WO2022175218A1 - Lait à teneur élevée en immunoglobulines - Google Patents

Lait à teneur élevée en immunoglobulines Download PDF

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Publication number
WO2022175218A1
WO2022175218A1 PCT/EP2022/053570 EP2022053570W WO2022175218A1 WO 2022175218 A1 WO2022175218 A1 WO 2022175218A1 EP 2022053570 W EP2022053570 W EP 2022053570W WO 2022175218 A1 WO2022175218 A1 WO 2022175218A1
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WO
WIPO (PCT)
Prior art keywords
active
milk
slga
igg
protein concentrate
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PCT/EP2022/053570
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English (en)
Inventor
Jeroen Margot Leon Heck
Maria Helena Petronella Wilhelmina Visker
Christine COUNET
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Frieslandcampina Nederland B.V.
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Application filed by Frieslandcampina Nederland B.V. filed Critical Frieslandcampina Nederland B.V.
Priority to CN202280013924.8A priority Critical patent/CN116829002A/zh
Priority to EP22705061.4A priority patent/EP4294214A1/fr
Publication of WO2022175218A1 publication Critical patent/WO2022175218A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/19—Dairy proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/04—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from milk
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/12—Immunoglobulins specific features characterized by their source of isolation or production isolated from milk
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype

Definitions

  • the present invention relates to milk and milk-derived products with a high immunoglobulin content and high slgA/lgG weight ratio; and to uses thereof.
  • the invention also relates to processes for providing such milk and milk-derived products.
  • Ig immunoglobulins
  • IgA immunoglobulins
  • IgM immunoglobulins
  • IgG can be subdivided in IgGi and lgG2
  • IgA can be subdivided in serum IgA and secretory IgA (slgA).
  • Human breast milk contains about 85-90 wt% IgA, about 2-3 wt% IgG, and about 8-10 wt% IgM (J.A. Cakebread et al. , J. Agric. Food Chem., 63 (2015) 7311 -7316).
  • the major Ig is IgG.
  • IgG the far majority being IgGi
  • IgM the far majority being IgGi
  • 10 wt% is IgM
  • slgA the far majority being IgGi
  • the Ig-content in bovine colostrum is much higher than in mature bovine milk: 70-80 wt% of the total protein content of colostrum are Ig’s, whereas in mature bovine milk Ig’s only provide for 1-2 wt% of the total protein content.
  • the IgG/slgA ratio in bovine colostrum is even higher than in mature bovine milk.
  • Infant formulae are prepared by combining at least one source of whey protein, at least one source of casein protein, at least one source of lipids, at least one carbohydrate source, and vitamins and minerals.
  • Ruminant milk such as bovine or goat milk
  • Suitable whey protein sources - in addition to milk - are whey protein concentrate (WPC) and serum protein concentrate (SPC).
  • WPC whey protein concentrate
  • SPC serum protein concentrate
  • These products are the result of separating (skimmed) milk into a casein-rich and a whey protein-rich fraction; either by renneting (i.e. cheese making, leading to cheese whey), acidification (leading to acid whey), or microfiltration (leading to native whey).
  • Immunoglobulins exist in the milk serum phase, instead of the casein micelle phase, and are therefore considered whey proteins.
  • Whey protein concentrate is a product obtained by ultrafiltration and/or reverse osmosis, and optionally demineralization, of acid or cheese whey. By ultrafiltration, a large part of the water, lactose, and ash are removed from the product, thereby concentrating the whey proteins. Reverse osmosis can be used to remove water and to further concentrate the WPC.
  • Serum protein concentrate is also a concentrated whey protein product and differs from WPC in the origin of the whey fraction.
  • the whey proteins in SPC result from microfiltration of skimmed milk. Said microfiltration results in a concentrated casein retentate fraction and a serum fraction containing most of the whey proteins as the permeate. Conventionally, this permeate fraction is then subjected to ultrafiltration and/or reverse osmosis in order to remove lactose, ash, and water.
  • ruminant milk products - such as skimmed milk, WPC, and SPC - have to be heat-treated at least once before consumption and before their use as ingredient in infant formulae. All suitable heat treatments, also the relatively mild ones, negatively affect the active Ig content of the milk; and even more so the active slgA content, since slgA is more heat sensitive than IgG. Hence, heat-treatment will negatively affect the total active Ig content and reduce the active slgA/active IgG ratio.
  • safe ruminant (e.g. bovine) milk-based food products with a high active total immunoglobulin content and especially a high active slgA content it is desired to start with raw milk having a total Ig content and slgA/lgG ratio as high as possible.
  • colostrum In order to reach higher Ig contents, it is not an option to use ruminant colostrum instead of or in admixture with mature bovine milk.
  • the composition of colostrum e.g. its high concentration of whey proteins
  • colostrum is such that it negatively affects various industrial dairy processing steps: it tends to precipitate on the surface of heat exchangers and evaporators, causing problems in their cleaning and maintenance.
  • the produced quantities of colostrum are only a fraction of that of normal milk and colostrum is normally not collected from farms. So, using colostrum to adjust Ig levels would cause a significant price increase.
  • bovine products it would not help in increasing the slgA/lgG ratio because bovine colostrum has an even lower ratio than mature bovine milk.
  • Parity also called lactation number, is the number of times the ruminant has had offspring. Ruminants with a parity of 3 are thus in their third lactation cycle.
  • ruminants with a parity of 3 or more have a higher slgA/lgG ratio than the same ruminants with a lower parity.
  • the invention relates to the use of mature milk from ruminants with a parity of at least 3 in a process for producing dairy products with a high active immunoglobulin content and a high weight ratio active slgA / active IgG.
  • the invention relates to a use of mature milk from ruminants with a parity of at least 3 in a process for producing dairy products with an active immunoglobulin content of at least 418 pg/mL and a weight ratio active slgA / active IgG of at least 0.13, said process involving the steps of collecting mature milk from ruminants with a parity of at least 3, and optionally subjecting said milk to a pasteurization treatment; preferably the active immunoglobulin content is at least 430 pg/mL and the weight ratio active slgA / active IgG is at least 0.14, more preferably the active immunoglobulin content is at least 446 pg/mL and the weight ratio active slgA / active IgG is at least 0.15.
  • ruminant includes true ruminants, like cattle, sheep, and goats, and pseudo ruminants, like camels.
  • the preferred ruminants from which milk is selected according to the invention are cattle and goats, meaning that the preferred ruminant milk and ruminant WPC/SPC are bovine milk, bovine WPC, and bovine SPC and goat milk, goat WPC, and goat SPC.
  • the most preferred milk, WPC, and SPC to be produced according to the present invention are bovine milk, bovine WPC, and bovine SPC.
  • regular mature milk collected from healthy Holstein Friesian cows from various farms not selected by parity has a mean slgA/lgG ratio of about 0.13.
  • Selecting the raw milk from healthy cows with a parity of at least 3, resulted in a mean slgA/lgG ratio of at least 0.14, preferably at least 0.15.
  • the difference between the active slgA/active IgG weight ratios of milk from different parties increases, since slgA is more sensitive to heat treatment than IgG.
  • Healthy ruminants in the present application are defined as ruminants that do not suffer from mastitis. Mastitis results in an increase in Ig’s in the milk; but the milk has low quality.
  • the raw milk used in the processes of the present invention has a somatic cell count below 200,000 cells/mL, preferably below 150,000 cells/mL, and most preferably below 100,000 cells/mL as determined by microscopy according to ISO 13366-1 :1977 [Milk - Enumeration of somatic cells - Part 1 : Microscopic method (Reference method); International Organization for Standardization (ISO), Geneva, Switzerland]
  • the present invention relates to heat-treated mature ruminant milk comprising, after said heat treatment, at least 0.50 g, preferably at least 0.75 g, more preferably at least 0.90, and most preferably at least 1.00 g active immunoglobulins - defined as active slgA + active IgG - per 100 gram protein and a weight ratio active slgA / active IgG of at least 0.13 preferably wherein the heat-treatment is a pasteurisation process.
  • Active immunoglobulins are immunoglobulins that are in native, i.e. undenatured state.
  • at least 50 wt% of the total amount of immunoglobulins present in the heat-treated milk is active - i.e. in undenatured state - such as at least 60 wt% or 70 wt%, preferably at least 75 wt%, such as at least 80 wt% or 85 wt%, more preferably at least 90 wt%, even more preferably at least 95 wt%.
  • the active immunoglobulin content and active slgA / active IgG weight ratio are determined by use of the bovine ELISA quantitation set as described by R. L. Valk- Weeber, T. Eshuis-de Ruiter, L. Dijkhuizen, S.S. van Leeuwen ( International Dairy Journal, Volume 110, November 2020, 104814).
  • the total protein content in milk can be determined by either the well-known standard Kjeldahl method, with a nitrogen to protein conversion factor of 6.38, or by infrared spectroscopy using MilkoscanTM (complient with ISO 9622:2013). Since the first method is used to calibrate the latter, both methods will result in the same values for the same milk sample.
  • the subsequent heat treatment which is required for food safety purposes, should be mild in order to preserve as much active Ig’s - in particular slgA - as possible.
  • the heat treatment according to the present invention is therefore preferably a pasteurization process.
  • Milk is considered to be pasteurized if it can be classified as “phosphatase-negative” according to the alkaline phosphate test of ISO 118161 IDF 155.
  • Phosphatase is an enzyme that is naturally present in milk, but is destroyed at a temperature just near to the pasteurization temperature.
  • the alkaline phosphatase test is based on the principle that the alkaline phosphatase enzyme in raw milk liberates phenol from a disodium para-nitro phenyl phosphate and forms a yellow coloured complex at alkaline pH.
  • the intensity of the yellow colour is proportional to the activity of the enzyme.
  • the colour intensity is measured by direct comparison with standard colour discs in a Lovibond comparator.
  • Phosphatase-negative is defined as a color intensity corresponding to a phosphatase content below 350U/I.
  • suitable time and temperature combinations can be used to achieve the required pasteurization. Examples of suitable combinations are: 72-75°C for 15-20 seconds, 63-65°C for 30-40 minutes, and 80-85°C for 1-5 seconds.
  • the milk is obtained from cows or goats.
  • cow breeds are Holstein-Friesian, Holstein, Jersey, Brown Swiss cows, Fleckvieh (Simmental), Montbeliarde, Guernsey, and Ayrshire.
  • the preferred cow breed is Holstein-Friesian.
  • the present invention also relates to a whey protein concentrate or serum protein concentrate comprising at least 9 g active immunoglobulins per 100 gram protein and a weight ratio active slgA / active IgG of at least 0.10.
  • WPC and SPC can be produced with generally known processes, involving ultrafiltration of acid whey, cheese whey, or native whey, an optional demineralization step, concentration, and optionally (spray) drying.
  • at least one step of reducing the count of micro-organisms preferably at least one pasteurization step, is conducted: to the milk after skimming and/or before concentration and optional (spray) drying.
  • a pasteurization treatment is performed to the milk before separation of the whey/serum fraction.
  • the heat-treated milk and the whey protein concentrate according to the invention are particularly suitable for use as a nutritional product of as an ingredient of a nutritional composition.
  • formula milk An example of such a nutritional composition is formula milk.
  • the formula milk is selected from the group of infant formulas, follow-up formulas and growing-up formulas (also called young child formulas).
  • the invention further relates to a nutritional composition, typically a nutritional composition for a child, such as formula milk, in particular an infant formula, a follow-up formula, or a growing-up formula.
  • Other examples of nutritional compositions are compositions for adults, such as patients or frail elderly or anyone else desiring to boost their immune system.
  • infant formula, baby formula or just formula (American English) or baby milk, infant milk or first milk (British English), is a manufactured food designed and marketed for feeding to babies and infants under 12 months of age, usually prepared for bottle-feeding or cup-feeding from powder (mixed with water) or liquid (with or without additional water).
  • the U.S. Federal Food, Drug, and Cosmetic Act (FFDCA) defines infant formula as "a food which purports to be or is represented for special dietary use solely as a food for infants by reason of its simulation of human milk or its suitability as a complete or partial substitute for human milk".
  • the Codex Alimentarius international food standards (WFIO and FAO) defines infant formula as a breast-milk substitute specially manufactured to satisfy, by itself, the nutritional requirements of infants during the first months of life up to the introduction of appropriate complementary feeding.
  • the Codex Alimentarius describes the essential composition of an infant formula with amounts and specifications for the lipid source, protein source, carbohydrate source, vitamins and minerals.
  • the heat- treated milk according to the invention and/or the WPC or SPC according to the invention are combined with at least a lipid source, at least one carbohydrate source, and vitamins and minerals.
  • the nutritional composition comprising the heat-treated milk according to the invention and/or the whey protein concentrate and/or serum protein concentrate according to the invention, comprises at least 0.50 g active immunoglobulins per 100 gram protein and a weight ratio active slgA / active IgG of at least 0.10.
  • the invention relates to the use of the heat-treated milk according or the invention and/or the whey protein concentrate and/or serum protein concentrate according the invention in a nutritional composition.
  • the lipid source in the above-mentioned nutritional composition may be any lipid or fat suitable for use in formula milk.
  • Preferred fat sources include milk fat, safflower oil, egg yolk lipid, canola oil, olive oil, coconut oil, palm kernel oil, soybean oil, fish oil, palm oleic, high oleic sunflower oil and high oleic safflower oil, and microbial fermentation oil containing long-chain, polyunsaturated fatty acids.
  • anhydrous milk fat is used.
  • the lipid source may also be in the form of fractions derived from these oils such as palm olein, medium chain triglycerides, and esters of fatty acids such as arachidonic acid, linoleic acid, palmitic acid, stearic acid, docosahexaenoic acid, linolenic acid, oleic acid, lauric acid, capric acid, caprylic acid, caproic acid, and the like. Small amounts of oils containing high quantities of preformed arachidonic acid and docosahexaenoic acid such as fish oils or microbial oils may be added.
  • oils such as palm olein, medium chain triglycerides, and esters of fatty acids such as arachidonic acid, linoleic acid, palmitic acid, stearic acid, docosahexaenoic acid, linolenic acid, oleic acid, lauric acid, capric acid, caprylic acid, ca
  • the fat source preferably has a ratio of n-6 to n-3 fatty acids of about 5:1 to about 15:1 ; for example about 8:1 to about 10:1.
  • the infant formula comprises an oil mix comprising palmitic acid esterified to triacylglycerols, for example wherein the palmitic acid esterified in the sn-2 position of triacylglycerol is in the amount from 10% to 60% by weight of total palmitic acid and palmitic acid esterified in the sn-1/sn-3 position of triacylglycerol is in the amount of from 30% to 80% by weight of total palmitic acid.
  • vitamins and minerals that are preferably present in formula milk are vitamin A, vitamin B1 , vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorous, iodine, iron, magnesium, copper, zinc, manganese, chloride, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are usually added in salt form.
  • Suitable HMOs include 2 -FL, 3’-GL, 3’-SL, 6’-SL, LNT, LNnT, and combinations thereof.
  • FIMO’s are commercially available or can be isolated from milk in particular from human breast milk.
  • the nutritional composition may contain emulsifiers and stabilisers such as soy lecithin, citric acid esters of mono- and di-glycerides, and the like. It may also contain other substances which may have a beneficial effect such as lactoferrin, nucleotides, nucleosides, probiotics, and the like.
  • Suitable probiotics include Lactobacteria, Bifidobacterium lactis such as Bifidobacterium lactis Bb12, Streptococcus thermophilus, Lactobacillus johnsonii La1, Bifidobacterium longum BL999, Lactobacillus rhamnosus LPR, L rhamnosus GG, Lactobacillus reuteri, Lactobacillus salivarius.
  • Bifidobacterium lactis such as Bifidobacterium lactis Bb12, Streptococcus thermophilus
  • Lactobacillus johnsonii La1 Bifidobacterium longum BL999
  • Lactobacillus rhamnosus LPR Lactobacillus rhamnosus LPR
  • L rhamnosus GG Lactobacillus reuteri
  • Lactobacillus salivarius Such prebiotics are commercially available.
  • Formula milk is generally available as a spray-dried powder.
  • Spray-drying involves an additional heating step.
  • a dairy product according to the invention e.g. a WPC or SPC
  • it is preferably either dry blended with the other ingredients or mixed as liquid WPC or liquid SPC with other liquid ingredients; all in order to the reduce the number of heat treatments that may negatively affect the immunoglobulin content.
  • Raw milk samples were collected from 1998 cows on 21 Dutch farms. The cows were not suffering from mastitis and hence the milk had a somatic cell count below 200,000 cells/mL.
  • Fresh milk samples were analyzed for their protein content; stored frozen samples were used for analysis of active IgG and active slgA content.
  • the protein content was measured by infrared spectroscopy using a MilkoScanTM (ISO 9622, Qlip, Zutphen, the Netherlands). Contents of active IgG and active slgA were measured using the quantitative ELISA method as described by R. L. Valk-Weebera, T. Eshuis - de Ruiter, L. Dijkhuizen, S.S. van Leeuwen ⁇ International Dairy Journal, Volume 110, November 2020, 104814).
  • Table 1 shows the mean active Ig and total protein contents, per parity class and for the overall population not selected on parity.
  • Table 1 The milk samples listed in Table 1 were pasteurized under standard pasteurization conditions: 75°C, 20 seconds.
  • Table 2 shows the active slgA and active IgG levels and the weight ratio active slgA/active IgG for these heat treated milks.
  • Example 2 The heat-treated milk samples of Example 2 were used to prepare a whey protein concentrate with a protein content of 70 wt%, as follows.
  • cheese was produced from said milk samples and the resulting whey was collected.
  • the whey was subsequently concentrated to a protein content of 35 wt% using a 10 kD UF spiral wound membrane.
  • a second filtration was performed using a ceramic membrane with a pore size ranging from 0.5 to 2.0 pm in order to reduce the microbial count.
  • the resulting permeate was subjected to a second ultrafiltration in order to increase the protein content to 70 wt%.
  • the resulting whey protein concentrate was finally pasteurized at 75°C, 20 seconds.
  • the total protein content was determined using the Kjeldahl method (conversion factor 6.38).
  • the active slgA and active IgG contents were determined by the same method as in Examples 1 and 2.
  • Example 2 Milk from Example 2 was microfiltered and skimmed.
  • the microfiltered skimmed milk was standardized on a whey protein/casein weight ratio of 1.1. using Hiprotal® Milkserum 60 Liquid (FrieslandCampina, The Netherlands).
  • Hiprotal® Milkserum 60 Liquid (FrieslandCampina, The Netherlands).
  • the resulting blend was heat treated for 30 seconds at 75°C, and evaporated in a Mechanical Vapor Recompression evaporator at 60°C. Afterwards, a vegetable fat blend was added and the product was homogenized and spray-dried.
  • the resulting base powder was dry blended with the WPC of Example 3 in order to obtain a whey protein/total protein weight ratio of 0.64 and a whey protein/casein weight ratio of 1 .76 .
  • the final product (IFT) contained 10.2 g protein and 26 g fat per 100 g powder.
  • the active slgA and IgG levels and the ratio active slgA/active IgG in the final IFT formula product are summarized in Table 4.

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Abstract

La présente invention concerne un procédé de production de produits laitiers ayant une teneur élevée en immunoglobulines actives et un rapport pondéral IgAs active/IgG active élevé, comprenant les étapes consistant à collecter du lait mature de ruminants avec une parité d'au moins 3, et éventuellement à soumettre ledit lait à un traitement de pasteurisation ; et des utilisations de ce procédé. L'invention concerne également des produits laitiers spécifiques présentant un rapport pondéral IgAs active/IgG active élevé.
PCT/EP2022/053570 2021-02-16 2022-02-15 Lait à teneur élevée en immunoglobulines WO2022175218A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202280013924.8A CN116829002A (zh) 2021-02-16 2022-02-15 具有高免疫球蛋白含量的乳
EP22705061.4A EP4294214A1 (fr) 2021-02-16 2022-02-15 Lait à teneur élevée en immunoglobulines

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EP21157397 2021-02-16
EP21157397.7 2021-02-16

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WO2006119560A1 (fr) * 2005-05-10 2006-11-16 Murray Goulburn Co-Operative Co Limited Fraction d'immunoglobuline et procede afferent
EP2280999A1 (fr) * 2008-05-15 2011-02-09 W. Health L.P. Procédé pour la fabrication de fractions de lait riches en immunoglobulines sécrétées

Patent Citations (2)

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Publication number Priority date Publication date Assignee Title
WO2006119560A1 (fr) * 2005-05-10 2006-11-16 Murray Goulburn Co-Operative Co Limited Fraction d'immunoglobuline et procede afferent
EP2280999A1 (fr) * 2008-05-15 2011-02-09 W. Health L.P. Procédé pour la fabrication de fractions de lait riches en immunoglobulines sécrétées

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CN116829002A (zh) 2023-09-29
EP4294214A1 (fr) 2023-12-27

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