Methods for testing milk

Abstract

The disclosure is related generally to methods for testing mammary fluid (including milk) to establish or confirm the identity of the donor of the mammary fluid. Such methods are useful in the milk-bank business to improve safety.

Claims

1. A method for determining whether a donated .[.mammary fluid.]. .Iadd.human breast milk .Iaddend.was obtained from a specific subject.Iadd., wherein the specific subject is a qualified donor.Iaddend., the method comprising: (a) testing a donated biological sample from the specific subject to obtain at least one reference identity marker profile for at least one .Iadd.identity .Iaddend.marker.Iadd., wherein the specific subject has been identified as a qualified donor by testing the specific subject for absence of chronic illness.Iaddend.; (b) testing a sample of the donated .[.mammary fluid.]. .Iadd.human breast milk .Iaddend.to obtain at least one identity marker profile for the at least one marker in step (a); (c) comparing the identity marker profiles .Iadd.obtained in step (a) and (b).Iaddend., wherein.Iadd., if one or more identity markers in the biological sample match one or more identity markers in the donated human breast milk, such .Iaddend.a match .[.between the identity marker profiles.]. indicates that the .[.mammary fluid.]. .Iadd.donated human breast milk .Iaddend.was obtained from the .[.specific subject.]. .Iadd.qualified door.Iaddend.; and (d) processing the donated .[.mammary fluid.]. .Iadd.human breast milk from the qualified door .Iaddend.whose identity marker profile has been matched with a reference identity marker profile, wherein the processed donated .[.mammary fluid.]. .Iadd.human breast milk .Iaddend.comprises .Iadd.a caloric content of 20 Cal/ounce; .Iaddend.a human protein constituent of 11-20 mg/mL; a human fat constituent of 35-55 mg/mL; and a human carbohydrate constituent of 70-120 mg/mL.

.[.2. The method of claim 1, wherein the mammary fluid is human breast milk..].

3. The method of claim 1 wherein the processing comprises: (a) filtering the milk; (b) heat-treating the milk; (c) separating the milk into cream and skim; (d) adding a portion of the cream to the skim; and (e) pasteurizing.

.[.4. The method of claim 1, wherein the composition further comprises one or more constituents selected from the group consisting of: calcium, chloride, copper, iron, magnesium, manganese, phosphorus, potassium, sodium, and zinc..].

5. The method of claim 1, wherein processing comprises separating the milk into a cream portion and a skim portion, processing the cream portion, and pasteurizing the cream portion.

6. The method of claim 1, further comprising nucleic acid typing, wherein the nucleic acid typing comprises a method selected from the group consisting of: STR analysis, HLA analysis, multiple gene analysis, and a combination thereof.

7. The method of claim 1, wherein the donated .[.mammary fluid.]. .Iadd.breast milk .Iaddend.is frozen.

8. The method of claim 1, wherein the .[.mammary fluid.]. .Iadd.breast milk .Iaddend.sample comprises a mixture of one or more .[.mammary fluid.]. .Iadd.breast milk .Iaddend.samples.

9. The method of claim 1, wherein the donated biological sample is selected from a group consisting of: milk, saliva, buccal cell, hair root, and blood.

10. The method of claim 1, wherein steps (a) through (c) are carried out at a human breast milk donation center or at a milk processing facility.

11. The method of claim 1, wherein steps (a) and (b) are carried out at different facilities.

12. The method of claim 11, wherein step (a) is carried out at a human breast milk donation facility and step (b) is carried out at a milk processing facility.

13. A method for processing a donated human breast milk obtained from a specific.Iadd., qualified donor .Iaddend.subject comprising: (a) .Iadd.screening a donated biological sample from the subject for the absence of chronic illness; (b) .Iaddend.testing a donated biological sample from the specific subject to obtain at least one reference identity marker profile for at least one marker; (.[.b.]. .Iadd.c.Iaddend.) testing a sample of the donated human breast milk to obtain at least one identity marker profile for the at least one marker in step (.[.a.]. .Iadd.b.Iaddend.); (.[.c.]. .Iadd.d.Iaddend.) comparing the identity marker profiles of steps (.[.a.]. .Iadd.b.Iaddend.) and (.[.b.]. .Iadd.c.Iaddend.), wherein.Iadd., if one or more identity markers in the biological sample of step (b) match one or more identity markers in the donated human breast milk of step (c), such .Iaddend.a match .[.between the identity marker profiles.]. indicates that the donated human breast milk was obtained from the specific subject; (.[.d.]. .Iadd.e.Iaddend.) .Iadd.selecting for .Iaddend.processing the donated human breast milk .Iadd.from the subject screened for the absence of chronic illness and .Iaddend.whose identity marker profile has been matched with a reference identity marker profile.Iadd., wherein the selected subject is a qualified donor; and (f) processing the donated human breast milk.Iaddend., wherein the processing comprises: (i) filtering the donated human breast milk; (ii) heat treating the donated human breast milk; (iii) separating the donated human breast milk into cream and skim; (iv) adding a portion of the cream to the skim to form a human milk composition; and (v) pasteurizing the human milk composition to produce a processed human breast milk composition; and wherein the processed donated human breast milk comprises .Iadd.a caloric content of 20 Cal/ounce; .Iaddend.a human protein constituent of 11-20 mg/mL; a human fat constituent of 35-55 mg/mL; and a human carbohydrate constituent of 70-120 mg/mL.

14. A processed human milk composition suitable for administration to an infant made by the process of claim 13.

.[.15. The method of claim 13, wherein the method further comprises adding to the processed human breast milk one or more constituents selected from the group consisting of: calcium, chloride, copper, iron, magnesium, manganese, phosphorus, potassium, sodium, and zinc..].

16. The method of claim 13, wherein the testing of the donated human breast milk of step (.[.b.]. .Iadd.c.Iaddend.) and the testing of the donated biological sample of step (.[.a.]. .Iadd.b.Iaddend.) comprises nucleic acid typing selected from the group consisting of: STR analysis, HLA analysis, multiple gene analysis, and a combination thereof.

17. The method of claim 13, wherein the donated biological sample .Iadd.of step (a) and/or the donated biological sample of step (b) .Iaddend.is selected from a group consisting of: milk, saliva, buccal cell, hair root, and blood.

18. The method of claim 13, wherein steps (.[.a.]. .Iadd.b.Iaddend.) through (.[.c.]. .Iadd.e.Iaddend.) are carried out at a human breast milk donation center or at a milk processing facility.

19. The method of claim 13, wherein steps (.[.a.]. .Iadd.b.Iaddend.) and (.[.b.]. .Iadd.c.Iaddend.) are carried out at different facilities.

20. The method of claim 19, wherein step (.[.a.]. .Iadd.b.Iaddend.) is carried out at a human breast milk donation facility and step (.[.b.]. .Iadd.c.Iaddend.) is carried out at a milk processing facility.

.Iadd.21. The method of claim 1, wherein the human protein constituent comprises about 11-13 mg/mL. .Iaddend.

.Iadd.22. The method of claim 1, wherein the human carbohydrate constituent comprises about 80-105 mg/mL. .Iaddend.

.Iadd.23. The method of claim 1, wherein the processed donated breast milk of step (d) does not comprise added non-human derived nutritional components. .Iaddend.

.Iadd.24. The method of claim 1, wherein the method comprises pooling the donated breast milk of step (c) whose identity marker profile has been matched with the reference identity marker profile, with other donated breast milk samples that have been matched with the reference identity marker profile to obtain a pool of identity matched breast milk. .Iaddend.

.Iadd.25. The method of claim 24, wherein the pool of identity matched breast milk is in a volume of at least about 75 liters/lot to about 2,000 liters/lot. .Iaddend.

.Iadd.26. The method of claim 1, wherein the processed breast milk of step (d) has an osmolality of less than about 400 mOsm/kg H.sub.2O. .Iaddend.

.Iadd.27. The method of claim 1, wherein the processing of step (d) comprises filtering the donated breast milk by ultrafiltration. .Iaddend.

.Iadd.28. The method of claim 27, wherein the ultrafiltration filters out water, and wherein the processing step further comprises washing the filters used during the ultrafiltration with the water obtained by the ultrafiltration. .Iaddend.

.Iadd.29. The method of claim 26, wherein the processing of step (d) further comprises reducing the bioburden of the human donor milk. .Iaddend.

.Iadd.30. The method of claim 1, wherein the processing of step (d) comprises concentrating the nutrients in the donated breast milk. .Iaddend.

.Iadd.31. The method of claim 30, wherein the processing of step (d) further comprises reducing the bioburden of the human donor milk. .Iaddend.

.Iadd.32. The method of claim 13, wherein the processed donated human breast milk comprises no added non-human derived nutritional components. .Iaddend.

.Iadd.33. The method of claim 13, wherein the pool of identity matched human donor milk is in a volume of at least about 75 liters/lot to about 2,000 liters/lot. .Iaddend.

.Iadd.34. The method of claim 13, wherein the processed milk has on osmolality of less than about 400 mOsm/kg H.sub.2O. .Iaddend.

.Iadd.35. The method of claim 13, wherein the processing step comprises filtering the human donor milk. .Iaddend.

.Iadd.36. The method of claim 13, wherein the processing step comprises filtering the human donor milk by ultrafiltration. .Iaddend.

.Iadd.37. The method of claim 36, wherein the ultrafiltration filters out water, and wherein the processing step further comprises washing the filters used during the ultrafiltration with the water obtained by the ultrafiltration. .Iaddend.

.Iadd.38. The method of claim 13, wherein the processing step comprises concentrating the nutrients in the human donor milk. .Iaddend.

.Iadd.39. The method of claim 38, wherein the processing step further comprises reducing the bioburden of the human donor milk. .Iaddend.

.Iadd.40. The method of claim 35, wherein the processing step further comprises reducing the bioburden of the human donor milk. .Iaddend.

.Iadd.41. The method of claim 13, wherein the human protein constituent comprises about 11-13 mg/mL. .Iaddend.

.Iadd.42. The method of claim 13, wherein the human carbohydrate constituent comprises about 80-105 mg/mL. .Iaddend.

.Iadd.43. A method for making concentrated, bioburden reduced, identity-matched and health screened human donor milk from a qualified human milk donor, wherein the human donor milk is safe and provides standardized nutrition, the method comprising: (a) screening a subject for drug use and/or chronic illness by testing a biological sample from the subject; (b) selecting a qualified donor, wherein the subject is a qualified donor if the biological sample of step (a) did not test positive for drug use or chronic illness; (c) testing a donated biological sample from the qualified donor to obtain at least one DNA marker profile for at least one DNA marker; (d) testing a sample of donated human breast milk from the qualified donor to obtain at least one DNA marker profile for the at least one DNA marker in step (c); (e) comparing the DNA marker profiles obtained in (c) and (d), wherein, if DNA markers in the biological sample of step (c) match DNA markers in the donated human breast milk of step (d), such match indicates that the donated human breast milk was obtained from the specific, health screened, qualified donor, thereby obtaining identity matched, health screened donor milk from a qualified human milk donor; (d) pooling the identity matched, health screened donor milk obtained in step (d) with other identity matched, health screened donor milk from other qualified donors to obtain a pool of identity matched, health screened human donor milk from qualified donors, comprising at least about 75 liters, wherein the pool of identity matched, health screened donor milk is not nutritionally sufficient for preterm infants; (e) concentrating the pool of non-nutritionally sufficient, identity matched, health screened human donor milk from qualified donors to comprise a human protein constituent of 11-13 mg/mL; a human fat constituent of 35-55 mg/mL; a human carbohydrate constituent of 70-120 mg/mL; a caloric content of 20 Cal/ounce; and no added non-human derived vitamins or minerals to obtain a pool of concentrated identity matched donor milk from qualified donors with an osmolality of less than about 400 mOsm/Kg H.sub.2O; and (f) treating the pool of concentrated, identity-matched, health screened donor milk obtained in (e) to reduce the bioburden; thereby obtaining concentrated, bioburden reduced, identity-matched, health screened donor milk from qualified donors. .Iaddend.

.Iadd.44. The method of claim 43, wherein the nucleic acid typing comprises STR analysis, HLA analysis, multiple gene analysis, or a combination thereof. .Iaddend.

.Iadd.45. The method of claim 43, wherein the donated biological sample of step (a) and/or (c) is milk, saliva, buccal cell, hair root, or blood. .Iaddend.

.Iadd.46. A processed human milk composition suitable for administration to an infant made by the process of claim 43. .Iaddend.

.Iadd.47. The processed human milk composition of claim 46, wherein the infant is a preterm infant. .Iaddend.

.Iadd.48. The method of claim 43, wherein the concentrating step comprises filtering the human donor milk. .Iaddend.

.Iadd.49. The method of claim 48, wherein the filtering step comprises ultrafiltering the human donor milk. .Iaddend.

.Iadd.50. The method of claim 49, wherein the ultrafiltration filters out water, and wherein the processing step further comprises washing the filters used during the ultrafiltration with the water obtained by the ultrafiltration. .Iaddend.

.Iadd.51. The method of claim 1, wherein the one or more identity markers comprise DNA. .Iaddend.

.Iadd.52. The method of claim 51, wherein the one or more identity markers comprise multiple DNA markers. .Iaddend.

.Iadd.53. The method of claim 1, wherein the specific subject has been identified as a qualified donor by further screening for drug use. .Iaddend.

.Iadd.54. The method of claim 1, wherein the specific subject has been identified as a qualified donor by interview and/or by biological sample testing for viral contamination. .Iaddend.

Description

Example 1

Testing of a Human Breast Milk Donor

(1) A woman who wishes to donate her breast milk will provide a biological reference sample prior to (or at the time of) her first donation. The biological sample will include a convenient tissue type, e.g., blood, cheek cell, hair etc. The sample will be donated under supervision of another individual(s), e.g., bank milk personnel. The sample will be labeled for later reference. The reference sample will be tested for a specific marker profile, e.g., nucleic acid and/or peptide profile. The sample will be tested for one or more markers. Results of the tests will be stored, e.g., on a computer-readable medium for future reference. Any remaining sample will be stored. The woman can also be screened (using the reference sample or another sample) for, e.g., drug use, viruses, bacteria, parasites, and fungi etc., to determine her health. The woman will be given a label corresponding to the reference sample to keep with her and use with her donated milk.

(2) Alternatively, the sample will be stored without testing, and will be tested at a later date, for example, together with the donated breast milk.

(3) The woman will express her milk for donation and either forward the milk to the milk bank or processing facility or store the milk in her refrigerator, e.g., the freezer, for donation with other samples at a later date. The donated milk will be labeled with the label given to the woman and matching the reference sample.

(4) A sample of the donated milk that arrives at the milk bank or processing facility will be tested for at least one of the same markers as the reference sample. The marker profile of the reference sample will be compared to the marker profile of the donated milk sample. If the profiles will match, the identity of the donor will be confirmed. If the profiles will not match, the results will be an indication that the donated milk is contaminated with another woman's milk or that it does not come from the woman whose reference sample was taken.

(5) The milk whose provenance (i.e., origin) will be confirmed by the matched profiles will be further processed, e.g., pasteurized, e.g., into human milk fortifiers, standardized human milk compositions, and/or human lipid compositions. Such compositions will be administered to human infants, e.g., premature infants, whose mothers may not be able to provide them with adequate nutrition.

(6) The reference sample and/or results of the reference sample tests will be stored for any future donation by the corresponding mother.

(7) A number of embodiments of the invention have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.