nBos d 4 α-lactalbumin, Milk

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Cow's milk f2 

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Code: f76
Latin name: Bos domesticus
Source material: nBos d 4 is purified from milk extract

Cow’s milk allergen components

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Milk contains more than 40 proteins, and all of them may act as human species antigens.

Milk of ruminant species other than Cow (e.g., buffalo, Sheep, Goat, human, and many other species) is constituted from the same or very homologous proteins, which share the same structural, functional, and biological properties. However, human milk does not contain b-lactoglobulin (beta-lactoglobulin)(BLG) (2). Human and Bovine milk differ substantially in the ratio of Whey to Casein protein (approximately 60:40 in human milk and approximately 20:80 in Bovine milk) and in the proportions of specific proteins (3).

Milk composition changes during processing. Cow’s milk contains approximately 30 to 35 g/L (3-3.5%) of Cow’s milk proteins (CMPs), which can be divided into 2 main classes: Caseins (80%) and Whey proteins (20%) (4). Caseins are precipitated out by chymosin (rennin) or the acidification of the Milk to pH 4.6, forming the coagulum (curd). The Whey or Lactoserum remains soluble in the Milk serum. Lactoserum constitutes approximately 20% of the CMPs, and coagulum approximately 80% of the CMPs. Caseins and Whey proteins show very different physico-chemical properties.

Coagulum contains the Casein fraction, comprising 4 proteins: ?S1-, aS2-, a -, and b-caseins (alphaS1-, alphaS2-, beta -, and kappa-caseins). Lactoserum contains mainly globular proteins, b-lactoglobulin (beta-lactoglobulin)(BLG) and a-lactalbumin (alpha-lactalbumin)(ALA), followed by minor constituents such as Bovine serum albumin (BSA), Lactoferrin (LF), immuno-globulins (Ig) and proteosepeptone. BLG and ALA are the major ones and are synthesised in the mammary gland. Others, such as BSA, Lactoferrin, and immunoglobulins, come from the blood. Proteosepeptone is derived from Milk proteins through the action of indigenous enzymes, the most significant of which are the hydrolases, such as the lipoprotein lipase, plasmin, and alkaline phosphatise (5). In addition to the above-mentioned proteins, proteolytic fragments of Casein and fat globule membrane proteins have been reported to occur in this fraction (6).

The main characteristics of the major Milk proteins are presented in Table 1 (7-8).

It was classically accepted that the major allergen in Cow’s milk allergy was beta-lactoglobulin, but subsequent research has proved that sensitivity to the various Cow’s milk proteins is widely distributed (3,9,10).

There is a great variability in human IgE response to Cow’s milk, and no single allergen or particular structure can account for a major part of Milk allergenicity (2).

Studies of large populations of allergic patients show that most of the patients are sensitised to several proteins, including BLG (Bos d 5), Casein (Bos d 8), ALA (Bos d 4), BSA (Bos d 6), Lactoferrin, and immunoglobulins (Bos d 7). A great variability is observed in IgE antibody response. Both Casein and BLG, as well as ALA, are major allergens. However, proteins present in very low quantities, such as BSA, immunoglobulins, and especially Lactoferrin, also appear to be important, since 35% to 50% of patients are sensitised to those proteins and sometimes to those proteins only (2).


Lactoserum (Whey)

b-lactoglobulin (BLG)
BLG is the most abundant protein in Whey, accounting for 50% of total protein in the Lactoserum fraction. It has no homologous counterpart in human milk.

a-lactalbumin (ALA)
ALA is a monomeric globular calcium-binding protein representing about 25% of Lactoserum (Whey) proteins. It is a regulatory component of the enzymatic system of galactosyl transferase responsible in mammary secretory cells for the synthesis of lactose.

Bovine serum albumin (BSA)
BSA accounts for around 5% of the total Whey proteins. BSA is physically and immunologically very similar to human blood serum albumin. Its main role is the transport, metabolism and distribution of ligands and the protection from free radicals (7).

Lactoferrin (LF)
LF is a protein of mammary origin and is a Milk-specific iron-binding glycoprotein of the Transferrin family. It can be found in the Milk of most species at levels lower than 1%. LF is present in much higher concentrations in human breast milk, and particularly in colostrum, as compared to Bovine milk. Although it is present in very low concentrations in Cow’s milk, it has been shown to be an important allergen.

The Immunoglobulin (Ig) fraction, which includes IgG and IgE, accounts for about 1% of total Milk protein and 6% of Whey protein. The basic structures of Ig in Bos species are very similar to those in humans, possessing a basic “Y- shaped” unit composed of 4 polypeptide chains linked through intra- and intermolecular disulfide bonds (4). Three IgG classes in Cattle have been recognised as IgG1, IgG2 and IgG3 (11). Data on the potential allergenicity of Bovine immunoglobulins are very limited. However, some studies propose IgG as another Milk allergen due to the observation that IgE from CMA patients specifically binds Bovine IgG (23). Bovine IgG has been reported to be a major Beef allergen (12).

The proteose-peptone fraction represents about 1.1% of the total Milk protein. it is a heat-stable and acid-soluble protein fraction of Milk with important functional properties. This Milk component is derived mainly from the proteolysis of Beta-casein, and the enzymatic activity of plasmin can over time increase its concentration in Milk (4).



The coagulum consists of the whole Casein fraction (i.e., the solid fraction of proteins obtained after coagulation of Milk). It is subdivided into a number of families, of which the most important are aS1 -, aS2 -, b-, K-, Y-caseins (alphaS1 -, alphaS2 -, beta-, kappa-, gamma-caseins) (4). Each individual Casein represents a well-defined chemical compound, but they cross-link to form ordered aggregates (nanoclusters) (i.e., micelles) that assemble into larger structures, forming Casein micelles characterised by a central hydrophobic part and a peripheral hydrophilic layer in suspension in Lactoserum (Whey) (13-14). Their proportion in the micelles is relatively constant at approximately 37%, 13%, 37%, and 13%, respectively.

The main characteristics that should be emphasised are the multiplicity and diversity of proteins that are involved in Cow’s milk allergy (CMA). Polysensitisation to several proteins occurs most often, and all Milk proteins appear to be potential allergens (1). A great variability is observed in the affinity, specificity and magnitude of IgE responses in patients’ sera (15). Most Milk-allergic patients are sensitised to several proteins, including BLG (Bos d 5), Casein (Bos d 8), ALA (Bos d 4), BSA (Bos d 6), Lactoferrin, and Immunoglobulins (Bos d 7) (10,14,16-22). A great variability is observed in IgE response (1).

Casein and BLG, as well as ALA, are major allergens. However, proteins present in very low quantities, such as BSA, immunoglobulins, and especially lactoferrin, also appear to be important since 35% to 50% of patients are sensitized to those proteins and sometimes to those proteins only (19). In the last few years, sensitivity to Casein seems to have increased in terms of both frequency and intensity of IgE response (1). Sensitizations to Casein, BLG, and ALA are closely linked. In contrast, sensitivity to BSA appears to be completely independent, with 50% of the patients being sensitized to BSA regardless of their sensitivity to other Milk allergens (1).

The role of various Cow’s milk proteins (CMPs) in the pathogenesis of CMA is still controversial. Sera from 20 Milk-allergic subjects have been used for Cow’s milk major allergen identification. The prevalence of CMP allergens has been measured as the following: 55% Alpha(s1)-casein, 90% Alpha(s2)-casein, 15% Beta-casein, 50% Kappa-casein, 45% Beta-lactoglobulin, 45% BSA, 95% IgG-heavy chain, 50% Lactoferrin, and 0% Alpha-lactalbumin (23).

Allergens from Bos domesticus listed by IUIS*

Bos d 2 Bos d 3 Bos d 4
Bos d 5 Bos d 6 Bos d 7
Bos d 8    


*International Union of Immunological Societies (www.allergen.org) Jan. 2008.

Table 1. Main Characteristics of the Major Bovine Milk Proteins (1).

Milk ProteinsConcentration in Milj (g/L)Molecular weight (kDa)
20% Whey (approximately 5 g/L)    
10% BLG (Bos d 5) 3-4 18.3
5% ALA (Bos d 4) 1-1.5 14.2
3% Immunoglobulins Bos d 7 0.6-1.0 150
1% BSA (Bos d 6) 0.1-0.4 66.3
Traces of Lactoferrin 0.09 80
80% Whole Casein (Bos d 8) (approximately 30 g/L)    
32% aS1-casein 12-15 23.6
10% aS2-casein 3-4 25.2
28% B-casein 9-11 24.0


Abbreviations: ALA; a-lactalbumin, BLG; B-lactoglobulin, BSA; Bovine serum albumin.


f76 nBos d 4

Native protein purified from cow’s milk (Bos domesticus)

Biological function: a-lactalbumin
Mw: 14 kDa


Allergen description

a-lactalbumin (Alpha-lactalbumin) (ALA) is one of the major allergens in Cow’s milk and represents about 25% of Lactoserum (Whey) proteins and approximately 5% of Cow’s milk protein. (See Table 1, page 52.)

Human and Bovine milk differ substantially in the ratio of Whey to Casein protein (approximately 60:40 in human milk, and approximately 20:80 in Bovine milk) and in the proportions of specific proteins. Although current infant formulas closely mimic the ratio of total Whey to Casein in human milk, the concentration of ALA (the dominant whey protein in human milk) is relatively low in formula, whereas Beta-lactoglobulin, a protein not found in human milk, is the most dominant Whey protein in formula (3). During ALA’s digestion, peptides appear to be transiently formed that have antibacterial and immunostimulatory properties, thereby possibly aiding in the protection against infection. A novel folding variant (“molten globule state”) of multimeric ALA has recently been discovered that has anti-infective activity and enhances apoptosis, thus possibly affecting mucosal cell turnover and proliferation. Cow’s milk also contains ALA, albeit less than human milk (2-5% of total protein in Bovine milk), and protein fractions enriched with ALA may now be added to infant formula to provide some of the benefits of human ALA (3). Recently, Whey sources with elevated concentrations of ALA have become available, which has permitted the development of formulas with increased concentrations of this protein and decreased concentrations of Beta-lactoglobulin (3).

Bos d 4, Alpha-Lactalbumin, is a 14.2 kDa protein (4,28,37,86-101).

An isoform, Bos d 4.0101, has been characterized.

ALA is a monomeric globular calcium binding protein with a molecular weight of about 14 kDa and 4 disulfide bridges, representing about 25% of Lactoserum (Whey) proteins. The protein is stabilised by 4 disulfide bonds and contains 2 structural domains. One of these domains (the alpha-domain) is rich in alpha-helix. The other domain (the beta-domain) is rich in beta-sheet, has 2 disulfide bonds, and includes 1 calcium binding site (102).

ALA plays a central biochemical role in the mammary gland as the regulatory subunit of lactose synthase, and also plays a nutritional role for the rapidly growing neonate as the protein in highest concentration in human milk (103). It is a regulatory component of the enzymatic system of galactosyl transferase, responsible for the synthesis of lactose in mammary secretory cells. It interacts with the enzyme beta-1,4-galactosyltransferase to form the lactose synthase complex. ALA modifies the substrate specificity of beta-1,4-galactosyl-transferase, allowing the formation of lactose from glucose and UDP-galactose (5). In its role in the production of lactose, this protein plays a major role in regulating physiological functions in the mammary gland (104).

ALA possesses a high-affinity binding site for calcium, and this bond stabilises its secondary structure. The complete amino acid sequence of Bovine ALA shows extensive homology with Hen’s egg white lysozyme but also with human ALA (87-90). Some forms of ALA can induce apoptosis in tumour cells (105).

ALA is a simple model Ca2+ binding protein, which does not belong to the EF-hand proteins. It is a classical example of molten globule state. It has a strong Ca2+ binding site, which binds Mg2+, Mn2+, Na+, and K+, and several distinct Zn2+ binding sites. The binding of cations to the Ca2+ site increases protein stability against heat and various denaturing agents, while the binding of Zn2+ to the Ca2+-loaded protein decreases its stability. Some folding variants of Alpha-LA demonstrate bactericidal activity (106).

ALA is characterised by 4 disulfide bridges and is present in 2 variants. As Bovine ALA shows a 72% sequence identity to human ALA, it makes an ideal protein for the nutrition of human infants. Conformational epitopes are important for the allergenicity of the protein. However, in some patients reduced peptides exhibited a similar or even higher IgE-binding capacity than the native corresponding fragment, suggesting that linear epitopes also exist, located in hydrophobic regions, and are exposed as a consequence of protein denaturation (7,28). The significance of this was demonstrated in an investigation of IgE antibody binding capacity of native Bovine ALA and tryptic peptides, utilising sera of 19 patients with CMA; 58% reacted exclusively with intact ALA, while 42% also presented an allergen-specific IgE response to different tryptic peptides derived from ALA (28).

IgE binding to native ALA and to large peptides confirms the importance of conformational epitope(s). However, in some sera, peptides of reduced size, e.g., 59-94 kDa, exhibited a similar or a higher IgE-binding capacity than did the native corresponding fragments, suggesting the existence of sequential epitope(s) exposed through protein denaturation (4,28). Moreover, IgE-binding sequences were also located in hydrophobic regions of the ALA molecule, where antigenicity is very unlikely to be predicted, and/or within parts of the molecule having a very high sequence homology with human ALA (2).

Cross-reactivity between Bovine ALA and ALA from other animal sources is possible but has not been fully elucidated, and unexpected results may be possible. For example, in a report of Mare’s milk allergy in a 51-year-old woman who was able to tolerate Cow’s milk, skin test and serum IgE antibodies for Cow’s milk was negative but was positive for Mare’s milk. Further investigation demonstrated 2 allergen bands most likely representing ALA and Beta-lactoglobulin (106). Allergy to Mare’s milk is rare.

Antibodies to Beta-lactoglobulin show 10% cross-reactivity with Bovine ALA, both in its native and in its denatured form, which has been attributed to a continuous stretch of 4 amino acids common to ALA and Beta-lactoglobulin. Cross-reactivity between this antibody and Bovine serum albumin was negligible. No cross-reaction was seen with antibodies to ALA and to serum albumin (107).

As the deduced amino acid sequence of buffalo ALA differs at 1 position from the Bovine ALA sequence, cross-reactivity between these 2 is possible but has not been clinically investigated (108).


f77 nBos d 5

Native protein purified from cow’s milk (Bos domesticus)

Biological function: b-lactoglobulin
Mw: 18 kDa


Allergen description

b-lactoglobulin (Beta-lactoglobulin) (BLG) is one of the major allergens in Cow’s milk. BLG is the most abundant protein in Whey, accounting for 50% of total protein in the lactoserum fraction and approximately 10% of Cow’s milk. (See Table 1, page 52)

Although current infant formulae closely mimic the ratio of total Whey to Casein in human milk, the concentration of Alpha-lactalbumin is relatively low in formula, whereas BLG, a protein not found in human milk, is the dominant Whey protein in formula. Whey sources with elevated concentrations of Alpha-lactalbumin have been developed, which has permitted the provision of formulae with increased concentrations of this protein and decreased concentrations of BLG (3). BLG was measured in 7 different infant Cow’s milk protein Whey or Casein hydrolysated formulae. BLG levels in these formulae were 1/100 to 1/4,800,000 lower than in Cow’s milk. There was a great difference in the BLG levels between the partly and the extensively hydrolysed formulae; the amount of BLG was 40,000-fold higher in the partially hydrolysated vs. the extensively hydrolysated formulae. Nontheless, residual BLG or peptides (see below) may still be responsible for allergic reactions described in some children with Cow’s milk allergy who are receiving these formulae (109).

Bos d 5, Bovine beta-lactoglobulin, is a 18.3 kDa protein (4,70,86,95,97,100-101,110-115).

rBos d 5

Beta-lactoglobulin is the most abundant protein in Whey, accounting for 50% of total protein in the Lactoserum (Whey) fraction. BLG occurs naturally in the form of a 36 kDa dimer possessing 2 disulfide bridges and 1 free cysteine. This structure is responsible for the main physicochemical properties and also for interaction with Casein during heat treatments. It has no homologous counterpart in human milk; i.e., human milk does not contain BLG (2). The relative resistance of BLG to acid hydrolysis and gut proteases allows part of the protein to be absorbed intact through the intestinal mucosa. By resisting digestion in the stomach, BLG is believed to act as a transporter of vitamin A and retinol to the intestines (116).

BLG belongs to the lipocalin superfamily and is one of the best characterised lipid-binding proteins. As such, it is capable of binding a wide range of molecules, including retinol, beta-carotene, saturated and unsaturated fatty acids, and aliphatic hydrocarbons (117-118). Lipocalins have a high allergenic potential, and several allergens of animal origin belong to this family. They share well-conserved sequence homologies in their N-terminus moiety (119-126). Other lipocalin protein family members include several allergens of animal origin such as the major Mouse (and Rat) urinary proteins (mMUP), the major Horse allergen Equ c 1, and the major Cockroach allergen BIa g 4 (125).

The molecule possesses 2 disulfide bridges and 1 free cysteine. This structure is responsible for the relative resistance of BLG to acid hydrolysis, as well as to proteases, which allows some of the protein to remain intact after digestion and increases the probability that intact BLG as well as digested fragments will be absorbed as antigens (116). The 2 intramolecular disulfide bonds may be responsible for the allergic effects (19). BLG is present in several variants. There are 2 main isoforms of BLG, genetic variants A and B, which differ only by 2 point mutations on residues 64 and 118; these are aspartic acid and valine in BLG A, and glycine and alaninc in BLG B. Variant C is found only in the Jersey breed (127). BLG occurs naturally as a mixture of monomers and dimers, but the proportion of monomers increases after heating to 70 °C (128). It has been demonstrated that there are many allergenic epitopes spread over the BLG structure.

Although the structure of the 2 variants A and B is very similar, in animal models the intensity and duration of the IgE response varies (7). Cleavage of the intra-chain disulfide bonds within the BLG molecule, and consequently the loss of the conformation of the molecule, had little if any effect on its immunoreactivity, suggesting that linear epitopes are implicated (7).

Chemical and immunological studies of BLG have identified a continuous epitope recognised by human IgE (71). However, sensitisation involves many epitopes that are widely spread all along the BLG molecule. Some have short linear sequences, while other immunoreactive structures corresponded to quite large fragments that might encompass conformational epitopes or parts of epitopes. In a study aimed at mapping the major allergenic epitopes on BLG by using specific IgE from sera of 46 Milk-allergic patients, several peptides capable of specifically binding human IgE were identified. Three fragments appeared to be major epitopes recognised by 92, 97 and 89% of sera, while a second group with 2 fragments was recognised by 58 and 72% of the population. A third group of peptides was detected by more than 40% of sera. Thus, 3 peptides were identified as major epitopes, recognised by a large majority of human IgE antibodies. The authors concluded that numerous other epitopes are scattered all along the BLG sequence (27).

A number of the BLG epitopes were mentioned as markers for persistent CMA. In addition to B cell epitopes, T cell epitopes of BLG have also been described (129). The monitoring of BLG IgE concentrations and the calculation of a ratio of IgE to IgG antibodies could be useful in predicting which patients will ultimately lose clinical reactivity (52).

Heating of Beta-lactoglobulin results in changes in the degree of allergenicity of the allergen, but this is dependent on the extent of heating: a slight but significant decreased IgE binding was seen between unheated Beta-lactoglobulin solution and Beta-lactoglobulin solution heat-treated at 74 degrees C. A more pronounced decrease was found at 90°C. The inhibition of IgE binding of Milk after heat treatment at 90°C was also significantly decreased. However, at all heat treatments, a similar total amount of IgE antibodies could be inhibited at a sufficiently high concentration of Beta-lactoglobulin (130). BLG also resists pasteurisation (131). Furthermore, heat-denatured proteins can also present new antigenic sites, uncovered by the unfolding process or created by new chemical reactions with other molecules present in the food. Heat-denatured BLG has been reported to have at least 1 new epitope, not found in the native state (132). Similarly in a study evaluating the specificity of serum IgE to different fragments of BLG in a group of 19 individuals allergic to Cow’s milk, a large number of epitopes were shown to be recognised by allergen-specific IgE of human allergic sera, and there were differences in the specific determinants recognised, depending on the serum (31).

The IgE binding of Beta-lactoglobulin appears to also be significantly impaired in some fermented, acidified Milk products such as yogurt, as compared to  nonfermented Milk (130).

BLG chemical hydrolysates appeared to retain most of the immunoreactivity of the native protein. IgE antibodies from 10 patients with CMA recognised enzymatic digestion products of BLG from pepsin or pepsin + trypsin (10 patients out of 10); the recognition of peptides was even better than that of the intact molecule in 4 of 10 patients. Researchers concluded that the digestive processes unmask new allergenic epitopes (40). It has been confirmed that cleavage may allow the presentation of determinants that, on the whole native protein, were not accessible to the antibodies (27).

BLG may be found in house dust. In a study of house dust, the amount of BLG ranged from < 16 to 71 ng/g dust, compared with Ovomucoid which ranged from 170 to 6,300 ng/g dust (133).

Bovine BLG seems to share structures with corresponding Milk proteins from other species.

Anti-bovine BLG antibodies show 10% cross-reactivity with Bovine alpha-lactalbumin, both in its native and in its denatured form, which appears to be a result of a continuous stretch of 4 amino acids common to Alpha-lactalbumin and BLG (107).

Crossreactivity between Cow’s milk and Mare’s milk has previously been demonstrated in inhibition studies, but this is contradicted by an earlier study in which an individual allergic to Mare’s milk was not allergic to Cow’s milk. Two allergenic proteins of 16 and 18 kDa were detected, and were thought to most likely represent Alpha-lactalbumin and BLG, but it was suggested that these 2 are not cross-reactive with Bovine equivalents (106).

Cross-reactivity has been suggested between Reindeer BLG and Bovine BLG. In a study of Reindeer milk-allergic patients, the patterns of Bovine BLG-specific IgE to Reindeer BLG varied among patients, suggesting only partial cross-reactivity (134).


e204 nBos d 6

Native protein purified from cow’s milk (Bos domesticus)

Common name: BSA
Biological function: Serum albumin
Mw: 67 kDa


Allergen description

Serum albumin is the main protein in mammalian blood tissue. It plays a very important role in the transport of nutritional substances into the system by virtue of its ability to bind with a large number of molecules. Beef also contains bovine serum albumin (BSA) and gamma globulin. These are heat-labile proteins found also in Cow’s milk. BSA is a distinct Milk allergen comprising approximately 1% of the total Milk protein.

BSA may be obtained from Bovine plasma collected in slaughterhouses, which is then highly purified and used in biochemistry, immuno-chemistry, haematology and microbiology, in all countries where these sciences are practiced. It is most often employed in the production of diagnostic test systems, as a growth medium for bacteria, and as a cell culture.

It is used in the manufacture of antiwrinkle skin-tightener and is a basic protein for biological reactants. It may be used as a medium for in vitro fertilisation techniques.

BSA, a 67 kDa, heat-labile protein, is a major allergen in Beef and a minor allergen in Milk (2,37,101,167-174).

In Cow’s milk, BSA accounts for around 5% of the total Whey proteins. BSA is physically and immunologically very similar to human blood serum albumin (HSA). Its main role is the transport, metabolism and distribution of ligands and the protection from free radicals (127). Its tertiary structure is quite stable, even under denaturing conditions. A reduction of disulfide bonds results in a complete abolishment of binding with anti-BSA antibodies. IgE antibodies specific for BSA from sera of allergic children were shown to be able to cross-react with albumins from Sheep and Pig, but they did not recognise those of Horse, Rabbit and Chicken (41).

Heating reduces sensitisation to Beef and to Bovine serum albumin but does not abolish reactivity to BSA under home conditions. However, industrially heat-treated and sterilised homogenised Beef and freeze-dried Beef may not be allergenic (169). Heat treatment and chemical denaturation are not able to decrease BSA’s capacity to bind BSA-specific IgE antibodies (175). Directly heated UHT Milks suffer less heat damage than indirectly heated Milk. During storage, BSA in directly heat-treated Milks decreased significantly, unlike Alpha-lactalbumin and Beta-lactoglobulin, in which changes were not statistically significant (176). Pepsin incubation at pH 4.0 was shown to result in a decreased hydrolysis and enhanced residual antigenicity of BSA (177). Research indicates that serum albumin antigenicity is only partially correlated to its native 3-dimensional structure (175).

There is a great variability in human IgE response to Cow’s milk, and no single allergen or particular structure can account for a major part of Milk allergenicity (2).

Studies of large populations of allergic patients show that most of the patients are sensitised to several proteins, including BLG (Bos d 5), Casein (Bos d 8), ALA (Bos d 4), BSA (Bos d 6), Lactoferrin, and immuno-globulins (Bos d 7). A great variability is observed in IgE antibody response. Both Casein and BLG, as well as ALA, are major allergens. However, proteins present in very low quantities, such as BSA, immuno-globulins, and especially Lactoferrin, also appear to be important, since 35% to 50% of patients are sensitised to those proteins and sometimes to those proteins only (2). Bovine BLG is a major Cow’s whey allergen, which together with a-lactalbumin is regarded as a major allergen in Cow’s milk. It is the main Whey protein, without any counterpart in human Milk.

Bovine serum albumin occurs as a major allergen in Beef, and a minor allergen in Cow’s milk. Beef-allergic individuals are at risk of being allergic to Cow’s milk and vice versa (36).

In a study evaluating the cross-reactivity between Lamb and Beef and the role of BSA and Ovine serum albumin (OSA) as allergens in Beef-allergic children, it was found that BSA and OSA are important Beef and Lamb allergens. They have similar amino acid sequences and allergenic properties (178-180). Considering that the major Beef allergen is BSA and that Beef-sensitive children are also sensitised to Ovine serum albumin, as well as to other serum albumins, the use of alternative meats in Beef-allergic children must be carefully evaluated on an individual basis (168).

There is a high degree of homology between the primary structures of human Milk protein serum albumin and the corresponding Bovine serum albumin (identity 76.6%), which has resulted in the hypothesis that there may be cross-reactivity between the bovine and the human albumin, since IgE antibodies from Birch profilin-allergic individuals have been reported to cross-react with human profilin where the identity between the 2 proteins is only 34% (159).

Previous reports have suggested that allergy to animal epithelia, possibly even sub-clinical allergy, may predispose towards sensitisation to mammalian meat as a result of sensitisation to BSA (159,180). And patients with persistent Milk allergy and specific IgE antibodies to BSA have a greater risk of rhinoconjunctivitis and asthma because of animal dander (181).

The aim of a study was to prove the cross-reactivity between serum albumin of different mammals in Milk, meat, and epithelia, and to determine whether heat treatment of meats decreases the allergenicity of albumins. All the patients’ sera, with the exception of 1, recognised serum albumin in different meats (Beef, Lamb, Deer, and Pork), epithelia (Dog, Cat, and Cow), and Cow’s milk. Some patients were sensitised only to serum albumin in meat and epithelia. Patients with allergy only to dander were sensitised to other proteins in epithelia but not to serum albumin. No patients reacted to serum albumin from heated meat extracts. Therefore, serum albumin appears to be an important allergen involved in Milk, meat, and epithelia allergy. The authors suggest that sensitisation first occurred to BSA in Cow’s milk and thereafter was developed to epithelial serum albumin, even though no direct contact with animals had been made; and that patients with both BSA and Cow’s milk allergy must avoid raw meats and furry pets (182).

Thiomucase (a mucopolysaccharidase obtained from Ovine tissues that is used mainly to facilitate the diffusion of local anaesthetics and in the treatment of cellulitis) is partially cross-reactive with BSA, Cat dander and Sheep dander (182).


f78 nBos d 8

Native protein purified from cow’s milk (Bos domesticus)

Biological function: Casein
Mw: 19-25 kDa


Allergen description

Casein is a major allergen in Milk (19) and the main protein constituent of cheese. Casein makes up about 75-80% of all Milk protein and is heat-stable. (See Table 1, page 52).

Casein is found in Milk and dairy products, especially cheese, and in other foods containing Milk. Even highly hydrolysed Milk-derived infant formulas may contain allergenic Casein residues (32,69,135-136).

Casein may occur in “Milk-free” products as undegraded residual Milk proteins or as contamination from previous productions of food containing Milk. Casein may be a cause of allergic reactions in patients eating so-called “non-dairy” products (48).

Casein and caseinates are used as extenders and tenderisers in sausages, loaves, soups and stews. They are often used to nutritionally fortify foods and as supplements because of the large amount of high-quality protein they contain, their low level of lactose, and their bland flavour. Such nutritionally fortified foods include high-protein beverage powders, fortified cereals, infant formula and nutrition bars. Casein is often an ingredient in coffee whiteners, sauces, ice cream, salad dressing, formulated meats, bakery glazes, and whipped toppings.

Bos d 8, Casein consists of a range of proteins varying in size from 19 to 25 kDa (4,29,37,70,95,97,99,11,137-140).

The coagulum consists of the whole Casein fraction (i.e., the solid fraction of proteins obtained after coagulation of Milk). It is subdivided into a number of families, of which the most important are aS1 -, aS2 -, b-, k-, and g-caseins (a = alpha, b = beta, k = kappa,
g = gamma) (5).

Each individual Casein among the types aS1 -, aS2 -, b-, k- represents a well-defined chemical compound, but they cross-link to form ordered aggregates (nanoclusters or micelles) that assemble into larger structures, forming Casein micelles characterised by a central hydrophobic part and a peripheral hydrophilic layer in suspension in Lactoserum (Whey) (2,13-14). Their proportion in the micelles is relatively constant at approximately 37%, 13%, 37%, and 13%, respectively.

Their distribution is not uniform within these micelles, which comprise a central hydrophobic part and a peripheral hydrophilic layer, where major sites of phosphorylation that contain phosphoserine residues are presented in relation to the calcium-binding and transfer properties of Caseins (2). aS1-, b-, aS2-, and k-casein have little primary structure homology. Their functional properties also differ, since 3 of them, aS1-, aS2-, and b-casein, appear to be calcium-sensitive, whereas k-casein is not. However, the 4 Caseins display common features that are unusual, which means that they differ greatly from other Milk proteins. They are phosphorylated proteins (2). Casein is rapidly and extensively degraded by proteolytic enzyme during digestion. Caseins are not significantly affected by severe heat treatments but are very susceptible to all proteinases and exopeptidases. Multisensitisations to the different Caseins occur most often in patients sensitised to the whole Casein fraction (2,138).

Casein is thermostable, whereas BLG is thermolabile, but it may be protected through interaction with Casein. Thermostability of Cow’s milk proteins depends not only on temperature and time spent heated but also on interactions within the food matrix. Heat denaturation, which leads to the loss of organised protein structures, does not always result in a decreased allergenic potential: formation of aggregates may increase the allergenicity of the heated product. When the treatment results in a decrease in the allergenicity, the decrease is always limited. Boiling of Milk for a few minutes (2, 5, or 10 minutes) results either in no difference or in a reduction of approximately 50% to 66% in positive reactions, compared to reactions to raw Milk; similar observations have been reported with raw vs. pasteurised or homogenised and pasteurised Milk (2). The Caseins are heat-stable, and even high pasteurisation (121 °C for 20 minutes) only reduces and does not eliminate the allergenicity of the Caseins (141).

aS1-casein represents up to 40% of the Casein fraction in Cow’s milk. aS1-casein consists of major and minor components; both are single-chain polypeptides with the same amino-acid sequence, differing only in their degree of phosphorylation (5,142). Variants A, B, C, D, F, G, H have been identified as characteristic of different cattle breeds.

The aS2-casein family accounts for 12.5% of the Casein fraction in Cow’s milk and comprises the most hydrophilic of all Caseins. aS2-casein consists of 2 major and several minor components. A post-translational modification occurring in this protein is the formation of disulfide bonds that do not participate in the interaction with other Caseins (5).

Further studies have confirmed that a-casein largely lacks a tertiary structure and therefore also lacks conformational epitopes (29,143). Indeed, Casein appears to preferentially have linear epitopes (35). The sequential epitopes are exposed even in denatured Casein, resulting in an apparent stability of the allergen to denaturing conditions, e.g., heat. In fact, some of the major epitopes already characterised on alpha-S-caseins are continuous epitopes that have also been located in hydrophobic regions of the molecule, where they are not accessible to antibodies unless the Casein is denatured or degraded, such as for instance, during digestion (144). This may explain the apparent difference in epitope recognition among patients with different natural histories of CMA (144).

The b-casein family accounts for 35% of the Casein fraction and is quite complex because of the action of the native Milk protease plasmin. Plasmin cleaves the b-casein, generating g1-, g2-, and g3-casein fragments. b-casein is the most hydrophobic component of the total Casein fraction. There are 10 genetic variants (5).

k-Casein accounts for 12.5% of the total Casein fraction. k-Casein consists of a major carbohydrate-free component and a minimum of 6 minor components. It is isolated from Milk as a mixture of disulfide-bonded polymers ranging from dimers to octamers. There are 2 common and 9 other genetic variants. The k-casein group plays an important role in the stability and coagulation properties of Milk. Hydrolysis by chymosin in rennet produces para-k-casein and a caseinomacropeptide that is important for the first stage of the cheesemaking process (145).

Anionic regions contain clusters of aS1-, aS2- and b-casein, and these clusters are able to chelate Ca2+ and other metal ions, including Zn2+ and Fe3+ (5).

Caseins, although less ordered in structure and more flexible than the typical globular Whey proteins, have significant amounts of secondary and, probably, tertiary structure (146). Howerver, Caseins appear to preferentially have linear epitopes. A study carried out on sera of 15 Milk-allergic children showed that 6 major and 3 minor IgE-binding epitopes as well as 8 major and 1 minor IgG-binding regions were identified on b-casein, while 2 major and 2 minor IgG-binding epitopes were found for k-casein. In another study, overlapping synthetic peptides were used to identify major IgE- and IgG-binding regions of aS1-casein in patients with CMA. Six major and 3 minor IgE-binding regions, and 5 major and 1 minor IgG-binding epitopes were identified (35). The implication of linear epitopes is that denaturing does not affect this protein to the same extent as those in which conformational epitopes are relevant.

A number of studies have demonstrated that most patients allergic to Casein are sensitised to each of the 4 major Caseins, and that there is great variability in the specificity and intensity of IgE response to these Casein fractions, which indicates, among other things, the presence of distinct epitopes on the individual Casein molecules (138,147). The intensity of the IgE responses appears to be closely related to the proportion of the 4 Caseins in Milk, and sensitisation probably occurs after the disruption of the Casein micelles during the digestive process (126,138). However, cross-sensitisation mechanisms also occurred through common or closely related epitopes. Importantly, Casein “allergenic epitopes” may be present in Whey (138). Therefore, polysensitisaton appears to be due both to cross-sensitisation and to common or closely related epitopes.

Partial hydrolysis of a fraction of the Casein, e.g., Beta-casein, occurring naturally due to endogenous enzymes such as plasmin, which are normally present in Milk, gives rise to Gamma-caseins, and to smaller fragments called proteoses-peptones, corresponding to the N-terminal part of the Beta-casein molecule. These peptides are soluble and remain in the Lactoserum (2). Similarly, the limited proteolysis due to the action of chymosin during clotting of Milk splits k-casein into 2 peptides: hydrophobic para-k-casein and a highly polar caseino-macro-peptide, which is soluble and remains in the Whey. Some proteoses-peptones are still allergenic, as is the caseino-macro-peptide, which explains why reactions may be observed after ingestion of Whey protein; hydrolysates in babies have serum-specific IgE to Casein but negative to Whey proteins (4).

Furthermore, the balance between Caseins and Whey proteins appears to play an important role in the sensitisation capacity of Cow’s milk (148). Also, a reduced Casein content and poorer renneting properties of Milk may occur in late summer, which may result in differences in the frequencies of sensitisation to Cow’s milk proteins (149). The formation of Casein monomers into a high-molecular-mass fraction to which CMA individuals display reactivity has been described (19).

It has been supposed that the majority of linear IgE epitopes in Caseins could contribute to persistent allergy (150). Milk-allergic children with persistent symptoms have significantly higher levels of specific IgE antibodies to linear epitopes from aS1-(AA69-78) Casein and b-Casein than children who have achieved tolerance (151). Five IgE-binding discriminative epitopes (2 on aS1- casein, 1 on aS2-casein, and 2 on k-casein) have been shown to be exclusively recognised by patients with persistent CMA (152).

A high degree of cross-reactivity occurs between Cow’s, Sheep and Goat’s milk as a result of the high sequence homology between their Caseins. Goat and Sheep Milk allergy may involve the Casein fraction and not Whey proteins (38,42,153). The high degree of cross-reactivity between these three Caseins appears to be as a result of alpha-caseins which share more than 85% identical amino acids homology (38).

Furthermore, multi-sensitisation to the different Caseins most often occurs in patients sensitised to the whole Casein fraction. It has been suggested that conserved regions shared by both Bovine and human Beta-caseins, and particularly those comprising clusters of phosphorylated seryl residues, are responsible for IgE cross-reactivity (138).

Twenty patients allergic to Cow’s milk proteins and with high levels of IgE antibodies directed against Bovine whole Casein were selected to evaluate the reactivity of their IgE antibodies to human Beta-casein. Seven sera contained IgE directed against human Beta-casein. Inhibition studies using native human and Bovine beta-caseins as well as Bovine beta-casein-derived peptides demonstrated that, depending on the sera, 1 or several common epitopes located in different parts of the molecule were shared by the 2 homologous proteins (154).

In a study evaluating the Alpha-caseins from Bovine, Ovine, and Goat’s milk sharing more than 85% identical amino acids, sera from 17 children with immediate-type allergy to Cow’s milk were compared with sera from non-CMA-allergic individuals. The sera of Cow’s milk-allergic children showed a significantly higher IgE and IgG binding to Alpha-caseins from all 3 species than did the sera of the other groups. All groups showed an increased antibody binding to Bovine alpha-casein, as compared to the Sheep and Goat proteins, but the differences were significant only in the groups of atopic children and of healthy controls. Inhibition of the IgE binding to Bovine alpha-casein with Alpha-casein from Cow, Goat, and Sheep revealed that the Alpha-caseins from these species are highly cross-reactive, on the basis of the small differences in their primary structures (38).

Structural homologies in Caseins of different species can share common epitopes for IgE of CMA patients, suggesting that prevention of Cow’s milk allergy cannot be achieved by using Milk from other species as substitutes. In A study of sera from 58 CMA individuals to determine the specificity of their IgE response to the whole Casein fraction of Milk from different ruminant and nonruminant species (e.g. Cow, Sheep, Goat, Rabbit and Rat), co-and/or cross-sensitisations to Caseins of the different species occurred extensively, though IgE responses to Sheep and Caprine casein appeared to be lower than those obtained with Casein from Cow, and in terms of specificity and intensity, the IgE response to Caseins demonstrated a great variability (155).

However, although many children who are allergic to Cow’s Milk cannot tolerate Goat’s or Sheep’s milk either, there are instances of patients who are allergic to Sheep and/or Goat’s milk and not to Cow’s milk Caseins (42,153). In a report on Goat and Sheep milk-allergic children who were not allergic to Cow’s milk, IgE specificity and affinity was high to Goat and Sheep milk, and lower to Cow’s milk caseins despite their marked sequence homology (42). It has also been shown that Sheep casein shows a high degree of cross-reactivity with Goat casein but not with Cow casein (153,156). These results may indicate sensitisation to Casein per se but not to the alpha-Casein fraction, which may contribute mostly to the cross-reactivity usually seen.

Adverse reactions have been reported in Milk-allergic patients fed Soy-based formulae as Cow’s milk substitutes. A 30-kDa, glycinin-like protein from Soybean that cross-reacts with Cow’s milk casein has been isolated and partially sequenced. The results of this study indicate that Soy-based formula, that contains the A5-B3 glycinin molecule could be involved in allergic reactions observed in Cow’s milk-allergic patients exposed to Soy-containing foods (44).


f334 nBos d lactoferrin

Native protein purified from cow’s milk (Bos domesticus)

Biological function: Bovine lactoferrin                 
Mw: 76 kDa

Allergen description

Lactoferrin is a major allergen in Milk. Lactoferrin is an allergen of the whey fraction of Milk and can be found in the Milk of most species at levels lower than 1%. (See Table 1, page 52)

Lactoferrin is a non-heme-iron-binding globular multifunctional glycoprotein with antimicrobial activity, produced during lactation and by epithelial cells at mucosal surfaces. The protein is a prominent component of the first line of mammalian host defence, and its expression is up-regulated in response to inflammatory stimuli. Lactoferrin may act as a potent anti-inflammatory protein at local sites of inflammation, including the respiratory and gastrointestinal tracts (157). Human colostrum has the highest concentration, followed by human milk, then Cow’s milk.

Lactoferrin appears to play several biological roles. Owing to its iron-binding properties, Lactoferrin is thought to play a role in iron uptake by the intestinal mucosa of the neonate.

Besides in Cow’s milk, the topic of this review, Lactoferrin is found in many mucosal secretions such as tears, saliva, bile, pancreatic juice, and genital and nasal secretions. Lactoferrin is released from neutrophil granules during inflammation and is also secreted by some acinar cells.

As Bovine milk-derived Lactoferrin is known to be an effective natural antimicrobial, it is used as a spray, applied electrostatically to raw Beef carcasses to detach bacteria adhering to the surface, in order to reduce microbial contamination. It is used only on Beef carcasses (not on subprimals or finished cuts) at a level not to exceed 0.20 ml of formulation per kg of Beef. An assessment of its use found that its application to Beef carcasses is in the range of existing background exposures of Lactoferrin, because Lactoferrin is found naturally in Beef, and that this potentially small incremental increase in Lactoferrin is safe (i.e., there is no reasonable expectation that the substance will become an allergen under the conditions of its intended use) (158).

Bos d Lactoferrin, a 76.1 kDa protein, has been characterized (8,97,127,159).

Lactoferrin (LF) is an allergen of the whey fraction of Cow’s milk (21). It is a protein of mammary origin and is a Milk-specific iron-binding glycoprotein of the transferrin family. It can be found in the Milk of most species at levels lower than 1% (160). LF is present in much higher concentrations in human breast milk (ie, 1 g/l), particularly in colostrum (2). Although it is present in very low concentrations in Cow’s milk, it has been shown to be an important allergen (8).

LF consists of a single polypeptide chain folded into 2 globular lobes. The molecular weight of this protein varies depending on the extent of its glycosylation. The LF content is species-dependent, with significantly higher levels in human milk and colostrums compared to Bovine milk, whereas the sequence homology and structure are very similar, with human and Bovine lactoferrin having an amino acid sequence homology of 69%, and structural similarity (2,5). Lactoferrin is partially heat-stable and relatively stable to enzymatic degradation by gut proteases and remains partly unchanged during digestion (2).

Lactoferrin is a multifunctional member of the transferrin family of nonheme, iron-binding glycoproteins. Lactoferrin is found at the mucosal surface, where it functions as a prominent component of the first line of host defence against infection and inflammation (161-162).

Its main role is to defend the organism against infections and inflammations through its ability to sequester iron from the environment and thereby remove this essential nutrient for bacterial growth, and to act as an antioxidant and a scavenger for free radicals, thus providing protection against oxidative stress (5,161). It also has antibacterial properties and has been shown to stimulate cellular immune defence of the organism against infections (2). Lactoferrin is also an abundant component of the specific granules of neutrophils and can be released into the serum upon neutrophil degranulation (161). Neutrophil lactoferrin has also been shown to inhibit tryptase released from mast cells (163). While the iron-binding properties were originally believed to be solely responsible for the host defence properties ascribed to this protein, it is now known that other mechanisms contribute to the broad-spectrum anti-infective and anti-inflammatory roles of this protein. Lactoferrin appears to function, collectively, as a key component of mammalian host defence at the mucosal surface (161). Recently, human lactoferrin was shown to be implicated in the pathophysiology of an asthma attack (164).

Bovine lactoferrin is able to form non-covalent complexes with Beta-lactoglobulin or Albumin, with Lactoferrin-protein molar ratios of 2:1 and 1:1 respectively. No association was detected with Alpha-lactalbumin (165).

Milk from related animals is important. Lactoferrin is present in human and Bovine milk, the proteins from the 2 species having about 70% homology. It is thus likely that the 2 proteins share common or similar epitopes, and it is thus possible that exposure and immunological reaction to Bovine lactoferrin during ingestion of Cow’s milk or Milk products in infancy could prime the immune system to subsequently react against human lactoferrin. A study testing this hypothesis concluded that there is evidence that development of anti-Lactoferrin autoantibodies in patients with anti-neutrophil cytoplasmic antibodies may be result, at least in part, from prior stimulation from Bovine lactoferrin in Cow’s milk (166).

Further support for potential cross-reactivity between Bovine and human lactoferrin is the evidence of a dose-dependent inhibition of serum IgE to Cow’s milk protein binding to human milk proteins, shown by denatured Bovine whey proteins, and vice versa, which suggests the presence of common epitopes (cross-reactivity) between Bovine and human milk proteins. A study investigating this hypothesis argued that lack of inhibition by native Bovine and human whey proteins suggested that such epitopes are probably linear (continuous) and should lie in the internal part of the molecules. The authors suggest that the rather high degree of homology between the primary structures of human milk proteins and the corresponding Bovine proteins (Serum albumin, identity 76.6%; Alpha-lactalbumin, identity 73.9%; Lactoferrin, identity 69.5%; Beta-casein, identity 56.5%) is congruent with cross-reactivity towards specific antibodies. In a similar way, IgE antibodies from Birch profilin-allergic individuals have been reported to cross-react with human profilin where the identity between the 2 proteins is only 34% (159).


Compiled by Dr Harris Steinman, harris@zingsolutions.com


  1. Milk. http://en.wikipedia.org/wiki/Cow’s_milk#Cow.27s_milk
  2. Wal JM. Bovine milk allergenicity.
    Ann Allergy Asthma Immunol 2004;93(5 Suppl 3):S2-11
  3. Lien EL. Infant formulas with increased concentrations of alpha-lactalbumin.
    Am J Clin Nutr 2003;77(6):1555S-8S
  4. Wal JM. Immunochemical and molecular characterization of milk allergens.
    Allergy 1998;53(46 Suppl):114-7
  5. Monaci L, Tregoat V, van Hengel AJ, Elke Anklam. Milk allergens, their characteristics and their detection in food: A review. Eur Food Research Tech 2006;223(2):149-79
  6. Mather IH. A review and proposed nomenclature for major proteins of the milk-fat globule membrane.
    J Dairy Sci 2000;83(2):203-47
  7. Wal JM. Cow’s milk allergens.
    Allergy 1998;53:1013-22
  8. Wal JM. Cow’s milk proteins/allergens.
    Aim Allergy Asthma lmmunol 2002;89 (Suppl 1):3-10
  9. Savilahti E, Kuitunen M. Allergenicity of cow milk proteins.
    J Pediatr 1992;121(5 Pt 2):S12-20
  10. Wal JM, Bernard H, Yvon M, Peltre G, David B, Creminon C, Frobert Y, Grassi J. Enzyme immunoassay of specific human IgE to purified cow’s milk allergens. Food Agric Immunol 1995;7:175-87
  11. Rabbiani H, Brown W, Butler J, Hammarstrom L. Immunogenetics 1997;46:326-31
  12. Ayuso R, Lehrer SB, Lopez M, Reese G, Ibanez MD, et al. Identification of bovine IgG as a major cross-reactive vertebrate meat allergen. Allergy 2000;55(4):348-54
  13. Holt C, Timmins PA, Errington N, Leaver J. A core-shell model of calcium phosphate nanoclusters stabilized by beta-Casein phosphopeptides, derived from sedimentation equilibrium and small-angle X-ray and neutron-scattering measurements.
    Eur J Biochem 1998;252:73-8
  14. Breiteneder H, Mills EN. Molecular properties of food allergens.
    J Allergy Clin Immunol 2005;115(1):14-23
  15. Wal JM, Bernard H, Creminon C, Hamberger C, David B, Peltre G. Cow’s milk allergy: the humoral immune response to eight purified allergens.
    Adv Exp Mod Biol 1995;371B:879-81
  16. Goldman AS, Anderson DW Jr, Sellers WA, Saperstein S, Kniker WT, Halpern SR. Milk allergy, I: oral challenge with milk and isolated milk proteins in allergic children. Pediatr 1963:32:425-43
  17. Goldman AS, Sellars WA, Halpem SR, Anderson DW Jr, Furlow TE, Johnson CH Jr. Milk allergy, II: skin testing of allergic and normal children with purified milk proteins. Pediatr 1963;32:572-9
  18. Gjesing B, Osterballe 0, Schwartz B, Wahn U, Lowenstein H. Allergen-specific IgE antibodies against antigenic components in cow milk and milk substitutes.
    Allergy 1986;41:51-6
  19. Docena GH, Femandez R Chirdo FG, Fossali CA. Identification of casein as (the major allergenic and antigenic protein of cow’s milk. Allergy 1996;51:412-6
  20. Kaiser C, Reibisch H, Folster-Holst R, Sick H. Cow’s milk-protein allergy: results of skin-prick test with purified milk proteins.
    Z Emahrungswiss 1990;29:122-8
  21. Host A, Husby S, Gjesing B, Larsen IN, Lowenstein H. Prospective estimation of IgG, IgG subclass and IgE antibodies to dietary proteins in infants with cow milk allergy: levels of antibodies to whole milk protein, BLG and ovalbumin in relation to repeated milk challenge and clinical course of cow milk allergy. Allergy 1992,47:218-29
  22. Stoger P, Wüthrich B. Type 1 allergy to cow milk proteins in adults: a retrospective study of 34 adult milk-and cheese-allergic patients. Int Arch Allergy Immunol 1993;102:399-407
  23. Natale M, Bisson C, Monti G, Peltran A, Garoffo LP, Valentini S, Fabris C, Bertino E, Coscia A, Conti A. Cow’s milk allergens identification by two-dimensional immunoblotting and mass spectrometry.
    Mol Nutr Food Res 2004;48(5):363-9
  24. Host A, Samuelsson EG. Allergic reactions to raw, pasteurized, and homogenized/pasteurized cow milk: a comparison. A double-blind placebo-controlled study in milk allergic children. Allergy 1988;43(2):113-8
  25. Werfel T, Ahlers G, Schmidt P, Boeker M, Kapp A, Neumann C. Milk-responsive atopic dermatitis is associated with a casein-specific lymphocyte response in adolescent and adult patients. J Allergy Clin Immunol 1997;99(1):124-33
  26. Norgaard A, Bernard H, Wal JM, Peltre G, Skov PS, et al. Allergenicity of individual cow milk proteins in DBPCFC-positive milk allergic adults.
    J Allergy Clin Immunol 1996,97:237
  27. Selo I, Clement G, Bernard H, Chatel J, Creminon C, Peltre G, Wal J. Allergy to bovine beta-lactoglobulin: specificity of human IgE to tryptic peptides. Clin Exp Allergy 1999;29(8):1055-63.
  28. Maynard F, Jost R, Wal JM. Human IgE binding capacity of tryptic peptides from bovine alpha-lactalbumin. Int Arch Allergy Immunol 1997;113(4):478-88.
  29. Spuergin P, Mueller H, Walter M, Schiltz E, Forster J. Allergenic epitopes of bovine a S1-casein recognized by human IgE and IgG. Allergy 1996;51(5):306-12
  30. Haddad ZH, Kalra V, Verma S. IgE antibodies to peptic and peptic-tryptic digests of betalactoglobulin: significance in food hypersensitivity.
    Ann Allergy 1979 Jun;42(6):368-71
  31. Selo I, Negroni L, Creminon C, Yvon M, Peltre G, Wal JM. Allergy to bovine beta-lactoglobulin: specificity of human IgE using cyanogen bromide-derived peptides. Int Arch Allergy Immunol 1998 Sep;117(1):20-8
  32. Oldaeus G, Björskten B, et al. Antigenicity and allergenicity of cow milk hydrolysates intended for infant feeding.
    Pediatr Allergy Immunol 1991;4:156-64
  33. Ragno V, Giampietro PG, et al. Allergenicity of milk protein hydrolysate formulae in children with cow’s milk allergy.
    Eur J Pediatr 1993;152(9):760-2
  34. de Boissieu D, Matarazzo P, Dupont C. Allergy to extensively hydrolyzed cow milk proteins in infants: identification and treatment with an amino acid-based formula. J Pediatr 1997 Nov;131(5):744-7
  35. Chatchatee P, Jarvinen KM, Bardina L, Vila L, Beyer K, Sampson HA. Identification of IgE and IgG binding epitopes on beta- and kappa- casein in cow’s milk allergic patients. Clin Exp Allergy 2001;31(8):1256-62
  36. Werfel S, Cooke SK, Sampson HA. Clinical reactivity to beef in children allergic to cow’s milk.
    J Allergy Clin Immunol 1997;99(3):293-300
  37. Szepfalusi Z, Ebner C, Urbanek R, Ebner H, Scheiner O, Boltz-Nitulescu G, Kraft D. Detection of IgE antibodies specific for allergens in cow milk and cow dander. Int Arch Allergy Immunol 1993;102:288-94
  38. Spuergin P, Walter M, Schiltz E, Deichmann K, Forster J, Mueller H.. Allergenicity of alpha -caseins from cow, sheep, and goat.
    Allergy 1997;52:293-8
  39. Dean TP, Adler BR, Ruge F, Warner JO. In vitro allergenicity of cow’s milk substitutes. Clin Exp Allergy 1993;23:205-10
  40. Bernad H, Creminon C, Negroni L, Peltre G, Wal JM. IgE cross-reactivity with caseins from different species in humans allergic to cow’s milk.
    Food Agric Immunol 1999;11(1):101-11
  41. Restani P, Gaiaschi A, Plebani A, Beretta B, Cavagni G, et al. Cross-reactivity between milk proteins from different animal species. Clin Exp Allergy 1999;29(7):997-1004
  42. Ah-Leung S, Bernard H, Bidat E, Paty E, Rance F, Scheinmann P, Wal JM. Allergy to goat and sheep milk without allergy to cow’s milk. Allergy 2006 Nov;61(11):1358-65
  43. Wüthrich B, Johansson SG. Allergy to cheese produced from sheep’s and goat’s milk but not to cheese produced from cow’s milk.
    J Allergy Clin Immunol 1995;96(2):270-3
  44. Rozenfeld P, Docena GH, Anon MC, Fossati CA. Detection and identification of a soy protein component that cross-reacts with caseins from cow’s milk.
    Clin Exp Immunol 2002;130(1):49-58
  45. Bousquet J, Chanez P, Michel F-B. The respiratory tract and food hypersensitivity. Food Allergy, Adverse Reactions to Foods and Food Additives. Metcalfe DD, Sampson HA, Simon RA. Boston, MA, USA: Blackwell Scientific Publications. 1991:139. ISBN: 0-86542-094-7
  46. Bahna SL. Milk allergy in infancy.
    Ann Allergy 1987;59 (5 pt 2):131-6
  47. Olalde S, Bensabat Z, Vives R, Fernandez L, Cabeza N, Rodriguez J. Allergy to cow’s milk with onset in adult life.
    Ann Allergy 1989;62:185a-5b
  48. Gern JE, Yang E, Evrard HM, Sampson HA. Allergic reactions to milk-contaminated “nondairy” products.
    New Eng J Med 1991;324:976-9
  49. Jones RT, Squillace DL, Yunginger JW. Anaphylaxis in a milk-allergic child after ingestion of milk-contaminated kosher-pareve-labeled “dairy-free” dessert.
    Ann Allergy 1992;68:223-7
  50. Okudaira H, Ito K, Miyamoto T, Wagatsuma Y, Matsuyama R, Kobayashi S, Nakazawa T, Okuda M, Otsuka H, Baba M, Iwasaki E, Takahashi T, Adachi M, Kokubu F, Nishima S, Shibata R, Yoshida H, Maeda K. Evaluation of new system for the detection of IgE antibodies (CAP) in atopic disease.
    Arerugi 1991;40(5):544-5
  51. Businco L, Benincori N, Cantani A. Epidemiology, incidence and clinical aspects of food allergy.
    Ann Allergy 1984;53:615-22
  52. James JM, Sampson HA. Immunologic changes associated with the development of tolerance in children with cow milk allergy.
    J Pediatr 1992;121:371-7
  53. Ahmed T, Sumazaki R, Nagai Y, Shibasaki M, Takita H. Immune response to food antigens: kinetics of food specific antibodies in the normal population.
    Acta Paediatr Jpn 1997;39:322-8
  54. Foucard T. Development of food allergies with special reference to cow’s milk allergy. Pediatrics 1985;75(1 pt 2):177-81
  55. Amlot PL, Kemeny DM, Zachary C, Parkes P, Lessof MH. Oral allergy syndrome (OAS): Symptoms of IgE-mediated hypersensitivity to food. Clin Exp Allergy 1987;17:33-42
  56. Businco L, Benincori N, Cantani A, Tacconi L, Picarazzi A. Chronic diarrhea due to cow’s milk allergy. A 4- to 10- year follow-up study. Ann Allergy 1985;55(6):844-7
  57. Høst A, Halken S. A prospective study of cow milk allergy in Danish infants during the first 3 years of life. Allergy 1990;45:587-96
  58. Stöger P, Wüthrich B. Type I allergy to cow milk proteins in adults. Int Arch Allergy Immunol 1993;102:399-407
  59. Joliat TL, Weber RW. Occupational asthma and rhinoconjunctivitis from inhalation of crystaline bovine serum albumin powder.
    Ann Allergy 1991;66:301-4
  60. Bernaola G, Echechipia S, Urrutia I, Fernandez E, Audicana M, Fernandez de Corres L. occupational asthma and rhinoconjunctivitis from inhalation of dried cow’s milk caused by sensitization to alpha-lactalbumin. Allergy 1994;49:189-91
  61. Lee EJ, Heiner DC. Allergy to cow milk-1985. Pediatrics in review 1986;7(7):195-203. ISSN: 0191-9601
  62. Pelto L, Salminen S, Lilius EM, Nuutila J, Isolauri, E. Milk hypersensitivity – key to poorly defined gastrointestinal symptoms in adults. Allergy 1998;53:307-10
  63. May CD, Remigio L, Feldman J, Bock SA, Carr RI. A study of serum antibodies to isolated milk proteins and ovalbumin in infants and children.
    Clin Exp Allergy 1977;7:583-95
  64. Halken S, Høst A, Hansen LG, Østerballe O. Effect of an allergy prevention programme on incidence of atopic symptoms in infancy. Allergy 1992;47:545-53
  65. Sigurs N, Hattevig G, Kjellman B. Maternal avoidance of eggs, cow’s milk, and fish during lactation.
    Pediatr 1992;89(4):735-9
  66. Casimir GJ, Duchateau J, Gossard B, Cuvelier P, Vandaele F. Atopic dermatitis: role of food and house dust mite allergens.
    Pediatr 1993;92:252-6
  67. Agata H, Kondo N, Fukutomi O, Shinoda S, Orii T. Effect of elimination diets on food-specific IgE antibodies and lymphocyte proliferative responses to food antigens in atopic dermatitis patients exhibiting sensitivity to food allergens.
    J Allergy Clin Immunol 1993;91(2):668-79
  68. Savilahti E, Kuitunen M. Allergenicity of cow milk protein. J Pediatr 1992;121:S12-20
  69. Chiancone E, Gattoni M, Giampietro PG, Ragno V, Businco L. Detectoion of undegraded b-lactoglobulins and evaluation of the molecular weight of peptides in hydrolysate cow’s milk formula. J Investig Allergol Clin Immunol 1995;5:228-33
  70. Gortler I, Urbanek R, Forster J. Characterization of antigens and allergens in hypo-allergenic infant formulae.
    Eur J Pediatr 1995;154:289-94
  71. Ball G, Shelton MJ, Walsh BJ, Hill DJ, Hoskings CS, Howden M. A major continuous epitope of bovine b-lactoglobulin recognized by human IgE binding.
    Clin Exp Allergy 1994;24:758-64
  72. Juvonen P, Jakobsen I, Lindberg T. Macromolecular absorbtion and cow’s milk allergy. Arch Dis Child 1991;66:300-303
  73. Maeda S, Morikawa A, Tokuyama K, Kuroume T. The concentration of bovine IgG in human breast milk measured using different methods. Acta Pediatr 1993;82:1012-6
  74. Malmheden Yman I, Eriksson A, Everitt G, Yman L, Karlsson T. Analysis of food proteins for verification of contamination or mislabelling.
    Food Agric Immunol 1994;6:167-72
  75. Axelsson I, Jakobsson I, Lindberg T, Benediktsson B. Bovine beta-lactoglobulin in the human milk. A longitudinal study during the whole lactation period.
    Acta Paediatr Scand 1986;75(5):702-7
  76. Machtinger S, Moss R. Cow’s milk allergy in breast-fed infants: the role of allergen and maternal secretory IgA antibody.
    J Allergy Clin Immunol 1986;77(2):341-7
  77. Host A, Husby S, Osterballe O. A prospective study of cow’s milk allergy in exclusively breast-fed infants. Incidence, pathogenetic role of early inadvertent exposure to cow’s milk formula, and characterization of bovine milk protein in human milk.
    Acta Paediatr Scand 1988;77(5):663-70
  78. Sorva R, Makinen-Kiljunen S, Juntunen-Backman K. Beta-lactoglobulin secretion in human milk varies widely after cow’s milk ingestion in mothers of infants with cow’s milk allergy.
    J Allergy Clin Immunol 1994;93(4):787-92
  79. Gerrard JW, Shenassa M. Food allergy: two common types as seen in breast and formula fed babies. Ann Allergy 1983;50:375-9
  80. Gerrard JW, Shenassa M. Sensitization to substances in breast milk: recognition, management and significance.
    Ann Allergy 1983;51(2 Pt 2):300-2
  81. Jakobsson I, Lindberg T. A prospective study of cow’s milk protein intolerance in Swedish infants.
    Acta Paediatr Scand 1979;68(6):853-9
  82. Laoprasert N, Wallen ND, Jones RT, Hefle SL, Taylor SL, Yunginger JW. Anaphylaxis in a milk-allergic child following ingestion of lemon sorbet containing trace quantities of milk. J Food Prot 1998;61(11):1522-4
  83. Fremont S, Kanny G, et al. Identification of a masked allergen, alpha-lactalbumin, in baby-food cereal flour guaranteed free of cow’s milk protein. Allergy 1996;51(10):749-54
  84. Nowak-Wegrzyn A, Shapiro GG, Beyer K, Bardina L, Sampson HA. Contamination of dry powder inhalers for asthma with milk proteins containing lactose. J Allergy Clin Immunol 2004;113(3):558-60
  85. Lonnerdal B, Lien EL. Nutritional and physiologic significance of alpha-lactalbumin in infants. Nutr Rev 2003;61(9):295-305
  86. Jarvinen KM, Chatchatee P, Bardina L, Beyer K, Sampson HA. IgE and IgG binding epitopes on alpha-lactalbumin and beta-lactoglobulin in cow’s milk allergy. Int Arch Allergy Immunol 2001;126(2):111-8
  87. Brew K, Castellino FJ, Vanaman TC, Hill RL. The complete amino acid sequence of bovine alpha-lactalbumin. J Biol Chem 1970:245:4570-82
  88. Browne WJ, North AC, Phillips DC, Brew K, Vanaman TC, Hill RL. A possible three-dimensional structure of bovine alpha-lactalbumin based on that of hen’s egg-while lysozyme. J Mol Biol 1969,42:65-86
  89. Nitta K, Sugai S. The evolution of lysozyme and alpha-lactalbumin.
    Eur J Biochem 1989;182:111-18
  90. Findlay JB, Brew K. The complete amino-acid sequence of human alpha-lactalbumin. Eur J Biochem 1972,27:65-86
  91. McKenzie HA, White FH Jr. Lysozyme and alpha-lactalbumin: structure, function, and interrelationships.
    Adv Protein Chem 1991;41:173-315
  92. Berliner LJ, Kaptein R, Koga K, Musci G. NMR studies of the structure and environment of the milk protein alpha-lactalbumin. Basic Life Sci 1990;56:231-53
  93. Nakata R. Alpha lactalbumin. [Japanese] Nippon Rinsho 2004;62 Suppl 11:288-90
  94. Nakata R. Alpha-lactalbumin. [Japanese] Nippon Rinsho 1999;57 Suppl:269-71
  95. Imafidon GI, Farkye NY, Spanier AM. Isolation, purification, and alteration of some functional groups of major milk proteins: a review. Crit Rev Food Sci Nutr 1997;37(7):663-89
  96. Anderson PJ, Brooks CL, Berliner LJ. Functional identification of calcium binding residues in bovine alpha-lactalbumin. Biochemistry 1997;36(39):11648-54
  97. Sawyer L, Holt C. The secondary structure of milk proteins and their biological function.
    J Dairy Sci 1993;76(10):3062-78
  98. Morr CV, Ha EY. Whey protein concentrates and isolates: processing and functional properties. Crit Rev Food Sci Nutr 1993;33(6):431-76
  99. Bordin G, Cordeiro Raposo F, de la Calle B, Rodriguez AR. Identification and quantification of major bovine milk proteins by liquid chromatography.
    J Chromatogr A 2001;928(1):63-76
  100. Kuitunen M, Savilahti E, Sarnesto A. Human alpha-lactalbumin and bovine beta-lactoglobulin absorption in infants.
    Allergy 1994;49(5):354-60
  101. Neyestani TR, Djalali M, Pezeshki M. Isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow’s milk using gel filtration and anion-exchange chromatography including evaluation of their antigenicity. Protein Expr Purif 2003;29(2):202-8
  102. Hendrix TM, Griko Y, Privalov P. Energetics of structural domains in alpha-lactalbumin. Protein Sci 1996;5(5):923-31
  103. Jackson JG, Janszen DB, Lonnerdal B, Lien EL, Pramuk KP, Kuhlman CF. A multinational study of alpha-lactalbumin concentrations in human milk.
    J Nutr Biochem 2004;15(9):517-21
  104. Brew K, Grobler J (1992) a-Lactalbumin. In: Fox P (ed) Advances in Dairy Chemistry, Elsevier Applied Sciences, New York, pp 191-229
  105. Permyakov EA, Berliner LJ. alpha-Lactalbumin: structure and function.
    FEBS Lett 2000;473(3):269-74
  106. Gall H, Kalveram CM, Sick H, Sterry W. Allergy to the heat-labile proteins alpha-lactalbumin and beta-lactoglobulin in mare’s milk.
    J Allergy Clin Immunol 1996;97(6):1304-7
  107. Baroglio C, Giuffrida MG, Cantisani A, Napolitano L, Bertino E, Fabris C, Conti A. Evidence for a common epitope between bovine alpha-lactalbumin and beta-lactoglobulin.
    Biol Chem 1998;379(12):1453-6
  108. Calderone V, Giuffrida MG, Viterbo D, Napolitano L, Fortunato D, Conti A, Acharya KR. Amino acid sequence and crystal structure of buffalo alpha-lactalbumin.
    FEBS Lett 1996;394(1):91-5
  109. Makinen-Kiljunen S, Sorva R. Bovine beta-lactoglobulin levels in hydrolysed protein formulas for infant feeding.
    Clin Exp Allergy 1993;23(4):287-91
  110. Bottaro G, Castellucci G, et al. Comparison of two methods to determine beta-lactoglobulin antibodies in children with cow’s milk protein intolerance. [Italian]
    Pediatr Med Chir 1992;14(1):27-30
  111. Restani P, Gaiaschi A, Plebani A, Beretta B, Velona T, et al. Evaluation of the presence of bovine proteins in human milk as a possible cause of allergic symptoms in breast-fed children. Ann Allergy Asthma Immunol 2000;84(3):353-60
  112. Juvonen P, Mansson M, Kjellman NI, Bjorksten B, Jakobsson I. Development of immunoglobulin G and immunoglobulin E antibodies to cow’s milk proteins and ovalbumin after a temporary neonatal exposure to hydrolyzed and whole cow’s milk proteins.
    Pediatr Allergy Immunol 1999;10(3):191-8
  113. Ehn BM, Ekstrand B, Bengtsson U, Ahlstedt S. Modification of IgE Binding during Heat Processing of the Cow’s Milk Allergen beta-Lactoglobulin.
    J Agric Food Chem 2004;52(5):1398-403
  114. Makinen-Kiljunen S, Palosuo T. A sensitive enzyme-linked immunosorbent assay for determination of bovine beta-lactoglobulin in infant feeding formulas and in human milk. Allergy 1992;47(4 Pt 2):347-52
  115. Chatel JM, Bernard H, Clement G, Frobert Y, Batt CA, Gavalchin J, Peltres G, Wal JM. Expression, purification and immunochemical characterization of recombinant bovine beta-lactoglobulin, a major cow milk allergen.
    Mol Immunol 1996;33(14):1113-8
  116. del Val G, Yee BC, Lozano RM, Buchanan BB, Ermel RW, Lee YM, Frick OL. Thioredoxin treatment increases digestibility and lowers allergenicity of milk.
    J Allergy Clin Immunol 1999;103(4):690-7
  117. Wal J. Milk allergens.
    Int Dairy J 1998;8:413-23
  118. Virtanen T. Zeller. Important animal allergens are lipocalin proteins: why are they allergenic?
    Int Arch Allergy Immunol 1999;120:247-58
  119. McKenzie IA, Ralston GB, Shaw DC. Location of sulfhydryl and disulfide groups in bovine b-lactoglobulins and effects of urea. Biochemistry 1972,11:4539-47
  120. Reddy IM, Kella NK, Kinsella JE. Structural and conformational basis of the resistance of b-lactoglobulin to peptic and chymotryptic digestion.
    J Agric Food Chem 1988:36:737-41
  121. Jakobsson I, Lindherg T, Benediktsson B, Hansson BG. Dietary bovine b-lactoglobulin is transferred to human milk. Acta Paediatr Scand 1985;74:342-45
  122. Godovac-Zimmermann J, Braunitzer G. Modern aspects of the primary structure and function of b-lactoglobulins. Milchwissenschaft 1987,42:294-7
  123. Papiz MZ, Sawyer L, Eliopoulos EE, North AC, Findlay JB, Sivaprasadarao R, Jones TA, Newcomer ME, Kraulis PJ. The structure of b-lactoglobulin and its similarity to plasma retinol-binding protein.
    Nature 1986,324:383-5
  124. Brownlow S, Morais Cabral JH, Cooper R, Flower DR, Yewdall SJ, Polikarpov I, North AC, Sawyer L. Bovine beta-lactoglobulin at 1.8 A resolution--still an enigmatic lipocalin. Structure 1997,5:481-95
  125. Flower DR. The lipocalin protein family: structure and function.
    Biochem J 1996;318:1-14
  126. Wal JM. Structure and function of milk allergens. Allergy 2001;56(Suppl 67):35-8
  127. Farrell HM Jr, Jimenez-Flores R, Bleck GT, Brown EM, Butler JE, Creamer LK, Hicks CL, Hollar CM, Ng-Kwai-Hang KF, Swaisgood HE. Nomenclature of the proteins of cows’ milk--sixth revision.
    J Dairy Sci 2004;87(6):1641-74
  128. Aymard P, Durand D, Nicolai T. The effect of temperature and ionic strength on the dimerisation of beta-lactoglobulin.
    Int J Biol Macromol 1996;19(3):213-21
  129. Inoue R, Matsushita S, Kaneko H, Shinoda S, Sakaguchi H, Nishimura Y, Kondo N. Identification of beta-lactoglobulin-derived peptides and class II HLA molecules recognized by T cells from patients with milk allergy.
    Clin Exp Allergy 2001;31(7):1126-34
  130. Ehn BM, Ekstrand B, Bengtsson U, Ahlstedt S. Modification of ige binding during heat processing of the cow’s milk allergen beta-lactoglobulin.
    J Agric Food Chem 2004;52(5):1398-403
  131. Rolfsen W, Tibell M, Yman L. Cow’s milk proteins as allergens and antigens. Allergol Immunolog Clinica (Madr) 1987;2:213
  132. Davis PJ, Williams SC. Protein modification by thermal processing.
    Allergy 1998;53(46 Suppl):102-5
  133. Witteman AM, van Leeuwen J, van der Zee J, Aalberse RC. Food allergens in house dust. Int Arch Allergy Immunol 1995;107(4):566-8
  134. Suutari TJ, Valkonen KH, Karttunen TJ, Ehn BM, Ekstrand B, Bengtsson U, Virtanen V, Nieminen M, Kokkonen J. IgE cross reactivity between reindeer and bovine milk beta-lactoglobulins in cow’s milk allergic patients. J Investig Allergol Clin Immunol 2006;16(5):296-302
  135. Sampson HA, James JM, Bernhisel-Broadbent J. Safety of an amino-acid derived infant formula in children allergic to cow milk. Pediatrics 1992;90:463-5
  136. Plebani A, Restani P, Naselli A, Galli CL, Meini A, Cavagni G, Ugazio AG, Poiesi C. Monoclonal and polyclonal antibodies against casein components of cow milk for evaluation of residual antigenic activity in “hypoallergenic” infant formulas.
    Clin Exp Allergy 1997;27:949-56
  137. Makinen-Kiljunen S, Mussalo-Rauhamaa H. Casein, an important house dust allergen. Allergy 2002;57(11):1084-5
  138. Bernard H, Creminon C, Yvon M, Wal JM. Specificity of the human IgE response to the different purified caseins in allergy to cow’s milk proteins. Int Arch Allergy Immunol 1998;115(3):235-44
  139. Bernard H, Meisel H, Creminon C, Wal JM Post-translational phosphorylation affects the IgE binding capacity of caseins.
    FEBS Lett 2000;467(2-3):239-44
  140. Ruiter B, Tregoat V, M’rabet L, Garssen J, Bruijnzeel-Koomen CA, Knol EF, Hoffen E. Characterization of T cell epitopes in alphas1-casein in cow’s milk allergic, atopic and non-atopic children.
    Clin Exp Allergy 2006;36(3):303-10
  141. Kilshaw PJ, Heppell LM, Ford JE. Effects of heat treatment of cow’s milk and whey on the nutritional quality and antigenic properties. Arch Dis Child 1982;57(11):842-7
  142. Mercier JC, Grosclaude F, Ribadeau-Dumas B. Primary structure of bovine s1 casein. Complete sequence.
    Eur J Biochem 1971;23(1):41-51
  143. Kohno Y, Honma K, Saito K, Shimojo N, Tsunoo H, Kaminogawa S, Niimi H. Preferential recognition of primary protein structures of alpha-casein by IgG and IgE antibodies of patients with milk allergy.
    Ann Allergy 1994;73(5):419-22
  144. Chatchatee P, Jarvinen KM, Bardina L, Beyer K, Sampson HA. Identification of IgE- and IgG-binding epitopes on alpha(s1)-casein: differences in patients with persistent and transient cow’s milk allergy. J Allergy Clin Immunol 2001;107(2):379-83
  145. Muller-Renaud S, Dupont D, Dulieu P. Quantification of beta casein in milk and cheese using an optical immunosensor.
    J Agric Food Chem 2004;52(4):659-64
  146. Swaisgood HE. Review and update of casein chemistry. J Dairy Sci 1993;76(10):3054-61
  147. Bernard H, Wal JM, Creminon C, Grassi J, et al. Sensitivities of cow’s milk allergic patients to caseins fraction of milks from different species. Allergy 1992;47:306
  148. Lara-Villoslada F, Olivares M, Xaus J. The balance between caseins and whey proteins in cow’s milk determines its allergenicity.
    J Dairy Sci 2005;88(5):1654-60
  149. Hermansen JE, Ostersen S, Justesen NC, Aaes O. Effects of dietary protein supply on caseins, whey proteins, proteolysis and renneting properties in milk from cows grazing clover or N fertilized grass.
    J Dairy Res. 1999;66(2):193-205
  150. Sicherer SH, Sampson HA Cow’s milk protein-specific IgE concentrations in two age groups of milk-allergic children and in children achieving clinical tolerance.
    Clin Exp Allergy 1999;29(4):507-12
  151. Vila L, Beyer K, Jarvinen KM, Chatchatee P, Bardina L, Sampson HA. Role of conformational and linear epitopes in the achievement of tolerance in cow’s milk allergy.
    Clin Exp Allergy 2001;31(10):1599-606
  152. Jarvinen KM, Beyer K, Vila L, Chatchatee P, Busse PJ, Sampson HA. B-cell epitopes as a screening instrument for persistent cow’s milk allergy. J Allergy Clin Immunol 2002;110(2 Pt 1):293-7
  153. Munoz Martin T, De La Hoz Caballer B, Maranon Lizana F, Gonzalez Mendiola R, Prieto Montano P, Sanchez Cano M. Selective allergy to sheep’s and goat’s milk proteins.
    Allergol Immunopathol (Madr) 2004;32(1):39-42
  154. Bernard H, Negroni L, Chatel JM, Clement G, Adel-Patient K, Peltre G, Creminon C, Wal JM. Molecular basis of IgE cross-reactivity between human beta-casein and bovine beta-casein, a major allergen of milk.
    Mol Immunol 2000;37(3):161-7
  155. Bernad H, Creminon C, Negroni L, Peltre G, Wal JM. IgE cross-reactivity with caseins from different species in humans allergic to cow’s milk.
    Food Agric Immunol 1999;11(1):101-11
  156. Umpierrez A, Quirce S, Maranon F, Cuesta J, Garcia-Villamuza Y, et al. Allergy to goat and sheep cheese with good tolerance to cow cheese. Clin Exp Allergy 1999;29(8):1064-8
  157. Conneely OM. Antiinflammatory activities of lactoferrin. J Am Coll Nutr 2001;20(5 Suppl):389S-95S
  158. Taylor S, Brock J, Kruger C, Berner T, Murphy M. Safety determination for the use of bovine milk-derived lactoferrin as a component of an antimicrobial beef carcass spray.
    Regul Toxicol Pharmacol 2004;39(1):12-24
  159. Cantisani A, Giuffrida MG, Fabris C, Bertino E, Coscia A, Oggero R, Monti G, Stroppiana P, Conti A. Detection of specific IgE to human milk proteins in sera of atopic infants.
    FEBS Lett 1997;412(3):515-7
  160. Schanbacher FL, Goodman RE, Talhouk RS. Bovine mammary lactoferrin: implications from messenger ribonucleic acid (mRNA) sequence and regulation contrary to other milk proteins.
    J Dairy Sci 1993;76(12):3812-31
  161. Ward PP, Uribe-Luna S, Conneely OM. Lactoferrin and host defense.
    Biochem Cell Biol 2002;80(1):95-102
  162. Hanson LA, Ahlstedt S, Andersson B, Carlsson B, Fällström SP, Mellander L, Porras O, Söderström T, Edén CS. Protective factors in milk and the development of the immune system. Pediatrics 1985;75(1 Pt 2):172-6
  163. Elrod KC, Moore WR, Abraham WM, Tanaka RD. Lactoferrin, a potent tryptase inhibitor, abolishes late-phase airway responses in allergic sheep. Am J Respir Crit Care Med 1997;156(2 Pt 1):375-81
  164. Tsokos M, Paulsen F. Expression of pulmonary lactoferrin in sudden-onset and slow-onset asthma with fatal outcome. Virchows Arch 2002;441(5):494-9
  165. Lampreave F, Pineiro A, Brock JH, Castillo H, Sanchez L, Calvo M. Interaction of bovine lactoferrin with other proteins of milk whey. Int J Biol Macromol 1990;12(1):2-5
  166. Brock JH, Lamont A, Boyle DJ, Holme ER, McSharry C, Bunn JE, Lonnerdal B. Antibodies to lactoferrin. A possible link between cow’s milk intolerance and autoimmune disease.
    Adv Exp Med Biol 1998;443:305-11
  167. Han GD, Matsuno M, Ito G, Ikeucht Y, Suzuki A. Meat allergy: investigation of potential allergenic proteins in beef. Biosci Biotechnol Biochem 2000;64(9):1887-95
  168. Fiocchi A, Restani P, Riva E. Beef allergy in children. Nutrition 2000;16(6):454-7
  169. Fiocchi A, Restani P, Riva E, Mirri GP, Santini I, Bernardo L, Galli CL. Heat treatment modifies the allergenicity of beef and bovine serum albumin.
    Allergy 1998;53(8):798-802
  170. Tanabe S, Shibata R, Nishimura T. Hypoallergenic and T cell reactive analogue peptides of bovine serum albumin, the major beef allergen.
    Mol Immunol 2004;41(9):9-890
  171. Tanabe S, Kobayashi Y, Takahata Y, Morimatsu F, Shibata R, Nishimura T. Some human B and T cell epitopes of bovine serum albumin, the major beef allergen.
    Biochem Biophys Res Commun 2002;293(5):1348-53
  172. Beretta B, Conti A, Fiocchi A, Gaiaschi A, Galli CL, Giuffrida MG, et al. Antigenic determinants of bovine serum albumin. Int Arch Allergy Immunol 2001;126(3):188-95
  173. Wahn U, Peters T Jr, Siraganian RP. Allergenic and antigenic properties of bovine serum albumin.
    Mol Immunol 1981;18(1):19-28
  174. Hilger C, Grigioni F, De Beaufort C, Michel G, Freilinger J, Hentges F. Differential binding of IgG and IgA antibodies to antigenic determinants of bovine serum albumin.
    Clin Exp Immunol 2001;123(3):387-94
  175. Restani P, Fiocchi A, Beretta B, Velona T, Giovannini M, Galli CL. Effects of structure modifications on IgE binding properties of serum albumins. Int Arch Allergy Immunol 1998;117(2):113-9
  176. Elliott AJ, Datta N, Amenu B, Deeth HC. Heat-induced and other chemical changes in commercial UHT milks.
    J Dairy Res 2005;72(4):442-6
  177. Schmidt DG, Meijer RJ, Slangen CJ, van Beresteijn EC. Raising the pH of the pepsin-catalysed hydrolysis of bovine whey proteins increases the antigenicity of the hydrolysates. Clin Exp Allergy 1995;25(10):1007-17
  178. Fiocchi A, Restani P, Riva E, Qualizza R, Bruni P, Restelli AR, Galli CL. Meat allergy: I--Specific IgE to BSA and OSA in atopic, beef sensitive children.
    J Am Coll Nutrition 1995;14(3):239-44
  179. San-Juan S, Lezaun A, Caballero ML, Moneo I. Occupational allergy to raw beef due to cross-reactivity with dog epithelium.
    Allergy 2005;60(6):839-40
  180. Fuentes Aparicio V, Sanchez Marcen I, Perez Montero A, Baeza ML, de Barrio Fernandez M. Allergy to mammal’s meat in adult life: immunologic and follow-up study. J Investig Allergol Clin Immunol 2005;15(3):228-31
  181. Vicente-Serrano J, Caballero ML, Rodriguez-Perez R, Carretero P, Perez R, Blanco JG, Juste S, Moneo I. Sensitization to serum albumins in children allergic to cow’s milk and epithelia. Pediatr Allergy Immunol 2007;18(6):503-7
  182. Caballero T, Alonso A, De Miguel S, Martin-Esteban M, Varga B, Pascual CY, Lopez-Serrano MC. IgE-mediated anaphylaxis to Thiomucase, a mucopolyssacharidase: allergens and cross-reactivity.
    Allergy 2002;57(3):254-7


As in all diagnostic testing, the diagnosis is made by the physican based on both test results and the patient history.