Methods for isolation, use and analysis of ferritin
This invention provides methods of isolating ferritin from plant and animal material. The isolated ferritin can be administered to humans or animals in need of iron, and can be used to treat or supplement iron deficiency. The isolated ferritin can be used in industrial applications, such as increasing the iron content in heat-processed food or beverages. The methods of the invention also include quantitation of iron derived from plant or animal ferritin.
1. A method for the isolation of ferritin from a legume, the method comprising the steps of:
(a) separation of the legume into soluble and insoluble legume fractions;
(b) addition of a neutral saline buffer to the insoluble legume fraction to make an insoluble legume solution;
(c) clarification of the insoluble legume solution of step (b) into soluble and insoluble legume solution fractions;
(d) enzymatic treatment with one or more glycosidase enzymes of the clarified soluble legume solution of step (c);
(e) fractionation of the enzymatically treated, clarified soluble legume solution of step (d) to remove non-ferritin components; and
(f) concentration of isolated ferritin from the fractionated soluble legume solution of step (e).
2. A method to isolate ferritin from plant material comprising the steps of:
(a) separation of the plant material into a soluble and insoluble fraction;
(b) enzymatic removal with one or more glycosidase enzymes of non-ferritin components from the soluble fraction of step (a), thereby isolating ferritin; and
(c) concentration of the isolated ferritin from step (b).
3. The method of claim 2 wherein the plant material comprises legumes.
4. The method of claim 2 wherein the plant material comprises the waste stream from processing of legumes.
5. The method of claim 2 further comprising the step of fractionation of the soluble fraction.
6. The method of claim 2 further comprising the step of treatment of the soluble fraction with a solvent.
7. The method of claim 6 wherein the solvent is an organic solvent.
8. The method of claim 3 wherein the legumes comprise soybeans.
9. The method of claim 2 wherein the plant material comprises the entire plant.
10. The method of claim 2 wherein the plant material comprises one or more of a seed, a bean, a stem, a fruit, a leaf, a root, and a flower.
11. The method of claim 2 wherein the step of soluble and insoluble separation comprises mechanical separation.
12. The method of claim 2 wherein concentration of the isolated ferritin comprises drying the soluble fraction.
13. The method of claim 2 wherein concentration of the isolated ferritin comprises ultrafiltration.
14. The method of claim 1 wherein the legume comprises soybeans.
15. A method for the isolation of ferritin from a legume processing stream, the method comprising the steps of:
(a) separation of the legume processing stream into soluble and insoluble legume fractions;
(b) addition of a neutral saline buffer to the insoluble legume fraction to make an insoluble legume solution;
(c) clarification of the insoluble legume solution of step (b) into soluble and insoluble legume solution fractions;
(d) enzymatic treatment with one or more glycosidase enzymes of the clarified soluble legume solution of step (c);
(e) fractionation of the enzymatically treated, clarified soluble legume solution of step (d) to remove non-ferritin components; and
(f) concentration of isolated ferritin from the fractionated soluble legume solution of step (e).