FIELD OF THE INVENTION

The present invention relates to spoonable nutritional compositions. In particular, the invention relates to spoonable nutritional compositions that are suitable for subjects with allergy-related conditions such as Cow's Milk Protein Allergy and Eosinophilic Esophagitis.

BACKGROUND TO THE INVENTION

Spoonable nutritional compositions provide a convenient way for delivering nutrition to a variety of subjects. Spoonable nutritional compositions may be formulated to provide for some or all of a subject's dietary needs while avoiding the use of allergens. Spoonable nutritional compositions may thus be formulated for subjects with allergy- related conditions.

Nutritional compositions for infants are commonly formulated with cow's milk protein. For example, bovine whey protein and/or casein are often used as the protein source in infant nutritional compositions. However, some infants exhibit an allergy to cow's milk proteins, making such compositions unsuitable. Allergies to cow's milk protein may be due to the differences between the proteins in cow's milk and those in human milk. The principal recognised cow's milk allergens are alpha-lactalbumin (aLA), beta- lactoglobulin (bLG) and bovine serum albumin (BSA).

To reduce allergenicity, cow's milk proteins may be hydrolysed (e.g. enzymatically) either partially or, in the case of products intended for the management of Cow's Milk Protein Allergy (CMPA), extensively. However, such proteins must be highly processed to provide sufficient hydrolysis to reduce the risk of an allergic reaction. Such processing may be viewed unfavourably with an increasing tendency to provide more natural diets, and a strong hydrolysis process also tends to have a negative impact on taste. In addition, the extensive processing increases the cost of the product formulas. Alternatives to cow's milk protein may be used in nutritional compositions, for example soy and rice proteins. However, soy-based nutritional compositions are not recommended by the European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) for infants under 12 months, because of the risk of a cross allergic response. Rice-based nutritional compositions require the addition of numerous free amino acids to provide an optimum amino acid profile for infant nutrition, due to the incomplete natural amino acid distribution in rice proteins. This increases cost and may provide the resulting nutritional composition with a less palatable taste.

Infant nutritional compositions may be formulated entirely from free amino acids for infants with severe cases of multiple allergies. However, ESPGHAN guidelines indicate that such compositions should not be used as a first line solution in the case of cow's milk protein allergic infants. Furthermore, overprescription of amino acid based nutritional compositions adds to the cost burden on national health systems as amino acid based nutritional compositions are even more expensive than extensively hydrolysed formulas.

Accordingly, there is a significant need for alternative infant nutritional compositions that comprise less potential allergens, and preferably which require minimal processing, have good taste and have low cost. In particular, there is a need for alternative nutritional compositions that are suitable for administration to infants with Cow's Milk Protein Allergy.

Eosinophilic Esophagitis (EoE) represents another condition where subjects would benefit from alternative nutritional compositions having reduced allergenicity.

EoE is a chronic allergic/immune esophageal disorder with an estimated prevalence of 0.06% in Europe and the USA. In EoE, the esophagus becomes inflamed or swollen, with an associated increase in tissue eosinophil numbers, which can lead to difficulty swallowing. The condition can lead to failure to thrive and nutritional deficits in adults and children. Current medical approaches for the treatment of EoE include an exclusively elemental diet, using an amino acid based formula. While this approach can have a high success rate, it is also associated with high cost and a reduced quality of life, and can lead to problems of food refusal especially in younger children. An alternative approach uses allergy test-directed elimination diets or an empiric elimination diet that excludes common 'trigger' foods. Since allergy tests may lead to false-positive or false-negative results the six-food elimination diet (SFED) excluding milk, wheat, egg, soy, peanuts/tree nuts and seafood has also been used.

However, a strict adherence to such diets may result in a dietary protein deficiency. Thus, alternative nutritional compositions with good taste properties and suitable for subjects having EoE would provide significant advantages.

SUMMARY OF INVENTION

The inventors have developed a spoonable nutritional composition based on potato protein as the major protein source, which is naturally absent in the major allergens found in milk and soy. Accordingly, the spoonable nutritional composition of the invention may provide a naturally hypoallergenic nutritional composition suitable for adult, child or infant subjects with allergy-related conditions such as CMPA and EoE.

With regard to infant nutrition, potato protein has been found to have a well balanced amino acid profile, which is closer to that of human milk than rice or soy protein. Accordingly, less or no addition of free amino acids is required to provide a composition with a nutritional profile close to that of human milk, which renders the resulting product more cost effective and gives it a more palatable taste.

Moreover, as a result of their lower allergen profile, the potato protein components do not require extensive hydrolysis, which provides significant benefits in terms of cost and for the development of infants, because the intact or slightly hydrolysed proteins facilitate improved gut maturation. Benefits are also provided for child or adult nutrition, as the absence of a need for extensive hydrolysis allows for the provision of nutritional compositions with improved taste properties compared to those formulated from free amino acids only. The inventors have also found that the use of potato protein as the major protein source advantageously increases the viscosity of the spoonable nutritional composition and provides improved gelling properties. The need for starches and/or hydrocolloids to be present in the spoonable nutritional compositions in order to increase viscosity and gelling is therefore reduced or removed.

Furthermore, the need for an emulsifier may be reduced or removed, because the potato protein itself may provide any necessary emulsifier properties.

In addition, use of potato protein provides for good acceptance, for example in terms of taste and texture of the spoonable nutritional composition.

In one aspect, the invention provides a spoonable nutritional composition comprising protein, carbohydrate and fat, wherein the major source of protein is potato protein.

In a preferred embodiment, the major source of protein in the spoonable nutritional composition is potato protein and the remaining protein is plant protein. The term "major source of protein is potato protein" means that the largest fraction of the total protein by weight in the nutritional composition originates from potato protein.

In one embodiment, at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%, preferably at least about 75%, by weight of the total protein in the spoonable nutritional composition is potato protein.

In one embodiment, 100% by weight of the total protein in the spoonable nutritional composition is potato protein. Thus, in one embodiment, the sole source of protein is potato protein.

In a preferred embodiment, the protein (in particular, the potato protein) is intact protein. Preferably, the protein has not been subjected to artificial hydrolysis.

In another embodiment, the protein (in particular, the potato protein) is partially hydrolysed protein. The spoonable nutritional composition does not comprise cow's milk protein.

In one embodiment, the spoonable nutritional composition further comprises free amino acids.

In one embodiment, the spoonable nutritional composition does not comprise a further emulsifier. The potato protein may provide sufficient function as an emulsifier.

In one embodiment, the spoonable nutritional composition does not comprise a starch and/or a hydrocolloid.

In one embodiment, the spoonable nutritional composition comprises probiotics. In an alternative embodiment, the spoonable nutritional composition does not comprise probiotics.

In one embodiment, the spoonable nutritional composition is suitable for a vegetarian diet. In a further embodiment, the spoonable nutritional composition is suitable for a vegan diet.

In one embodiment, the spoonable nutritional composition further comprises a natural sugar selected from sucrose, saccharose, all types of glucose syrup or fructose syrup. In certain embodiments, the spoonable nutritional composition may comprise up to about 15 wt.% of the natural sugar, for example up to about 2, about 5, or about 10 wt.%, or 1 .0 to 10.0 wt.%, or 1 .0 to 5.0 wt.%.

In one embodiment, the spoonable nutritional composition further comprises at least one natural or artificial high intensity sweetener selected from the list comprising stevia extracts, rebaudioside A, aspartame, neotame, acesulfame, sucralose, saccharin. The natural or artificial high intensity sweetener may be present in addition to a natural sugar or as an alternative to a natural sugar.

In one embodiment, the spoonable nutritional composition further comprises a flavouring agent. In certain embodiments, the spoonable nutritional composition may comprise up to about 5.0 wt.% of the flavouring agent, for example up to about 1 .0, about 2.0, about 3.0, or about 4.0 wt.%, or 0.1 to 4.0 wt.%, or 0.2 to 2.0 wt.%.

In one embodiment, the spoonable nutritional composition further comprises at least one probiotic.

In one embodiment, the spoonable nutritional composition is suitable for administration to a subject having Cow's Milk Protein Allergy and/or Eosinophilic Esophagitis. In one embodiment, the subject has Cow's Milk Protein Allergy and is an infant.

In one embodiment, the spoonable nutritional composition is an infant nutritional composition.

In another aspect, the invention provides a spoonable nutritional composition for use in feeding a subject having Cow's Milk Protein Allergy and/or Eosinophilic Esophagitis, wherein the spoonable nutritional composition is as defined above; preferably the subject is an infant.

DESCRIPTION OF THE DRAWINGS

Figure 1

Comparison of branched-chain amino acid (BCAA) levels between potato and rice protein, and whole milk.

Figure 2

Comparison of mean levels of combined aromatic amino acids between potato and rice protein, and whole milk.

Figure 3

Flowchart depicting example preparation process. DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a spoonable nutritional composition comprising protein, carbohydrate and fat, wherein the major source of protein is potato protein.

The term "nutritional composition" refers to a composition that provides nutrition. Such a composition may include protein, carbohydrate, fat and optionally other components (e.g. vitamins and minerals) useful for nutrition of a subject. The levels of the individual components in a nutritional composition may be selected so as to provide tailored nutritional intake for an individual.

By "spoonable" nutritional composition is meant a composition that may be readily consumed using a spoon.

An example of a spoonable nutritional composition is a yoghurt-type composition. Subject

A subject as described herein, for example a subject having Cow's Milk Protein Allergy or Eosinophilic Esophagitis, may be an adult or a child, e.g. an infant.

An infant may be a child under the age of 36 months, for example between 0 and 24 months, or between 0 and 12 months.

Allergy

The term "allergy" refers to a hypersensitivity of the immune system to a substance which is normally tolerated. The allergy may be an allergy detected by a medical doctor.

The term "food allergy" refers to an allergy with respect to a nutritional composition. Infant nutritional composition The term "infant nutritional composition" as used herein refers to a foodstuff such as a nutritional composition that is suitable for use in feeding an infant.

Protein

The term "protein" refers to polymers of amino acids, and includes polypeptides and peptides. The term "protein" does not encompass free amino acids, which may also be present in the spoonable nutritional compositions of the invention.

A spoonable nutritional composition of the invention may comprise between 1 and 10 wt.% protein, for example between 1 and 5 wt.%, or between 2 and 4 wt.%, or about 1 , about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9 or about 10 wt.%.

The spoonable nutritional composition of the invention comprises potato protein as the major protein source.

In one embodiment, at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% by weight of the total protein is potato protein.

The remaining protein in the spoonable nutritional composition of the invention may be any protein which is suitable for use in a spoonable nutritional composition.

In one embodiment, the major source of protein in the spoonable nutritional composition is potato protein and the remaining protein is plant protein.

Example plant proteins that may optionally be used in the spoonable nutritional compositions of the invention, in addition to the potato protein, include pea, rice, quinoa, oat, sunflower or coconut proteins, or combinations thereof.

In one embodiment, the major source of protein in the spoonable nutritional composition is potato protein and the remaining protein is rice protein. The spoonable nutritional compositions of the invention may be formulated to be entirely absent in meat or dairy products, which may be advantageous for nutritional, taste, ethical or religious reasons. For example, the spoonable nutritional compositions of the invention may be formulated to be suitable for vegetarian or vegan diets.

In a preferred embodiment, 100% by weight of the total protein is potato protein. Thus, in one embodiment, the sole source of protein in the spoonable nutritional composition is potato protein.

Potato protein for use in the spoonable nutritional compositions of the invention is readily accessible or available, for example as concentrates or isolates, for example from commercial sources of potato protein.

Potato protein may be extracted from potato tuber juice, which may itself be separated from potato solids by any of a number of suitable techniques known in the art. Chromatographic techniques may be used to purify potato proteins from the tuber juice in a similar manner to the isolation of milk proteins. Once isolated, the potato protein may be concentrated and subjected to temperature treatment and/or pH adjustment. Further steps may for example include removal of triglycoalkaloids, spray drying and/or UV treatment.

Suitable potato protein sources include complete potato protein extract (i.e. extract not subjected to fractionation by molecular mass); and potato protein fractionated by molecular mass, for example a high molecular mass fraction (e.g. greater than 35 kDa); or a low molecular mass fraction (e.g. less than 35 kDa). In one embodiment, the potato protein source is a low molecular mass potato protein fraction of less than 35 kDa.

The protein may be, for example, intact protein or hydrolysed protein (e.g. partially hydrolysed protein). Preferably, the protein is intact protein.

Hydrolysis of protein may in general be termed "partial" or "extensive" depending on the degree to which hydrolysis is carried out. Protein hydrolysates may have an extent of hydrolysis that is characterised by NPN/TN%, which refers to the non-protein nitrogen divided by the total nitrogen χ 100. The non-protein nitrogen refers to amino nitrogen that is free to react with a reagent such as trinitrobenzenesulfonic acid (TNBS). NPN/TN% may be measured as described in Adler-Nissen (Adler-Nissen, J. (1979) J. Agric. Food Chem. 27: 1256-1262).

The term "extensive hydrolysis" may refer to hydrolysis that provides protein that has a NPN/TN% greater than 95%. The term "partial hydrolysis" may refer to hydrolysis that provides protein that has a NPN/TN% in the range 70-85%.

In one embodiment, the protein has an NPN/TN% between 5-90%, 70-90% or 70- 85%, preferably between 70-85%. In another embodiment, the protein has an NPN/TN% between 25-55%, 25-50% or 50-55%.

In one embodiment, 60-70% of the protein population has a molecular mass of less than 5000 Da.

In another embodiment, the protein has an NPN/TN% greater than 95%. These are "extensive" hydrolysates.

In one embodiment, 60-70% of the protein population has a molecular mass of less than 3000 Da. In one embodiment, at least 95% of the protein population has a molecular mass of less than 3000 Da.

Proteins for use in the spoonable nutritional compositions of the invention may be hydrolysed by any suitable method known in the art. For example, proteins may enzymatically hydrolysed, for example using a protease.

For example, protein may be hydrolysed using alcalase (e.g. at an enzyme:substrate ratio of about 2-15% by weight and for a duration of about 1 -5 hours).

Free amino acids

The spoonable nutritional compositions of the invention may further comprise free amino acids. Such free amino acids provide a protein equivalent source. Free amino acids may be incorporated in the spoonable nutritional compositions of the invention to supplement the amino acids comprised in the protein. The levels of free amino acids may be chosen to provide an amino acid profile that is sufficient for infant nutrition, in particular an amino acid profile that satisfies nutritional regulations (e.g. European Commission Directive 2006/141 /EC).

Example free amino acids for use in the spoonable nutritional compositions of the invention include histidine, isoleucine, leucine, lysine, methionine, cysteine, phenylalanine, tyrosine, threonine, tryptophan, valine, alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, proline, serine, carnitine, taurine and mixtures thereof.

Carbohydrate

The carbohydrate may be any carbohydrate which is suitable for use in spoonable nutritional compositions.

A spoonable nutritional composition of the invention may comprise between 5 and 25 wt.% carbohydrate, for example between 10 and 20 wt.%, or between 10 and 15 wt.%, or about 5, about 10, about 15, about 20 or about 25 wt.%.

Example carbohydrates for use in the spoonable nutritional compositions of the invention include lactose, sucrose, saccharose, maltodextrin and starch. Mixtures of carbohydrates may be used.

In one embodiment, the carbohydrate comprises maltodextrin. In one embodiment, at least about 40%, 50%, 60% or 70% by weight of the total carbohydrate is maltodextrin. In one embodiment, the carbohydrate comprises maltodextrin and sucrose.

Fat

The fat may be any lipid or fat which is suitable for use in spoonable nutritional compositions. A spoonable nutritional composition of the invention may comprise between 1 and 10 wt.% fat, for example between 1 and 5 wt.%, or between 2 and 4 wt.%, or about 1 , about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9 or about 10 wt.%.

Example fats for use in the spoonable nutritional compositions of the invention include, sunflower oil, low erucic acid rapeseed oil, safflower oil, canola oil, olive oil, coconut oil, palm kernel oil, fish oil, palm oleic, high oleic sunflower oil and high oleic safflower oil, and microbial fermentation oil containing long chain, polyunsaturated fatty acids.

The fat 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, docosahexaeonic acid, linolenic acid, oleic acid, lauric acid, capric acid, caprylic acid, caproic acid, and the like.

Further example fats include structured lipids (i.e. lipids that are modified chemically or enzymatically in order to change their structure). Preferably, the structured lipids are sn2 structured lipids, for example comprising triglycerides having an elevated level of palmitic acid at the sn2 position of the triglyceride.

Oils containing high quantities of preformed arachidonic acid and/or docosahexaenoic acid, such as fish oils or microbial oils, may also be added.

Long chain polyunsaturated fatty acids, such as dihomo-Y-linolenic acid, arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid, may be added. Willemsen et al. showed that the addition of such fatty acids supported epithelial barrier integrity and reduced IL-4 mediated permeability (Willemsen, L.E. et al. (2008) Eur. J. Nutr. 47: 183-91 ).

Structured lipids may be added or may be omitted. Medium chain triglycerides may be added or may be omitted.

Further ingredients A spoonable nutritional composition of the invention may contain vitamins and minerals understood to be essential in the daily diet in nutritionally significant amounts. Minimum requirements have been established for certain vitamins and minerals.

Example vitamins, minerals and other nutrients for use in the spoonable nutritional compositions of the invention include 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, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine and L-carnitine.

Minerals are usually added in their salt form.

A spoonable nutritional composition of the invention may comprise at least one probiotic. The term "probiotic" refers to microbial cell preparations or components of microbial cells with beneficial effects on the health or well-being of the host (Salminen, S. et al. (1999) Trends Food Sci. Technol. 10: 107-10).

In particular, probiotics may improve gut barrier function (Rao, R.K. (2013) Curr. Nutr. Food Sci. 9: 99-107).

Preferred probiotics are those which as a whole are safe, are L(+) lactic acid producing cultures and have acceptable shelf-life for products that are required to remain stable and effective for up to 24 months.

In an embodiment the probiotics may be in the form of non-replicating probiotics. "Non- replicating" probiotics are further defined as probiotic bacteria which have been heat treated. This includes microorganisms that are inactivated, dead, non-viable and/or present as fragments such as DNA, metabolites, cytoplasmic compounds, and/or cell wall materials.

Examples of probiotic micro-organisms for use in the spoonable nutritional compositions of the invention include yeasts, such as Saccharomyces, Debaromyces, Candida, Pichia and Torulopsis; and bacteria, such as the genera Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus and Lactobacillus.

Specific examples of suitable probiotic microorganisms are: Saccharomyces cereviseae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei subsp. casei, Lactobacillus casei Shirota, Lactobacillus curvatus, Lactobacillus delbruckii subsp. lactis, Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus rhamnosus (Lactobacillus GG), Lactobacillus sake, Lactococcus lactis, Micrococcus varians, Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus acidilactici, Pediococcus halophilus, Streptococcus faecalis, Streptococcus thermophilus, Staphylococcus carnosus and Staphylococcus xylosus.

Preferred probiotic bacterial strains include Lactobacillus rhamnosus; Lactobacillus rhamnosus LPR (CGMCC 1 .3724); Bifidobacterium lactis BL818 (CNCM 1 -3446) sold inter alia by the Christian Hansen company of Denmark under the trade mark BB 12; and Bifidobacterium longum BL999 (ATCC BAA-999) sold by Morinaga Milk Industry Co. Ltd. of Japan under the trade mark BB536.

The spoonable nutritional compositions of the invention may also contain other substances which may have a beneficial effect such as prebiotics, lactoferrin, fibres, nucleotides, nucleosides and the like.

The spoonable nutritional compositions of the invention may comprise one or more natural sugars, for example sucrose, saccharose, all types of glucose syrup or fructose syrup. Additionally, or alternatively, the spoonable nutritional compositions of the invention may comprise at least one natural or artificial high intensity sweetener, for example stevia extracts, rebaudioside A, aspartame, neotame, acesulfame, sucralose, or saccharin. The spoonable nutritional compositions of the invention may comprise a flavouring agent.

Flavouring agents may be used alone or in any possible combination in connection with the present invention. Flavouring agents used in the present invention may be any natural flavouring agent or any artificial flavouring agent.

The flavouring agent may be in the form of a flavour and/or extracts and/or aromatic oils.

The flavouring agent may be derived from fruit and berries (e.g. banana, raspberry, strawberry, apricot, blueberry, apple, pear, mango, papaya, citrus, orange etc.), from nuts (e.g. hazel nuts, almonds, pine nuts, pistachio, cashew nuts, etc.), spices (e.g. chai, basil, cardamom, cinnamon, thyme, etc.), flowers (e.g. rose, elderflower, etc.), roots (e.g. liquorice/licorice), plants (e.g. vanilla, mint, fir, tea, etc.), as well as other sources (e.g. honey). Flavouring agents furthermore comprise ingredients/flavours such as cocoa, chocolate, coffee, mocha, caramel, caramel essence, nougat, nougat essence, marzipan, almond essence, oils, liquor, brandy, rum, port, whiskey, wine etc.)

In one embodiment, the flavour is vanilla flavour powder. In one embodiment, the flavour is cocoa powder. Method of manufacture

The spoonable nutritional compositions of the invention may be prepared in any suitable manner.

Preparation of a spoonable nutritional composition of the invention may comprise combining protein, carbohydrate and fat components in appropriate proportions. Water may be used to form a liquid mixture. The protein component may be mixed into water at a temperature of between 50 and 60°C, preferably about 55°C, to aid dispersion. Further dry ingredients may be added, followed by liquid ingredients, for example oils.

The pH of the liquid mixture may be adjusted, for example to between 6.0 and 7.0, or to or at least about 6.4, or at least about 6.6, or at least about 6.8, or at least about 7.0.

The pH of the liquid mixture may be adjusted using standard techniques known in the art. By way of example, the pH of the liquid mixture may be adjusted using citric acid (e.g. a 30% aqueous solution) or potassium hydroxide (e.g. a 30% aqueous solution).

The liquid mixture may be homogenised. Homogenisation may be carried out at a temperature of between 50 and 65°C, preferably between 50 and 60°C, and at a pressure of between 200 and 250 bar. Homogenisation may be carried out in two stages. A two-stage homogenisation may be carried out at two different pressures, for example a 200 bar first stage followed by a 50 bar second stage.

The liquid mixture may optionally be thermally treated to reduce bacterial loads, for example by rapidly heating the liquid mixture to a temperature between about 80°C and about 150°C for a duration between about 5 seconds and about 5 minutes, for example between about 10 seconds and about 1 minute. This may be carried out, for example, by means of steam injection, or using an autoclave or heat exchanger, for example a plate heat exchanger, or by indirect UHT

The liquid mixture may then be cooled, for example to a temperature of between 20 and 30°C, and subsequently aseptically filled into a suitable container.

The spoonable nutritional composition obtained may be suitable for shelf storage at room temperature or refrigerated storage.

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of chemistry, biochemistry, molecular biology, microbiology and immunology, which are within the capabilities of a person of ordinary skill in the art. Such techniques are explained in the literature. See, for example, Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press; Ausubel, F.M. et al. (1995 and periodic supplements) Current Protocols in Molecular Biology, Ch. 9, 13 and 16, John Wiley & Sons; Roe, B., Crabtree, J. and Kahn, A. (1996) DNA Isolation and Sequencing: Essential Techniques, John Wiley & Sons; Polak, J.M. and McGee, J.O'D. (1990) In Situ Hybridization: Principles and Practice, Oxford University Press; Gait, M.J. (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press; and Lilley, D.M. and Dahlberg, J.E. (1992) Methods in Enzymology: DNA Structures Part A: Synthesis and Physical Analysis of DNA, Academic Press. Each of these general texts is herein incorporated by reference.

Various preferred features and embodiments of the present invention will now be described by way of non-limiting examples.

EXAMPLES

Example 1

Nutritional comparison between potato protein and rice protein

Potato protein contains higher levels of the following essential amino acids compared to rice protein: valine, isoleucine, leucine, lysine, threonine and aromatic amino acids.

The concentrations of tryptophan and the sulphur-containing amino acids are similar between potato and rice proteins.

Overall, the essential amino acid concentrations in potato protein are better than rice protein, and may require lower levels of additional amino acid fortification.

Branched-chain amino acids (BCAA)

Branched-chain amino acids (BCAA; leucine, isoleucine and valine) have an important role in protein synthesis. Leucine is an activator of mTOR, and promotes protein synthesis and suppress protein catabolism, resulting in maintenance of muscle protein during restricted dietary intake. Children with food allergies follow dietary restrictions, therefore they are at risk of developing malnutrition, hence consumption of plant protein with high levels of BCAAs may help maintain muscle proteins.

Additionally, the best food sources of BCAAs are meat, fish, dairy products and eggs, which may not be consumed at all, or at least consumed in smaller amounts by infants and small children with food allergies. Figure 1 shows that the sum of BCAA (%AA) in potato is closer to that in milk and therefore provides an advantage to children with cow's milk protein allergy. Accordingly, providing a protein source with higher levels of BCAAs may benefit this paediatric population.

Lysine and threonine Lysine and threonine are the first and second most limiting amino acids, respectively, for protein synthesis in human subjects consuming a predominantly cereal-based diet such as wheat and rice. The main roles of lysine and threonine are in protein synthesis. Unlike other plant proteins sources such as rice and wheat proteins, potato protein has higher levels of these two amino acids.

The best food sources of threonine and lysine are soy, dairy products, nuts, and fish, beef or chicken. These food sources may not be consumed at all, or at least consumed in smaller amounts by infants and small children with food allergies. Therefore providing a non-animal source of protein with high concentrations of these two amino acids will benefit this paediatric population.

Aromatic amino acids

Phenylalanine is a precursor for tyrosine, the neurotransmitters dopamine, norepinephrine, and adrenaline, and the skin pigment melanin.

The best food sources of phenylalanine are eggs, chicken, liver, beef, milk and soybeans. These food sources may not be consumed at all, or at least consumed in smaller amounts by infants and small children with food allergies. However, the combined levels of phenylalanine and tyrosine in potato protein are similar to those in milk (Figure 2), which provides an advantage to infants and children with cow's milk protein allergy.

Example 2

10 different example spoonable nutritional compositions were prepared, with the ingredients of each set out in Tables 1 and 2, below. Also set out are the conditions used to prepare each example composition (Tables 3 and 4, below).

>35 kDa <35 kDa >35 kDa >35 kDa >35 kDa potato potato potato potato potato protein protein protein protein protein fraction fraction fraction fraction fraction pH neutral pH neutral pH neutral pH neutral pH neutral

Protein: 3.0 3.0 3.0 5.0 9.0

Carbohydrate 16.7 16.7 13.3 13.3 13.3

Fat 3.5 3.5 3.5 3.5 3.5

TS 23.8 23.8 20.3 22.4 26.7

Energy (kcal/100g) 110.4 110.4 96.7 104.6 120.8

Added sugar per lOOkcal 5.4 5.4 6.2 5.7 5.0

Material Description % % % % %

Water 74.99 74.99 78.59 76.39 71.89

Maltodextrin 8.00 8.00 8.00 8.00 8.00

Sugar 6.00 6.00 6.00 6.00 6.00

Protein Potato Isolate HMW 3.30 3.30 5.50 10.00

Protein Potato Isolate LMW 3.30

Canola Oil 3.70 3.70 3.70 3.70 3.70

Starch Mod. Potato 3.50 3.50

Gum Carob 0.10 0.10

Cocoa Powder

Tricalciumphosphate 0.42 0.42 0.42 0.42 0.42

Sum [%] 100.0000 100.0000 100.0000 100.0000 100.0000

Table 1 - compositions 1 to 5

Added sugar per lOOkcal 6.2 5.7 5.0 6.2 6.2

Material Description % % % % %

Water 78.59 76.39 71.89 77.14 77.14

Maltodextrin 8.00 8.00 8.00 8.00 8.00

Sugar 6.00 6.00 6.00 6.00 6.00

Protein Potato Isolate HMW 3.50 3.50

Protein Potato Isolate LMW 3.30 5.50 10.00

Canola Oil 3.70 3.70 3.70 3.70 3.70

Starch Mod. Potato

Cocoa Powder 1.25 1.25

Tricalciumphosphate fine 0.42 0.42 0.42 0.42 0.42

KOH

Sum [%] 100.0000 100.0000 100.0000 100.0000 100.0000

Table 2 - compositions 5-10

Mixing dry mix dry mix dry mix dry mix dry mix powders powders powders powders powders add to add to add to add to add to water and water and water and water and water and oil oil oil oil oil pH (evtl. ja nach pH) measure measure measure measure measure mix, adjust mix, adjust mix, adjust mix, adjust mix, adjust to pH > 6.8 to pH > 6.8 to pH > 6.8 to pH > 6.6 to pH > 6.6 swelling time 30 min 30 min 30 min 30 min 30 min

Homo (55-60°C) Evtl. 200- Evtl. 200- Evtl. 200- Evtl. 200- Evtl. 200- 250 bar 250 bar 250 bar 250 bar 250 bar

Pasteurization/UHT Bench 137 °C 137 °C 137 °C 137 °C 137 °C

Holding 33 s 33 s 33 s 33 s 33 s

Cooling 30°C 30°C 30°C 30°C 30°C

Storage 6 °C 6 °C 6 °C 6 °C 6 °C

Table 4 - preparation conditions for compositions 6-10

Example 3

A spoonable nutritional composition according to the present invention was prepared with ingredients as set out in Table 5 below. In this example composition, 80% of the total protein was potato protein. The spoonable nutritional composition produced was stable and with a pudding-like texture.

Table 5

Example 4 A spoonable nutritional composition of the invention was prepared as follows, with steps as shown in Figure 3.

Water was provided at a temperature of 55°C.

Potato protein and rice protein were added to the water and stirred continuously for 30 minutes at 55°C.

Sucrose, starch and cocoa powder were added to the mixture, followed by vegetable oil.

The mixture was heated to a temperature of 60°C before being homogenised at a pressure of 200 bar.

The homogenised mixture underwent a heat treatment to 137°C.

The heat treated mixture was cooled to 25°C and filled into containers using aseptic techniques.