Claims for Patent: 5,091,318

Summary for Patent: 5,091,318

Title:	Binding of allergens to a solid phase
Abstract:	A method for producing a binding assay device composed of antigens on a cellulose nitrate, cellulose nitrate/acetate or similar solid phase is described. The method involves applying to a solid phase a small amount of an allergen composition, or a pretreated allergen composition, containing a certain concentration of allergen and drying the solution. The device is used by contacting a patient test sample to the immobilized allergen and determining whether or not the test sample contains IgE antibodies for the allergen.
Inventor(s):	Anawis; Mark A. (Grayslake, IL), Lindberg; Roger E. (Libertyville, IL)
Assignee:	Abbott Laboratories (Abbott Park, IL)
Application Number:	07/509,255
Patent Claims:	1. A device, for detecting the presence or amount of IgE in a test sample, comprising: a) a solid phase comprising a material selected from the group consisting of: nitrocellulose, nitrocellulose derivatives, nitrocellulose compounds, or combinations thereof, and b) at least one allergen immobilized upon said solid phase, wherein said allergen was applied as an allergen composition and wherein said allergen composition is formed from the combination of said allergen with a pretreatment substance selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents; crosslinking agents; concentrated salt solutions; and combinations thereof. 2. The device according to claim 1, wherein said pretreatment substance is hydrochloric acid or acetic acid. 3. The device according to claim 1, wherein said pretreatment substance is tetrahydrofuran. 4. The device according to claim 1, wherein said pretreatment substance is a concentrated sodium chloride solution. 5. The device according to claim 1, wherein said pretreatment substance is formaldehyde, glutaraldehyde or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. 6. The device according to claim 1, where said solid phase is selected from the group consisting of cellulose, cellulose derivatives, silica, fiberglass, a porous polymer matrix, porous gels, polymeric films, agarose and porous fibrous matrixes. 7. The device according to claim 1, where said solid phase is selected from the group consisting of cellulose acetate, nitrocellulose and cellulose acetate/nitrate mixed ester cellulose. 8. The device according to claim 1, further comprising a protein blocking reagent on said solid phase. 9. A method for producing a device according to claim 1, comprising the steps of: a) forming the allergen composition by pretreating the allergen with a substance selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents; crosslinking agents; concentrated salt solutions; and combinations thereof b) applying said allergen composition to said solid phase, and c) drying said allergen composition on said solid phase, thereby immobilizing said allergen upon said solid phase. 10. The method according to claim 9, further comprising the step of applying a protein blocking reagent to said solid phase. 11. The method according to claim 10, wherein said protein blocking reagent is selected from the group consisting of equine serum albumin, bovine serum albumin, fish gelatin and casein. 12. The method according to claim 11, wherein said pretreatment substance is hydrochloric acid or acetic acid. 13. The method according to claim 11, wherein said pretreatment substance is tetrahydrofuran. 14. The method according to claim 11, wherein said pretreatment substance is a concentrated sodium chloride solution. 15. The method according to claim 11, wherein said pretreatment substance is formaldehyde, glutaraldehyde or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. 16. The method according to claim 9, where said solid phase is selected from the group consisting of cellulose, cellulose derivatives, silica, fiberglass, a porous polymer matrix, porous gels, polymeric films, agarose and porous fibrous matrixes. 17. The method according to claim 9, where said solid phase is selected from the group consisting of cellulose acetate, nitrocellulose and cellulose acetate/nitrate mixed ester cellulose. 18. The device of claim 1, wherein the allergen composition contains, on the basis of allergen protein in water, i) from 0.05 to 4.0 mg/mL of Altenaria allergen, ii) from 0.05 to 50 mg/mL of Aspergillus allergen, iii) from 0.8 to 81.6 mg/mL of Bermuda grass allergen, iv) from 0.1 to 6.0 mg/mL of birch allergen, v) from 0.6 to 20.6 mg/mL of cat allergen, vi) from 0.04 to 4.5 mg/mL of mountain cedar allergen, vii) from 0.1 to 20.5 mg/mL of Japanese cedar allergen, viii) from 0.05 to 38.4 mg/mL of Cladosporium allergen, ix) from 1.3 to 38.4 mg/mL of dog allergen, x) from 0.7 to 22.4 mg/mL of D. farinase allergen, xi) from 0.6 to 84.2 mg/mL of D. pteronyssinus allergen, xii) from 0.1 to 146.0 mg/mL of elm allergen, xiii) from 0.02 to 0.2 mg/mL of feather allergen, xiv) from 0.2 to 148.2 mg/mL of giant ragweed allergen, xv) from 0.4 to 100 mg/mL of house dust allergen, xvi) from 0.05 to 21.8 mg/mL of June/Kentucky bluegrass allergen, xvii) from 0.2 to 47.0 mg/mL of lamb's quarters allergen, xviii) from 0.1 to 166.3 mg/mL of maple allergen, xix) from 0.3 to 90.4 mg/mL of mugwort allergen, xx) from 0.1 to 12 mg/mL of mulberry allergen, xxi) from 0.2 to 29.2 mg/mL of oak allergen, xxii) from 0.1 to 66.8 mg/mL of olive allergen, xxiii) from 1.0 to 40.0 mg/mL of Parietaria allergen, xxiv) from 1.7 to 130.4 mg/mL of plantain allergen, xxv) from 0.1 to 4.8 mg/mL of Penicillium allergen, xxvi) from 0.05 to 17.3 mg/mL of perennial rye allergen, xxvii) from 0.2 to 151.6 mg/mL of short ragweed allergen, or xxviii) from 0.05 to 6.6 mg/mL of timothy allergen,. 19. An allergen composition, comprising: a) a solvent; and b) an allergen solubilized in said solvent, thereby forming an allergen solution; and c) wherein said allergen solution is combined with a pretreatment substance to form an allergen composition; wherein said pretreatment substance is selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents; crosslinking agents; concentrated salt solutions; and combinations thereof; the allergen composition is used for the in vitro detection of the presence or amount of IgE in a test sample. 20. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of Altenaria allergen, birch allergen and dog allergen; and b) said pretreatment substance is a formaldehyde solution. 21. The allergen composition according to claim 20, wherein said pretreatment substance is about 10 to about 20 microliters of a 37 percent formaldehyde solution. 22. The allergen composition according to claim 20, wherein said pretreatment substance is about 12.5 microliters of a 37 percent formaldehyde solution. 23. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of Bermuda grass allergen; Japanese cedar allergen; June/Kentucky bluegrass allergen; perennial rye allergen; and timothy allergen; and b) said pretreatment substance is tetrahydrofuran. 24. The allergen composition according to claim 23, wherein said pretreatment substance is about 10 to about 50 microliters of tetrahydrofuran. 25. The allergen composition according to claim 23, wherein said pretreatment substance is about 25 microliters of tetrahydrofuran. 26. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of mountain cedar allergen; oak allergen and olive allergen; and b) said pretreatment substance comprises tetrahydrofuran and a formaldehyde solution. 27. The allergen composition according to claim 26, wherein said pretreatment substance comprises about 10 to about 50 microliters of tetrahydrofuran and about 10 to about 20 microliters of a 37 percent formaldehyde solution. 28. The allergen composition according to claim 26, wherein said pretreatment substance comprises about 25 microliters of tetrahydrofuran and about 15 microliters of a 37 percent formaldehyde solution. 29. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of Cladosporium allergen and feather allergen; and b) said pretreatment substance is about 0.5M to about 10M sodium chloride solution. 30. The allergen composition according to claim 29, wherein said pretreatment substance is about 10 to about 20 microliters of 5M sodium chloride. 31. The allergen composition according to claim 29, wherein said pretreatment substance is about 12 microliters of 5M sodium chloride. 32. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of Dermatophagoides farinae allergen and D. pteronyssinus allergen; and b) said pretreatment substance comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and sodium borohydride. 33. The allergen composition according to claim 32, wherein said pretreatment substance comprises about 5.0 to about 15 microliters of 50 mg/mL 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and about 1.0 to about 5.0 microliters of 20 mg/mL sodium borohydride. 34. The allergen composition according to claim 32, wherein said pretreatment substance comprises about 10 microliters of 50 mg/mL 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and about 2.0 microliters of 20 mg/mL sodium borohydride. 35. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of lamb's quarters allergen and mulberry allergen; and b) said pretreatment substance comprises acetic acid, buffered to about a pH of 7. 36. The allergen composition according to claim 35, wherein said pretreatment substance comprises about 5.0 to about 30 microliters of acetic acid and 6N aqueous sodium hydroxide at about a pH of 7. 37. The allergen composition according to claim 35, wherein said pretreatment substance comprises about 12.5 microliters of acetic acid and 6N aqueous sodium hydroxide to a pH of 7. 38. The allergen composition according to claim 19, wherein: a) said allergen is selected from the group consisting of Parietaria allergen and Penicillium allergen; and b) said pretreatment substance comprises hydrochloric acid, buffered to about a pH of 7. 39. The allergen composition according to claim 38, wherein said pretreatment substance comprises about 6.0 to about 30 microliters of 6N hydrochloric acid and 6N aqueous sodium hydroxide at about a pH of 7. 40. The allergen composition according to claim 38, wherein said pretreatment substance comprises about 24 microliters of 6N hydrochloric acid and 6N aqueous NaOH at about a pH of 7. 41. An allergen composition, comprising: a) a solvent; b) an allergen solubilized in said solvent, thereby forming an allergen solution; and c) a pretreatment substance selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents; crosslinking agents; concentrated salt solutions; and combinations thereof; wherein said allergen solution is combined with said pretreatment substance to form an allergen composition, and wherein the allergen composition is used for the in vitro detection of the presence or amount of IgE in a test sample. 42. A method for determining the presence or amount of IgE in a test sample, comprising the steps of: a) providing at least one allergen immobilized upon a solid phase comprising a material selected from the group consisting of nitrocellulose, nitrocellulose derivatives, nitrocellulose compounds, and combinations thereof, wherein said allergen is applied to said solid phase as an allergen composition; said allergen composition being formed from combining an allergen with a pretreatment substance; wherein said pretreatment substance is selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents; cross-linking agents, concentrated salt solutions; and combinations thereof containing, on the basis of allergen protein in a solvent, i) from 0.05 to 4.0 mg/mL of Altenaria allergen, ii) from 0.05 to 50 mg/mL of Aspergillus allergen, iii) from 0.8 to 81.6 mg/mL of Bermuda grass allergen, iv) from 0.1 to 6.0 mg/mL of birch allergen, v) from 0.6 to 20.6 mg/ml of cat allergen, vi) from 0.04 to 4.5 mg/ml of mountain cedar allergen, vii) from 0.1 to 20.5 mg/ml of Japanese cedar allergen, viii) from 0.05 to 38.4 mg/ml of Cladosporium allergen, ix) from 1.3 to 38.4 mg/ml of dog allergen, x) from 0.7 to 22.4 mg/ml of D. farinase allergen, xi) from 0.6 to 84.2 mg/ml of D. pteronyssinus allergen, xii) from 0.1 to 146.0 mg/ml of elm allergen, xiii) from 0.02 to 0.2 mg/ml of feather allergen, xiv) from 0.2 to 148.2 mg/ml of giant ragweed allergen, xv) from 0.4 to 100 mg/ml of house dust allergen, xvi) from 0.05 to 21.8 mg/ml of June/Kentucky bluegrass allergen, xvii) from 0.2 to 47.0 mg/ml of lamb's quarters allergen, xviii) from 0.1 to 166.3 mg/ml of maple allergen, xix) from 0.3 to 90.4 mg/ml of mugwort allergen, xx) from 0.1 to 12 mg/ml of mulberry allergen, xxi) from 0.2 to 29.2 mg/ml of oak allergen, xxii) from 0.1 to 66.8 mg/ml of olive allergen, xxiii) from 1.0 to 40.0 mg/ml of Parietaria allergen, xxiv) from 1.7 to 130.4 mg/ml of plantain allergen, xxv) from 0.1 to 4.8 mg/ml of Penicillium allergen, xxvi) from 0.05 to 17.3 mg/ml of perennial rye allergen, xxvii) from 0.2 to 151.6 mg/ml of short ragweed allergen, or xxviii) from 0.05 to 6.6 mg/ml of timothy allergen, and b) contacting said test sample to said solid phase, thereby immobilizing allergen-specifific IgE antibody from the test sample upon said solid phase by forming allergen/antibody complexes; and c) detecting said immobilized allergen-specific antibody to determine the presence or amount of the antibody in the test sample. 43. The method according to claim 42, wherein step c) comprises contacting said solid phase with an indicator reagent to determine the presence or amount of IgE in the test sample. 44. The method according to claim 43, wherein said indicator reagent comprises a label conjugated to a binding member specific for a member selected from the group consisting of allergen, IgE and an ancillary specific binding member. 45. The method according to claim 44, wherein said indicator reagent comprises a label conjugated to an anti-IgE antibody or anti-IgE antibody fragment. 46. The method according to claim 45, wherein free or bound labeled anti-IgE antibody is detected to determine the presence or amount of IgE in the test sample. 47. The method according to claim 44, wherein said label is a member selected from the group consisting of chromogens, catalysts, fluorescent compounds, chemiluminescent compounds, radioactive isotopes, colloidal metallic particles, colloidal selenium particles, dye particles, enzymes, substrates, organic polymer latex particles and liposomes or other vesicles containing signal producing components. 48. The method according to claim 43, further comprising the step of washing unbound indicator reagent from said solid phase prior to detecting the presence or amount of IgE in the test sample. 49. The method according to claim 42, wherein said solid phase is a member selected from the group consisting of cellulose, cellulose derivatives, silica, fiberglass, a porous polymer matrix, porous gels, polymeric films, agarose and porous fibrous matrixes. 50. A method for determining the presence or amount of IgE in a test sample, comprising the steps of: a) providing at least one allergen immobilized upon a solid phase, wherein said solid phase comprises a material selected from the group consisting of nitrocellulose, nitrocellulose derivatives, nitrocellulose compounds, and combinations thereof, wherein said allergen is applied to said solid phase as an allergen composition comprising.: i) a solvent; ii) an allergen solubilized in said solvent, thereby forming an allergen solution; and iii) a pretreatment substance selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents, crosslinking agents; concentrated salt solutions, and combinations thereof, wherein said allergen solution is combined with said pretreatment substance to form an allergen composition; b) contacting the test sample to said solid phase, thereby immobilizing allergen-specific IgE antibody from the test sample upon said solid phase by forming allergen/antibody complexes; and c) detecting said immobilized allergen-specific antibody to determine the presence or amount of the antibody in the test sample. 51. A kit for determining the presence or amount of IgE in a test sample, comprising: a) an allergen immobilized upon a solid phase comprising a material selected from the group consisting of nitrocellulose, nitrocellulose derivatives, nitrocellulose compounds, and combinations thereof, wherein said allergen is applied to said solid phase as an allergen composition comprising: i) a solvent; ii) an allergen solubilized in said solvent, thereby forming an allergen solution; and iii) a pretreatment substance selected from the group consisting of: denaturants excluding organic solvents and concentrated salt solutions; organic solvents; crosslinking agents; concentrated salt solution,; and combinations thereof, wherein said allergen solution is combined with said pretreatment substance to form an allergen composition; and b) an indicator reagent in a container, wherein said indicator reagent is used to determine the presence or amount of IgE in the test sample, wherein said indicator reagent comprises a label conjugated to a binding member specific for a member selected from the group consisting of the allergen, IgE and an ancillary specific binding member.

Details for Patent 5,091,318

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Greer Laboratories, Inc.	N/A	insects (whole body), mite dermatophagoides farinae	Injection	101834	09/15/1958		2040-01-28
Allermed Laboratories, Inc.	N/A	pollens - weeds and garden plants, ragweed, short ambrosia artemisiifolia	Injection	103113	03/13/1974		2040-01-28