WO2003055502A1

WO2003055502A1 - Immunoglobulin composition

Info

Publication number: WO2003055502A1
Application number: PCT/NZ2002/000293
Authority: WO
Inventors: Peter Graeme Hobman; Charles Edward Williams; Joanne Campbell Hutchinson
Original assignee: Fonterra Co-Operative Group Limited
Priority date: 2001-12-24
Filing date: 2002-12-24
Publication date: 2003-07-10
Also published as: AU2002367161A1

Abstract

A protein powder composition having an antibody binding activity effect against Candida albicans and against a range of other antigens.

Description

IMMUNOGLOBULIN COMPOSITION

TECHNICAL FIELD

The invention relates to protein powder composition having a broad antibody binding activity. The invention also relates to a method of producing such a protein powder composition, compositions containing the protein powder composition and methods of treating infections using such compositions.

BACKGROUND ART

IgA is a secretory antibody that is present in mucosal systems in the body. It is the predominant antibody in the human body, while in cows IgG is the predominant antibody. An elevation in the amount of IgA in a dairy product is beneficial as it can be used to fortify the mucosal immunity of the animal and also consumer products produced to have immunoprotective effects.

One method of increasing the level. of IgA in milk from cows is to specifically immunise for Candida albicans as discussed in WO98/54226 to AgResearch Limited. This increases the levels of Candida albicans specific IgA in the cow and hence in the milk. Milk produced by such an immunised cow will have levels of antibody binding activity, resulting from higher IgA levels, on the human gastro-intestinal tract that will assist in combating the effects of Candida albicans.

This effect is specific and it would be desirable to be able to extend the antibody binding activity effect beyond the single antigen to a broader range of antigens in both the animal and the animal's products.

It will be clearly understood that reference to the prior art publication referred to herein does not constitute an admission that this document forms part of the common general knowledge in the art, in New Zealand or in any other country. SUMMARY OF THE INVENTION

In a first aspect the invention provides a protein powder composition having an antibody binding activity effect against Candida albicans and against a range of other antigens.

In a second aspect the invention provides a milk protein powder composition that is a milk protein concentrate having increased levels of IgA and IgG, and which has elevated antibody binding activity against C. albicans and a number of other antigens.

Preferably the amount of IgA in the powder composition is at least about 0.15%, preferably about 0.15% to 0.35% more preferably about 0.2%.

Preferably the amount of IgG in the powder composition is at least about

1.5% of the composition and more preferably at least about 2.0%.

Preferably the composition includes pharmaceutically acceptable carriers or excipients.

Preferably the amount of IgA in the powder composition is elevated as a result of an immunisation protocol targeted at Candida albicans.

Preferably the IgG effect is enhanced against any one or more of Candida albicans, Enterobacter aerogenes, Staphylococcus epidermidis, E. coli,

Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

Preferably there is a synergistic antigenic effect between the IgA and IgG in the powder composition resulting in increased antigenic activity against any one of a number of antigens.

In another aspect the invention provides a composition for the treatment of pathogen related infections of mucosal membranes, the composition including a milk protein concentrate powder having elevated levels of IgA and IgG. Preferably the amount of IgA in the powder composition is at least about 0.15%, preferably about 0.15% to 0.35% more preferably about 0.2%.

Preferably the amount of IgG in the powder composition is at least about 1.5% of the composition and more preferably at least about 2.0%.

Preferably the mucosal membrane is the gastrointestinal tract, (including the oral cavity and throat), skin, nasal passages and/or the vagina.

In another aspect the invention provides a method of producing a powder composition having a broad antibody binding activity effect, the method including the steps of completing an immunisation protocol targeted at a specific antigen in a lactating mammal, collecting the milk produced by the mammal, and concentrating the milk into a powder composition.

Preferably the lactating mammal is a cow, goat or sheep.

Preferably the targeted antigen is Candida albicans and the powder composition has an elevated antibody binding activity effect against Candida albicans and against any one or more of Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri,

Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

In another aspect the invention provides a method of treatment of a number of pathogen related conditions in the mucosal membranes of the body of a mammal by administration of a dairy protein composition having elevated levels of IgA and IgG.

Preferably the pathogens creating the conditions are Candida albicans, Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, and Corynebacterium ovis, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

Preferably the dairy protein composition is produced from a milk product taken from a cow that has been subjected to an immunisation protocol targeted at Candida albicans.

Preferably the mucosal membrane is in the gastrointestinal tract, (including the oral cavity and throat), skin, nasal passages and/or the vagina.

In another aspect, the invention provides a method of treating, or reducing the occurrence of, conditions resulting from the presence of Candida albicans, and any one or more of Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle in the mucosal membranes of the body of a mammal, by the administration of a dairy protein composition or milk product that has elevated levels of IgA and IgG as a result of a specific immunisation protocol targeting Candida albicans.

Preferably the mucosal membrane is the gastrointestinal tract (including the oral cavity and throat), skin, nasal passages and/or the vagina.

Preferably the conditions treated result from C. albicans and Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis.

In another aspect of the invention provides a method of immunising a lactating mammal against a range of pathogens by specifically immunising the mammal against a specific pathogen to increase the levels of IgA specific to that pathogen in the mammal.

Preferably the mammal is a cow, goat or sheep and the targeted pathogen is Candida albicans.

Preferably the levels of both IgA and IgG in the mammal are elevated. Preferably the mammal is a cow which has increased resistance to Candida albicans and to Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, and Corynebacterium ovis.

In another aspect the invention provides the use of a dairy protein composition or milk product having increased levels of IgA and IgG as a result of a specific immunisation protocol targeting Candida albicans in the manufacture of a medicament for the treatment of conditions in the mucosal membrane of the body of. a mammal resulting from the presence of Candida albicans, and any one or more of Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

Preferably the conditions treated result from C. albicans and Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori.

Preferably the mammal is a human, cow, goat or sheep.

DRAWINGS

Figures 1 - 7: show ELISA assays of IgG levels against a series of pathogens. Figure 8: shows blot assays showing inhibition of C. albicans adhesion to human salivary cells in-vitro.

DETAILED DESCRIPTION OF THE INVENTION

In general terms the invention relates to a milk protein powder which combines two immunoglobulins, IgA and IgG in elevated proportions in comparison to a standard milk protein powder (or concentrate) The amount of IgA in a milk protein powder can be increased by a variety of methods including specific immunisation protocols targeting specific pathogens, such as Candida albicans. A methodology to elevate the levels of IgA in milk is disclosed in WO 98/54226. The disclosure of this document is incorporated herein by way of reference. However, this document does not disclose the preparation of a protein powder from the milk, nor does it disclose elevated levels of IgG, nor does it disclose the increased antibody binding activity effect against a broad array of antigens, nor does it disclose an increased immune effect in the animal.

It has now been surprisingly found that such an immunisation protocol not only increases the levels of IgA but also the levels of IgG. It has also been surprisingly found that the antibody binding activity effect of the concentrate formed from the milk product has a broad range of activity against a number of pathogens other than that specifically targeted in the immunisation protocol. When Candida albicans is targeted, other pathogens affected include Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

That this effect occurs from a single immunisation target protocol is surprising as a targeted immunisation protocol should result in an increased response only against the targeted antigen.

It is unclear at this stage whether the increased antibody binding activity effect observed is due solely to the higher IgG levels or whether there is some form of synergistic effect occurring between the IgA and the IgG. The invention may therefore be seen to include a synergistic combination of IgA and IgG in these elevated proportions.

By boosting the antibody binding activity effect of IgG in the protein composition, an immune boost supplementation of compositions including IgG and IgA can be achieved.

To this extent, the invention can be broadly seen to be directed to a composition that has increased levels of IgA and IgG and which has a broad antibody binding activity effect in relation to a number of pathogens.

The invention can also be seen to be a method of immunising a mammal, and in particular a ruminant mammal, more particularly a bovine mammal (although sheep or goats could also be targeted), against a broad range of pathogens by following an immunisation protocol targeted at a single pathogen. In particular Candida albicans is a preferred immunisation target.

For example, targeting Candida albicans has the result of raising both IgA and IgG levels in the cow's system thus increasing resistance to the broad range of pathogens listed earlier. Thus the overall health of the animal benefits in an unexpected manner.

The elevated levels of IgA will preferably be in the range of about 0.15% to 0.35% of the composition and most preferably above about 0.2%. The amount of IgG in the composition will preferably be about 1.5% of the composition and preferably above about 2.0%.

The results in Example 1 and Figures 1 to 7 show the occurrence of increased antibody binding activity effects of a product produced according to the invention method. The results also show that the subject mammals' immune systems has been boosted to produce an immune response that is reflected in the antibody binding activity effects of the concentrate product.

Thus the invention can be seen to provide a method of treating conditions created by a variety of pathogens in a mammal via the use of a dairy protein composition having elevated levels of IgA and IgG resulting from an immunisation protocol targeting a specific antigen, in particular Candida albicans. The method treats pathogens in mucosal membranes in the mucosal systems of the body, predominantly this will be in the gastrointestinal tract (that includes the oral cavity and throat) but will also include the nasal passages, skin and vagina. More specifically the invention can be seen to be a method of treating or reducing the incidence of conditions resulting from Candida albicans and any one or more of Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, and/or Shigella flexneri, Corynebacterium ovis Helicobacter pylori, and Clostridum spp. including Clostridium difficle in a mammal via the use of a dairy protein composition or milk product having elevated levels of IgA and IgG created by an immunisation protocol specifically targeting Candida albicans.

Such pathogens adhere to the cells in the mucosal membranes in the mucosal systems of the body. It is hypothesised that treatment with a composition according to the present invention interferes with that adhesion at the adhesion site.

With reference to Example 2, it can be seen that results indicate activity in- vivo against C. albicans but not C. difficle. It is hypothesised that this is due to the effect of digestive enzymes on the active part of the composition, preventing activity at the site of the C. difficle infection.

C. difficle is usually present in the large bowel and this may therefore be a limitation on using the composition and method of the invention without some form of protective coating on the composition to allow passage to the large bowel while avoiding the impact of the digestive enzymes.

To this extent the invention will include a composition including a dairy protein powder concentrate having elevated IgA and IgG levels together with a suitable protective coating (eg an enteric coating). The composition could take the form of a capsule for example. For infections of the upper gastrointestinal tract, such requirements are unlikely to the needed as the activity against C. albicans clearly shows in the in-vivo tests of Example 2.

By way of further option, if the area to be treated is in the nasal passages, the composition could be formulated as a spray or like product as will, again, be known to the skilled person. For other mucosal membranes at the mouth, skin and vagina, similar application issues will arise relevant to the respective areas. The composition could be formulated as on oral solution, a skin or vaginal cream or a douche, as desired, although such options are not intended to be limiting.

All of the bacterial species listed earlier tend to be present in the lower gastrointestinal tract of humans - with the exception of:

^■ Staphylococcus epidermidis - this is often a topical infection of mucosal membranes (not unlike Candida), which may become systemic;

■ Corynebacterium ovis - this can present in the gastrointestinal tract of some animals (not humans), and it is a common cause of abscesses, especially in goats and sheep;

■ Helicobacter pylori - this is present in the upper gastrointestinal tract - mainly in the stomach.

Therefore, for those bacterial species in the lower gastrointestinal tract, some form of protective shield may be needed to ensure activity. The same may be the case for others as needed but such issues are well within the knowledge of the skilled person to determine and to overcome using known methods. Another option would be the use of enemas to target infections in the lower gastro-intestinal tract.

The invention allows the use of such a composition or food product containing a dairy protein composition in the manufacture of a composition for the treatment of conditions arising from the presence of Candida albicans, Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle. Such conditions are, at least in part, listed in Table 1. The immunisation protocol is conducted on the source of the dairy product. While reference to cows is made herein, similar results can be expected for goats or sheep. The manufacture of the composition itself can be according to known formulation principles as will be known to a skilled person in this art. Table 1

The mammal treated with the resultant powder composition will preferably be a human and the treatment can be directed at existing symptoms created by the presence of the pathogens in the mucosal membranes (such as the gastrointestinal tract, (including the mouth and throat), nasal passages, skin and the vagina) and can be a prophylactic treatment to prevent or restrict the ability of the pathogen taking hold in the cells in the mucosal membranes. As will be apparent, the composition/medicament will preferably be taken orally or topically applied to the mouth, skin or vagina in a suitable form as will be known to the skilled person. The amount of IgG and IgA in the composition as a whole when taken orally or topically should be: IgA - at least about 0.15%, preferably about 0.15% to about 0.35%, and more preferably about 0.2%; IgG - at least about 1.5% and more preferably at least about 2.0%.

The amount of IgA and IgG in a composition can be varied as desired. It may be in some circumstances that higher concentrations of IgA/lgG will be used. To this extent, the ranges of IgA and IgG given are preferences only.

The invention can therefore also be seen to include a method of treating C. albicans related infections of mucosal membranes of the body. For example, treatment of the skin, gastro-intestinal tract (including the mouth and throat), nasal passages and vagina can be undertaken using compositions including the dairy protein composition having increased IgA and IgG described herein. As has been stated earlier, such mucosal membrane treatment compositions can be formulated by any known means and will include known carriers and excipients suitable for use at the relevant treatment site. The amount of IgA and IgG in the mucosal treatment composition should reflect the preferred ranges discussed previously herein, but could be varied if needed.

As can be seen from Example 3, the composition significantly inhibits binding of C.albicans to salivary proteins in in-vitro models. The three most common forms of C.albicans infection in humans are oral, vaginal and cutaneous (skin) thrush (Candidosis). The mucous membranes in all sites are subject to the same type of infection, and therefore the same type of treatment. Treatment of Candidosis at mucosal sites using a composition or method according to the present invention is therefore another aspect of the present invention.

Examples:

Example 1

A dairy protein powder containing elevated levels of IgA and standard levels of IgG was prepared by the following method:

The method used to manufacture a protein concentrate with an elevated level of IgA (specific activity against Candida albicans), and an elevated level of activity against other pathogens involves collection of milk from immunised cows, pasteurisation to industry standard, separation, concentration and spray drying. This process is monitored throughout to ensure high quality is maintained in the final product. The following diagram illustrates this process:

MILK COLLECTION

I

PASTEURISATION Milk heated to 72° for 15 minutes

SEPARATION Milk skimmed to remove fat

CONCENTRATION

I

EVAPORATION Milk concentrated further using low temperature

^

SPRAY DRYING

Using the powder composition thus created a series of ELISA assays for the effect of IgG against a variety of antigens was run. The dairy powder was compared with a skim milk powder (SMP) and also with a milk protein concentrate (MPC) of similar gross composition to the dairy powder of the present invention, but manufactured from standard miik.

A typical IgA content in MPC is 0.09%.

A typical IgG content in MPC is about 1.2%.

The IgA content in SMP is typically undetectable.

A typical IgG content in SMP is about 0.4%.

A dairy protein powder according to the present invention is represented in the Figures 1 to 7 which illustrate the elevation of antibody binding activity effect observed using an ELISA assay for IgG. The powder composition of the present invention produced as described previously is represented by "IgA" and has been compared with a skim milk powder (SMP) and also with a milk protein concentrate (MPC) of similar gross composition to the "IgA" powder composition (but without the elevated IgA and IgG levels), but manufactured from standard milk. Comparison is also made with commercially available hyperimmune SMP and MPC.

As can be seen from Figures 1 to 7 there is a clear elevation of effect for IgG against a variety of antigens in the powder composition of the present invention.

In conclusion, dairy protein powder according to the present invention has been found to have increased activity in binding at least the following pathogens: Candida albicans, Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis. Efficacy against these pathogens, and hence conditions resulting from their presence, forms the basis for treatments of conditions resulting from pathogen infection.

Example 2

A randomised, two way crossover safety study of anti-Cancf/^'cfa albicans IgA enhanced (IgA MPC) milk protein concentrate and standard milk protein concentrate (standard MPC) was carried out using chronic and acute dosing in healthy male and female volunteers at Zenith Technology Clinical Site, 2^nd floor, 248 Cumberland Street, Dunedin, New Zealand.

The main objective of the study was to evaluate the safety of IgA MPC versus standard MPC in order to compare any side effects of the two products.

30 healthy male (19) and female (1 1) subjects, 18 to 53 years of age (mean: 27, SD: 9 yr) weighing 52 to 105 kg (mean 80.9, SD:15.1 kg) and a body height of 1.58 to 1.95 m (mean 1.76, SD: 0.1 1 m) were enrolled and randomised with 29 subjects completing the study in accordance with the protocol.

Each subject received 20 grams in 200 ml of BP purified water of either the test or reference product once daily for 12 days during the chronic dosing. Each subject received 200 grams in 2000 ml of BP purified water with either the test or reference product over three occasions within a 10 hour period on day 13 of each phase during acute dosing.

Male and females participants in the study were diagnosed as healthy (based on extensive protocol defined pre-study evaluation) and were aged between 18 and 55 years, with a body weight within +/- 15% of Metropolitan Life Tables. Informed consent was obtained from each participant and normal clinical and laboratory results were required for each participant prior to the trial.

Table 2

Efficacy Results

Subjects were required to provide a stool sample for the determination of (i) Clostridium difficle and (ii) Candida albicans levels before the study, after phase one and after phase two.

The results indicate that there is minimal effect on Clostridium difficle following the administration of either formulation ie: Sum of scores IgA enhanced MPC (Test) (2 +/- 0.40); Sum of scores Standard MPC (Reference) (0 +/- 0.40).

The results do indicate that IgA enhanced MPC (test) show a marked reduction in the stool Candida albicans growth levels (4.0 +/- 1.3) compared to that of the Standard MPC (Reference) (26+/- 1.7).

These results indicated that for pathogens present in the upper gastrointestinal tract (eg Candida albicans) use of the composition directly will suffice. For pathogens in the lower gastrointestinal tract (eg Candida difficle) protection from the digestive enzymes or the choice of an alternative administration route will be required.

Statistical Results - Toxicitv

Evaluation of the side effects reported resulting from the administration of IgA enhanced MPC (test) and Standard MPC (Reference) were analysed for variance in crossover design using the sum of scores. No significant difference was detected between the IgA enhanced and the Standard Milk Protein concentrate for the side effects of headache, diarrhoea, nausea, vomiting and rash following chronic dosing, ie all P-values for > 0.05; Headache = 0.7142, Diarrhoea = 0.0972, Vomiting = 0.3432, Nausea = 0.2907 and Rash = 0.4652.

Chronic Dosing: Table 3

No significant difference was detected in the acute dosing for side effects: headache, diarrhoea, vomiting and rashes, ie headache having a p-value of 0.1936 and diarrhoea having a p-value of 0.1854. There were no reported effects for vomiting and rashes. Standard milk protein concentrate however, was found to have significantly higher nausea effect that that of the IgA concentrate, ie a p-value of 0.0065.

The usefulness of the dairy protein composition with elevated IgA and IgG to treat humans for conditions stemming from the variety of pathogens listed earlier can be seen from the results of the efficiency and toxicity tests above.

Example 3

The objective of this experiment was to compare the ability of the two IgA

MPC products (IgA MPC-A and IgA MPC-B) with standard MPC to prevent adherence of C. albicans to human salivary cells (ie relating to treatment of a mucosal membrane).

10g of each of the IgA MPC-A, IgA MPC-B and standard MPC powders were tested by the Department of Oral Sciences and Orthodontics, Otago University in June 2002. Normal MPC from non immunised cows was included as a negative control.

Samples were reconstituted in KCL buffer to 5%. From this stock solution further dilutions of 0.05% and 0.5% were made. A control using KCI buffer only was used as the 0% concentration in the assay. Samples were tested in duplicate. The ability of MPC to inhibit the binding of ³⁵S-radiolabelled Candida albicans to salivary proteins was determined. Briefly, salivary proteins were separated by electrophoresis and transferred to a nylon membrane. The membranes were incubated with a suspension of ³⁵S- radiolabelled Candida albicans that had been pre-incubated with MPC. Membranes were washed, dried and exposed to X-ray film to detect the binding of ³⁵S-radiolabelled Candida albicans to salivary proteins. The adhesion was quantified using NIH image analysis software and the percentage inhibition of adhesion (relative to the no product control) was calculated.

Figure 8 shows one of the duplicate sets of overlay blots comparing the IgA MPC-A and IgA MPC-B with standard MPC. Both IgA MPC products inhibited adherence of ^3SS-radiolabelled Candida albicans to salivary proteins at concentration as low as 0.05%.

Greater than 90% inhibition of adherence was observed over the whole concentration range tested for both the IgA MPC-A and IgA MPC-B. The inhibition of adherence by 0.05% IgA MPC was clearly more that twice that of non-specific inhibition by standard MPC at 0.05%.

In conclusion, both the IgA MPC-A and IgA MPC-B products inhibited adherence of Candida albicans to human salivary proteins compared with the standard MPC. Thus the IgA products could be used as actives in compositions for treating at least Candida based infections, in the oral mucosal membrane. Treatment in other mucosal membranes (eg skin, vagina), by blocking adherence of C. albicans, can therefore also be infered

The foregoing describes the invention including preferred forms thereof. Alterations and modifications that would be obvious to a person skilled in this particular art are intended to be included within the spirit and scope of the invention as defined in the attached claims.

Claims

1. A protein powder composition having an antibody binding activity effect against Candida albicans and against a range of other antigens.

2. A milk protein powder concentrate composition having increased levels of IgA and IgG, and elevated antibody binding activity against Candida albicans and a number of other antigens.

3. A composition according to claims 1 or 2 wherein the amount of

IgA in the powder composition is at least about 0.15% of the composition.

4. A composition according to claim 1 , 2 or 3 wherein the amount of IgG in the powder composition is at least about 1.5% of the composition.

5. A composition according to any one of the preceding claims wherein the composition also includes pharmaceutically acceptable carriers or excipients.

6. A composition according to any one of the preceding claims wherein the amount of IgA in the powder composition is elevated as a result of an immunisation protocol targeted at Candida albicans.

7. A composition according to any one of the preceding claims wherein the IgG effect is enhanced against any one or more of Candida albicans, Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri,

8. A composition according to any one of the preceding claims wherein there is a synergistic antigenic effect between the IgA and IgG in the powder composition resulting in increased antigenic activity against any one of a number of antigens.

9. A composition for the treatment of pathogen related infections of mucosal membranes, the composition including a milk protein concentrate powder having elevated levels of IgA and IgG.

10. A composition according to claim 9 wherein the amount of IgA in the powder composition is at least about 0.15% of the composition.

11. A composition according to claim 9 or 10 wherein the amount of IgG in the powder composition is at least about 1.5% of the composition.

12. A composition according to any one of claims 9 to 11 wherein the composition includes pharmaceutically acceptable carriers or excipients.

13. A composition according to any one of claims 9 to 12 wherein the mucosal membrane is the gastrointestinal tract (including the oral cavity and throat), skin, nasal passages and/or the vagina.

14. A method of producing a powder composition having a broad antibody binding activity effect, the method including the steps of completing an immunisation protocol targeted at a specific antigen in a lactating mammal, collecting the milk produced by the mammal, and concentrating the milk into a powder composition.

15. A method according to claim 14 wherein the lactating mammal is a cow, goat or sheep.

16. A method according to claims14 or 15 wherein the targeted antigen is Candida albicans and the powder composition has an elevated antibody binding activity effect against Candida albicans and against any one or more of Enterobacter aerogenes,

Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

17. A method of treatment of a number of pathogen related conditions in the mucosal membranes of the body of a mammal by administration of a dairy protein composition having elevated levels of IgA and IgG.

18. A method according to claim 17 wherein the pathogens creating the conditions are Candida albicans, Enterobacter aerogenes,

Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, and Corynebacterium ovis, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle.

19. A method according to claim 18 wherein the dairy protein composition is produced from a milk product taken from a cow that has been subjected to an immunisation protocol targeted at Candida albicans.

20. A method according to any one of claims 17 to 19 wherein the mucosal membrane is the gastrointestinal tract (including the oral cavity and throat), skin, nasal passages and/or the vagina.

21. A method of treating, or reducing the occurrence of, conditions resulting from the presence of Candida albicans, and any one or more of Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori, and Clostridum spp. including Clostridium difficle in the mucosal membranes of the body of a mammal, by the administration of a dairy protein composition or milk product that has elevated levels of IgA and IgG as a result of a specific immunisation protocol targeting Candida albicans.

22. A method according to claim 21 wherein the mucosal membrane is the gastrointestinal tract (including the oral cavity and throat), skin, nasal passages and/or the vagina.

23. A method according to claim 21 or 22 wherein the conditions treated result from C. albicans and Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis.

24. A method of immunising a lactating mammal against a range of pathogens by specifically immunising the mammal against a specific pathogen to increase the levels of IgA specific to that pathogen in the mammal.

25. A method according to claim 24 wherein the mammal is a cow, goat or sheep and the targeted pathogen is Candida albicans.

26. A method according to claim 24 or 25 wherein the levels of both IgA and IgG in the mammal are elevated.

27. A method according to claim 26 wherein the mammal is a cow which has increased resistance to Candida albicans and to

Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, and Corynebacterium ovis.

28. The use of a dairy protein composition or milk product having increased levels of IgA and IgG as a result of a specific immunisation protocol targeting Candida albicans in the manufacture of a medicament for the treatment of conditions in the mucosal membrane of the body of a mammal resulting from the presence of Candida albicans, and any one or more of Enterobacter aerogenes, Staphylococcus epidermidis, E. coli,

29. The use of a dairy protein composition or milk product according to claim 28 wherein the mucosal membrane is the gastrointestinal tract (including the oral cavity and throat), skin, nasal passages and/or the vagina.

30. The use of a dairy protein composition or milk product according to claim 28 or 29 wherein the conditions treated result from C. albicans and Enterobacter aerogenes, Staphylococcus epidermidis, E. coli, Proteus vulgaris, Shigella flexneri, Corynebacterium ovis, Helicobacter pylori.

31. The use of a dairy protein composition or milk product according to claim 30 wherein the mammal is a human, cow, goat or sheep.

32. A composition according to any one of claims 9 to 3 wherein the composition is in the form of a capsule, solution or cream.