PURIFIED HUMAN MILK OLIGOSACCHARIDES COMPOSITIONS
The present invention relates to purified and concentrated human milk oligosaccharide compositions and methods of making and using the same.
1 . A method of making a purified human milk oligosaccharide composition comprising:
(a) mixing human milk permeate with lactase under conditions suitable for the digestion of lactose in the human milk permeate to create a permeate/lactases mixture;
(b) removing the lactase from the permeate/lactase mixture; and
(c) filtering the mixture to purify and concentrate human milk oligosaccharides.
2 . The method of claim 1 wherein the pH of the permeate is adjusted to a pH of about 4.3 to about 4.7 prior to or concurrent with the addition of the lactase.
3 . The method of claim 2 wherein the pH is adjusted to 4.5 prior to the addition of the lactose.
4 . The method of claim 1 wherein the permeate is heated to a temperature of about 45° C. to about 55° C. prior to or concurrent with the addition of the lactase.
5 . The method of claim 4 wherein the permeate is heated to about 50° C. prior to adding the lactase.
6 . The method of claim 1 wherein lactase is added at about 0.1% to about 0.5% weight/weight.
7 . The method of claim 6 wherein lactase is added at about 0.1% weight/weight.
8 . The method of claim 1 further comprising cooling the mixture to a temperature of about 20° C. to about 30° C. prior to the removal of lactase.
9 . The method of claim 8 wherein the temperature is about 25° C.
10 . The method of claim 1 further comprising clarifying the lactase digested permeate through a depth filter.
11 . The method of claim 10 wherein the depth filter is about 1 to about 5 micron filter.
12 . The method of claim 1 wherein the filtering comprises filtering the mixture of step (b) through a membrane having a pore size of about 50,000 Dalton to create a 50,000 Dalton filtered mixture.
13 . The method of claim 12 further comprising filtering the 50,000 Dalton filtered mixture through a membrane having a pore size of about 2,000 Dalton to about 3,000 Dalton to create a 2,000 to 3,000 Dalton filtered mixture.
14 . The method of claim 12 further comprising filtering the 50,000 Dalton filtered mixture through a membrane having a pore size of ≤600 Dalton.
15 . The method of claim 1 wherein the concentration of human milk oligosaccharides after purification in step (c) is about 1% to about 5% weight/weight.
16 . The method of claim 15 wherein the concentration of human milk oligosaccharides is about 5% weight/weight.
17 . A purified human milk oligosaccharide (HMO) composition made according to the method of claim 1 .
18 . The purified human milk oligosaccharide composition of claim 17 comprising ≤5% lactose.
19 . The purified human milk oligosaccharide composition of claim 17 as shown in FIG. 5E and FIG. 5F .
20 . A method of preventing NEC in a subject in need thereof comprising administering to the subject an effective amount of purified HMO composition of claim 17 .
21 . A method for decreasing systemic inflammation in a subject in need thereof comprising administering to the subject an effective amount of the purified HMO composition of claim 17 .
22 . A method for treating or preventing an infection in a subject in need thereof comprising administering to the subject an effective amount of the purified HMO composition of claim 17 .