Methods for separating soluble selenoglycoproteins

• US 8,263,752 B2
• Filed: 03/18/2011
• Issued: 09/11/2012
• Est. Priority Date: 03/18/2010
• Status: Active Grant

• Current Assignee: Alltech Incorporated (Standard Industries Holdings, Inc.)
• Original Assignee: Alltech Incorporated (Standard Industries Holdings, Inc.)
• Inventors: Kwiatkowski, Stefan, Power, Ronan, Matney, Clayton, Ostertag, Eric M., Ghoroghchian, Paiman P.
• Primary Examiner(s): NOAKES, SUZANNE MARIE
• Application Number:
  US13/051,646
• Publication Number:
  US 20110280948A1
• Time in Patent Office: 543 Days
• Field of Search: None
• US Class Current: 530/418
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 9/1273 Polymersomes; Liposomes with polymerisable or polymerised bilayer-forming substances polymers grafted or coated on phosphatidyl liposomes A61K9/1271, on non-phosphatidyl liposomes A61K9/1272
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C07K 14/39 from yeasts

Abstract

The invention relates to soluble selenium compositions and methods of production, separation and purification thereof. In particular the present invention provides methods of preparing water soluble selenoglycoproteins (e.g., via extracting selenoglycoproteins from selenium enriched yeast), methods of supplementing a selenium deficient composition via admixing water soluble selenoglycoproteins with the selenium deficient composition, compositions comprising the water soluble selenoglycoproteins and methods of administering the same.

Claims

1. A method for preparation of soluble selenoglycoproteins comprising:

a) providing selenium enriched yeast;

b) exposing the selenium enriched yeast to acidic conditions followed by centrifugation to generate

i) a pellet comprising acid insoluble material; and

ii) a liquid phase comprising the extract of selenium enriched yeast soluble under the acidic conditions;

c) precipitating selenoglycoproteins from the liquid phase comprising the extract of selenium enriched yeast soluble under the acidic conditions via raising the pH of the liquid phase; and

d) separating the precipitated selenoglycoproteins from the liquid phase.

2. The method of claim 1, wherein the acidic conditions are obtained using acidic buffer or the addition of an acid.

3. The method of claim 2, wherein the acid is hydrochloric acid.

4. The method of claim 1, wherein exposing the selenium enriched yeast to acidic conditions comprises exposing the selenium enriched yeast to a pH between 1.5 and pH 6.5.

5. The method of claim 4, wherein the pH is 1.5.

6. The method of claim 1, wherein the selenium enriched yeast are exposed to acidic conditions for between one and twenty-four hours.

7. The method of claim 1, wherein the selenium enriched yeast are exposed to acidic conditions for between five and ten hours.

8. The method of claim 1, wherein the selenium enriched yeast are exposed to acidic conditions for about 8 hours.

9. The method of claim 1, wherein exposing the selenium enriched yeast to acidic conditions occurs at a temperature greater than room temperature.

10. The method of claim 9, wherein the temperature is between 50°

C. and 100°

C.

11. The method of claim 9, wherein the temperature is between 70°

C. and 85°

C.

12. The method of claim 9, wherein the temperature is 80°

C.

13. The method of claim 1, wherein selenoglycoproteins are precipitated from the liquid phase at a variety of pH conditions to generate multiple pH dependent fractions of soluble selenoglycoproteins.

14. The method of claim 13, wherein the multiple pH dependent fractions of soluble selenoglycoproteins are generated at pH values of 1.85, 3.0, 4.0 and 6.0.

15. The method of claim 1, wherein separating the precipitated selenoglycoproteins from the liquid phase comprises centrifugation to form a pellet of the precipitated selenoglycoproteins followed by removal of the liquid phase from the precipitated selenoglycoproteins.

16. The method of claim 1, wherein the method is used to create a fraction of soluble selenoglycoproteins precipitated at a value selected from the group consisting of pH of 1.85, 3.0, 4.0 and 6.0.

17. The method of claim 1, wherein the selenium enriched yeast are dried, nonviable selenium-enriched yeast containing 2% or less inorganic selenium.