METHOD FOR THE INACTIVATION AND INACTIVATION TESTING OF XENO ANTIGENS IN FOODS OF ANIMAL ORIGIN, PARTICULARLY FOR MILK AND DERIVATIVES, AND IN FOODS OF PLANT ORIGIN, PARTICULARLY FOR MILK SUBSTITUTES BASED ON SOY AND/OR RICE

• US 20190317069A1
• Filed: 11/14/2017
• Published: 10/17/2019
• Est. Priority Date: 11/16/2016
• Status: Active Grant

• Current Assignee: Biocompatibility Innovation Srl
• Original Assignee: Biocompatibility Innovation Srl
• Inventors: Naso, Filippo, Gandaglia, Alessandro, Stefanelli, Ugo
• Granted Patent: US 11,366,090 B2
• Time in Patent Office: Days
• CPC Class Codes: A23C 7/04 Removing unwanted substances other than lactose or milk proteins from milk
• CPC Class Codes: A23L 33/00 Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
• CPC Class Codes: A23L 5/20 Removal of unwanted matter, e.g. deodorisation or detoxification
• CPC Class Codes: A23V 2002/00 Food compositions, function of food ingredients or processes for food or foodstuffs
• CPC Class Codes: A61K 39/35 Allergens
• CPC Class Codes: G01N 33/025 Fruits or vegetables
• CPC Class Codes: G01N 33/04 Dairy products
• CPC Class Codes: G01N 33/08 Eggs, e.g. by candling
• CPC Class Codes: G01N 33/12 Meat; Fish
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q

Abstract

A method for the inactivation and inactivation testing of xenoantigens in foods of vegetable and animal origin, comprising the following steps: making up a solution with a food of vegetable or animal origin as a solvent and one or more phenolic compounds, polyphenolic compounds and derivatives thereof, comprising phenylpropanoids, as a solute, for the inactivation of at least part of the xenogeneic epitopes from said food, incubating samples of the food of vegetable or animal origin with the addition of an antibody aimed at a xenoantigen epitope that is present in the food, separating the resulting immune complex created owing to the bond between antigen and antibody, preparing a well plate for the E.L.I.S.A. test with coating with xenoantigen epitope, adding, in the wells, supernatant taken from the samples, the supernatant containing the part of antibody that has not bonded with epitopes, a column of wells being adapted to define a reference value that corresponds to the maximum signal between antibody and epitopes, completing the plate with a secondary antibody conjugated with an enzyme, or other molecule, adapted to chromatically highlight any presence of anti-xenoantigen antibody, reading the plate, determining the presence of anti-xenoantigen antibody that has remained free in the solutions of the samples, comparing the absorbance values detected in the reference column with the values in the other columns of samples of the plate.

Claims

1. A method for the inactivation and inactivation testing of xenoantigens in foods of vegetable and animal origin, characterized in that it comprises the following steps:

making up a solution with a food of vegetable or animal origin as a solvent and one or more phenolic compounds, polyphenolic compounds and derivatives thereof, comprising phenylpropanoids, as a solute, for the inactivation of at least part of the xenogeneic epitopes from said food;

incubating samples of said food of vegetable or animal origin with the addition of an antibody aimed at a xenoantigen epitope that is present in said food;

separating the resulting immune complex created owing to the bond between antigen and antibody;

preparing a well plate for the E.L.I.S.A. test with coating comprising said xenoantigen epitope,

adding, in said wells, supernatant taken from said samples, said supernatant containing the part of anti-xenoantigen antibody that has not bonded with epitopes, a column of wells being adapted to define a reference value that corresponds to the maximum signal between antibody and epitopes,

completing said plate with a secondary antibody conjugated with an enzyme, or other molecule, adapted to chromatically highlight any presence of anti-xenoantigen antibody,

reading said plate, determining the presence of anti-xenoantigen antibody that has remained free in the solutions of the samples,

comparing the absorbance values detected in the column that defines the reference values with the values detected in the other columns of samples of the plate.

2. The method according to claim 1, characterized in that said food is cow'"'"'s milk, or soy milk, or rice milk, said xenoantigen epitope is the alpha-Gal epitope, and said anti-xenoantigen antibody is an antibody specifically aimed at the alpha-Gal epitope.

3. The method according to claim 1, characterized in that said vegetable extract with high content of phenylpropanoids, for the inactivation of at least part of said xenogeneic epitopes from said food, is constituted by vegetable extracts with a high content of one or both of verbascoside and teupolioside.

4. The method according claim 3, characterized in that said vegetable extracts with high content of phenylpropanoids contain verbascoside titrated in the 50-93% range.

5. The method according to claim 1, characterized in that said vegetable extracts titrated at 93% verbascoside are present in a solution in a quantity not lower than 0.005±

0.001% w/v.

6. The method according to claim 1, characterized in that said vegetable extracts with high content of phenylpropanoids contain teupolioside titrated in the 50-85% range.

7. The method according to claim 1, characterized in that said vegetable extracts titrated at 50% teupolioside are present in a solution at a concentration of 0.5±

0.05% w/v.

8. The method according to claim 1, characterized in that said one or more phenolic compounds, for removing at least part of said xenogeneic epitopes from said food, comprise at least one between a phenyl derivative of cinnamic acid and a phenyl derivative of tannin.

9. The method according to claim 8, characterized in that said phenyl derivative of cinnamic acid is constituted by caffeic acid.

10. The method according to claim 1 characterized in that said caffeic acid is present in a solution with a concentration of 0.5±

0.05% w/v.

11. The method according to claim 8, characterized in that said phenyl derivative of tannin is constituted by tannic acid.

12. The method according to claim 11, characterized in that said tannic acid is present in a solution at a concentration of 0.5±

0.05% w/v.

13. The method according to claim 1, characterized in that said one or more phenolic compounds, polyphenolic compounds and derivatives thereof, comprising phenylpropanoids, for the inactivation of at least part of the xenogeneic epitopes from said food, comprise:

vegetable extracts titrated at 93% verbascoside,

vegetable extracts titrated at 50% teupolioside,

at least one phenyl derivative of cinnamic acid,

at least one phenyl derivative of tannin.

14. The method according to claim 1, characterized in that said at least one phenyl derivative of cinnamic acid is constituted by caffeic acid.

15. The method according to claim 1, characterized in that at least one phenyl derivative of tannin is constituted by tannic acid.

16. A mixture of phenolic compounds, polyphenolic compounds and derivatives thereof, comprising phenylpropanoids, for the inactivation of at least part of said xenogeneic epitopes from foods of vegetable and animal origin.

17. Milk of animal origin, characterized in that it comprises at least part of the animal component constituted by the antigen in inactivated form.