Use of specific properties of allergens, allergens from animal or botanical sources and methods for their isolation

• US 5,667,979 A
• Filed: 01/05/1990
• Issued: 09/16/1997
• Est. Priority Date: 01/05/1989
• Status: Expired due to Term

• Current Assignee: Laboratorios Lesvi, S.L.
• Original Assignee: Laboratories LEO SA
• Inventors: Berrens, Lubertus
• Primary Examiner(s): Saunders, David
• Application Number:
  US07/461,642
• Time in Patent Office: 2,811 Days
• Field of Search: 435/7.4, 435/23, 435/24, 435/18, 436/513, 436/518
• US Class Current: 435/7.4
• CPC Class Codes: A61K 39/35 Allergens
• CPC Class Codes: C12N 9/00 Enzymes; Proenzymes; Compositions thereof preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113; Processes for preparing, activating, inhibiting, separating or purifying enzymes preparation of malt C12C1/00
• CPC Class Codes: C12N 9/14 Hydrolases (3)
• CPC Class Codes: C12Q 1/34 involving hydrolase

Abstract

It was found that atopic allergens have enzymatic properties, in particular the properties to hydrolyze amide and/or ester linkages. These properties may be used for various purposes, e.g. for analysis of samples, standardization of pharmaceutical compositions and also for the preparation of the allergens in a pure form.

Claims

1. A method of hydrolyzing amide and/or ester linkages in a substrate, comprising:

contacting a substantially purified form of an atopic allergen having protease properties with a substrate having amide and/or ester linkages to thereby hydrolyze said linkages, wherein said allergen is isolated from mammalian and non-mammalian excrement sources, and is capable of hydrolyzing proteins and split substrates specific for enzyme active sites.

2. Method according to claim 1, wherein the allergen originates from mammals and is isolated from the group consisting of saliva, sweat, dry skin scales, and urine, said allergen being capable of splitting low-molecular weight substrates specific for tissue kallikreins.

3. Method according to claim 1, wherein the substrates are peptide derivatives comprising an amino acid or amino acid sequence designed to fit the active site of proteases with the substrate-specificity of human and bovine trypsin and chymotrypsin, or designed to fit the active site of proteases with the substrate-specificity of animal glandulary kallikreins.

4. Method according to claim 3, wherein the peptide derivatives are the naphthylamides of monoamino, dipeptide, tripeptide or oligopeptide compounds.

5. Method according to claim 3, wherein the substrates are alkylesters of naphthyl, p-benzoyl or p-tosyl chromogenic conjugates of monoamino, dipeptide, tripeptide or oligopeptide derivatives comprising an amino acid or amino acid sequence designed to fit the active site of proteases with the substrate-specificity of human and bovine trypsin and chymotrypsin or with the substrate-specificity of animal glandulary kallikreins respectively.

6. Method according to claim 1, wherein the enzymatic substrates have been conjugated to a fluophoric group.

7. Method of establishing the allergenic potency of environmental dust samples or individual allergenic preparations, comprising:

preparing an aqueous extract of an environmental allergen or allergen preparation, and quantitatively measuring the protease activity of said aqueous extract towards different specific substrates, said substrates having amide and/or ester linkages.

8. A kit for establishing the allergenic presence of environmental dust samples or individual allergenic preparations, comprising:

a carrier containing a substrate having amide and/or ester linkages, said carrier adapted to detect the protease activity of an aqueous extract of an environmental allergen, or of an allergenic preparation.

9. A kit according to claim 8, wherein the carrier is in the form of a strip.