 30, approximately 40, approximately 50, approximately 60, approximately 70, approximately 80, approximately 90, approximately 100, approximately 110, approximately 120, approximately 130, approximately 140, approximately 150, approximately 200, or approximately 250 amino acids. In some embodiments, the antigen-binding protein comprises a maximum of 5, a maximum of 10, a maximum of 20, a maximum of 30, a maximum of 40, a maximum of 50, a maximum of 60, a maximum of 70, a maximum of 80, a maximum of 90, a maximum of 100, a maximum of 110, a maximum of 120, a maximum of 130, a maximum of 140, a maximum of 150, a maximum of 200, or a maximum of 250 amino acids. In some embodiments, the antigen-binding protein comprises at least 5, at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 110, at least 120, at least 130, at least 140, at least 150, at least 200, or at least 250 amino acids.In some embodiments, the antigen-binding protein comprises a number of amino acids between two values ​​described above, for example, between about 5 and about 250, about 10 and about 200, about 20 and about 150, about 30 and about 140, about 50 and about 130, about 60 and about 120, about 70 and about 110, about 80 and about 100, or about 90 and about 250 amino acids. In some embodiments, the antigen-binding protein comprises a number of amino acids between two values ​​described above, for example, between 5 and 250, 10 and 200, 20 and 150, 30 and 140, 50 and 130, 60 and 120, 70 and 110, 80 and 100, or 90 and 250 amino acids. In some embodiments, the antigen-binding protein comprises 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 200, or 250 amino acids.

[250] In some embodiments, the nanobody may be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 nm in length. In some embodiments, the nanobody may be at most 1, at most 2, at most 3, at most 4, at most 5, at Petition 870250113096, dated 09 / 12 / 2025, p. 53 / 183 46 / 119 maximum 6, maximum 7, maximum 8, maximum 9, or maximum 10 nm in length. In some embodiments, the nanobody may have at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 nm in length. In some embodiments, the nanobody may have between two values ​​described above, for example, between about 1 and about 10, about 2 and about 9, about 3 and about 8, about 4 and about 7, or about 5 and about 6 nm in length. In some embodiments, the nanobody may have between two values ​​described above, for example, between 1 and 10, 2 and 9, 3 and 8, 4 and 7, or 5 and 6 nm in length. In some embodiments, the nanobody can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nm in length.

[251] In some embodiments, the nanobody may have a molecular weight of about 9, 11, 13, 15, 17, 19, or 21 kDa. In some embodiments, the nanobody may have a molecular weight of at most 9, 11, 13, 15, 17, 19, or 21 kDa. In some embodiments, the nanobody may have a molecular weight of at least 9, 11, 13, 15, 17, 19, or 21 kDa. In some embodiments, the nanobody may have a molecular weight between two values ​​described above, for example, between about 9 and 21, about 11 and 19, about 13 and 17, about 15 and 21, or about 12 and 15 kDa. In some embodiments, the nanobody may have between two of the values ​​described above, for example, between 9 and 21, 11 and 19, 13 and 17, 15 and 21, or 12 and 15 kDa in molecular weight. In some embodiments, the nanobody may have 9, 11, 13, 15, 17, 19, or 21 kDa in molecular weight.

[252] In some embodiments, the nanobody comprises about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, or about 150 amino acids. In some embodiments, the nanobody comprises at most 70, at most 80, at most 90, at most 100, at most 110, at most 120, at most 130, at most 140, or at most 150 amino acids. In some embodiments, the nanobody comprises Petition 870250113096, dated 09 / 12 / 2025, p. 54 / 183 47 / 119 at least 70, at least 80, at least 90, at least 100, at least 110, at least 120, at least 130, at least 140 or at least 150 amino acids. In some embodiments, the nanobody comprises a number of amino acids between two values ​​described above, for example, between about 70 and about 150, about 80 and about 140, about 90 and about 130, about 100 and about 120 or about 110 and about 150 amino acids. In some embodiments, the nanobody comprises a number of amino acids between two values ​​described above, for example, between 70 and 150, 80 and 140, 90 and 130, 100 and 120 or 110 and 150 amino acids. In some embodiments, the nanobody comprises 70, 80, 90, 100, 110, 120, 130, 140, or 150 amino acids.

[253] In some embodiments, the antigen-binding protein comprises a multispecific protein. In some embodiments, the antigen-binding protein may be conjugated with a protein or peptide.

[254] In some embodiments, the small peptide or protein linker comprises a modified amino acid or an unnatural amino acid, or a modified unnatural amino acid, or a combination thereof. In some embodiments, the modified amino acid or a modified unnatural amino acid comprises a post-translational modification.In some embodiments, the small peptide comprises a modification including, but not limited to, acetylation, acylation, ADP-ribosylation, amidation, flavin covalent linkage, heme moiety covalent linkage, nucleotide or nucleotide derivative covalent linkage, lipid or lipid derivative covalent linkage, phosphatidylinositol covalent linkage, crosslinking, cyclization, disulfide bond formation, demethylation, covalent crosslinking, cystine formation, pyroglutamate formation, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing. Petition 870250113096, dated 09 / 12 / 2025, page 55 / 183 48 / 119 phosphorylation, prenylation, racemization, selenoylation, sulfation, RNA-mediated transfer of amino acids to proteins, such as arginylation and ubiquitination. Modifications are made anywhere on the small peptide, including the peptide structure, amino acid side chains, and the terminus.

[255] In some embodiments, the allergen-binding protein comprises at least one modified amino acid. In some embodiments, the allergen-binding protein comprises about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 60, about 65 or about 70 modified amino acids. In some embodiments, the allergen-binding protein comprises a maximum of 1, a maximum of 2, a maximum of 3, a maximum of 4, a maximum of 5, a maximum of 6, a maximum of 7, a maximum of 8, a maximum of 9, a maximum of 10, a maximum of 11, a maximum of 12, a maximum of 13, a maximum of 14, a maximum of 15, a maximum of 20, a maximum of 25, a maximum of 30, a maximum of 35, a maximum of 40, a maximum of 45, a maximum of 50, a maximum of 60, a maximum of 65, or a maximum of 70 modified amino acids.In some embodiments, the allergen-binding protein comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 60, at least 65, or at least 70 modified amino acids. In some embodiments, the allergen-binding protein comprises about 1 to about 70, about 2 to about 65, about 3 to about 60, about 4 to about 50, about 5 to about 45, about 6 to about 40, about 7 to about 35, about 8 to about. Petition 870250113096, dated 09 / 12 / 2025, p. 56 / 183 49 / 119 of 30, approximately 9 to approximately 25, approximately 10 to approximately 20, or approximately 11 to approximately 15 modified amino acids. In some embodiments, the allergen-binding protein comprises 1 to 70, 2 to 65, 3 to 60, 4 to 50, 5 to 45, 6 to 40, 7 to 35, 8 to 30, 9 to 25, 10 to 20, or 11 to 15 modified amino acids. In some embodiments, the allergen-binding protein comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, 50, 60, 65, or 70 modified amino acids.

[256] In some embodiments, the modified amino acid comprises a non-canonical amino acid. In some embodiments, the non-canonical amino acid is selected from the group consisting of para-benzoylphenylalanine, 3,4-dihydroxyphenylalanine, a tetrazine, a chloroctene, homopropargylglycine, paraproparglyoxyphenylalanine, para-azidophenylalanine, phenylalanine paraisothiocyanate, para-benzoylphenylalanine, paracyanophenylalanine, para-nitrophenylalanine, a halogenated m-tyrosine analog, a halogenated proline analog, a halogenated tryptophan analog, and a halogenated leucine analog. In some embodiments, the non-canonical amino acid is para-benzoylphenylalanine. In some embodiments, the non-canonical amino acid is 3,4-dihydroxyphenylalanine. In some embodiments, the non-canonical amino acid is a tetrazine. In some embodiments, the non-canonical amino acid is chloroctene. In some embodiments, the non-canonical amino acid is homopropargylglycine.In some embodiments, the non-canonical amino acid is para-proparglyoxyphenylalanine. In some embodiments, the non-canonical amino acid is para-azidophenylalanine. In some embodiments, the non-canonical amino acid is phenylalanine paraisothiocyanate. In some embodiments, the non-canonical amino acid is para-benzoylphenylalanine. In some embodiments, the non-canonical amino acid is para-cyanophenylalanine. In some embodiments, the non-canonical amino acid is para-nitrophenylalanine. In some embodiments, the non-canonical amino acid is an analog. Petition 870250113096, dated 09 / 12 / 2025, page 57 / 183 50 / 119 of tyrosine m-halogenated. In some embodiments, the non-canonical amino acid is a halogenated proline analog. In some embodiments, the non-canonical amino acid is a halogenated tryptophan analog. In some embodiments, the non-canonical amino acid is a halogenated leucine analog.

[257] In some embodiments, the solubility of the allergen-binding protein is increased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, about 500%, about 600%, about 700%, about 800%, about 900%, approximately 1000% or more compared to an unmodified allergen-binding protein.In some embodiments, the solubility of the allergen-binding protein is increased by a maximum of 5%, a maximum of 10%, a maximum of 15%, a maximum of 20%, a maximum of 25%, a maximum of 30%, a maximum of 35%, a maximum of 40%, a maximum of 45%, a maximum of 50%, a maximum of 55%, a maximum of 60%, a maximum of 65%, a maximum of 70%, a maximum of 75%, a maximum of 80%, a maximum of 90%, a maximum of 100%, a maximum of 150%, a maximum of 200%, a maximum of 250%, a maximum of 300%, a maximum of 350%, a maximum of 400%, a maximum of 450%, a maximum of 500%, a maximum of 600%, a maximum of 700%, a maximum of 800%, a maximum of 900%, a maximum of 1000% or more compared to an unmodified allergen-binding protein.In some embodiments, the solubility of the allergen-binding protein is increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%. Petition 870250113096, dated 09 / 12 / 2025, page 58 / 183 51 / 119 at least 500%, at least 600%, at least 700%, at least 800%, at least 900%, at least 1000% or more compared to an unmodified allergen-binding protein. In some embodiments, the solubility of the allergen-binding protein is increased by about 5% to about 1000%, about 10% to about 900%, about 15% to about 800%, about 20% to about 700%, about 25% to about 600%, about 30% to about 500%, about 35% to about 400%, about 40% to about 300%, about 45% to about 200%, about 50% to about 100%, about 55% to about 90%, or about 60% to about 80% compared to an unmodified allergen-binding protein.In some embodiments, the solubility of the allergen-binding protein is increased by 5% to 1000%, 10% to 900%, 15% to 800%, 20% to 700%, 25% to 600%, 30% to 500%, 35% to 400%, 40% to 300%, 45% to 200%, 50% to 100%, 55% to 90%, or 60% to 80% compared to an unmodified allergen-binding protein. In some embodiments, the solubility of the allergen-binding protein is increased by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, 1000% or more compared to an unmodified allergen-binding protein.

[258] In some embodiments, the binding affinity of the allergen-binding protein is increased by about 10%, about 15%, about 20%, about 25%, about 50%, about 75%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, about 500%, about 550%, about 600%, about 650%, about 700%, about 750%, about 800%, about 850%, about 900%, about 950%, about 1000%, about 1500%, about 2000%, about 3,000% or more compared to an unmodified allergen-binding protein. In some embodiments, the maximum binding affinity of the allergen-binding protein is... Petition 870250113096, dated 09 / 12 / 2025, p. 59 / 183 52 / 119 increased by a maximum of 10%, a maximum of 15%, a maximum of 20%, a maximum of 25%, a maximum of 50%, a maximum of 75%, a maximum of 100%, a maximum of 150%, a maximum of 200%, a maximum of 250%, a maximum of 300%, a maximum of 350%, a maximum of 400%, a maximum of 450%, a maximum of 500%, a maximum of 550%, a maximum of 600%, a maximum of 650%, a maximum of 700%, a maximum of 750%, a maximum of 800%, a maximum of 850%, a maximum of 900%, a maximum of 950%, a maximum of 1000%, a maximum of 1500%, a maximum of 2000%, a maximum of 3000% or more compared to an unmodified allergen-binding protein.In some embodiments, at least the binding affinity of the allergen-binding protein is increased by at least 10%, at least 15%, at least 20%, at least 25%, at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, at least 600%, at least 650%, at least 700%, at least 750%, at least 800%, at least 850%, at least 900%, at least 950%, at least 1000%, at least 1500%, at least 2000%, at least 3000% or more compared to an unmodified allergen-binding protein.In some embodiments, the binding affinity of the allergen-binding protein is increased by approximately 10% to approximately 3,000%, approximately 15% to approximately 2,000%, approximately 20% to approximately 1,500%, approximately 25% to approximately 1,000%, approximately 50% to approximately 950%, approximately 75% to approximately 900%, approximately 100% to approximately 850%, approximately 150% to approximately 800%, approximately 200% to approximately 750%, approximately 250% to approximately 700%, approximately 300% to approximately 650%, approximately 350% to approximately 600%, approximately 400% to approximately 550%, or approximately 450% to approximately 500%. compared to an unmodified allergen-binding protein. In some embodiments, the binding affinity of the allergen-binding protein is increased by 10% to 3,000%, 15% to 2,000%, 20% to 1,500%, 25% to 1,000%, 50% to 950%, 75% to 900%, 100% to 850%, 150% to 800%, 200% to 750%, 250% to 700%, 300% to 650%, 350% to 600%, 400% to 550%, or 450% to 500% compared to a protein. Petition 870250113096, dated 09 / 12 / 2025, p. 60 / 183 53 / 119 of unmodified allergen binding. In some embodiments, the binding affinity of the allergen-binding protein is increased by 10%, 15%, 20%, 25%, 50%, 75%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 550%, 600%, 650%, 700%, 750%, 800%, 850%, 900%, 950%, 1000%, 1500%, 2000%, 3000% or more compared to an unmodified allergen-binding protein.

[259] In some embodiments, the thermal stability of the allergen-binding protein is increased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, about 500%, about 600%, about 700%, about 800%, about 900%, approximately 1000% or more compared to an unmodified allergen-binding protein. In some embodiments, the thermal stability of the allergen-binding protein is increased by a maximum of 5%. maximum 10%, maximum 15%, maximum 20%, maximum 25%, maximum 30%, maximum 35%, maximum 40%, maximum 45%, maximum 50%, maximum 55%, maximum 60%, maximum 65%, maximum 70%, maximum 75%, maximum 80%, maximum 90%, maximum 100%, maximum 150%, maximum 200%, maximum 250%, maximum 300%, maximum 350%, maximum 400%, maximum 450%, maximum 500%, maximum 600%, maximum 700%, maximum 800%, maximum 900%, maximum 1000% or more compared to a unmodified allergen-binding protein. In some embodiments, the thermal stability of the allergen-binding protein is increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, by Petition 870250113096, dated 09 / 12 / 2025, page 61 / 183 54 / 119 less 75%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 600%, at least 700%, at least 800%, at least 900%, at least 1000% or more compared to an unmodified allergen-binding protein. In some embodiments, the thermal stability of the allergen-binding protein is increased by about 5% to about 1000%, about 10% to about 900%, about 15% to about 800%, about 20% to about 700%, about 25% to about 600%, about 30% to about 500%, about 35% to about 400%, about 40% to about 300%, about 45% to about 200%, about 50% to about 100%, about 55% to about 90%, or about 60% to about 80% compared to an unmodified allergen-binding protein.In some embodiments, the thermal stability of the allergen-binding protein is increased by 5% to 1000%, 10% to 900%, 15% to 800%, 20% to 700%, 25% to 600%, 30% to 500%, 35% to 400%, 40% to 300%, 45% to 200%, 50% to 100%, 55% to 90%, or 60% to 80% compared to an unmodified allergen-binding protein. In some embodiments, the thermal stability of the allergen-binding protein is increased by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, 1000% or more compared to an unmodified allergen-binding protein.

[260] In some embodiments, the allergen-binding protein is multivalent. In some embodiments, the allergen-binding protein is bivalent. In some embodiments, the allergen-binding protein is trivalent. In some embodiments, the allergen-binding protein is quadrivalent. In some embodiments, the solubility of the allergen-binding protein is increased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, Petition 870250113096, dated 09 / 12 / 2025, p. 62 / 183 55 / 119 approximately 40%, approximately 45%, approximately 50%, approximately 55%, approximately 60%, approximately 65%, approximately 70%, approximately 75%, approximately 80%, approximately 90%, approximately 100%, approximately 150%, approximately 200%, approximately 250%, approximately 300%, approximately 350%, approximately 400%, approximately 450%, approximately 500%, approximately 600%, approximately 700%, approximately 800%, approximately 900%, approximately 1000% or more compared to a monovalent allergen-binding protein.In some embodiments, the solubility of the allergen-binding protein is increased by a maximum of 5%, a maximum of 10%, a maximum of 15%, a maximum of 20%, a maximum of 25%, a maximum of 30%, a maximum of 35%, a maximum of 40%, a maximum of 45%, a maximum of 50%, a maximum of 55%, a maximum of 60%, a maximum of 65%, a maximum of 70%, a maximum of 75%, a maximum of 80%, a maximum of 90%, a maximum of 100%, a maximum of 150%, a maximum of 200%, a maximum of 250%, a maximum of 300%, a maximum of 350%, a maximum of 400%, a maximum of 450%, a maximum of 500%, a maximum of 600%, a maximum of 700%, a maximum of 800%, a maximum of 900%, a maximum of 1000% or more compared to a monovalent allergen-binding protein.In some embodiments, the solubility of the allergen-binding protein is increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 600%, at least 700%, at least 800%, at least 900%, at least 1000% or more compared to a monovalent allergen-binding protein.In some embodiments, the solubility of the allergen-binding protein is increased by about 5% to about 1000%, about 10% to about 900%, about 15% to about 800%, about 20% to about 700%, about 25% to about 600%, about 30% to about 500%, about 35% to about 400%, about 40% to about 300%, about. Petition 870250113096, dated 09 / 12 / 2025, p. 63 / 183 56 / 119 from 45% to about 200%, about 50% to about 100%, about 55% to about 90%, or about 60% to about 80% compared to a monovalent allergen-binding protein. In some embodiments, the solubility of the allergen-binding protein is increased by 5% to 1000%, 10% to 900%, 15% to 800%, 20% to 700%, 25% to 600%, 30% to 500%, 35% to 400%, 40% to 300%, 45% to 200%, 50% to 100%, 55% to 90%, or 60% to 80% compared to a monovalent allergen-binding protein. In some embodiments, the solubility of the allergen-binding protein is increased by 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 600%, 700%, 800%, 900%, 1000% or more compared to a monovalent allergen-binding protein.

[261] In some embodiments, the short peptide may be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 28, about 30, about 32 or about 35 kDa in molecular weight. In some embodiments, the small peptide may have a maximum molecular weight of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28, 30, 32, or 35 kDa.In some embodiments, the small peptide may have at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least. Petition 870250113096, dated 09 / 12 / 2025, page 64 / 183 57 / 119 minus 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 28, at least 30, at least 32 or at least 35 kDa in molecular weight. In some embodiments, the small peptide may be between two values ​​described above, for example, between about 1 to about 35, about 2 to about 34, about 3 to about 33, about 4 to about 32, about 5 to about 31, about 6 to about 30, about 7 to about 29, about 8 to about 28, about 9 to about 27, about 10 to about 26, about 11 to about 25, about 12 to about 24, about 13 to about 23, about 14 to about 21, about 15 to about 20, about 16 to about 19, about 17 to about 18 kDa in molecular weight. In some embodiments, the small peptide may have a molecular weight of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28, 30, 32, or 35 kDa.

[262] In some embodiments, the small peptide comprises about 5, about 10, about 20, about 60, about 120, about 140, about 160, about 180, about 200, about 220, about 240, about 260, about 280, about 300, about 320, about 340, about 360, about 380 or 400 amino acids in length. In some embodiments, the small peptide comprises a maximum of 5, a maximum of 10, a maximum of 20, a maximum of 60, a maximum of 120, a maximum of 140, a maximum of 160, a maximum of 180, a maximum of 200, a maximum of 220, a maximum of 240, a maximum of 260, a maximum of 280, a maximum of 300, a maximum of 320, a maximum of 340, a maximum of 360, a maximum of 380, or a maximum of 400 amino acids in length.In some embodiments, the small peptide comprises at least 5, at least 10, at least 20, at least 60, at least 120, at least 140, at least 160, at least 180, at least 200, at least 220, at least 240, at least 260, at least 280, at least 300, at least 320, at least 340, at least 360, at least 380, or at least 400 amino acids. Petition 870250113096, dated 09 / 12 / 2025, page 65 / 183 58 / 119 in length. In some embodiments, the small peptide comprises a number of amino acids between two values ​​described above, for example, between about 5 and about 400, about 10 and about 380, about 20 and about 360, about 60 and about 340, about 120 and about 320, about 140 and about 300, about 160 and about 280, about 180 and about 260, or about 200 and about 240 amino acids in length. In some embodiments, the small peptide comprises a number of amino acids between 5 and 400, 10 and 80, 20 and 360, 60 and 340, 120 and 320, 140 and 300, 160 and 280, 180 and 260, or 200 and 240 amino acids in length. In some embodiments, the small peptide comprises approximately 5, 10, 20, 60, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, or 400 amino acids in length.

[263] The allergen-binding protein disclosed herein may be formulated in any suitable physical form. Suitable forms may include, but are not limited to, an aerosol, a liquid, a gel, a semisolid, a solid, or a powder. In some embodiments, the allergen-binding protein may be formulated as a tonic, a cream, an emulsion, a lotion, an ointment, a paste, a gel, a suspension, a serum, an oil, a spray, a shampoo, a foam, a cleanser, a mousse, an aerosol, or a powder to be resuspended in a solvent such as water.

[264] In some embodiments, allergen-binding protein can be formulated to coat a food. In some embodiments, allergen-binding protein can be mixed with any type of food for an animal. In some embodiments, allergen-binding protein can be provided as food or added to it. In some embodiments, allergen-binding protein can be provided as a supplement to the animal's fluid intake, including its drinking water. In some embodiments, allergen-binding protein can be formulated as a food product. Petition 870250113096, dated 09 / 12 / 2025, p. 66 / 183 59 / 119

[265] In some embodiments, the allergen-binding protein can be formulated for spraying, misting, or brushing onto an animal or a household surface. In some embodiments, the allergen-binding protein can be formulated for a humidifier and an air filter. In some embodiments, the allergen-binding protein can be applied to a surface in the environment, for example, by spraying, misting, depositing, cleaning, or other suitable application method. In some embodiments, the surface can be the surface of an animal that is a source of the allergen.

[266] In some embodiments, the allergen-binding protein disclosed herein can reduce, minimize, or prevent at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the allergen-binding protein disclosed herein can reduce at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the allergen-binding protein disclosed herein can minimize at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the allergen-binding protein disclosed herein can prevent at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the allergen-binding protein can come into contact with an environmental allergen, bind to the allergen, and prevent the allergen from inducing an allergic reaction in the susceptible individual or the individual suffering from allergies caused by the allergen.In some embodiments, the allergen-binding protein may come into contact with an environmental allergen. In some embodiments, the allergen-binding protein may bind to the allergen.

[267] In some embodiments, the allergen-binding protein may be resuspended or be in solution at a concentration of about 0.01 milligrams per milliliter (mg / ml) to about 500 mg / ml. In some embodiments, the binding protein Petition 870250113096, dated 09 / 12 / 2025, p. 67 / 183 60 / 119 allergens may be in a concentration of about 0.001, about 0.005, about 0.01, about 0.05, about 0.1, about 0.5, about 1, about 5, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 120, about 140, about 160, about 180, about 200, about 220, about 240, about 260, about 280, about 300, about 320, about 340, about 360, about 380, about 400, about 420, about 440, about 460, about 480, about 500, about 520, about 540, about 560, about 580 or about 600 mg / ml.In some embodiments, the allergen-binding protein may be at a concentration of at most 0.001, at most 0.005, at most 0.01, at most 0.05, at most 0.1, at most 0.5, at most 1, at most 5, at most 10, at most 20, at most 30, at most 40, at most 50, at most 60, at most 70, at most 80, at most 90, at most 100, at most 120, at most 140, at most 160, at most 180, at most 200, at most 220, at most 240, at most 260, at most 280, no. maximum 300, maximum 320, maximum 340, maximum 360, maximum 380, maximum 400, maximum 420, maximum 440, maximum 460, maximum 480, maximum 500, maximum 520, no maximum 540, maximum 560, maximum 580 or maximum 600 mg / ml. In some embodiments, the allergen-binding protein may be at a concentration of at least 0.001, at least 0.005, at least 0.01, at least 0.05, at least 0.1, at least 0.5, at least 1, at least 5, at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 120, at least 140, at least 160, at least 180, at least 200, at least 220, at least 240, at least 260, at least 280, at least 300, at least 320, at least 340, at least 360, at least 380, at least 400, at least 420, at least 440, at least 460, at least 480, at least 500, by Petition 870250113096, dated 09 / 12 / 2025, page 68 / 183 61 / 119 minus 520, at least 540, at least 560, at least 580 or at least 600 mg / ml. In some embodiments, the allergen-binding protein may be at a concentration of approximately 0.001 to approximately 600, approximately 0.005 to approximately 580, approximately 0.01 to approximately 560, approximately 0.05 to approximately 540, approximately 0.1 to approximately 520, approximately 0.5 to approximately 500, approximately 1 to approximately 480, approximately 5 to approximately 460, approximately 10 to approximately 440, approximately 20 to approximately 420, approximately 30 to approximately 400, approximately 40 to approximately 380, approximately 50 to approximately 360, approximately 60 to approximately 340, approximately 70 to approximately 320, approximately 80 to approximately 300, approximately 90 to approximately 280, about 100 to about 260, about 120 to about 240, about 140 to about 220, about 160 to about 200, or about 180 to about 500 mg / ml.In some embodiments, the allergen-binding protein may be at a concentration of 0.001 to 600, 0.005 to 580, 0.01 to 560, 0.05 to 540, 0.1 to 520, 0.5 to 500, 1 to 480, 5 to 460, 10 to 440, 20 to 420, 30 to 400, 40 to 380, 50 to 360, 60 to 340, 70 to 320, 80 to 300, 90 to 280, 100 to 260, 120 to 240, 140 to 220, 160 to 200 or 180 to 500 mg / ml. In some embodiments, the allergen-binding protein may be at a concentration of 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580 or 600 mg / ml.

[268] In some embodiments, the allergen may induce an allergic reaction in a human. In some embodiments, the allergen may comprise an environmental allergen. In some embodiments, the allergen may comprise an animal allergen. In some embodiments, the allergen may comprise a pet allergen. In some embodiments, the allergen may comprise a cat, puppy, rabbit, mouse, or cockroach allergen. In some embodiments, the allergen may be a cat allergen. In some embodiments, the allergen may be an allergen. Petition 870250113096, dated 09 / 12 / 2025, p. 69 / 183 62 / 119 of dog. In some forms, the allergen may be a rabbit allergen. In some forms, the allergen may be a mouse allergen. In some forms, the allergen may be a cockroach allergen.

[269] In some forms, the allergen may comprise a dust allergen.

[270] In some embodiments, the allergen may comprise a plant allergen or plant pollen allergen. In some embodiments, the plant may comprise trees, grasses, or weeds. In some embodiments, the plant allergen or pollen allergen may be selected from the group consisting of Bet v1, Phl p 5, Phl p 1, Poa p 1, Cyn d 1, Bet v 2, Ole e 1, Amb a 1, Amb a 11, and Art v 1. In some embodiments, the plant allergen or pollen allergen may be Bet v1. In some embodiments, the plant or pollen allergen may be Phl p 5. In some embodiments, the plant or pollen allergen may be Phl p 1. In some embodiments, the plant or pollen allergen may be Poa p 1. In some embodiments, the plant or pollen allergen may be Cyn d 1. In some embodiments, the plant or pollen allergen may be Bet v 2. In some embodiments, the plant or pollen allergen may be Ole e 1.In some modalities, the plant or pollen allergen may be Amb a 1. In some modalities, the plant or pollen allergen may be Amb a 11. In some modalities, the plant or pollen allergen may be Art v 1.

[271] In some embodiments, the allergen comprises a mold allergen. In some embodiments, the mold allergen may be selected from the group consisting of Alt a 1, Asp f 1, Asp f 2, Cla h 8, Pen ch 13 and Pen ch 18. In some embodiments, the mold allergen may be Alt a 1. In some embodiments, the mold allergen may be Asp f 1. In some embodiments, the mold allergen may be Asp f 2. In some embodiments, the mold allergen may be Cla h 8. In some embodiments, the Petition 870250113096, dated 09 / 12 / 2025, p. 70 / 183 63 / 119 mold allergen may be Pen ch 13. In some forms, the mold allergen may be Pen ch 18.

[272] In some embodiments, the allergen comprises a food allergen. In some embodiments, the food allergen may be selected from the group consisting of Pen a 1, Ara h 1, Ara h 3. In some embodiments, the food allergen may be Pen a 1. In some modalities, the food allergen may be Ara h 1. In some modalities, the food allergen may be Ara h 1. In some modalities, the food allergen may be Ara h 3.

[273] In some forms, the allergen may include Fel d 1, Fel d 2, Fel d 3, Fel d 4, Can f1, Can f2, Can f4, Can f7, Der P1, Der P2, Ory C1, Mus M1, Bla G2, Bet v 1, Phl p 5 or a combination thereof. In some embodiments, the allergen may comprise Fel d 1. In some embodiments, the allergen may comprise Fel d 2. In some embodiments, the allergen may comprise Fel d 3. In some embodiments, the allergen may comprise Fel d 4. In some embodiments, the allergen may comprise Can f1. In some embodiments, the allergen may comprise Can f2. In some embodiments, the allergen may comprise Can f4. In some embodiments, the allergen may comprise Can f7. In some embodiments, the allergen may comprise Der P1. In some embodiments, the allergen may comprise Der P2. In some embodiments, the allergen may comprise Ory C1. In some embodiments, the allergen may comprise Mus M1. In some formulations, the allergen may comprise Bla G2. In some formulations, the allergen may comprise Bet v 1. In some formulations, the allergen may comprise Phl p 5. Composition

[274] In one particular aspect, this disclosure provides a composition comprising an allergen-binding protein disclosed herein. In some embodiments, the composition may be a Petition 870250113096, dated 09 / 12 / 2025, page 71 / 183 64 / 119 pharmaceutical composition. In some embodiments, the composition may additionally comprise a carrier. In some embodiments, the carrier may be a pharmaceutically acceptable carrier. The carrier may include, but is not limited to, a solvent, a stabilizing agent, a diluent, a dispersion medium, and a coating. In some embodiments, the carrier may comprise silica. In some embodiments, the composition may be in dry form.

[275] In some embodiments, the carrier may comprise water. In some embodiments, the composition may comprise a liquid. In some embodiments, the composition may comprise a buffer comprising a phosphate buffer or saline buffer. In some embodiments, the composition may comprise phosphate-buffered saline (PBS), citric acid buffer, carbonic acid-bicarbonate buffer, sodium hydrogen phosphate-citrate buffer solution, citric acid-sodium citrate buffer, sodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution, or acetic acid-sodium acetate buffer. In some embodiments, the pH of the composition may be approximately 2, approximately 3, approximately 4, approximately 5, approximately 6, approximately 7, approximately 8, approximately 9, approximately 10, or approximately 11. In some embodiments, the pH of the composition may be at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, or at least 11.In some embodiments, the pH of the composition may be a maximum of 5, a maximum of 6, a maximum of 7, a maximum of 8, a maximum of 9, a maximum of 10, or a maximum of 11. In some embodiments, the pH of the composition may be from about 2 to about 11, from about 3 to about 10, or from about 4 to about 9. In some embodiments, the pH of the composition may be from 2 to 11, from 3 to 10, or from 4 to 9. In some embodiments, the pH of the composition may be 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11.

[276] Non-limiting examples of the carrier may include Petition 870250113096, dated 09 / 12 / 2025, p. 72 / 183 65 / 119 water, dimethyl sulfoxide (DMSO), alpha-thujene, alpha-pinene, camphene, sabinene, beta-pinene, alpha-terpinene, limonene, peltay2-carene, trans-sabinene hydrate, terpinolene, 3-cyclohexen-1-ol, terpinene-4-ol, 1,2-benzenediol, linalyl acetate, borneol, bornyl acetate, alpha-thujone, terpinyl acetate, isolongifolene, epithi-bicyclosesquiphellandrene, alpha-humulene, guaiol, elemol, cedrol, beta-eudesmol, rosifoliol, rimuene, hexadecanoic acid, cembrene, verticelol, totarol, totara-1,9-octadecenamide, tatarol, 2-(hexylthiol)decanal and a combination thereof.

[277] In some embodiments, the composition may also contain a preservative. In some embodiments, the preservative may contain potassium sorbate. In some embodiments, the composition may contain one or more preservatives.Preservatives may include, but are not limited to, benzyl alcohol, methylparaben, ethylparaben, propylparaben, butylparaben, isobutylparaben, glycerin, ethylhexylglycerin, phenoxyethanol, sodium benzoate, ethylenediaminetetraacetic acid (EDTA), benzoic acid, phenoxyethanol, maltol, potassium sorbate, imidazolidinyl urea, diazolidinyl urea, sorbic acid, methylisothiazolinone, chlorhexidine digluconate, polyaminopropyl biguanide, sodium dehydroacetate, grapefruit seed extract, salicylic acid, DMDM ​​hydantoin, formaldehyde, chlorpheniramine, triclosan, dehydroacetic acid, quaternium-15, stearalkonium chloride, zinc pyrithione, sodium metabisulfite, 2-bromo-2-nitropropane, chloride of benzalkonium, sodium sulfite, sodium salicylate, citric acid, neem oil, essential oils, lactic acid, vitamin E (tocopherol), and a combination thereof.

[278] The composition disclosed herein may optionally comprise at least one excipient. The at least one excipient may be a pharmaceutically acceptable excipient. In some embodiments, the at least one excipient may be selected from Petition 870250113096, dated 09 / 12 / 2025, p. 73 / 183 66 / 119 group consisting of: animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, zinc oxide, lactose, talc, silicic acid, aluminum hydroxide, calcium silicates, polyamide powder and a combination thereof. In some embodiments, at least one excipient may be naturally occurring. In other embodiments, at least one excipient may be non-naturally occurring.

[279] The composition disclosed herein may optionally comprise at least one additive. Non-limiting examples of the at least one additive may include a fatty substance, an organic solvent, a solubilizing agent, a thickener, a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a flavoring, a surfactant, water, an ionic or non-ionic emulsifying agent, a filler, a sequestering agent, a chelating agent, a preservative, vitamins, a blocker, a moisturizing agent, an essential oil, a colorant, a pigment, a hydrophilic or hydrophobic activator, a lipid vesicle, antiseptics, stabilizing agents, moisturizing agents, emulsifying promoters or salts and / or buffers for osmotic control and a combination thereof. In some embodiments, the at least one additive may be naturally occurring. In other embodiments, the at least one additive may be non-naturally occurring.In some embodiments, at least one additive may comprise other useful substances. In additional embodiments, at least one additive may further comprise absorption enhancers, permeation enhancers, thickening agents, viscosity enhancers, pH adjusting and / or maintaining agents, osmotic pressure adjusting agents, preservatives, surfactants, buffers, salts, suspending agents, dispersing agents, solubilizing agents. Petition 870250113096, dated 09 / 12 / 2025, page 74 / 183 67 / 119 stabilizers and / or tonicity agents.

[280] In some embodiments, the composition may further comprise a stabilizing or thickening agent. In some embodiments, the stabilizing or thickening agent may comprise dextrin, maltodextrin, glycerol, glucose, sucrose or trehalose. In some embodiments, the composition may further comprise an isotonic agent. In some embodiments, the stabilizing agent may be glucose, sucrose, glycerol or trehalose.

[281] In some embodiments, the carrier may comprise maltodextrin, dextrin, potassium sorbate, silica, water or a combination thereof.

[282] In other embodiments, the composition of the present invention may comprise a pharmaceutically acceptable carrier. Non-limiting examples of pharmaceutically acceptable carriers may include a solvent, a dispersion medium, a coating, an adjuvant, a stabilizing agent, a diluent, a preservative, an antibacterial and antifungal agent, an isotonic agent, or a combination thereof. In some embodiments, the diluent may comprise water, saline solution, dextrose, ethanol, glycerol, or a combination thereof. In some embodiments, the isotonic agent may comprise sodium chloride, dextrose, mannitol, sorbitol, lactose, or a combination thereof.

[283] In some embodiments, the carrier may be at a concentration of, at most, about 80% by weight per volume (w / v, grams per milliliter of solvent) of the allergen-binding protein. In some embodiments, the carrier may be at a concentration of about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90 or about 95% by w / v of the composition. In some embodiments, the carrier may be at a concentration of, at most 50, at most 55, at most 60, at most 65, at most 70, at most Petition 870250113096, dated 09 / 12 / 2025, p. 75 / 183 68 / 119 75%, at most 80%, at most 85%, at most 90%, or at most 95% by w / v of the composition. In some embodiments, the carrier may be at a concentration of at least 0.01%, at least 0.05%, at least 0.1%, at least 0.5%, at least 1%, at least 2%, at least 5%, at least 10%, or at least 20% by w / v of the composition. In some embodiments, the carrier may be at a concentration of about 0.01% to about 95%, about 0.1% to about 90%, about 0.5% to about 85%, about 1% to about 80%, about 5% to about 75%, about 10% to about 70%, or about 20% to about 65% by w / v of the composition. In some formulations, the carrier may be present in a concentration of 0.01 to 95%, 0.1 to 90%, 0.5 to 85%, 1 to 80%, 5 to 75%, 10 to 70%, or 20 to 65% by w / v of the composition. In some formulations, the carrier may be present in a concentration of 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% by w / v of the composition.

[284] In some embodiments, the composition comprises one or more allergen-binding proteins disclosed herein. In some embodiments, the composition comprises two allergen-binding proteins disclosed herein. In some embodiments, the composition comprises three or more allergen-binding proteins disclosed herein.

[285] The composition may be formulated in any suitable physical form. Suitable forms may include, but are not limited to, aerosol, liquid, gel, semi-solid, solid or powder. In some embodiments, the composition may be formulated as a tonic, cream, emulsion, lotion, ointment, paste, gel, suspension, serum, oil, spray, shampoo, foam, cleanser, mousse, aerosol or powder to be resuspended in a solvent such as water.

[286] In some embodiments, the composition may be formulated to coat a feed. In some embodiments, the composition may be edible. In some embodiments, the composition may be mixed with any type of feed for an animal. In some embodiments, the composition may be provided as or added to the normal intake of feed. In some Petition 870250113096, dated 09 / 12 / 2025, p. 76 / 183 In some embodiments, the composition can be provided as a supplement to the animal's fluid intake, including its drinking water. In some embodiments, the composition may be formulated as a food product. In some embodiments, the food is a pet food. In some embodiments, the allergen is a pet allergen. In some embodiments, the allergen-binding protein is formulated to be applied to the food as a coating. In some embodiments, the allergen-binding protein is formulated to be mixed into the food.

[287] In some embodiments, the composition may be formulated for spraying, misting or brushing onto an animal or a household surface. In some embodiments, the composition may be formulated for a humidifier and an air filter. In some embodiments, the composition may be applied to a surface in the environment, for example, by spraying, misting, depositing, rubbing or other suitable method of application. In some embodiments, the surface may be the surface of an animal that is a source of the allergen.

[288] In some embodiments, the composition disclosed herein may reduce, minimize, or prevent at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the composition disclosed herein may reduce at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the composition disclosed herein may minimize at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the composition disclosed herein may prevent at least one symptom of an allergic response to an environmental or animal allergen. In some embodiments, the composition may come into contact with an environmental allergen, bind to the allergen, and prevent the allergen from inducing an allergic reaction in a susceptible individual or an individual suffering from allergies caused by the allergen. In some embodiments, the Petition 870250113096, dated 09 / 12 / 2025, p. 77 / 183 The 70 / 119 composition may come into contact with an environmental allergen. In some embodiments, the allergen-binding protein may bind to the allergen. Methods

[289] In certain aspects, the present disclosure provides a method for reducing, minimizing, or preventing at least one symptom of an allergic response to an environmental allergen. In some embodiments, the present disclosure provides a method for reducing at least one symptom of an allergic response to an environmental allergen. In some embodiments, the present disclosure provides a method for minimizing at least one symptom of an allergic response to an environmental allergen. In some embodiments, the present disclosure provides a method for preventing at least one symptom of an allergic response to an environmental allergen. The method may include neutralizing an allergen.Non-limiting examples of the symptoms of an allergic response may include congestion, itching of the nose or throat, sneezing, runny nose and itchy eyes, watery eyes, pain or tenderness around the cheeks, eyes or forehead, cough, wheezing or shortness of breath, itchy skin or raised rash (hives), feeling sick, swelling of the eyes, lips, mouth or throat, weak and rapid pulse, nausea, vomiting or diarrhea, dizziness or fainting, or a combination thereof. In some embodiments, the composition comprising the allergen-binding protein may come into contact with an environmental allergen, bind to the allergen and prevent the allergen from inducing an allergic reaction in the susceptible individual or in an individual suffering from allergies caused by the allergen. In some embodiments, the method comprises administering to an individual an allergen-binding protein, as disclosed herein.In some embodiments, the method involves administering to the individual a composition comprising an allergen-binding protein, as disclosed herein. Petition 870250113096, dated 09 / 12 / 2025, p. 78 / 183 71 / 119

[290] In some embodiments, the method may comprise spraying or nebulizing the composition disclosed herein or contacting the surface with any of the allergen-binding proteins or compositions disclosed herein after resuspending the allergen-binding protein or composition in a solvent disclosed herein. In some embodiments, the contact may comprise coating or brushing. In some embodiments, the surface may comprise the allergen. In some embodiments, the surface may comprise the surface of an air filter or a humidifier. In some embodiments, the surface may comprise food. In some embodiments, the food may be consumed by an animal that is the source of the antigen.

[291] In some forms, the composition can neutralize the allergen on the surface.

[292] In some embodiments, the surface may comprise a pet accessory (for example, a collar or brush), an air filter or an area / product where an animal can sit or walk (for example, a dog bed).

[293] In some forms, nebulization can be carried out by a humidifier or a sprayer.

[294] In some embodiments, about 0.01 milliliter (ml) to about 20 ml of the sprayed composition per about 1 square meter of area can neutralize the allergenicity of the antigen. In some modalities, the neutralization of the antigen's allergenicity can be achieved by approximately 0.001, approximately 0.005, approximately 0.01, approximately 0.05, approximately 0.1, approximately 0.5, approximately 1, approximately 2, approximately 3, approximately 4, approximately 5, approximately 6, approximately 7, approximately 8, approximately 9, approximately 10, approximately 11, approximately 12, approximately 13, approximately 14, approximately 15, approximately 16, approximately 17, approximately 18, approximately 19, approximately 20, approximately 21, approximately 22, approximately 23, approximately 24, approximately 25, approximately 26, approximately 27, approximately 28, approximately 29, or approximately 30 ml of the solution. Petition 870250113096, dated 09 / 12 / 2025, page 79 / 183 72 / 119 composition sprayed over an area of ​​approximately 1 square meter. In some formulations, the neutralization of the antigen's allergenicity can be achieved by a maximum of 0.001, a maximum of 0.005, a maximum of 0.01, a maximum of 0.05, a maximum of 0.1, a maximum of 0.5, a maximum of 1, a maximum of 2, a maximum of 3, a maximum of 4, a maximum of 5, a maximum of 6, a maximum of 7, a maximum of 8, a maximum of 9, a maximum of 10, a maximum of 11, a maximum of 12, a maximum of 13, a maximum of 14, a maximum of 15, a maximum of 16, a maximum of 17, a maximum of 18, a maximum of 19, a maximum of 20, a maximum of 21, a maximum of 22, a maximum of 23, a maximum of 24, a maximum of 25, a maximum of 26, a maximum of 27, a maximum of 28, a maximum of 29, or a maximum of 30 ml of the composition sprayed for approximately... 1 square meter in area.In some embodiments, the neutralization of the antigen's allergenicity can be achieved by at least 0.001, at least 0.005, at least 0.01, at least 0.05, at least 0.1, at least 0.5, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29 or at least 30 ml of the sprayed composition per approximately 1 square meter in area.In some embodiments, the neutralization of the antigen's allergenicity can be achieved by approximately 0.001 to approximately 30 ml, approximately 0.005 to approximately 29 ml, approximately 0.01 to approximately 28 ml, approximately 0.05 to approximately 27 ml, approximately 0.1 to approximately 26 ml, approximately 0.5 to approximately 25 ml, approximately 1 to approximately 24 ml, approximately 2 to approximately 23 ml, approximately 3 to approximately 22 ml, approximately 4 to approximately 21 ml, approximately 5 to approximately 20 ml, approximately 6 to approximately 19 ml, approximately 7 to approximately 18 ml, approximately 8 to approximately 17 ml, approximately 9 to approximately 16 ml, approximately 10 to approximately 15 ml, approximately 11 to approximately 14 ml, or approximately 12 to approximately 13 ml of the sprayed composition per approximately 12 ml. 1 square meter of area. In some. Petition 870250113096, dated 09 / 12 / 2025, p. 80 / 183 73 / 119 modalities, the neutralization of the antigen's allergenicity can be achieved by approximately 0.001 to 30, 0.005 to 29, 0.01 to 28, 0.05 to 27, 0.1 to 26, 0.5 to 25, 1 to 24, 2 to 23, 3 to 22, 4 to 21, 5 to 20, 6 to 19, 7 to 18, 8 to 17, 9 to 16, 10 to 15, 11 to 14 or 12 to 13 ml of the sprayed composition per approximately 1 square meter of area. In some formulations, the neutralization of the antigen's allergenicity can be achieved by spraying 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 ml of the composition per square meter of area.

[295] In certain aspects, the present disclosure provides a method for preparing the allergen-binding protein disclosed herein. In some embodiments, the method may comprise collecting the allergen-binding protein from an engineered microbe or from a secretion of the microbe. In some embodiments, the microbe may comprise a heterologous nucleic acid encoding the allergen-binding protein. In some embodiments, the microbe may comprise a yeast or bacterium. In some embodiments, the microbe may comprise a bacterium. In some embodiments, the microbe may comprise a bacterium comprising E. coli. In some embodiments, the microbe may comprise a yeast comprising Pichia pastoris. In some embodiments, the allergen-binding protein may be purified or concentrated from a secretion of the microbe.In some embodiments, the allergen-binding protein can be secreted by the microbe via a secretion tag and subsequently purified. In some embodiments, the allergen-binding protein can be purified by affinity purification. In some embodiments, the allergen-binding protein can be purified by hyperfiltration. In some embodiments, the purification or concentration comprises... Petition 870250113096, dated 09 / 12 / 2025, page 81 / 183 74 / 119 a filtration step. In some embodiments, the filtration step comprises the passage of the secretion through a filter with dimensions of approximately 0.01 nm, 0.1 nm, 1 nm, 5 nm or more. In some embodiments, the filter has dimensions of approximately 0.0001 nm, approximately 0.0005 nm, approximately 0.001 nm, approximately 0.005 nm, approximately 0.01 nm, approximately 0.05 nm, approximately 0.1 nm, approximately 0.5 nm, approximately 1 nm, approximately 2 nm, approximately 3 nm, approximately 4 nm, approximately 5 nm, approximately 6 nm, approximately 7 nm, approximately 8 nm, approximately 9 nm, approximately 10 nm or more in size. In some configurations, the filter has a maximum size of 0.0001 nm, a maximum size of 0.0005 nm, a maximum size of 0.001 nm, a maximum size of 0.005 nm, a maximum size of 0.001 nm, a maximum size of 0.05 nm, a maximum size of 0.1 nm, a maximum size of 0.5 nm, a maximum size of 1 nm, a maximum size of 2 nm, a maximum size of 3 nm, a maximum size of 4 nm, a maximum size of 5 nm, a maximum size of 6 nm, a maximum size of 7 nm, a maximum size of 8 nm, a maximum size of 9 nm, a maximum size of 10 nm or more.In some embodiments, the filter has at least 0.0001 nm, at least 0.0005 nm, at least 0.001 nm, at least 0.005 nm, at least 0.01 nm, at least 0.05 nm, at least 0.1 nm, nm, at least 0.5 nm, at least 1 nm, at least 2 nm, at least 3 nm, at least 4 nm, at least 5 nm, at least 6 nm, at least 7 nm, at least 8 nm, at least 9 nm, at least 10 nm or more in size. In some embodiments, the filter size may be between two values ​​described above, for example, between about 0.0001 nm to about 10 nm, about 0.0005 nm to about 9 nm, about 0.001 nm to about 8 nm, about 0.005 nm to about 7 nm, about 0.01 nm to about 6 nm, about 0.05 nm to about 5 nm, about 0.1 nm to about 4 nm, about 0.5 nm to about 3 nm, about 1 nm to about 2 nm.In some embodiments, the filter size may be 0.0001 nm, 0.0005 nm, 0.001 nm, 0.005 nm, 0.01 nm, 0.05 nm, 0.1 nm, nm, 0.5 nm, 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, or 10 nm. In some embodiments, the method does not include a centrifugation step.

[296] In some modalities, the method may include Petition 870250113096, dated 09 / 12 / 2025, p. 82 / 183 75 / 119 additionally the incorporation of the heterologous nucleic acid encoding the allergen-binding protein into a cell-free protein expression system. In some embodiments, the cell-free protein expression system comprises ribosomes.

[297] A method for treating an allergy in an individual is also provided herein. In some embodiments, the method comprises administering to the individual a therapeutically effective amount of a microbe designed to produce any of the allergen-binding proteins disclosed herein. In some embodiments, the method comprises administering to the individual a therapeutically effective amount of a microbe designed to produce any of the compositions comprising an allergen-binding protein disclosed herein. In some embodiments, the microbe may be a yeast. In some embodiments, the microbe may be a bacterium. In some embodiments, the microbe may comprise a bacterium. In some embodiments, the microbe may comprise a bacterium comprising E. coli. In some embodiments, the microbe may comprise a yeast comprising Pichia pastoris.In some modalities, the microbe can be administered to the individual orally. In some modalities, the microbe can be ingested. In some modalities, the microbe produces the allergen-binding protein or compound when ingested.

[298] A method for neutralizing an allergen is also provided herein. In some embodiments, the method comprises aerosolizing any of the allergen-binding proteins described herein into a mist and contacting the mist with a surface containing an allergen. In some embodiments, the method comprises aerosolizing any of the compositions containing an allergen-binding protein described herein into a mist and contacting the mist with a surface containing an allergen. In some embodiments, the Petition 870250113096, dated 09 / 12 / 2025, p. 83 / 183 76 / 119 Aerosolization can be performed by an aerosolization machine. In some embodiments, the aerosolization machine can be used by an individual. In some embodiments, the surface comprises an accessory for pets. In some embodiments, the surface comprises an area / product where an animal can sit or walk.

[299] A method for neutralizing an allergen is also provided herein. In some embodiments, the method comprises aerosolizing any of the allergen-binding proteins described herein into a mist and contacting the mist with a food. In other embodiments, the method comprises aerosolizing any of the compositions comprising an allergen-binding protein described herein into a mist and contacting the mist with a food. In some embodiments, the food is a pet food. In some embodiments, the allergen is a pet allergen. In some embodiments, the mist is applied over the food as a coating. In some embodiments, the mist is mixed into the food.

[300] A method for neutralizing an allergen is also provided herein. In some embodiments, the method comprises contacting any of the allergen-binding proteins described herein with a food. In some embodiments, the method comprises contacting any of the compositions comprising an allergen-binding protein described herein with a food. In some embodiments, the food is pet food. In some embodiments, the allergen is a pet allergen. In some embodiments, the allergen-binding protein is applied to the food as a coating. In some embodiments, the allergen-binding protein is mixed into the food. In some embodiments, the composition is applied to the food as a coating. In some embodiments, the composition is mixed into the food. Petition 870250113096, dated 09 / 12 / 2025, p. 84 / 183 77 / 119

[301] A method for treating an allergy in an individual in need thereof is also provided herein. In some embodiments, the method comprises aerosolizing any of the allergen-binding proteins described herein into a mist. In some embodiments, the method comprises aerosolizing any of the compositions comprising an allergen-binding protein described herein into a mist. In some embodiments, the aerosolization may be performed by an aerosolizing machine. In some embodiments, the aerosolizing machine may be used by an individual. In some embodiments, the method further comprises inhalation of the mist by the individual. In some embodiments, the individual may be a human being. In some embodiments, the individual may be a non-human animal.

[302] A method for identifying allergen-binding proteins is also provided herein. In some embodiments, the method comprises transfecting target cells with the human Fc epsilon I receptor (FceR1). In some embodiments, the method comprises contacting target cells with an IgE isolated from an individual allergic to pets. In some embodiments, the method comprises contacting target cells with a candidate allergen-binding protein and a pet allergen. In some embodiments, the individual allergic to pets is an individual allergic to cats. In some embodiments, the pet allergen is a cat allergen. In some embodiments, the cat allergen is FelD1. In some embodiments, the method comprises measuring histamine release by target cells. In some embodiments, the method comprises measuring betahexosaminidase release by target cells. Kits

[303] Kits for the use of the compositions described here are revealed here in some forms. In some forms, the Petition 870250113096, dated 09 / 12 / 2025, page 85 / 183 78 / 119 kits disclosed here can be used to reduce, minimize, or prevent at least one symptom of an allergic response to an environmental allergen. In some embodiments, the kits may comprise a set of materials or components in addition to the composition. In some embodiments, the kits may require additional external materials, including water.

[304] Instructions for use may be included in the kit. In some embodiments, the kit may include instructions for administering the composition to an individual who needs it. In some embodiments, the kit may include instructions for measuring the viability of the restored compositions in order to ensure efficacy for the intended purpose (e.g., therapeutic efficacy if used to treat an individual).

[305] Optionally, the kit may also include other useful components, such as diluents, buffers, pharmaceutically acceptable carriers, syringes, catheters, applicators or measuring instruments, or other useful paraphernalia. The materials or components assembled in the kit may be provided to the professional, stored in any convenient and appropriate manner that preserves their operability and usefulness. For example, the components may be dissolved, dehydrated or lyophilized; they may be provided at room temperature, refrigerated or frozen. The components may typically be packaged in suitable packaging material(s). Definitions

[306] Unless otherwise defined, all technical terms, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by someone skilled in the subject matter to which the claimed subject matter relates. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions Petition 870250113096, dated 09 / 12 / 2025, p. 86 / 183 79 / 119 here should not necessarily be interpreted as representing a substantial difference from what is generally understood in the art.

[307] Throughout this application, various forms may be presented in range format. It should be understood that the range description is merely for convenience and brevity and should not be interpreted as an inflexible limitation on the scope of disclosure. Consequently, the description of a range should be considered as having specifically disclosed all possible sub-ranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered as having specifically disclosed sub-ranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the range width.

[308] As used in the descriptive report and claims, the singular forms a, an and the include plural references, unless the context clearly indicates otherwise. For example, the term a sample includes a plurality of samples, including mixtures thereof.

[309] Whenever the term at least, greater than, or greater than or equal to precedes the first numerical value in a series of two or more numerical values, the term at least, greater than, or greater than or equal to applies to each of the numerical values ​​in that series of numerical values. For example, greater than or equal to 1, 2, or 3 is equivalent to greater than or equal to 1, greater than or equal to 2, or greater than or equal to 3.

[310] Whenever the term not more than, less than, or less than or equal to precedes the first numerical value in a series of two or more numerical values, the term not more than, less than, or less than or equal to applies to each of the values Petition 870250113096, dated 09 / 12 / 2025, p. 87 / 183 80 / 119 are numerical values ​​in this series. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1.

[311] The expressions “at least one of A and B” and “at least one of A or B” can be interpreted as meaning at least A, at least B, or at least A and B (i.e., a set comprising A and B, this set possibly including one or more additional elements). The term “A and / or B” can be interpreted as meaning only A, only B, or both A and B.

[312] The expressions “at least about A, B and C” and “at least about A, B or C” can be interpreted as meaning at least about A, at least about B or at least about C. The expressions “at most about A, B and C” and “at most about A, B or C” can be interpreted as meaning at most about A, at most about B or at most about C.

[313] The expression “between approximately A and B, C and D, and E and F” can be interpreted as meaning between approximately A and approximately B, between approximately C and approximately D, and between approximately E and approximately F. The expression “between approximately A and B, C and D, or E and F” can be interpreted as meaning between approximately A and approximately B, between approximately C and approximately D, or between approximately E and approximately F.

[314] The expression “about A to B and C to D” can be interpreted as meaning between about A and about B and between about C and about D. The expression “about A to B or C to D” can be interpreted as meaning between about A and about B or between about C and about D.

[315] The term “exemplary,” as used herein, means “to serve as an example, instance, or illustration.” Any modality described herein as “exemplary” must not be Petition 870250113096, dated 09 / 12 / 2025, p. 88 / 183 81 / 119 interpreted as preferential or advantageous in relation to other modalities.

[316] The terms determine, measure, assess, estimate, test, and analyze are frequently used interchangeably in this document to refer to forms of measurement. The terms include determining whether an element is present or not (e.g., detection). These terms may include quantitative, qualitative, or quantitative and qualitative determinations. Assessing may be relative or absolute. Detecting the presence of may include determining the quantity of something present, as well as determining whether it is present or absent, depending on the context.

[317] The terms subject, individual, or patient are frequently used interchangeably in this document. A subject may be a biological entity containing expressed genetic material. The biological entity may be a plant, an animal, or a microorganism, including, for example, bacteria, viruses, fungi, and protozoa. The individual may be tissues, cells, and their progeny from a biological entity obtained in vivo or cultured in vitro. The individual may be a mammal. The mammal may be a human being. The individual may be diagnosed with or suspected of being at high risk for a disease. In some cases, the individual is not necessarily diagnosed with or suspected of being at high risk for the disease.

[318] The term pharmaceutically acceptable refers to a product approved or approvable by a federal or state government regulatory agency or listed in the U.S. Pharmacopoeia (USP) or other generally recognized pharmacopoeia for use in animals, including humans. A pharmaceutically acceptable excipient refers to an excipient that can be administered to an individual along with an active fraction (i.e., an allergen-binding protein) and that does not destroy its pharmacological activity and is nontoxic when administered in doses sufficient to provide a therapeutic amount of the fraction. Petition 870250113096, dated 09 / 12 / 2025, page 89 / 183 82 / 119 The term pharmaceutically acceptable carrier refers to any non-toxic substance that can be safely administered to a patient and does not interfere with the effectiveness of the biological activity of the active ingredients. The term pharmaceutically acceptable carrier may refer to a biologically compatible carrier, which is a substance that does not interfere with the binding capacity of an allergen-binding protein and is non-toxic for human or animal use. The carrier may be solid, liquid, or gaseous and assists in delivering the active ingredient of the drug to the target area of ​​the body. The pharmaceutically acceptable carrier may be chosen based on its compatibility with the active ingredient(s) and its suitability for the route of administration. It may include substances that serve to stabilize, solubilize, emulsify, suspend, or otherwise facilitate the functional dispersion of the active ingredient(s) in the pharmaceutical composition.Examples include, but are not limited to, binders, fillers, diluents, solvents, buffers, preservatives, surfactants, and the like, which are commonly used in pharmaceutical formulation techniques.

[319] The term therapeutically effective amount refers to the amount of a compound, composition, or pharmaceutical agent that, when administered to an individual or patient, is sufficient to produce a beneficial therapeutic response over time. This response may include, but is not limited to, the relief of one or more symptoms, the modification or halting of disease progression, or the complete elimination of the pathological condition. A therapeutically effective amount may vary based on several factors, such as the specific condition being treated, the specific compound, composition, or pharmaceutical agent being used, the severity of the condition, the patient's age and weight, and the route of administration. Determining a therapeutically effective amount is within the capacity Petition 870250113096, dated 09 / 12 / 2025, page 90 / 183 83 / 119 from a technician in the field, frequently employing a dosage administration regimen adjusted over time.

[320] The term “in vivo” is used to describe an event that occurs in an individual’s body.

[321] The term “ex vivo” is used to describe an event that occurs outside the body of an individual. An ex vivo assay is not performed on an individual. Instead, it is performed on a separate sample from the individual. An example of an ex vivo assay performed on a sample is an “in vitro” assay.

[322] The term in vitro is used to describe an event that occurs within a container for storing laboratory reagents, in such a way that it is separated from the biological source from which the material is obtained. In vitro assays can encompass cell-based assays, in which live or dead cells are used. In vitro assays can also encompass a cell-free assay, in which no intact cells are used.

[323] As used herein, the term about a number refers to that number plus or minus 10% of that number. The term about a range refers to that range minus 10% of its smallest value and plus 10% of its largest value.

[324] As used herein, the terms treatment or treat are used in reference to a pharmaceutical regimen or other intervention regimen to achieve beneficial or desired results in the recipient. Beneficial or desired results include, but are not limited to, a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to the eradication or improvement of symptoms or of an underlying disorder being treated. In addition, a therapeutic benefit may be achieved by eradicating or improving one or more physiological symptoms associated with the underlying disorder, such that an improvement is observed in the individual, notwithstanding the fact that the individual may still be affected by the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the Petition 870250113096, dated 09 / 12 / 2025, page 91 / 183 84 / 119 appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, stopping or reversing the progression of a disease or condition, or any combination thereof. For prophylactic benefit, an individual at risk of developing a specific disease, or an individual reporting one or more physiological symptoms of a disease, may undergo treatment even if a diagnosis of that disease has not been made.

[325] The term antibody may include fully assembled antibodies, antibody fragments that can bind to antigens, for example, Fab, F(ab')2, Fv, single-chain antibodies (scFv), diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, camelid single VHH domains (also known as nanobodies) and the like.

[326] The term complementarity-determining region or CDR is a segment of the variable region of an antibody or allergen-binding protein that is complementary in structure to the epitope to which the antibody binds and is more variable than the rest of the variable region. Consequently, a CDR is sometimes referred to as a hypervariable region. A variable region comprises three CDRs. CDR peptides can be obtained by constructing genes that encode the CDR of an antibody of interest. Such genes are prepared, for example, using the polymerase chain reaction to synthesize the variable region from the RNA of antibody-producing cells. See, for example, Larrick et al., Methods: A Companion to Methods in Enzymology 2: 106 (1991); Courtenay-Luck, Genetic Manipulation of Monoclonal Antibodies, in Monoclonal Antibodies: Production, Engineering and Clinical Application, Ritter et al. (eds.), pages 166-179 (Cambridge University Press 1995); and Ward et al., Genetic Manipulation and Expression of Antibodies, in Monoclonal Antibodies: Principles and Applications, Birch et al., (eds.), pages 137-185 (Wiley-Liss, Inc. 1995). Petition 870250113096, dated 09 / 12 / 2025, page 92 / 183 85 / 119

[327] The term Fab refers to a protein containing the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab fragments differ from Fab' fragments by the addition of some residues at the carboxy-terminal of the CH1 domain of the heavy chain, including one or more cysteines from the hinge region of the antibody. Fab'-SH is the designation used here for Fab' in which the cysteine ​​residue(s) of the constant domains contain a free thiol group. Fab' fragments are produced by reduction of the disulfide bridge of the heavy chain of the F(ab')2 fragment. Other chemical couplings of antibody fragments are also known.

[328] A single-chain variable fragment (scFv) is a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of an antibody, connected to a short linker peptide of ten to about 25 amino acids. The linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility, and may connect the N-terminus of the VH to the C-terminus of the VL, or vice versa. This protein retains the specificity of the original antibody despite the removal of the constant regions and the introduction of the linker. scFv antibodies are, for example, described in Houston, JS, Methods in Enzymol. 203 (1991) 46-96.Furthermore, antibody fragments comprise single-chain polypeptides with the characteristics of a VH domain, i.e., being able to assemble with a VL domain, or a VL domain, i.e., being able to assemble with a VH domain at a functional antigen-binding site, thus providing the antigen-binding property of full-length antibodies.

[329] A nanobody is a single-domain antibody or part of a single-domain antibody, usually derived from alpacas, llamas, or other camelids. The nanobody may refer to a single variable domain in an antibody heavy chain (VHH domain). A nanobody can be modified to increase the Petition 870250113096, dated 09 / 12 / 2025, page 93 / 183 86 / 119 solubility, stability, efficacy or any other important characteristics.

[330] A monobody is a synthetic binding protein derived from mutation of the fibronectin type III (FN3) domain. The binding interface for the FN3 domain is mutated in a monobody to develop protein linkers that bind to a protein of interest.

[331] A DARPin is a genetically engineered antibody mimetic protein that stands for engineered ankyrin repeat proteins.

[332] As used herein, the terms polypeptide, peptide, and protein may be used interchangeably in reference to a polymer of amino acid residues. A protein may refer to a full-length polypeptide, translated from an open reading frame of coding, or processed to its mature form, while a polypeptide or peptide may refer to a degradation or processing fragment of a protein that nevertheless uniquely or identifiably maps to a specific protein. A polypeptide may be a single linear polymeric chain of amino acids linked together by peptide bonds between the carboxyl and amino groups of adjacent amino acid residues. Polypeptides may be modified, for example, by the addition of carbohydrate, phosphorylation, etc.

[333] As used herein, the terms fragment or portion or equivalent terms may refer to a portion of a protein that is less than the total length of the protein and optionally retains the function of the protein.

[334] The term allergen refers to any natural protein or mixture of proteins that induce allergic reactions, that is, IgE-mediated reactions, after repeated exposure to an individual.

[335] The section titles used here are for informational purposes only. Petition 870250113096, dated 09 / 12 / 2025, page 94 / 183 87 / 119 organizational and should not be interpreted as limiting the subject matter described. Examples

[336] The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention. Example 1: ELISA binding assay for anti-Fel D1 nanobodies

[337] 2 micrograms per milliliter (g / mL) of target antigen are coated in a 96-well plate and the ELISA assay is performed after incubation with candidate VHHs. The binding EC50 of each clone is calculated using Prism GraphPad. The positive control antibody acts as an internal control. Example 2: Functional Blocking Assay of Fel D1

[338] Human FceRl (Fc epsilon RI) is stably transfected into RBL-2H3 cells and the resulting stable cells, human IgE from the cat-allergic population, cat saliva, positive control antibody and candidate VHH are used to assess the function of the candidate VHH. The EC50 of each clone is calculated using Prism GraphPad. The functional assay is schematically illustrated in Figure 1. Example 3: Using the Composition to Relieve Allergy Symptoms

[339] A person allergic to cat or dog hair obtains the composition disclosed here with nanobodies that neutralize pet allergens. The composition is mixed with water in a bottle with an attached sprayer. The water can be tap water. The composition is well suspended in the water by shaking the bottle. The composition suspended in the water is sprayed into the air, around the house and on various household surfaces, including pet beds, personal bedding, air filters, sofa, pillows, walls or floors. The person finds that the composition relieves the allergy symptoms. Example 4: Identification of VHH anti-Der p2

[340] To obtain a unique variable domain in the heavy chain, Petition 870250113096, dated 09 / 12 / 2025, page 95 / 183 88 / 119, i.e., VHH, specific for the dust mite allergen Der p2, a recombinant Der p2 labeled with His was prepared, which achieved high purity, as shown via Western blotting SDS-Page in Figure 2. To confirm this, an ELISA was performed. Briefly, Der p2-HIS was coated onto an ELISA plate and a known Der p2 antibody (clone 6OY4) was added. A secondary detection antibody capable of binding to the Der p2 antibody was then added. The secondary detection antibody was conjugated to a horseradish peroxidase, which can be used to produce a colorimetric reaction. Then, the optical density was measured. As summarized in Table 1, the recombinant Der p2-His was effectively bound by the antibody. Table 1: Der p2-His Binding ELISA Assay Recombinant Concentration of Der p2-His ^g / mL) Replication 1 (OD at 450 nm) Replication 2 (OD at 450 nm) 10 4.61 4.57 2 4.06 4.02 0.4 1.31 1.37 0.08 0.38 0.35 0.016 0.11 0.12 0.0032 0.09 0.10 0.00064 0.07 0.09 Blank 0.11 0.06

[341] Recombinant Der p2 was then administered to two alpacas in four rounds. The alpacas received 250 μg of recombinant Der p2 on days 0, 14, 28, and 49. An ELISA using titrated blood serum was performed to detect anti-Der p2 HHV levels. The results are summarized in Table 2. Each measurement was performed in duplicate. Negative serum was collected from uninfected alpacas to be used as a negative control. Table 2. Serum Titration of Alpaca Immunized with Der p2 Alpaca 1 Petition 870250113096, dated 09 / 12 / 2025, page 96 / 183 89 / 119 Negative Serum Titration OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1:1K 0.246 0.255 3.574 3.468 3.697 3.739 1:2K 0.149 0.154 3.486 3.365 3.679 3.623 1:4K 0.119 0.116 3.126 3.020 3.215 3.231 1:8K 0.104 0.099 2.341 2.308 2.460 2.383 1:16K 0.099 0.093 1.484 1.521 1.641 1.601 1:32k 0.099 0.081 0.929 0.887 1.045 1.063 1:64K 0.085 0.081 0.505 0.491 0.589 0.648 PBS 0.124 0.089 0.095 0.084 0.091 0.100 Alpaca 2 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1:1K 0.201 0.190 0.474 0.509 3.628 3.739 1:2K 0.152 0.150 0.297 0.310 2.916 3.120 1:4k 0.113 0.113 0.203 0.201 2.198 2.230 1:8K 0.101 0.103 0.151 0.147 1.480 1.452 1:16K 0.091 0.089 0.103 0.107 0.927 0.899 1:32k 0.082 0.082 0.105 0.107 0.603 0.554 1:64K 0.071 0.079 0.090 0.087 0.359 0.327 PBS 0.093 0.079 0.095 0.091 0.105 0.084

[342] After the third immunization, peripheral blood mononuclear cells (PBMCs) were obtained from the immunized alpacas, RNA was extracted from the PBMCs, and the RNA was reverse transcribed into cDNA. VHH sequences were then amplified from the cDNA using single-domain antibody cloning primer combinations and subcloned into the pYDisplay yeast display vector, which was electrotransformed into competent EYB100 cells to construct single-domain antibody yeast display libraries.

[343] The sequences of 50 randomly selected clones Petition 870250113096, dated 09 / 12 / 2025, page 97 / 183 90 / 119 are shown in Figure 3A, which demonstrated low overlap, indicating a diverse library. Individual clones were subjected to a flow cytometry-based assay, in which biotinylated recombinant Der p2 was incubated with the VHH clone and, after a washing step, the detection of bound Der p2 identified the clone as a VHH candidate. The VHH candidates were then constructed into prokaryotic expression vectors.

[344] The isolated VHH clones were subsequently validated by performing an ELISA assay as described in Example 1. Thermostability metrics including melting temperature (Tm), onset temperature (Tinitial), and aggregation temperature (Tagg) were also tested with increasing temperature in the range of 20°C - 95°C at a rate of 1°C / min. These measurements were performed using a Nanotemper DSF. Where applicable, Tm1 refers to the first observed melting point, Tm2 refers to the second observed melting point, etc. Sequence information for the VHH clones and the linkage assay results are summarized in Table 3. Individual linkage curves are shown for each candidate in Figures 3B-3I. Each clone demonstrated strong linkage, indicated by low EC50 values. Table 3. Validation of VHH anti-Der p2 Clone SEQ ID NO: Sequência EC50 (pg / mL) Tagg Tm SDAB221 11-1-D3 1 QLQLVESGGGLVQPGGSLRLSCVVSGRTL SGHNMGWFRQAPGKEREFVARINSNGGTT RYADAVKGRFTISRDNAKNTAYLQMNSLK SEDTAVYYCRLVGPAGYWGQGTQVTVSS 8,407 42,45 Tm1: 53,78 Tm2 : 69,76 Petition 870250113096, on 12 / 09 / 2025, page. 98 / 183 91 / 119 SDAB221 11-1-D6 2 DVQLVESGGGLVQPGGSLRLSCVASGLTF SYYDMKWHRQAPGKERE LVAAIIE GGRTM YADSVKGRFTISRDSPKKSVYLQMNNLKS EDTAVYYCNAVRARFRGLGNDDAWGQGTQ VTVSS 9, 656 41,57 Tm1: 49, 02 Tm2 : 62,55 SDAB221 11-1-C1 3 QVQLVESGGGLVQPGGSLRLSCAASASFV GAYAMYWFRQATGKQRELVAFITSGNSTN YADSVKGRFTISRDYTKNTVYLQMNKLKP EDTAVYSCNLVTSRGDYWGQGTQVTVSS 1, 114 43,53 Tm1: 54,52 Tm2 : 66, 51 SDAB221 11-1- F9-B03 4 EVQLVESGGGLVQPGGSLRLSCAASGRTF SSYAMGWFRQAPGKEREFVAGISRRGSNI YYADSVKGRFTISRDNAKNTVYLQMNSLK PEDTAVYYCNARHPRISTRLYWGQGTQVT VSS 15, 94 40, 04 71,38 SDAB221 11-1- H12 5 QVQLVESGGGLVQPGGSLRLSCAASRSID RFPAMGWYRQAPGKKRELVAAISSANTRT KYADSVKGRFTISRDNAENTMYLQMNSLK PEDTAVYYCNVFPRPVQGYWGQGTQVTVS S 2,668 43,37 Tm1: 51, 93 Tm2 : 59, 13 Tm3 : 68,62 SDAB221 11-1-F9 6 QVQLVESGGGLVQPGGSLRLSCAASGRIF RPDGMAWYRQAPGKQREFVAGITRGGRTT YADSVKGRFTISRDNAENTVYLQMNSLKP EDTAVYYCNAKPSAWTLPRYRYDYWGQGT QVTVSS 4,953 SDAB221 11-1-B1 7 DVQLVESGGGLVQPGGSLRLSCAASGFTF ALYSMRWYRQAPGKERELVAAITSGRSTN YADSVKGRFTISRDNAKNTVYLQMNSLKP EDTAVYHCNARHPIATLGNYWGQGTPVTV SS 8.231, Petition 870250113096, dated 09 / 12 / 2025, p. 99 / 183 92 / 119 SDAB221 11-1- E10 8 QLQLVESGGGMVQPGGSLRLSCAASGTLF STALMGWYRQAPGKQRTLVASITDGGSTN YVDSVKGRFTISRDNEKNTVYLQMNSLKF EDTAVYYCNAHWGSYDYWGQGTQVVVSS 6, 569 Example 5: Identification of VHH anti-Can f1

[345] To obtain a unique variable domain in the heavy chain, i.e., VHH, specific for the canine allergen Can f1, a recombinant Can f1 labeled with His was prepared, which achieved high purity, as shown by Western blotting SDS-Page in Figure 4. To confirm this, an ELISA was performed measuring the optical density when recombinant Can f1-His was incubated with a biotinylated anti-His antibody. As summarized in Table 4, the recombinant Can f1-His was effectively bound to the antibody. Table 4: Can f1-His Binding ELISA Assay Recombinant Concentration of Can f1-His ^g / mL) Replicate 1 (OD at 450 nm) Replicate 2 (OD at 450 nm) 10 2.84 2.92 2 0.95 1.03 0.4 0.21 0.24 0.08 0.15 0.16 0.016 0.09 0.12 0.0032 0.09 0.11 0.00064 0.07 0.11 Blank 0.09 0.08

[346] Recombinant Can f1 was then administered to two alpacas in four rounds. The alpacas received 250 μg of recombinant Can f1 on days 0, 14, 28, and 49. An ELISA using titrated blood serum was performed to detect anti-Can f1 VHH levels. The results are summarized in Table 5. Each measurement was performed in duplicate. Negative serum was collected Petition 870250113096, dated 09 / 12 / 2025, pages 100 / 183 93 / 119 of uninfected alpacas to be used as a negative control. Table 5. Serum Titration of Alpaca Immunized with Can fl Alpaca 1 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1:1K 0.294 0.342 3.972 4.008 4.040 4.079 1:2K 0.182 0.198 3.917 3.911 4.055 4.088 1:4K 0.137 0.147 3.505 3.495 3.962 3.989 1:8K 0.116 0.117 2.724 2.799 3.873 3.775 1:16K 0. 101 0, 110 1,925 2,018 3,329 3,249 1: 32k 0, 102 0, 107 1,243 1,279 2, 684 2, 606 1: 64K 0, 103 0, 103 0,765 0,737 1, 881 1, 812 PBS 0, 139 0, 111 0, 106 0, 108 0, 105 0, 106 Alpaca 2 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1: 1K 0,228 0,218 3,521 3.597 4.285 4.157 1:2K 0.185 0.179 3.522 3.437 4.112 4.104 1:4k 0.122 0.122 2.835 2.863 3.985 4.020 1:8K 0.108 0.111 2.268 2.339 3.879 3.892 1:16K 0.101 0.098 1.515 1.558 3.583 3.610 1:32k 0.091 0.093 1.018 1. 044 2,897 2.942 1:64K 0.088 0.087 0.600 0.607 2.038 2.122 PBS 0.112 0.100 0.123 0.108 0.125 0.141

[347] After the third immunization, peripheral blood mononuclear cells (PBMCs) were obtained from the immunized alpacas, RNA was extracted from the PBMCs, and the RNA was reverse transcribed into cDNA. VHH sequences were then amplified from the cDNA using single-domain antibody cloning primer combinations and subcloned into the pYDisplay yeast display vector, which was electrotransformed into cells. Petition 870250113096, dated 09 / 12 / 2025, pp. 101 / 183 94 / 119 competent EYB100 for building single-domain antibody yeast display libraries.

[348] The sequences of 50 randomly selected clones are shown in Figure 5A, which demonstrated low overlap, indicating a diverse library. Individual clones were subjected to a flow cytometry-based assay, in which biotinylated recombinant Can f1 was incubated with the VHH clone and, after a washing step, the detection of bound Can f1 identified the clone as a VHH candidate. The VHH candidates were then constructed into prokaryotic expression vectors.

[349] The isolated VHH clones were subsequently validated by performing an ELISA assay as described in Example 1. Thermostability metrics, including melting temperature (Tm), onset temperature (Tinitial), and aggregation temperature (Tagg), were also tested with increasing temperature in the range of 20°C - 95°C at a rate of 1°C / min. Sequence information for the VHH clones and the linkage assay results are summarized in Table 6. Individual linkage curves are shown for each candidate in Figures 5B-5H. Each clone demonstrated strong linkage, indicated by low EC50 values. Table 6. Validation of VHH anti-Can f1 Clone SEQ ID NO: Sequence EC50 (pg / mL) Tagg Tm SDAB221 12-2- B06 67 QVQLVESGGGLVQPGGSLRLSCAA SGFTFRLAAMGWYRQAPEKEREWV ASITGPGTDTNYADSVKGRFTVSR DNAKNT VYLQMNSLKP EDTAVY YC RGMGYWGKGTLVTVSS 0.1442 48.00 Tm1: 56.38 Tm2: 63.74 Tm3: 70.13 Petition 870250113096, dated 09 / 12 / 2025, p. 102 / 183 95 / 119 SDAB221 12-1- D12 68 AVQLVDSGGGLVQAGGSLRLACAA SGRTFDTYAVGWFRQAPGKERDVV ASITWTSGSTWYADFVKGRFTISK DNAKNT VYLQMNNLSP EDTAVY YC GARNQIYTRWDSWGQGTQVTVSS 4,008 49, 15 57,78 SDAB221 12-2- D08 69 QVKLEESGGGLVQAGGSLRLSCVA SGRTFSRWHTGWYRQAPGREREFV ATLRASGGDTYYADSVKGRFTISR DNAKNT VYLQMNSLKP EDTAVY YC NVWANWGAPPSDFSSWGQGTQVTV SS 5,845 SDAB221 12-1- C10 70 QLQLVESGGGLVQPGGSLRLSCAA SGFTFSSYSMSWYRQAPGKERELV ATIDTDGRTNYADSVKGRFTISRD NAKNSVYLQMNSLKPEDTAVYYCN RRQLGVDYWGQGTQVTVS S 9, 401 SDAB221 12-1- D12-3 71 QVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYGMSWVRQAPGKGLEWV SDVNSSGSRRFYVDSVKGRFTISR DNAKNTVFLQMNSLKPEDSAVYYC VKLAEAGTLIHVGSWGQGTRVTVS S 10,39 SDAB221 12-1-C9 72 QLQLVESGGGLVQPGGSLRLSCVG SGFTFSLASMGWYRQAPGKEREWV ASISSLDASTNYADSVKGRFTISR DNAKRMVYLQMNNL KS EDTAVYYC KAMNYWGKGTQVTVSS 13,14 Petition 870250113096, on 12 / 09 / 2025, page. 103 / 183 96 / 119 SDAB221 12-1- B6-B05 73 QLQLVESGGGLVQPGGSLRLSCAA SRSIFSSYVMAWYRRAPGKKRELV ASIANVGSNTDYAS FAKGRFTISR DDDKSRDNDKITVYLQMNSLNPED TAVYYCNAWLGAGSDYWGQGTQVT VSS 8, 418 Example 6: Identification of VHH anti-Can f2

[350] To obtain a unique variable domain in the heavy chain, i.e., VHH, specific for the canine allergen Can f2, a recombinant Can f2 labeled with His was prepared, which achieved high purity, as shown by Western blotting SDS-Page in Figure 6. To confirm this, an ELISA was performed measuring the optical density when recombinant Can f2-His was incubated with a biotinylated anti-His antibody. As summarized in Table 7, the recombinant Can f2-His was effectively bound to the antibody. Table 7: Can f2-His Binding ELISA Assay Recombinant Concentration of Can f2-His ^g / mL) Replication 1 (OD at 450 nm) Replication 2 (OD at 450 nm) 10 1.12 1.07 2 0.57 0.65 0.4 0.22 0.19 0.08 0.12 0.13 0.016 0.10 0.11 0.0032 0.11 0.11 0.00064 0.10 0.11 Blank 0.11 0.07

[351] Recombinant Can f2 was then administered to two alpacas in four rounds. The alpacas received 250 µg of recombinant Can f2 on days 0, 14, 28, and 49. An ELISA using titrated blood serum was performed, detecting VHH anti-Can f2 levels. The results are summarized in Table 8. This was also repeated with alpacas immunized with a sumomilated Can f2-His, Petition 870250113096, dated 09 / 12 / 2025, pp. 104 / 183 97 / 119, the results of which are found in Table 9. Suomo markers were used to test their effectiveness in increasing solubility. Each measurement was performed in duplicate. Negative serum was collected from uninfected alpacas to be used as a negative control. Table 8. Serum Titration of Alpaca Immunized with Can f2His Alpaca 1 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1:1K 0.128 0.133 2.171 1.809 2.466 2.207 1:2K 0.093 0.099 1.737 1.522 2.156 1.893 1:4K 0.077 0.075 1.460 1.375 1.723 1.774 1:8K 0.076 0.076 1.233 1.245 1.681 1.559 1:16K 0.075 0. 076 0.783 0.809 1.220 1.245 1:32k 0.069 0.062 0.562 0.530 0.874 0.891 1:64K 0.064 0.069 0.359 0.340 0.626 0.584 PBS 0.083 0.088 0.096 0.092 0.084 0.111 Alpaca 2 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1:1K 0.227 0.241 2.001 1.985 2.604 2.638 1:2K 0.143 0.133 1.497 1.404 2.056 2.060 1:4k 0.108 0.101 1.098 1.081 1.626 1.578 1:8K 0.099 0.084 0.772 0.678 1.263 1.252 1:16K 0.080 0.075 0.454 0.464 0.867 0.799 1:32k 0.075 0.068 0.290 0.302 0.573 0.530 1: 64K 0.070 0.062 0.194 0.195 0.363 0.334 PBS 0.080 0.072 0.073 0.080 0.100 0.081 Table 9. Serum Titration of Alpaca Immunized with Can f2sumo-His Petition 870250113096, dated 09 / 12 / 2025, page 105 / 183 98 / 119 Alpaca 1 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1:1K 0.119 0.111 4.028 4.074 4.120 4.205 1:2K 0.088 0.085 3.755 3.792 3.855 4.047 1:4K 0.071 0.072 3.438 3.392 3.828 3.973 1:8K 0.072 0.072 2.976 2.909 3.690 3.817 1:16K 0. 071 0, 071 2,190 2,215 3,212 3,487 1: 32k 0, 056 0, 057 1,560 1,557 2,570 2,883 1: 64K 0, 057 0, 058 1, 011 0, 999 1,923 2,109 PBS 0, 078 0, 080 0, 088 0, 079 0, 078 0, 114 Alpaca 2 Serum Titration Negative OD at 450 nm Day 14 (2nd Immunization) OD at 450 nm Day 28 (3rd Immunization) 1: 1K 0, 186 0, 181 4, 060 3.970 4.053 4.084 1:2K 0.119 0.113 3.318 3.461 3.804 3.846 1:4k 0.090 0.081 2.915 2.860 3.576 3.564 1:8K 0.083 0.079 2.267 2.290 3.192 3.196 1:16K 0.074 0.081 1.506 1.520 2.591 2.598 1:32k 0.071 0.061 1.004 1, 009 1,957 1.957 1:64K 0.070 0.057 0.638 0.616 1.319 1.322 PBS 0.084 0.067 0.064 0.068 0.066 0.075

[352] After the third immunization, peripheral blood mononuclear cells (PBMCs) were obtained from the immunized alpacas, RNA was extracted from the PBMCs, and the RNA was reverse transcribed into cDNA. VHH sequences were then amplified from the cDNA using single-domain antibody cloning primer combinations and subcloned into the pYDisplay yeast display vector, which was electrotransformed into competent EYB100 cells to construct single-domain antibody yeast display libraries. Petition 870250113096, dated 09 / 12 / 2025, pp. 106 / 183 99 / 119

[353] The sequences of 50 randomly selected clones are shown in Figure 7A, which demonstrated low overlap, indicating a diverse library. Individual clones were subjected to a flow cytometry-based assay, in which biotinylated recombinant Can f2 was incubated with the VHH clone and, after a washing step, the detection of bound Can f2 identified the clone as a VHH candidate. The VHH candidates were then constructed into prokaryotic expression vectors.

[354] The isolated VHH clones were subsequently validated by performing an ELISA assay as described in Example 1. Thermostability metrics, including melting temperature (Tm), onset temperature (Tinitial), and aggregation temperature (Tagg), were also tested with increasing temperature in the range of 20°C - 95°C at a rate of 1°C / min. Sequence information for the VHH clones and the linkage assay results are summarized in Table 10. Individual linkage curves are shown for each candidate in Figures 7B-7P. Each clone demonstrated strong linkage, indicated by low EC50 values. Table 10. Validation of VHH anti-Can f2 Clone SEQ ID NO: Sequence EC50 (pg / mL) Tagg Tm SDAB221 18-1- F04 9 QVQLVESGGGLVQPGGSLRLSCAYS GFTLDNNVIGWFRQAPGKEREFVAA ISRSGAFTHYTESVQGRFTISRDNA KNTVYLQMNSLKPEDTAVYYCAAGA ILLPTERRYDYWGQGTQVTVSS 0.1644 44.54 55.17 Petition 870250113096, dated 09 / 12 / 2025, page 107 / 183 100 / 119 SDAB221 18-1- D05 11 QVQLVESGGGLVQAGGSLGLSCAAS GRT FNNYVMGWFRQAPGKERE FVAA ISRSGSFTHYAEAVQGRFTISRDNA KITVYLQMNSLKPEDTAVYYCAAGA ILMPTERTYDYWGQGTQVTVSS 0,0762 39,54 Tm1: 48,35 Tm2 : 58,58 Tm3 : 64,58 SDAB221 18-1- D08 12 EVQLVESGGGLVQPGGSLRLSCVVS GSIFSDNAMGWYRQAPGKQREMVAI ISSVGTTNNVDSVNGRFTISRDNAK NTVYLQMNSLKPEDTAVYYCKDFSA PRYWGQGTQVTVSS 0,04863 40, 82 Tm1: 38,89 Tm2 : 43, 62 Tm3 : 54, 69 SDAB221 18-2- G01 13 AVQLVDSGGGLVQAGDSLRLSCVAS GRTFSSYVMGWFRQAPGKERLIVAT ISKSGSLTHYADSVEGRFTISRDNA KNTVYLQMNSLEPEDTAVYYCTPVS DLTGRRLGSYWGQGTQVTVSS 0,07356 38,93 43,71 SDAB221 18-3- B02 14 AVQLVDSGGGLVQPGGSLRLSCAAS GRTFSYYAMGWFRQALGKEREFVAA SSRTGRVTNYADSVKGRFTISRDNA KNTVYLQMNSLKPEDTAIYYCAADD RFYGGDNPAFYNSWGQGTQVTVSS 0,007586 SDAB221 18-3- D01 15 QVQLVESGGGLVQAGDSLRLSCAAS GRSFSNYVMNWFRQAPGKEREFVAA ISRSGRSTFYADSVKGRFTISRDNP KATVYLQMNSLVIEDTAVYFCAAAS DITSMRSLAAITSWGQGTQVIVSS 0,4291 SDAB221 18-3- G05 16 AVQLVESGGGLVQAGGSLRLSCAAS GRTFNNSVMGWFRQAPGKEREFVAA ISRSGAFAHYAESVEGRFTISRDNA KNTVYLQMNSLKPEDTAVYYCAAGA ILLPTERRYDYWGQGTQVTVSS 0.1494, Petition 870250113096, dated 09 / 12 / 2025, pp. 108 / 183 101 / 119 SDAB221 18-3- F08 17 QVQLVESGGGLVQPGESLRLSCAAS GSIFSIYRMGWYRQAPGEQREHVAT VTSGGGTGYADSVKGRFTIYRDNAK NTVYLQMNSLKPEDTAVYSCYAIPK SWSRTSEYSWGQGTQVTVSS 0,05955 SDAB221 18-189- 192-1- H03 18 EVQLVESGGGLVQARGSLRLSCAAS GRT FNNYAMGWFRQAPGKE REFVAA IASNTGTTYYAGSVKGRFAISRDNA KNTVDLHMNSLKPEDTAVYYCALGP LRLGWWYDSRPYDYWGQGTQVTVSS 0,0575 SDAB221 18-189- 192-1- C05 19 QVQLVESGGGGVQAGGSLRLSCVAS QSHFGYNVMGWYRQAPGRQRELVAT IISSGGTNYADSVKGRFTISRDNAK NTVHLSMDSLNVEDTAVYFCYAKGV WLGREYWGQGTQVTVSS 0,03249 SDAB221 18-189- 192-1- C10 20 EVQLVESGGGLVQAGGSLRLSCAAS GRTFSEYIMGWFRQAPGKERVFVST ISKSGAITNYADSVQGRFTISRDNA KNTVYLQMNSLKPEDSAVYYCAAGP LLSPAGRQYDYWGQGTQVTVSS 0,02166 SDAB221 18-189- 192-1- E08 21 AVQLVESGGGLVQPGGSLRLSCVAS GSSSRNYAMGWYRQAPGNEREFVAV ITSRGYTHYANSVSGRFTISRDHAK DTAYLQMNSLKPEDTAVYYCNTQAG LLPFMTVYDWGTGTQVTVSS 0,00411 SDAB221 18-189- 192-1- E09 22 EVQLVESGGGLVQPGGSLRLSCAAS GTIFLINRMGWYRQAPGKQRELVAT SFTSGNSTMYADSVKGRFTISRDNA KKTVYLQMNSLKPEDTAVYYCKARI SRRYGNWDDYWGQGTQVTVSS 0,05708 Petition 870250113096, dated 09 / 12 / 2025, page 109 / 183 102 / 119 SDAB221 18-189- 192-1- E10 23 AVQLVDSGGGLVQTGGSLRLSCAAS GTIFSINRMAWYRQAPGKQRELVAS IFFSGGITHYADFVKGRFTISRDNA KNTMYLQMNSLKPEDTAVYYCKGVI APNYRRTSNFQDYWGQGTQVTVSS 0.04267 Example 7: Identification of VHH anti-Der pl

[355] To obtain a unique variable domain in the heavy chain, i.e., VHH, specific for the dust mite allergen Der p1, a recombinant Der p1 labeled with His was prepared, which achieved high purity, as shown by Western blotting SDS-Page in Figure 8. To confirm this, an ELISA was performed measuring the optical density when the recombinant Der p1-His was incubated with a known antibody against Der p1 (clone 10B9). As summarized in Table 11, the recombinant Der p1-His was effectively bound to the antibody. Table 11: Der p1-His Binding ELISA Assay Recombinant Concentration of Der pl-His ^g / mL) Replication 1 (OD at 450 nm) Replication 2 (OD at 450 nm) 10 4.37 4.40 2 4.03 4.31 0.4 1.80 1.87 0.08 0.58 0.49 0.016 0.20 0.17 0.0032 0.25 0.12 0.00064 0.13 0.12 Blank 0.13 0.11

[356] Recombinant Der p1 was then administered to two alpacas in four rounds. The alpacas received 250 μg of recombinant Der p1 on days 0, 14, 28, and 49. A fifth round of immunization was completed in one of the alpacas on day 56. An ELISA using titrated blood serum was performed to detect anti-Der p1 HHV levels. The results are summarized in Petition 870250113096, dated 09 / 12 / 2025, page 110 / 183 103 / 119 Table 12. Each measurement was performed in duplicate. Negative serum was collected from uninfected alpacas to be used as a negative control. Table 12. Serum Titration of Alpaca Immunized with Der p1 Alpaca 1 Serum Titration Negative OD at 450 nm Day 28 (3rd Immunization) OD at 450 nm Day 49 (4th Immunization) 1:1K 0.216 0.191 2.770 3.005 4.021 3.960 1:2K 0.145 0.124 1.847 2.003 3.407 3.369 1:4K 0.105 0.089 1.178 1.268 2.625 2.613 1:8K 0.100 0.083 0.757 0.756 1.711 1.695 1:16K 0.085 0.073 0.447 0.460 1.118 1.091 1:32k 0.094 0.073 0.265 0.278 0.722 0.745 1:64K 0.082 0.061 0.163 0.133 0.455 0.470 PBS 0.083 0.079 0.078 0.065 0.062 0.065 Alpaca 2 Serum Titration Negative OD at 450 nm Day 49 (4th Immunization) OD at 450 nm Day 56 (5th Immunization) 1:1K 0.387 0.399 3.119 3. 021 4. 135 0. 159 0. 652 0.719 1.309 1.275 1:16K 0. 104 0. 123 0.385 0.394 0.775 0.772 1: 32k 0. 104 0. 103 0.244 0.243 0.480 0.485 1: 64K 0.097 0.097 0.119 0.110 0.350 0.335 PBS 0.089 0.084 0.110 0.111 0.112 0.108

[357] After the 4th and 5th immunizations, peripheral blood mononuclear cells (PBMCs) were obtained from the two immunized alpacas, respectively. RNA was then extracted from the PBMCs and reverse transcribed into cDNA. VHH sequences were then amplified from the cDNA using the Petition 870250113096, dated 09 / 12 / 2025, page 111 / 183 104 / 119 single-domain antibody cloning primer combinations and subclones in the pYDisplay yeast display vector, which was electrotransformed into competent EYB100 cells to construct single-domain antibody yeast display libraries.

[358] Sequences from 50 randomly selected clones showed low overlap, indicating a diverse library. Individual clones were subjected to a flow cytometry-based assay, in which biotinylated recombinant Der p1 was incubated with the VHH clone and, after a washing step, the detection of bound Der p1 identified the clone as a VHH candidate. The VHH candidates were then constructed into prokaryotic expression vectors.

[359] The isolated VHH clones were subsequently validated by performing an ELISA assay, as described in Example 1. Sequence information for the VHH clones and the linkage assay results are summarized in Table 13. Individual linkage curves are shown for each candidate in Figures 9A-9K. Each clone demonstrated strong linkage, indicated by low EC50 values. Table 13. Validation of VHH anti-Der p1 Clone SEQ ID NO: Sequência EC50 (pg / mL) Tagg Tm SDAB221 29-55- 35-46- 49-1- B02 24 QLQLVESGGGLVQSGGSLRLSCVAKG GTNHPYPIGWFRQAPGKEQEGVLCIA SGGQTQDSAHNPNWADSVKGRFTISR DDATHTVYLEMNNLKADDTAVYFCAA PREIYIDSRCATYKYEYWGQGTQVTV SP 0.01926 Petition 870250113096, on 12 / 09 / 2025, page. 112 / 183 105 / 119 SDAB221 29-55- 35-46- 49-1- C12 25 QVQLVESGGGLVQAGGSLRLSCAASG RTFSSYAMGWFRQAPGKEREFVAIIS NTGGLTDYAHSVKGRFTISRDNAKNT GYLQMNSLKPEDTALYYCAADFLGPK SWPSYWGQGTQVTVSS 4, 948 SDAB221 29-55- 35-46- 49-1- D04 26 QLQLVESGGGLVQPGGSLRLSCVISG GTFHPYPIGWFREAPGKEREGVLCIN SGGQSEISANSAHDPKYADSVKGRFT ISRDNAANTVYLQMNNLEPGDTAVYY CAAPREIYVDPYCPTYAYEYWGQGTQ VTVSA 0,06209 SDAB221 29-55- 35-46- 49-1- D01 27 QVQLVESGGGLVQAGGSLRLSCAASG RTFSRYAMGWFRQAPGKEREFVAAIT GSGSRTYYADSIQGRFTISRDNAKNT VYLQMNSLKPEDTAVYYCVQGWAEAT MTSLGEDYDYWGQGTQVTVSS 0,01468 SDAB221 29-55- 35-46- 49-1- G07 28 QVQLVESGGGLVQAGGSLRLSCAASG RTFSSYAMGWFRQAPGKERELVATIS WSGGSTYYADSVKGRFTISRDNAKNT GFLQMNSLKPEDTAVYYCAADFASTV GTPLTRPAYWGQGTQVTVSS 0,4846 SDAB221 29-55- 35-46- 49-1- G09 29 QVKLEESGGGAVQPGGSLRLSCTASG QTFSNYIISWFRQAPGKERE FVVGIS KSGGRTYYADSAKGRFTISRDNAKNT VYLQMSSLKPEDTAVYYCAADGLVLT AAAAEYDYWGQGTQVTVSS 0,1543 SDAB221 29-55- 35-46- 49-1- A03 30 QVQLVESGGGLVQAGGSLRLSCAVSG RTVSRNVMGWFRQAPGKEREFVAGIG FSGGSTYYADSVKGRFTISRDNAKNT VYLEMNRLQPEDTAVYYCAAPSLPLL TSDLHDYDYWGQGTQVTVSS 0.02911, Petition 870250113096, dated 09 / 12 / 2025, page 113 / 183 106 / 119 SDAB221 29-55- 35-46- 49-1- B12 31 QVQLVESGGGLVQAGGPLRLSCAASG RTFSSYAMGWFRQAPGKEREFVAAIS WSGDSTYYADSVKGRFTLSRDNAKNT VYLQMNSLKPEDTAVYYCAVPSSGRG TYYYTLSAYEYWGQGTQVTVFS 1,767 SDAB221 29-55- 35-46- 49-1- G02 32 QLQLVESGGGLVQAGGSLRLSCTASG RTFRNFGMGWFRQAPGKERKLVASIS YVGGNTDYADSVKGRFTISMDNAKNT VVLQMNSLKPEDTAMYYCAARNPNRN EYPWWGQGTQVTVSS 4,207 SDAB221 29-55- 35-46- 49-1- A04 33 QLQLVESGGGLVQPGGSLRLSCAASG RVYNSWTMAWFRQAPGKEREFVGAFS LLDSGTRYADSLKDRVAISRDDAANT QWLQLSALKPEDTAVYYCAAKSGTIR ATSEGQYNYWGQGIQVTVSS 3,408 SDAB221 29-55- 35-46- 49-1- H08 34 EVQLVESGGGLVQAGGSLTLSCGRTT SIWGMGWFRQGRGKEREFVAAITPSG SITFYSDHVKGRFTVSRDNALNTVYL QMNSLKPEDTAVYYCARRGSSGSYW AGSYEAWGQGTQVTVSS 16, 49

[360] The cysteine ​​protease activity of Der p1 contributes significantly to its allergenicity, through the increase of total IgE and the synthesis of specific IgE 1 for Der p1. To screen for VHH candidates capable of blocking this enzymatic activity and, consequently, the induced IgE synthesis, VHH candidates were incubated with Der p1 and cysteine ​​solution, and the cysteine ​​protease activity of Der p1 was measured in a continuous rate assay with the fluorogenic substrate. Der p1 was pre-activated with 5 mM cysteine ​​(Sigma Chemical Co.) to regenerate its thiol group, which oxidizes during purification. The catalytic activity of Der p1 was measured in a continuous rate (kinetic) assay using the fluorogenic peptide substrate N-tert-butoxycarbonyl (Boc)-Gln-Ala-Arg-7-amino Petition 870250113096, dated 09 / 12 / 2025, pp. 114 / 183 107 / 119 4-methylcoumarin (AMC), which was conducted in 50 mM sodium phosphate buffer, pH 7.0, containing 2.5 mM EDTA and 2.5 mM dithiothreitol (DTT) at 37°C in a total volume of 1 ml. The hydrolysis of AMC substrates was monitored using a Hitachi F-2000 fluorescence with Xex = 380 nm and Xem = 460 nm. As summarized in Figure 10, many VHHs demonstrated blocking effects on Der p1 enzymatic activity. These candidates were subsequently tested in titration experiments to confirm their activity. Each VHH was diluted three times by five times in sodium phosphate buffer, starting with 0.6 mg / ml, as shown in Table 14. Its cysteine ​​protease blocking activity of Der p1 was measured. The titration results for each VHH are shown in Figures 11A to 11H and summarized in Table 15. Table 14. Titration Concentrations Candidate antibody concentration (µg / ml) 600.00 200.00 66.67 22.22 7.41 Final candidate antibody concentration (µg / ml) 200.00 66.67 22.22 7.41 2.47 Table 15. Effect of VHH Blocking on Der p1 Activity Maximum Clone Blocking Rate 1-B12 95.38 1-G07 93.34 1-H08 92.83 1-C12 91.59 1-G02 85.17 2-F04 79.36 1-B02 75.36 1-D01 72.95 Example 8. Identification of Der p1-Binding Peptides

[361] The recombinant Der p1 obtained in Example 7 was used to generate a phage display library to screen 7-amino acid peptides that can bind to Der p1. This Petition 870250113096, dated 09 / 12 / 2025, pages 115 / 183 108 / 119 was performed similarly to the display libraries described in the previous examples, using nucleic acids encoding the peptides expressed in phages. Shown in Figure 12, several peptides were identified and demonstrated a high degree of diversity. The sequences of these peptides are summarized in Table 16. Table 16. Der p1-Binding Peptides Name SEQ ID NO: Sequence SDAB22110-2-B2 35 GIHMALM SDAB22110-2-B7 36 GAIIAMK SDAB22110-2-E9 37 GLLVSDW SDAB22110-2- B10 38 VVPRPSF SDAB22110-2-G1 39 VVAFGIS SDAB22110-2-F3 40 VIFNHVP SDAB22110-2-D3 41 HSGPECG SDAB22110-2-G4 42 HQPQGVW SDAB22110-2-C3 43 YPTWAYR SDAB22110-2-C4 44 FTFQGMD SDAB22110-2-D4 45 SLHRIPS SDAB22110-2-D9 46 LTGYGMS SDAB22110-2-E4 47 TSSVIQL SDAB22110-2-F1 48 AVVSFYP

[362] An ELISA was used to validate the identified peptides. Der p1 was attached to the bottom of a plate. Petition 870250113096, dated 09 / 12 / 2025, pp. 116 / 183 109 / 119 Different concentrations of the selected (biotinylated) peptides were added and then washed. Next, HRP-streptavidin (which binds to biotin) was added and an OD was measured at 450 nm. The results are summarized in Table 17. Duplicate measurements are separated by commas. An exemplary binding curve for the 2-D3 peptide is shown in Figure 13. Table 17. Peptide Binding Assay to Der p1 Peptide Concentration Peptide: 2-B2 (OD450) Peptide: 2-C3 (OD450) Peptide: 2-D3 (OD450) Peptide: 2-D9 (OD450) Peptide: 2-F1 (OD450) Peptide: 2-G1 (OD450) 30 0.536, 0.537 0.684, 0.748 4.097, 4.131 0.751, 0.771 0.938, 0.805 0.666, 0.629 10 0.275, 0.289 0.346, 0.376 3.550, 3.571 0.374, 0.357 0.479, 0.516 0.323, 0.293 3.333 0.132, 0.136 0.162, 0.191 2.770, 2.875 0.154, 0.154 0.221, 0.209 0.147, 0.145 1.111 0.089, 0.089 0.104, 0.102 2.158, 2.250 0.095, 0.102 0.131, 0.125 0.102, 0.109 0.370 0.083, 0.071 0.072, 0.066 1.372, 1.358 0.068, 0.078 0.076, 0.073 0.088, 0.082 0.123 0.078, 0.066 0.065, 0.065 0.866, 0.906 0.067, 0.068 0.073, 0.075 0.082, 0.073 0.041 0.070, 0.061 0.058, 0.057 0.468, 0.463 0.061, 0.059 0.079, 0.066 0.057, 0.062 0.000 0.074, 0.071 0.066, 0.063 0.063, 0.067 0.062, 0.067 0.062, 0.070 0.070, 0.065 Example 9: Identification of anti-Fel D1 VHH

[363] To obtain a single variable domain in the heavy chain, i.e., VHH, specific for the cat allergen Fel D1, a recombinant Fel D1 labeled with His was prepared, which achieved high purity, as shown by Western blotting SDS-Page in Figure 14. To confirm this, an ELISA was performed measuring the optical density when the recombinant Fel d1-His is incubated with two known Fel D1 antibodies that are summarized in Table 18. As summarized in Table 19, the recombinant Fel d1-His was effectively bound by both antibodies. Petition 870250113096, dated 09 / 12 / 2025, pp. 117 / 183 110 / 119 Table 18. Positive Control Antibodies Fel D1 Name SEQ ID NO: Sequence Name REGN19 62 Heavy Chain Variable Region EVQLVESGGGLVQPGGSLRLSCA ASGFTFSSYAMSWVRQAPGKGLE WVSAISGRGYNADYADSVKGRFT ISRDNSKNTLYLQMNSLRAEDTA VYYCAKLEYFDYWGQGTLVTVSS 09 63 Light Chain Variable Region DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKL LIYKASSLESGVPSRFSGSGSGT DFTLTISSLRPEDFATYYCQQYN SYPLTFGGGTKVEIK 370VH+ 37 8VL 64 Heavy Chain Variable Region EVQLVESGGGLAQPGGSLRLSCA ASGFTFNNYAMTWVRQAPGKGLD WVSAISDSGRSTFSADSVKGRFT ISRDNSKNTLYLQMDSLRAEDTA LYYCAKHRNWNYPVFDYWGQGTL VTVSS 370VH+ 37 8VL 65 Variable Region of the Light Chain DIQLTQSPSFLSASVGDRVTITC WASQGISSYLAWYQQKPGKAPKL LIYSASTLQSGVPSRFSGSGSGT EFTLTISSLQPEDFATYYCQQLN SYPFTFGPGTKVDIK 18VH+3 7 8VL 66 Variable Region of the Heavy Chain EVQLVESGGGLVKPGGSLRLSCA ASGFTFRNYNINWVRQAPGKGLE WVSLISGSSSYIYYADSVKGRFT VSRDNAKNSLYLQMNSLRAEDTA VYYCARRTLSYYVMDVWGQGTTV TVSS 18VH+3 7 8VL 65 Variable Region of the Light Chain DIQLTQSPSFLSASVGDRVTITC WASQGISSYLAWYQQKPGKAPKL Petition 870250113096, dated 09 / 12 / 2025, pages 118 / 183 111 / 119 LIYSASTLQSGVPSRFSGSGSGT EFTLTISSLQPEDFATYYCQQLN SYPFTFGPGTKVDIK Table 19. Recombinant Fel D1 His Binding Assay Volume of Fel D1-His (μL) (concentration ~1μ9 / μL) OD450 using positive control: 18VH+378VL 100 4.60 4.60 50 4.55 4.53 10 4.47 4.44 Negative Control 0.16 0.09 Volume of Fel D1-His (μL) OD450 using positive control: 370VH+378VL 100 4.60 4.60 50 4.55 4.53 10 4.47 4.44 Negative Control 0.16 0.09

[364] Four distinct epitopes of Fel D1 were selected for further identification. These epitopes are summarized in Table 20. Peptides were synthesized for the four epitopes for VHH identification. Table 20. Fel D1 epitopes used SEQ ID NO: Name Sequence Positive Control Ab 58 SDAB22110- 2 25-38AA VAQYKALPVVLENA N / A 59 SDAB221103 15-28AA- 1 FAVANGNELLLDLS REGN1909 60 SDAB221104 46-59AA AKMTEEDKENALS N / A 61 SDAB221101 15-28AA- 2 ENARILKNCVDAKM 370VH+378VL, 18VH+378VL

[365] Recombinant Fel D1 was then administered to two alpacas in four rounds. The alpacas received 250 μg of recombinant Fel D1 on days 0, 14, 28, and 49. A fifth round of immunization was completed in one of the alpacas on day 56. An ELISA Petition 870250113096, dated 09 / 12 / 2025, pp. 119 / 183 112 / 119 using titrated blood serum was performed to detect VHH anti-Fel D1 levels. The results are summarized in Table 21. Each measurement was performed in duplicate. Negative serum was collected from uninfected alpacas to be used as a negative control. Table 21. Serum Titration of Alpaca Immunized with Fel D1 Alpaca 1 Serum Titration Negative OD at 450 nm Day 28 (3rd Immunization) OD at 450 nm Day 49 (4th Immunization) 1:1K 0.258 0.289 3.438 3.300 4.137 3.971 1:2K 0.167 0.176 2.630 2.550 3.858 3.806 1:4K 0.132 0.138 1.793 1.702 3.654 3.648 1:8K 0.128 0.115 1.139 1.065 3.046 3.033 1:16K 0. 103 0, 103 0.727 0.668 2.319 2.266 1:32k 0.106 0.115 0.447 0.416 1.625 1.612 1:64K 0.107 0.108 0.278 0.257 1.018 1.041 PBS 0.135 0.112 0.107 0.098 0.112 0.103 Alpaca 2 Serum Titration Negative OD at 450 nm Day 49 (4th Immunization) OD at 450 nm Day 56 (5th Immunization) 1:1K 0.248 0.220 3.705 3.586 3.957 4.115 1:2K 0.151 0.155 2.904 2.741 3.859 3.971 1:4k 0.120 0.119 2.131 2.040 3.808 4.009 1: 8K 0.114 0.117 1.434 1.330 3.768 3.917 1:16K 0.096 0.098 0.803 0.830 3.219 3.406 1:32k 0.105 0.108 0.496 0.533 2.456 2.510 1: 64K 0.092 0.101 0.297 0.290 1.623 1.548 PBS 0.106 0.104 0.111 0.119 0.119 0.144

[366] After the 4th and 5th immunizations, peripheral blood mononuclear cells (PBMCs) were obtained from the two immunized alpacas, respectively. Then, RNA was extracted from the Petition 870250113096, dated 09 / 12 / 2025, pages 120 / 183 113 / 119 PBMCs and RNA were reverse transcribed into cDNA. VHH sequences were then amplified from the cDNA using single-domain antibody cloning primer combinations and subcloned into the pYDisplay yeast display vector, which was electrotransformed into competent EYB100 cells to construct single-domain antibody yeast display libraries.

[367] Sequences from 50 randomly selected clones showed low overlap, indicating a diverse library, as shown in Figure 15A. Individual clones were subjected to a flow cytometry-based assay, in which biotinylated recombinant Fel D1 was incubated with the VHH clone and, after a washing step, the detection of bound Fel D1 identified the clone as a VHH candidate. The VHH candidates were then constructed into prokaryotic expression vectors.

[368] The isolated VHH clones were subsequently validated by performing an ELISA assay as described in Example 1. The clones were incubated with each of the epitopes listed in Table 20. The binding results are shown in Figure 15B-15D for the epitope with SEQ ID NO: 61. The binding results are shown in Figure 16 for the epitope with SEQ ID NO: 59. The binding results are shown in Figures 17A-17E for the epitope with SEQ ID NO: 60. The binding results are shown in Figure 18 for the epitope with SEQ ID NO: 58. Sequence information for the VHH clones and the binding assay results are summarized in Table 22. Each clone demonstrated strong binding, indicated by low EC50 values. Table 22. Validation of VHH anti-Fel dl Clone SEQ ID NO: Recognized Epitope Sequence EC50 (pg / mL) Tabb Tm Petition 870250113096, dated 09 / 12 / 2025, pp. 121 / 183 114 / 119 SEQ ID NO: 61 VAQYKALPVVL ENA (SEQ ID NO: 58) 0,352 58,93 66, 50 SDAB2 2110- 1-1- B9-1 50 QVQLVESGGGLVQPGGSL RLSCAASGFTFRLAAMGW YRQAPEKEREWVASITGP GTDTNYADSVKGRFTVSR DNAKNTVYLQMNSLKPED TAVYYCRGMGYWGKGTLV TVSS ENARILKNCVD AKM (SEQ ID NO: 61) 14,440 41,73 Tm1: 54,03 Tm2: 63,20 Possíveis agregados SDAB2 2110- 1-1- E4 51 AVQLVDSGGGLVQAGGSL RLACAASGRTFDTYAVGW FRQAPGKERDVVASITWT SGSTWYADFVKGRFTISK DNAKNTVYLQMNNLS PED TAVYYCGARNQIYTRWDS WGQGTQVTVSS ENARILKNCVD AKM (SEQ ID NO: 61) 3,305 42,78 Tm1: 41,13 Tm2: 52,88 Possíveis agregados SEQ ID NO: 59) 12,940 42,30 47,50 SDAB2 2110- 3-2- H08 53 QLQLVESGGGLVQPGGSL RLSCAASGFTFSSYSMSW YRQAPGKERELVATIDTD GRTNYADSVKGRFTISRD NAKNSVYLQMNSLKPEDT AVYYCNRRQLGVDYWGQG TQVTVSS AKMTEEDKENA LS (SEQ ID NO: 60) 5, 024 35, 68 42,43, Petition 870250113096, on 12 / 09 / 2025, page. 122 / 183 115 / 119 SEQ ID NO: 60 12,200 40, 93 46, 83 SDAB2 2110- 4-3- E05 55 QLQLVESGGGLVQPGGSL RLSCVGSGFTFSLASMGW YRQAPGKEREWVASISSL DASTNYADSVKGRFTISR DNAKRMVYLQMNNLKSED TAVYYCKAMNYWGKGTQV TVSS AKMTEEDKENA LS (SEQ ID NO: 60) 1,719 31,74 Tm1: 37,85 Tm2: 47,65 Tm3: 62,39 Possíveis agregados SDAB2 2110- 4-3- G08 56 QLQLVESGGGLVQPGGSL RLSCAASRSIFSSYVMAW YRRAPGKKRELVASIANV GSNTDYASFAKGRFTISR DDDKSRDNDKITVYLQMN SLNPEDTAVYYCNAWLGA GSDYWGQGTQVTVSS AKMTEEDKENA LS (SEQ ID NO: 60) 2,215 42,29 58,58 SDAB2 2110- 4-3- H02 57 QLQLVESGGGLVQPGGSL RLSCEASGFTFKYYTMSW YRQAPGKERELVATITNG DRTNYADSVKGRFTISRD NAKNTLYLQMNSLKPYDT AVYYCNRHLLPLQIWGQG TQVTVSS AKMTEEDKENA LS (SEQ ID NO: 60) 12,940 42,30 47,50

[369] The detailed thermostability plots used to determine Tm, Tinício and Tagg are illustrated in Figures 19A19I and the raw data are summarized in Table 23. Any subsequent observed melting points (Tm1, Tm2, ​​Tm3, etc.) are separated by commas. Table 23. Thermostability Results for VHH AntiFel d1 Tm T x ·*· start T xagg Petition 870250113096, dated 09 / 12 / 2025, pages 123 / 183 116 / 119 Clone: ​​DAB22110- 1-1-C3 0 (mean) σ (standard deviation) 0 σ 0 σ Mean 66.50 0.01 58.06 0.16 58.93 0.12 Replicate 1 66.50 58.17 59.02 Replicate 2 66.51 57.94 58.85 Clone: ​​SDAB22110- 1-1-B9-1 Tm T χ ·*· start T xagg 0 σ 0 σ 0 σ Mean 54.03, 63.20 0.11, 0.08 42.07 0.05 41.73 0.00 Replicate 1 53.95, 63.26 42.11 41.73 Replicate 2 54.10, 63.15 42.04 41.73 Clone: ​​SDAB22110- 1-1-E4 Tm T x ·*· initial T xagg 0 σ 0 σ 0 σ Average 50.69 0.05 40.64 0.07 39.61 0.00 Replicate 1 50.73 40.60 39.62 Replicate 2 50.66 40.64 0.07 39.61 Clone: ​​SDAB22110- 3-1-D9-1 T xm T - ·*· initial T xagg 0 σ 0 σ 0 σ Average 41.13, 52.88 0.17, 0, 03 32.74 0.23 42.78 0.03 Replicate 1 41.01, 52.86 32.57 42.76 Replicate 2 41.25, 52.91 32.90 42.80 T xm T - ·*· start T xagg Petition 870250113096, dated 09 / 12 / 2025, pp. 124 / 183 117 / 119 Clone: ​​SDAB22110- 3-2-H08 0 σ 0 σ 0 σ Average 47.50 0.05 37.18 0.22 42.30 0.16 Replicate 1 47.46 37.03 42.41 Replicate 2 47.53 37.34 42.19 Clone: ​​SDAB22110- 4-3-B02 Tm T χ ·*· inxcio T xagg 0 σ 0 σ 0 σ Average 42.43 0, 12 33.78 0.00 35.68 0.19 Replicate 1 42.52 33.78 35.55 Replicate 2 42.34 33.78 35.82 Clone: ​​SDAB22110- 4-3-E05 Tm T x ·*· start T xagg 0 σ 0 σ 0 σ Average 46.83 0.27 35.67 0.08 40.93 0.00 Replicate 1 47.02 35.61 40.93 Replicate 2 46.64 35.73 40.93 Clone: ​​SDAB22110- 4-3-G08 T xm T - ·*· start T xagg 0 σ 0 σ 0 σ Average 37.85, 47.65, 62.92 0.10, 0.16, 0.75 27.92 0.00 31.74 0.04 Replicate 1 37.77,47.76, 61.85 27.93 31.71 Replicate 2 37.92, 47.53, 62.92 27.92 31.77 T xm T - ·*· start T xagg Petition 870250113096, dated 09 / 12 / 2025, pages 125 / 183 118 / 119 Clone: ​​SDAB22110- 4-3-H02 0 σ 0 σ 0 σ Average 58.58 0.02 47.54 0.31 42.29 0.08 Replicate 1 58.60 47.33 42.24 Replicate 2 58.57 47.76 42.35

[370] Functional validation was also performed to test the ability of an exemplary clone to neutralize Fel D1. Fel D1 is naturally a weaker tetramer that causes allergic reactions by cross-linking neighboring IgEs in immune cells (mast cells, etc.). Size Exclusion Chromatography (SEC) was performed, in which proteins were passed through a mesh where larger proteins do not flow into the mesh as easily and therefore elute earlier. In SEC, there are two distinct peaks for pure Fel D1: the earlier peak represents the tetramer and the weaker peak represents the monomer (Figure 20A). The conditions for this SEC plot are (Column: Zenix-C SEC80, 7.8x300mm, 3um, 80A; Sample: Fel D1; Sample Volume: 30 μL (25ug); Mobile Phase: 150 mM phosphate buffer, pH 7.0; Flow Rate: 0.7 mL / min; Detection: UV 280 nm; System: Thermo Vanquish Flex.

[371] When clone C3 was analyzed, it was observed that it was smaller than the Fel D1 monomer, with a main peak around 14 minutes (Figure 20B). The conditions for this SEC plot are (Column: Zenix-C SEC-80, 7, 8x300mm, 3um, 80A; Sample: 1C3; Sample Volume: 30 pL; Mobile Phase: 150 mM phosphate buffer, pH 7.0; Flow Rate: 0.7 mL / min; Detection: UV 280 nm; System: Thermo Vanquish Flex.

[372] When clone C3 and Fel D1 were incubated together for 4 hours at 37°C and then analyzed, the two peaks of Fel D1 were no longer detected, only the C3-Fel D1 complex was detected. Notably, this complex appears at a later time than the Fel D1 tetramer, demonstrating that the Petition 870250113096, dated 09 / 12 / 2025, pp. 126 / 183 119 / 119 nanobody C3 is preventing the formation of the Fel D1 tetramer (Figure 20C). The conditions for this SEC plot are (Column: Zenix-C SEC-80, 7.8x300mm, 3µm, 80A; Sample: 30µg of Fel D1 protein and 30µg of 1-C3 were pre-incubated at 37°C for 4 hours in a 60µl incubation system; Sample Volume: 30 µL (25µg); Mobile Phase: 150 mM phosphate buffer, pH 7.0; Flow Rate: 0.7 mL / min; Detection: UV 280 nm; System: Thermo Vanquish Flex).

[373] Although preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided only as examples. Numerous variations, alterations, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in the practice of the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents are covered by them.

Claims

1. Allergen-binding protein that binds to or neutralizes an allergen, characterized in that the allergen-binding protein comprises a nanobody, a monobody, a DARPin, or a small peptide with a molecular weight of less than about 25 kilodaltons or a length of less than about 300 amino acids.

2. Allergen-binding protein that binds to or neutralizes an allergen, characterized in that the allergen-binding protein comprises a nanobody or small peptide with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 1-57 and 67-73.

3. Allergen-binding protein, according to claim 1 or 2, characterized in that the allergen-binding protein is formulated to coat a food or to spray, spray or brush onto an animal or household surface.

4. Allergen-binding protein, according to claim 3, characterized in that the food is a pet food.

5. Allergen-binding protein, according to claim 4, characterized in that the allergen is a pet allergen.

6. Allergen-binding protein, according to any one of claims 3 to 5, characterized in that the allergen-binding protein is formulated to be applied to food as a coating.

7. Allergen-binding protein, according to any one of claims 3 to 5, characterized in that the allergen-binding protein is formulated to be mixed into food. Petition 870250094198, dated 10 / 15 / 2025, page 12 / 38 2 / 25 8. Allergen-binding protein, according to any one of claims 1 to 7, characterized in that the allergen-binding protein is at a concentration of about 0.01 milligrams per milliliter (mg / ml) to about 500 mg / ml in solution or after suspension.

9. Allergen-binding protein, according to any one of claims 1 to 8, characterized in that the allergen induces an allergic reaction in a human.

10. Allergen-binding protein, according to any one of claims 1 to 9, characterized in that the allergen comprises an environmental allergen.

11. Allergen-binding protein, according to any one of claims 1 to 10, characterized in that the allergen comprises an animal allergen.

12. Allergen-binding protein, according to any one of claims 1 to 11, characterized in that the allergen comprises a pet allergen.

13. Allergen-binding protein, according to any one of claims 1 to 12, characterized in that the allergen comprises a cat allergen, a dog allergen, a rabbit allergen, a mouse allergen, or a cockroach allergen.

14. Allergen-binding protein, according to claim 13, characterized in that the allergen is selected from the group consisting of Fel d 1, Fel d 2, Fel d 3, Fel d 4, Can f 1, Can f 2, Can f 4, Can f 7, Ory C 1, Mus M 1 and Bla G 2.

15. Allergen-binding protein, according to claim 14, characterized in that the allergen is Fel d 1.

16. Allergen-binding protein, according to claim 15, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID Nos: 49-57.

17. Allergen-binding protein, according to claim 16, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO:

49.

18. Allergen-binding protein, according to claim 17, characterized in that the allergen-binding protein comprises a nanobody with a SEQ ID sequence NO:

49.

19. Allergen-binding protein, according to claim 16, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO:

57.

20. Allergen-binding protein, according to claim 19, characterized in that the allergen-binding protein comprises a nanobody with a sequence SEQ ID NO:

57.

21. Allergen-binding protein, according to claim 14, characterized in that the allergen is Can f 1.

22. Allergen-binding protein, according to claim 21, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, Petition 870250094198, 10 / 15 / 2025, page 14 / 38 4 / 25 at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID Nos: 67-73.

23. Allergen-binding protein, according to claim 22, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

67.

24. Allergen-binding protein, according to claim 23, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

67.

25. Allergen-binding protein according to claim 14, characterized in that the allergen is Can f 2.

26. Allergen-binding protein, according to claim 25, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 9-23.

27. Allergen-binding protein, according to claim 26, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

16.

28. Allergen-binding protein, according to claim 27, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence consisting of SEQ ID NO:

16.

29. Allergen-binding protein, according to any one of claims 1 to 10, characterized in that the allergen comprises a dust allergen.

30. Allergen-binding protein, according to claim 29, characterized in that the dust allergen comprises Der p1 or Der p2.

31. Allergen-binding protein, according to claim 30, characterized in that the dust allergen is Der p1.

32. Allergen-binding protein, according to claim 31, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 24-34.

33. Allergen-binding protein, according to claim 32, characterized in that the allergen-binding protein comprises a peptide having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOS: 3548.

34. Allergen-binding protein, according to claim 33, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

27.

35. Allergen-binding protein, according to Petition 870250094198, dated 10 / 15 / 2025, page 16 / 38 6 / 25, claim 34, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

27.

36. Allergen-binding protein, according to claim 33, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

28.

37. Allergen-binding protein, according to claim 36, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

28.

38. Allergen-binding protein, according to claim 30, characterized in that the dust allergen is Der p2.

39. Allergen-binding protein, according to claim 38, characterized in that the allergen-binding protein comprises a nanobody with at least 80%, at least 85%, at least 90% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 1-8.

40. Allergen-binding protein, according to claim 39, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

3.

41. Allergen-binding protein, according to claim 40, characterized in that the allergen-binding protein comprises a nanobody with a sequence Petition 870250094198, dated 10 / 15 / 2025, page 17 / 38 7 / 25 consisting of SEQ ID NO:

3.

42. Allergen-binding protein, according to any one of claims 1 to 10, characterized in that the allergen comprises a plant allergen or plant pollen allergen.

43. Allergen-binding protein, according to claim 42, characterized in that the plant or pollen allergen is selected from the group consisting of Bet v1, Phl p 5, Phl p 1, Poa p 1, Cyn d 1, Bet v 2, Ole e 1, Amb a 1, Amb a 11 and Art v 1.

44. Allergen-binding protein, according to any one of claims 1 to 10, characterized in that the allergen comprises a mold allergen.

45. Allergen-binding protein, according to claim 44, characterized in that the mold allergen is selected from the group consisting of Alt a 1, Asp f 1, Asp f 2, Cla h 8, Pen ch 13 and Pen ch 18.

46. ​​Allergen-binding protein, according to any one of claims 1 to 10, characterized in that the allergen is a food allergen.

47. Allergen-binding protein, according to claim 46, characterized in that the food allergen is selected from the group consisting of Pen a 1, Ara h 1, Ara h 3.

48. Allergen-binding protein, according to any one of claims 1 to 47, characterized in that the allergen-binding protein comprises at least one modified amino acid.

49. Allergen-binding protein, according to any one of claims 1 to 48, characterized in that the allergen-binding protein comprises about 1, about 5, about 10, about 15, about 20, about 25 or about 30 modified amino acids. Petition 870250094198, dated 10 / 15 / 2025, p. 18 / 38 8 / 25 50. Allergen-binding protein, according to claim 48 or 49, characterized in that the modified amino acid comprises a non-canonical amino acid.

51. Allergen-binding protein, according to claim 50, characterized in that the non-canonical amino acid is selected from the group consisting of parabenzoylphenylalanine, 3,4-dihydroxyphenylalanine, a tetrazine, a chloroctene, homopropargylglycine, paraproparglyoxyphenylalanine, para-azidophenylalanine, phenylalanine paraisothiocyanate, para-benzoylphenylalanine, paracyanophenylalanine, para-nitrophenylalanine, a halogenated m-tyrosine analog, a halogenated proline analog, a halogenated tryptophan analog, and a halogenated leucine analog.

52. Allergen-binding protein, according to any one of claims 48 to 51, characterized in that the solubility of the allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600% or more compared with an unmodified allergen-binding protein.

53. Allergen-binding protein, according to any one of claims 48 to 51, characterized in that the binding affinity of the allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to an unmodified allergen-binding protein.

54. Allergen-binding protein, according to any one of claims 48 to 51, characterized in that the thermal stability of the allergen-binding protein is increased by about 10%, 25%, 50%, 75%, 100%, 200%, 300% or more compared to an unmodified allergen-binding protein.

55. Allergen-binding protein, according to any Petition 870250094198, dated 10 / 15 / 2025, page 19 / 38 9 / 25 one of claims 1 to 54, characterized in that the allergen-binding protein is a multivalent allergen-binding protein.

56. Allergen-binding protein according to claim 55, characterized in that the multivalent allergen-binding protein is a bivalent allergen-binding protein.

57. Allergen-binding protein, according to claim 55 or 56, characterized in that the binding affinity of the multivalent allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to a monovalent allergen-binding protein.

58. Allergen-binding protein, according to any one of claims 1 to 57, characterized in that it further comprises a carrier.

59. Allergen-binding protein, according to claim 58, characterized in that the carrier comprises a solvent, a diluent, a dispersion medium or a coating.

60. Allergen-binding protein according to claim 59, characterized in that the carrier comprises silica.

61. Allergen-binding protein, according to any one of claims 1 to 60, characterized in that the allergen-binding protein is in a dry form.

62. Allergen-binding protein according to claim 58, characterized in that the carrier comprises water.

63. Allergen-binding protein, according to any one of claims 1 to 62, characterized in that the allergen-binding protein comprises a liquid. Petition 870250094198, dated 10 / 15 / 2025, page 20 / 38 10 / 25 64. Allergen-binding protein, according to any one of claims 1 to 63, characterized in that the allergen-binding protein additionally comprises a preservative.

65. Allergen-binding protein, according to claim 64, characterized in that the preservative comprises potassium sorbate, EDTA, benzoic acid, phenoxyethanol or maltol.

66. Allergen-binding protein, according to any one of claims 1 to 65, characterized in that the allergen-binding protein further comprises a stabilizing or thickening agent.

67. Allergen-binding protein, according to claim 66, characterized in that the stabilizing or thickening agent comprises dextrin, maltodextrin, glycerol, glucose, sucrose or trehalose.

68. Allergen-binding protein, according to any one of claims 1 to 67, characterized in that the allergen-binding protein additionally comprises an isotonic agent.

69. Allergen-binding protein, according to claim 1 or 2, characterized in that the allergen-binding protein is suspended or resuspended in a solvent.

70. Allergen-binding protein according to claim 69, characterized in that the solvent comprises water.

71. Composition characterized in that it comprises: an allergen-binding protein that binds to or neutralizes an allergen, wherein the allergen-binding protein comprises a nanobody, a monobody, a DARPin or a small peptide with a molecular weight of less than about 25 kilodaltons or a length of less than about 300 amino acids.

72. Composition characterized in that it comprises: an allergen-binding protein that binds to or neutralizes an allergen, wherein the allergen-binding protein comprises a nanobody or small body with a molecular weight of less than about 25 kilodaltons or a length of less than about 300 amino acids, wherein the allergen-binding protein comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs: 1-57 and 67-73.

73. Composition according to claim 71 or 72, characterized in that the composition is formulated to coat a food or to spray, mist or brush onto an animal or domestic surface.

74. Composition according to claim 73, characterized in that the food is a pet food.

75. Composition according to claim 74, characterized in that the allergen is a pet allergen.

76. Composition, according to any one of claims 73 to 75, characterized in that the allergen-binding protein is formulated to be applied to the food as a coating.

77. Composition, according to any one of claims 73 to 75, characterized in that the allergen-binding protein is formulated to be mixed into food.

78. Composition, according to any one of claims 71 to 73, characterized in that the allergen-binding protein is in a concentration of about 0.01 milligrams per milliliter (mg / ml) to about 500 mg / ml in solution or after suspension.

79. Composition, according to any one of claims 71 to 74, characterized in that the allergen induces an allergic reaction in a human.

80. Composition, according to any one of claims 71 to 79, characterized in that the allergen comprises an environmental allergen.

81. Composition, according to any one of claims 71 to 80, characterized in that the allergen comprises an animal allergen.

82. Composition, according to any one of claims 71 to 81, characterized in that the allergen comprises a pet allergen.

83. Composition, according to any one of claims 71 to 82, characterized in that the allergen comprises a cat allergen, a dog allergen, a rabbit allergen, a mouse allergen or a cockroach allergen.

84. Composition according to claim 83, characterized in that the allergen is selected from the group consisting of Fel d 1, Fel d 2, Fel d 3, Fel d 4, Can f 1, Can f 2, Can f 4, Can f 7, Ory C 1, Mus M 1 and Bla G 2.

85. Composition according to claim 84, characterized in that the allergen is Fel d 1.

86. Composition according to claim 85, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 49-57.

87. Composition according to claim 86, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO:

49. Petition 870250094198, dated 10 / 15 / 2025, p. 23 / 38 13 / 25 88. Composition according to claim 86, characterized in that the allergen-binding protein comprises a nanobody with a sequence SEQ ID NO:

49.

89. Composition according to claim 86, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence of SEQ ID NO:

57.

90. Composition according to claim 86, characterized in that the allergen-binding protein comprises a nanobody with a sequence SEQ ID NO:

57.

91. Composition according to claim 84, characterized in that the allergen is Can f 1.

92. Composition, according to claim 91, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 67-73.

93. Composition according to claim 92, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

67.

94. Composition according to claim 92, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

67.

95. Composition, according to claim 84, Petition 870250094198, dated 10 / 15 / 2025, page 24 / 38 14 / 25 characterized in that the allergen is Can f 2.

96. Composition, according to claim 95, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 9-23.

97. Composition according to claim 95, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

16.

98. Composition according to claim 97, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence consisting of SEQ ID NO:

16.

99. Composition, according to any one of claims 71 to 80, characterized in that the allergen comprises a dust allergen.

100. Composition according to claim 99, characterized in that the dust allergen comprises Der p1 or Der p2.

101. Composition according to claim 100, characterized in that the dust allergen is Der p1.

102. Composition, according to claim 101, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group Petition 870250094198, dated 10 / 15 / 2025, page 25 / 38 15 / 25 consisting of SEQ ID Nos: 24-34.

103. Composition according to claim 101, characterized in that the allergen-binding protein comprises a peptide with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs 35-48.

104. Composition according to claim 101, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

27.

105. Composition according to claim 104, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

27.

106. Composition according to claim 101, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

28.

107. Composition according to claim 104, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

28.

108. Composition according to claim 100, characterized in that the dust allergen is Der p2.

109. Composition, according to claim 106, characterized in that the allergen-binding protein Petition 870250094198, 10 / 15 / 2025, page 26 / 38 16 / 25 comprises a nanobody with at least 80%, at least 85%, at least 90% or 100% sequence identity with a sequence selected from the group consisting of SEQ IDs NOS: 1-8.

110. Composition according to claim 107, characterized in that the allergen-binding protein comprises a nanobody comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% or 100% sequence identity with a sequence established in SEQ ID NO:

3.

111. Composition according to claim 108, characterized in that the allergen-binding protein comprises a nanobody with a sequence consisting of SEQ ID NO:

3.

112. Composition, according to any one of claims 71 to 80, characterized in that the allergen comprises a plant allergen or a plant pollen allergen.

113. Composition according to claim 112, characterized in that the plant or pollen allergen is selected from the group consisting of Bet v1, Phl p 5, Phl p 1, Poa p 1, Cyn d 1, Bet v 2, Ole e 1, Amb a 1, Amb a 11 and Art v 1.

114. Composition, according to any one of claims 71 to 80, characterized in that the allergen comprises a mold allergen.

115. Composition according to claim 114, characterized in that the mold allergen is selected from the group consisting of Alt a 1, Asp f 1, Asp f 2, Cla h 8, Pen ch 13 and Pen ch 18.

116. Composition, according to any one of claims 72 to 80, characterized in that the allergen is a food allergen.

117. Composition, according to claim 116, characterized in that the food allergen is selected from the group consisting of Pen a 1, Ara h 1, Ara h 3.

118. Composition, according to any one of claims 71 to 117, characterized in that the allergen-binding protein comprises at least one amino acid modification.

119. Composition according to claim 118, characterized in that the allergen-binding protein comprises about 1, about 5, about 10, about 15, about 20, about 25 or about 30 amino acid modifications.

120. Composition according to claim 118 or 119, characterized in that the amino acid modification comprises the introduction of a non-canonical amino acid.

121. Composition according to claim 120, characterized in that the non-canonical amino acid is selected from the group consisting of para-benzoylphenylalanine, 3,4-dihydroxyphenylalanine, a tetrazine, chloroctenes, homopropargylglycine, para-proparglyoxyphenylalanine, paraazidophenylalanine, para-isothiocyanate phenylalanine, parabenzoylphenylalanine, para-cyanophenylalanine, paranitrophenylalanine, a halogenated m-tyrosine analog, a halogenated proline analog, a halogenated tryptophan analog and a halogenated leucine analog.

122. Composition, according to any one of claims 118 to 121, characterized in that the solubility of the composition is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600% or more compared with an unmodified allergen-binding protein.

123. Composition, according to any one of claims 118 to 121, characterized in that the binding affinity of the allergen-binding protein is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared with an unmodified allergen-binding protein.

124. Composition, according to any one of claims 118 to 121, characterized in that the thermal stability of the allergen-binding protein is increased by about 10%, 25%, 50%, 75%, 100%, 200%, 300% or more compared with an unmodified allergen-binding protein.

125. Composition, according to any one of claims 71 to 124, characterized in that the allergen-binding protein is a multivalent allergen-binding protein.

126. Composition according to claim 125, characterized in that the multivalent allergen-binding protein is a bivalent allergen-binding protein.

127. Composition according to claim 124 or 125, characterized in that the binding affinity of the multivalent allergen-binding protein composition is increased by about 25%, 50%, 75%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 1500% or more compared to a monovalent allergen-binding protein.

128. Composition, according to any one of claims 71 to 127, characterized in that it further comprises a carrier.

129. Composition, according to any one of claims 71 to 128, characterized in that the carrier comprises a solvent, a diluent, a dispersion medium or a coating.

130. Composition according to claim 128, characterized in that the carrier comprises silica.

131. Composition, according to any one of claims 71 to 130, characterized in that the composition is in a dry form.

132. Composition, according to claim 128, Petition 870250094198, dated 10 / 15 / 2025, pp. 29 / 38 19 / 25 characterized by the fact that the carrier comprises water.

133. Composition, according to any one of claims 71 to 132, characterized in that the composition comprises 134. Composition, a liquid, according to any one of claims 71 to 133, characterized in that the composition comprises 135. Composition, additionally a preservative, according to claim 134, characterized in that the preservative comprises potassium sorbate, EDTA, benzoic acid, phenoxyethanol or maltol.

136. Composition, according to any one of claims 71 to 135, characterized in that the composition further comprises a stabilizing or thickening agent.

137. Composition according to claim 136, characterized in that the stabilizing or thickening agent comprises dextrin, maltodextrin, glycerol, glucose, sucrose or trehalose.

138. Composition, according to any one of claims 71 to 137, characterized in that the composition further comprises an isotonic agent.

139. Composition according to claim 71 or 72, characterized in that the composition is suspended or resuspended in a solvent.

140. Composition according to claim 139, characterized in that the solvent comprises water.

141. A method characterized in that it comprises: spraying or spraying the allergen-binding protein, as defined in any one of claims 3 to 70, or the composition, as defined in any one of claims 73 to 140, or placing the surface in contact with the composition, as defined in any one of claims 73 to 140. Petition 870250094198, dated 10 / 15 / 2025, pp. 30 / 38 20 / 25 142. Method according to claim 141, characterized in that the contact comprises coating or brushing.

143. Method according to claim 141 or 142, characterized in that the surface comprises the surface of an air filter or a humidifier.

144. Method, according to any one of claims 141 to 143, characterized in that the surface comprises the allergen.

145. Method, according to any one of claims 141 to 144, characterized in that the composition neutralizes the allergen on the surface.

146. Method, according to any one of claims 141 to 145, characterized in that the surface comprises food.

147. Method according to claim 146, characterized in that the food is consumed by an animal that is the source of the antigen.

148. A method according to any one of claims 141 to 145, characterized in that the surface comprises a pet accessory (e.g., a collar or brush) or an area / product where an animal can sit or walk (e.g., a dog bed).

149. Method, according to any one of claims 141 to 148, characterized in that the nebulization is carried out by a humidifier or a sprayer.

150. Method, according to any one of claims 141 to 149, characterized in that about 0.01 milliliter (ml) to about 20 ml of the composition sprayed per about 1 square meter of area neutralizes the allergenicity of the antigen.

151. Method for preparing allergen-binding protein, as defined in any one of claims 1 to 140, or the composition, as defined in any one of claims 71 to 140, characterized in that Petition 870250094198, dated 10 / 15 / 2025, page 31 / 38 21 / 25, comprises: harvesting allergen-binding protein from an engineered microbe or from a secretion of the microbe, wherein the microbe comprises a heterologous nucleic acid encoding the allergen-binding protein.

152. Method according to claim 151, characterized in that it further comprises incorporating the heterologous nucleic acid encoding the allergen-binding protein into a cell-free protein expression system.

153. Method according to claim 151, characterized in that the microbe comprises a yeast or bacterium.

154. Method according to claim 153, characterized in that the microbe comprises a bacterium and comprises E. coli.

155. Method according to claim 153, characterized in that the microbe comprises a yeast and comprises Pichia pastoris.

156. Method according to claim 151, characterized in that the allergen-binding protein is purified or concentrated from a secretion by the microbe.

157. Method according to claim 154, characterized in that the purification or concentration comprises a filtration step.

158. Method according to claim 157, characterized in that the filtration step comprises passing the secretion through a filter of about 0.01 nm, 0.1 nm, 1 nm, 5 nm or more in size.

159. Method, according to any one of claims 151 to 158, characterized in that the method does not comprise a centrifugation step.

160. Method for treating an allergy in an individual in need, the method characterized in that it comprises administering to the individual the allergen-binding protein, Petition 870250094198, dated 10 / 15 / 2025, page 32 / 38 22 / 25 as defined in any one of claims 1 to 70 or the composition, as defined in any one of claims 71 to 140.

161. Method for treating an allergy in an individual in need, the method characterized in that it comprises (i) administering to the individual a therapeutically effective amount of a microbe designed to produce the allergen-binding protein as defined in any one of claims 1 to 70 or the composition as defined in any one of claims 71 to 140.

162. Method according to claim 161, characterized in that the microbe is a yeast or bacterium.

163. Method according to claim 162, characterized in that the microbe comprises a bacterium and comprises E. coli.

164. Method according to claim 162, characterized in that the microbe comprises a yeast and comprises Pichia pastoris.

165. Method according to claim 161 or 162, characterized in that the microbe is administered to the individual orally.

166. A method according to any one of claims 161 to 163, characterized in that the microbe produces the allergen-binding protein or composition when ingested.

167. Method for neutralizing an allergen, the method characterized in that it comprises (i) aerosolizing the allergen-binding protein, as defined in any one of claims 3 to 70, or the composition, as defined in any one of claims 73 to 140, into a mist and (ii) bringing the mist into contact with a surface comprising an allergen.

168. Method, according to claim 167, characterized in that the aerosolization is carried out by a machine of Petition 870250094198, of 10 / 15 / 2025, page 33 / 38 23 / 25 aerosolization.

169. Method according to claim 168, characterized in that the aerosolization machine is used by an individual.

170. A method according to any one of claims 167 to 169, characterized in that the surface comprises a pet accessory or an area / product where an animal can sit or walk.

171. Method for neutralizing an allergen, the method characterized in that it comprises (i) aerosolizing the allergen-binding protein, as defined in any one of claims 3 to 70, or the composition, as defined in any one of claims 73 to 140, into a mist and (ii) bringing the mist into contact with a food.

172. Method according to claim 171, characterized in that the food is a pet food.

173. Method according to claim 171, characterized in that the allergen is a pet allergen.

174. A method according to any one of claims 171 to 173, characterized in that the mist is applied over the food as a coating.

175. Method, according to any one of claims 171 to 173, characterized in that the mist is mixed with the food.

176. A method for neutralizing an allergen, the method characterized in that it comprises contacting the allergen-binding protein, as defined in any one of claims 1 to 70, or the composition, as defined in any one of claims 71 to 140, with a food.

177. Method according to claim 176, characterized in that the food is a pet food. Petition 870250094198, dated 10 / 15 / 2025, pp. 34 / 38 24 / 25 178. Method according to claim 176, characterized in that the allergen is a pet allergen.

179. A method according to any one of claims 176 to 178, characterized in that the allergen-binding protein or composition is applied to the food as a coating.

180. Method, according to any one of claims 176 to 178, characterized in that the allergen-binding protein or composition is mixed into the food.

181. A method for treating an allergy in an individual in need, the method characterized in that it comprises aerosolizing the allergen-binding protein, as defined in any one of claims 3 to 7, or the composition, as defined in any one of claims 73 to 140, into a mist.

182. Method according to claim 171, characterized in that the aerosolization is carried out by an aerosolization machine.

183. Method according to claim 171 or 182, characterized in that an aerosolization machine is used.

184. Method according to claim 183, characterized in that the aerosolization machine is used by the individual.

185. A method, according to any one of claims 171 to 184, characterized in that the method further comprises the individual inhaling the mist.

186. Method, according to any one of claims 171 to 185, characterized in that the individual is a human.

187. Method, according to any one of claims 171 to 185, characterized in that the individual is a non-human animal. Petition 870250094198, dated 10 / 15 / 2025, pp. 35 / 38 25 / 25 188. Pharmaceutical composition characterized in that it comprises (i) the allergen-binding protein, as defined in any one of claims 1 to 70 or the composition, as defined in any one of claims 71 to 140 and (ii) a pharmaceutically acceptable excipient.

189. Allergen-binding protein characterized in that it comprises a sequence of either of the following SEQ ID numbers: 1-57 or 67-73.

190. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO:

3.

191. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO:

16.

192. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO:

27.

193. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO:

28.

194. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO:

49.

195. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO:

57.

196. Allergen-binding protein characterized in that it comprises a sequence with SEQ ID NO: 67.

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