Major Milk Allergen Research Articles

CRISPR-mediated editing of β-lactoglobulin (BLG) gene in buffalo

Milk is a good source of nutrition but is also a source of allergenic proteins such as α-lactalbumin, β-lactoglobulin (BLG), casein, and immunoglobulins. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas technology has the potential to edit any gene, including milk allergens. Previously, CRISPR/Cas has been successfully employed in dairy cows and goats, but buffaloes remain unexplored for any milk trait. In this study, we utilized the CRISPR/Cas9 system to edit the major milk allergen BLG gene in buffaloes. First, the editing efficiency of designed sgRNAs was tested in fibroblast cells using the T7E assay and Sanger sequencing. The most effective sgRNA was selected to generate clonal lines of BLG-edited cells. Analysis of 15 single-cell clones, through TA cloning and Sanger sequencing, revealed that 7 clones exhibited bi-allelic (−/−) heterozygous, bi-allelic (−/−) homozygous, and mono-allelic (−/+) disruptions in BLG. Bioinformatics prediction analysis confirmed that non-multiple-of-3 edited nucleotide cell clones have frame shifts and early truncation of BLG protein, while multiple-of-3 edited nucleotides resulted in slightly disoriented protein structures. Somatic cell nuclear transfer (SCNT) method was used to produce blastocyst-stage embryos that have similar developmental rates and quality with wild-type embryos. This study demonstrated the successful bi-allelic editing (−/−) of BLG in buffalo cells through CRISPR/Cas, followed by the production of BLG-edited blastocyst stage embryos using SCNT. With CRISPR and SCNT methods described herein, our long-term goal is to generate gene-edited buffaloes with BLG-free milk.

Allergenicity evaluation of fermented milk prepared by co-fermentation of Lactobacillus plantarum 7-2 and commercial starters after in vitro digestive

Milk allergy is one of the most common food allergies, in which αS-casein is the major milk allergen. Under optimized conditions, mixed starter (containing Lactobacillus plantarum 7-2 and commercial starter) effectively degraded αS-casein of skimmed milk and reduced the pressure of stomach digestion. The fermented milk prepared by mixed starter was determined by ELISA, the antigenicity of αS-casein was reduced by 77.53%. Compared with the fermented milk prepared by commercial starter, label-free quantitative proteomics demonstrated that the mixed starter more efficiently degraded the epitopes of major milk allergens and influenced the digestion pattern of the fermented milk. Therefore, L. plantarum 7-2 shows positive potential in reducing the antigenicity of αS-casein and others. In addition, this study predicted that the new epitopes produced in the fermentation process could induce immunity using molecular simulation.

Glutamine and lysine as common residues from epitopes on α-lactalbumin and β-lactoglobulin from cow milk identified by phage display technology

Cow milk is an important source of food protein for children; however, it could lead to allergy, especially for infants. α-Lactalbumin (α-LA) and β-lactoglobulin (β-LG) from whey protein make up a relatively high proportion of milk proteins and have received widespread attention as major allergens in milk. However, few studies have identified the epitopes of both proteins simultaneously. In this study, ImmunoCAP and indirect ELISA were first used for detection of sIgE to screen sera from allergic patients with high binding capacity for α-LA and β-LG. Subsequently, the mimotopes was biopanned by phage display technology and bioinformatics and 17 mimic peptide sequences were obtained. Aligned with the sequences of α-LA or β-LG, we identified one linear epitope on α-LA at AA 11-26 and 5 linear epitopes on β-LG at AA 9-29, AA 45-57, AA 77-80, AA 98-101, and AA 121-135, respectively. Meanwhile, the 8 conformational epitopes and their distributions of α-LA and β-LG were located using the Pepitope Server. Finally, glutamine and lysine were determined as common AA residues for the conformational epitopes both on α-LA and β-LG. Moreover, we found the addition of mouse anti-human IgE during the biopanning process did not significantly affect the identification of the epitopes.

Recent advances in selective allergies to mammalian milk proteins not associated with Cow's Milk Proteins Allergy.

Cow's milk proteins allergy (CMA) is an atypical immune system response to cow's milk and dairy products. It's one of the most common food allergies in children affecting 8% of the total pediatric population pediatric population. This comprehensive review examines recent studies in CMA, especially regarding mammalian milk allergies such as goat's, sheep's, buffalo's, camel's, mare's and donkey's milk allergies in order to increase awareness of these selective allergies and to reduce allergy risks for those who have them. The consumption of other mammalian milk types is not recommended because of the significant homology between milk proteins from cow, sheep, goat and buffalo resulting in clinical cross-reactivity. However, camel's, mare's or donkey's milk may be tolerated by some allergic patients. Selective mammalian milk allergies are unusual and rare disorders characterized by severe symptoms including angio-oedema, urticaria, respiratory manifestations and anaphylaxis. Based on the reported allergic cases, cheese products including Ricotta, Romano, Pecorino and Mozzarella, are considered as the most common source of allergens especially in goat's, sheep's and buffalo's milk allergies, while the major allergens in donkey's and mare's milk seems to be whey proteins including lysozyme, α-lactalbumin and β-lactogloblin due to the low casein/whey proteins ratio in equine's milk.

Safe food through better labelling; a robust method for the rapid determination of caprine and bovine milk allergens.

Accidental milk cross-contamination is one of the most common causes for costly food recalls. Yet, quantifying trace-levels of allergen is time-consuming and current methods are not adapted for routine analyses making quality control for trace-level allergen content impractical. This perpetuates voluntary "may-contain" statements that are unhelpful for people suffering from food allergies. Here, we developed a rapid LC-MS method enabling milk allergen quantification by comparing all tryptic-peptides of major milk allergens. The bovine-specific αS-2 casein peptide and allergen-epitope NAVPITPTLNR provided excellent performance in sensitivity (LOD 1mg.kg-1; LOQ 2mg.kg-1) across various dairy products, good recovery rates in baked croissants (77% with a 10% inter-day RSD) and a linear range of 2-2,000mg.kg-1. The method can be used for routine determination of trace-contamination with bovine milk allergen and the adulteration of high-value caprine dairy products with lower-value bovine milk products, protecting consumer trust and the growing population suffering from food allergies.

Cross-reactivity analysis of milk proteins from different goat breeds with cow's milk allergens using a proteomic approach.

Goat's milk thought to be a good substitute for cow's milk protein allergic (CMPA) individuals. However, there is growing evidence that their proteins have cross-reactivities with cow's milk allergens. This study aimed to profile and compare milk proteins from different goat breeds that have cross-reactivity to cow's milk allergens. Proteomics was used to compare protein extracts of skim milk from Saanen, Jamnapari, and Toggenburg. Cow's milk was used as a control. IgE-immunoblotting and mass spectrometry were used to compare and identify proteins that cross-reacted with serum IgE from CMPA patients (n=10). The analysis of IgE-reactive proteins revealed that the protein spots identified with high confidence were proteins homologous to common cow's milk allergens such as α-S1-casein (αS1-CN), β-casein (β-CN), κ-casein (κ-CN), and beta-lactoglobulin (β-LG). Jamnapari's milk proteins were found to cross-react with four major milk allergens: α-S1-CN, β-CN, κ-CN, and β-LG. Saanen goat's milk proteins, on the other hand, cross-reacted with two major milk allergens, α-S1-CN and β-LG, whereas Toggenburg goat's milk proteins only react with one of the major milk allergens, κ-CN. These findings may help in the development of hypoallergenic goat milk through cross-breeding strategies of goat breeds with lower allergenic milk protein contents.

Insight into the molecular-level details of αs1 casein interactions with IgG: Combining with LC-MS/MS and molecular modelling techniques

αs1-Casein (αs1-CN) is a major cow milk allergen, while the tertiary structure of αs1-CN and conformational epitopes of αs1-CN have not been clarified. Here, a reasonable three-dimensional structure of αs1-CN was established using ab initio methods, and hot-spot residues and epitopes were investigated by combining molecular dynamics simulation, peptides synthesis, and ELISA. Obtained results demonstrated that the binding mechanism between αs1-CN and IgG was located on three main regions: a helical structure zone (E77-Q97), the flexible loop zone (Y154-T174), and a flexible C-terminal (N190-L198), mainly connecting via hydrogen bond and ionic bonds. The hydrolysates produced by papain with lowest antigenicity (12.43%), which could considerably destroy the essential epitopes of αs1-CN confirmed by epitope synthesis, and LC-MS/MS. The results reported herein would provide novel insights into the interface interactions between αs1-CN and IgG, and prove valuable for developing hypoallergenic infant-formula and peptide vaccines for allergen-specific immunotherapy.

Development of a light-initiated chemiluminescence assay for the quantitative detection of cow's milk allergen-specific IgE

BackgroundCow's milk allergy is a common food allergy in children. Clinically, cow's milk-specific IgE (CM-sIgE) antibody test is often used to diagnose milk allergy. An inexpensive light-initiated chemiluminescence assay (LICA), with fast detection speed and small sample volume demand, has application prospects in the field of quantitative detection of CM-sIgE. MethodsChemibeads coated with five major milk allergens, serum samples, biotinylated anti-human IgE antibodies, and streptavidin-coated sensibeads constitute a system to establish a LICA method for the quantitative detection of CM-sIgE. A series of experiments were performed to optimize its reaction conditions and evaluated its performance. ResultsThe optimal conditions for LICA were 10:4 mass ratio of chemibeads to milk allergen, 20 μg/mL chemibeads, 1.0 μg/mL biotinylated anti-human IgE antibodies and a 1/10 dilution of serum for 30-min incubation. The limit of Quantitation (LoQ) was 0.22 kUA/L. For repeatability, the CV ranged from 3.71% to 8.11%. For intermediate precision, the CV ranged from 4.08% to 14.71%. It was linear within 0.20–18.20 kUA/L. This method did not interfere with common interfering substances and total IgE in serum, and there was no obvious cross-reaction with milk-specific IgG and non-milk-specific IgE. ConclusionWe have established a method to quantitatively detect CM-sIgE based on light-initiated chemiluminescence assay, which has good analytical performance and could meet the needs of clinical laboratories.

Pressure assisted thermal pasteurization (PATP) of hypoallergenic and low protein ready-to-feed (RTF) infant formula fortified with bioactives: A novel approach

Infant Milk Formula (IMF) is an instant alternative to breastfeeding which is being investigated continuously to mimic the composition of human milk. Hypoallergenic IMF is recommended for infants who cannot tolerate cow milk due to allergy. Paediatric obesity resulted in burgeoning demand for low protein infant formula, especially in Asia. Beta-lactoglobulin (β-Lg), a major cow milk allergen to infants, is present in cow milk as 3–4 g/L, whereas, interestingly, it is absent in human milk. Besides, human milk contains Fe3+ binding lactoferrin (LF) and Ca2+ binding alpha-lactalbumin (α-Lac) about 10 and 3 times, respectively, higher than cow milk. Therefore, a mechanism to formulate a hypoallergenic IMF with low protein content could be the addition of LF and α-Lac and reducing β-Lg at the same time using an appropriate processing method. Hence, as a pragmatic approach, this study considered a pasteurized form of ready-to-feed (RTF) by applying Pressure Assisted Thermal Pasteurization (PATP) achieved from HPP of 600 MPa for 15 s applied at 57.1 °C to attain 72.3 °C after pressurization. In parallel, conventional HTST was also applied at 72 °C for both 15 and 30 s to compare PATP's effect. HTST and PATP retained >90% and > 75% LF respectively whereas PATP induced higher retention of α-Lac compared to HTST. However, HTST retained >60% β-Lg while PATP induced a high denaturation of β-Lg with 28% retention only. Moreover, both PATP and HTST induced around 5 log reduction of E. coli (ATCC 8739) in IMF. Therefore, the current investigation potentially indicates a route to develop a PATP-processed hypoallergenic and low protein RTF formula, a niche product.

Simultaneous Detection of Three Major Milk Allergens in Food by UPLC-MS/MS

Simultaneous Detection of Three Major Milk Allergens in Food by UPLC-MS/MS

Enhanced Uptake of Processed Bovine β-Lactoglobulin by Antigen Presenting Cells: Identification of Receptors and Implications for Allergenicity.

Scopeβ‐lactoglobulin (BLG) is a major cow milk allergen encountered by the immune system of infants fed with milk‐based formulas. To determine the effect of processing on immunogenicity of BLG, this article characterized how heated and glycated BLG are recognized and internalized by APCs. Also, the effect of heat‐induced structural changes as well as gastrointestinal digestion on immunogenicity of BLG is evaluated.Methods and resultsThe binding and uptake of BLG from raw cow milk and heated either alone (BLG‐H) or with lactose/glucose (BLG‐Lac and BLG‐Glu) to the receptors present on APCs are analyzed by ELISA and cell‐binding assays. Heated and glycated BLG is internalized via galectin‐3 (Gal‐3)and scavenger receptors (CD36 and SR‐AI) while binding to the receptor for advanced glycation end products (R AGE) does not cause internalization. Receptor affinity of BLG is dependent on increased hydrophobicity, β‐sheet exposure and aggregation. Digested glycated BLG maintained binding to sRAGE and Gal‐3 but not to CD36 and SR‐AI, and is detected on the surface of APCs. This suggests a mechanism via which digested glycated BLG may trigger innate (via RAGE) and adaptive immunity (via Gal‐3).ConclusionsThis study defines structural characteristics of heated and glycated BLG determining its interaction with APCs via specific receptors thus revealing enhanced immunogenicity of glycated versus heated BLG.

Bos d 11 in baked milk poses a risk for adverse reactions in milk-allergic patients.

Patients are commonly challenged with foods containing baked milk, for example muffins, yet little is known about the specific allergen content of muffins used in milk challenges or of the effect that baking has on allergenicity. Our objective was to compare the levels of major milk allergens in uncooked and baked muffins using monoclonal immunoassays and IgE antibody binding before and after baking. Uncooked and baked muffins were prepared using recipes from Mount Sinai and Imperial College. Allergen levels were compared by ELISA for Bos d 5 (β-lactoglobulin) and Bos d 11 (β-casein). IgE reactivity was assessed using sera from milk-sensitized donors in direct binding and inhibition ELISA. Bos d 5 was reduced from 680µg/g in uncooked muffin mix to 0.17µg/g in baked muffins, representing a >99% decrease after baking. Conversely, Bos d 11 levels in baked muffin remained high and only decreased by 30% from a mean of 4249µg/g in uncooked muffin mix to 2961µg/g when baked (~181mg Bos d 11 per muffin). Baked muffins retained ~70% of the IgE binding to uncooked muffin mix. Baked muffin extract inhibited IgE binding to uncooked muffin mix by up to 80%, demonstrating retention of in vitro IgE reactivity. High levels of Bos d 11 in baked muffins pose a risk for adverse reactions, especially in patients who have high anti-casein IgE antibodies.

Potential allergenicity assessment after bovine apo-α-lactalbumin binding to calcium ion.

Bovine α-lactalbumin (α-LA) is recognized as a major milk allergen. Generally, α-LA in the natural state combines with a calcium ion, however, some studies have shown that calcium ions can binding the other metal binding sites in α-LA as well. In our study, the optimal condition of calcium ion binding to α-LA and the change of structure and allergenicity were explored. By optimizing the conditions, the maximum calcium binding amounts of apo-α-LA were obtained in a ratio of 1:4. The structure of α-LA after removal of calcium obviously changed by the spectroscopic detection. For the digestive stability, there was no obvious change in three forms of α-LA. While the allergenic properties were characterized by IgG/IgE inhibition ELISA and the human basophil KU812 degranulation assay. The results showed that IgG and IgE binding decreased, and the degranulation capacity of basophils weakened. Based on these results, calcium binding to apo-α-LA can reduce the potential allergenic properties. PRACTICAL APPLICATIONS: By optimizing the conditions, bovine apo-α-LA can obtain the most calcium binding amount. And calcium binding to apo-α-LA can reduce the potential allergenic properties. Compared with α-LA in the natural state, calcium binding to α-LA not only can reduce the allergenic properties, but also play a role in calcium supplementation. It might be used to guide the development of hypoallergenic α-LA and provide a method to reduce the potential allergenic properties of α-LA.

Beta-Lactoglobulin Sensor Based on ISFET with CеO2 Gate Dielectric

In this article sensor with improved measurement technique for the detection of milk allergens was developed. β-lactoglobulin (BLG) protein was used as the biomarker for cow milk detection. The structure of the sensor with nanocrystalline cerium oxide is proposed with an advanced measurement method, which consists of using a micro-flow system to provide a constant volume of the substance to be tested for the dynamic analysis of the beta-lactoglobulin protein, the major milk allergen. The influence of illumination on the experiment and the degree of cleaning of the sensor surface between each measurement are analyzed. Because the existing ELISA method is too long (1.5 hours), and the proposed method allows to identify the presence of beta-lactoglobulin in a few minutes. U gs = µA

Astrogliosis Associated With Behavioral Abnormality in a Non-anaphylactic Mouse Model of Cow's Milk Allergy.

Etiology of neuropsychiatric disorders is complex, involving multiple factors that can affect the type and severity of symptoms. Although precise causes are far from being identified, allergy or other forms of hypersensitivity to dietary ingredients have been implicated in triggering or worsening of behavioral and emotional symptoms, especially in patients suffering from depression, anxiety, attention-deficit hyperactivity, and/or autism. Among such ingredients, cow's milk, along with wheat gluten, is commonly suspected. However, the contributory role of cow's milk in these disorders has not been elucidated due to insufficient pathophysiological evidence. In the present study, we therefore investigated neuroinflammatory changes that are associated with behavioral abnormality using a non-anaphylactic mouse model of cow's milk allergy (CMA). Male and female C57BL/6J mice were subjected to a 5-week oral sensitization procedure without or with a major milk allergen, beta-lactoglobulin (BLG). All mice were then later challenged with BLG, and their anxiety- and depression-associated behaviors were subsequently assessed during the 6th and 7th weeks. We found that BLG-sensitized male mice exhibited significantly increased anxiety- and depression-like behavior, although they did not display anaphylactic reactions when challenged with BLG. Female behavior was not noticeably affected by BLG sensitization. Upon examination of the small intestines, reduced immunoreactivity to occludin was detected in the ileal mucosa of BLG-sensitized mice although the transcriptional expression of this tight-junction protein was not significantly altered when measured by quantitative RT-PCR. On the other hand, the expression of tumor necrosis factor alpha (TNFα) in the ileal mucosa was significantly elevated in BLG-sensitized mice, suggesting the sensitization had resulted in intestinal inflammation. Inflammatory responses were also detected in the brain of BLG-sensitized mice, determined by the hypertrophic morphology of GFAP-immunoreactive astrocytes. These reactive astrocytes were particularly evident near the blood vessels in the midbrain region, resembling the perivascular barrier previously reported by others in experimental autoimmune encephalitis (EAE) mouse models. Interestingly, increased levels of COX-2 and TNFα were also found in this region. Taken together, our results demonstrated that BLG sensitization elicits inflammatory responses in the intestine and brain without overt anaphylactic signs of milk allergy, signifying food allergy as a potential pathogenic factor of neuropsychiatric disorders.

Effect of thermal and microwave processing on secondary structure of bovine β-lactoglobulin: A molecular modeling study.

Milk allergy is known to cause severe allergic reactions in hypersensitive patients, especially in infants and children. β-Lactoglobulin is one of the major allergens in bovine milk. The influence of thermal and microwave processing on the structural deviations of β-lactoglobulin protein have been studied using molecular modeling techniques. The structural deviations are studied using root mean square deviations, radius of gyration, dipole moment, and solvent accessible surface area. STRIDE analysis showed significant changes in the β-lactoglobulin, especially when oscillating electric fields were applied along with heat. Root mean square fluctuations (RMSF) has been assessed for known epitopes in the β-lactoglobulin molecule. This showed that when the protein is exposed to certain thermal stress, it compacts by burying hydrophobic residues in the core. However, few allergic epitope residues also exhibit increased RMSF leading to higher reactive sites on the surface of the protein molecule. PRACTICAL APPLICATIONS: This study showed that molecular modeling can be used to gain valuable insights regarding the structural changes during processing. In the future, with more computational capacity, it can be used to make comparison between results obtained from simulations and real-time experiments. The current techniques used in food industries such as Nuclear Magnetic Resonance Imaging, Fourier Transformation Infrared Spectroscopy, X-ray diffraction can analyze pre- and post-processing effects. Hence, it become necessary to understand the changes that takes place during the processing techniques. Molecular dynamic simulation could be a useful technique in analyzing the changes occurring during the processing.

A rapid immobilized trypsin digestion combined with liquid chromatography – Tandem mass spectrometry for the detection of milk allergens in baked food

Cow's milk allergy is the most common food allergy, and caseins are the major allergens in milk. A quantitative proteomics method based on liquid chromatography coupled to mass spectrometry has been widely used in allergen analysis. However, the long period of digestion has limited its use in routine analysis. PHMN-Trypsin (trypsin immobilized on hairy polymer-chain hybrid magnetic nanoparticles), a new type of immobilized trypsin, was used to shorten the digestion time and enhance the digestion efficiency in this study. We developed a rapid digestion method using PHMN-Trypsin to detect milk allergens in baked food by ultrahigh-performance liquid chromatography – tandem mass spectrometry (UPLC-MS/MS). The PHMN-Trypsin digestion was completed within 15 min, with higher or equal sequence coverage compared to 12–16 h of conventional free trypsin digestion. αs1-casein, αs2-casein, β-casein, and κ-casein were monitored as the allergen proteins. For each target allergen, two or three specific and signature peptides were selected for the UPLC-MS/MS strategy in view of the species specificity, mass response intensity as well as the stability. The limits of quantification of the four proteins in baked foods were 0.38–0.83 μg/g, and their recoveries ranged from 65.2% to 86.1%, with relative standard deviations of less than 11.0%. The method was applied to investigate allergens in commercial baked foods, and the results indicate that this method would be helpful in food allergen screening.

A novel electrochemical mast cell-based paper biosensor for the rapid detection of milk allergen casein

Developing low-cost, portable and simple analysis tools is of vital importance for food safety point-of-care testing. Therefore, herein, a new low-cost, simple to fabricate, disposable, electrochemical mast cell-based paper sensor is proposed and developed to sensitively determine the major milk allergen casein. Then, a graphene (GN)/carbon nanofiber (CN)/ Gelatin methacryloyl (GelMA) composite material with high conductivity and good biocompatibility was modified on the cell-based paper sensor to improve the electrical conductivity and provide a sensing recognition interface for the immobilization of rat basophilic leukemia (RBL-2H3) mast cells. The cyclic voltammetry and differential pulse voltammetry measurement of the mast cells in the paper sensor revealed an irreversible anodic peak, whose peak current is proportional to the number of cells in the range from 1 × 102 to 1 × 108 cells/mL. For the milk allergen detection tests, mast cells exposed to the casein caused a significant reduction in the current signal, displaying an inverse dose-dependent relationship. The developed cell sensor exhibited a range of linearity between 1 × 10−7 and 1 × 10−6 g/mL of casein with a detection limit of 3.2 × 10−8 g/mL and a great reproducibility and selectivity. The electrochemical responses obtained using the cell-based paper sensor were well consistent with the conventional detection assay, with good stability and reproducibility. Therefore, a simple and novel electrochemical method for food allergens detection was developed, demonstrating its potential application in the food safety determination and evaluation.

Production of hypoallergenic milk from DNA-free beta-lactoglobulin (BLG) gene knockout cow using zinc-finger nucleases mRNA

The whey protein β-lactoglobulin (BLG) is a major milk allergen which is absent in human milk. Here, we for the first time generated DNA-free BLG bi-allelic knockout cow by zinc-finger nuclease (ZFNs) mRNA and produced BLG-free milk. According to the allergenicity evaluation of BLG-free milk, we found it can trigger lower allergic reaction of Balb/c mice including the rectal temperature drop and the allergen-specific immunoglobulin IgE production; BLG free-milk was easily digested by pepsin at 2 min, while BLG in control milk was still not completely digested after 60 min, and the binding of IgE from cow’s milk allergy (CMA) patients to BLG free-milk was significantly lower than that to the control milk. Meanwhile, the genome sequencing revealed that our animal is free of off-target events. Importantly, editing animal genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to foods produced by genome edited animals. Finally, the ZFNs-mediated targeting in cow could be transmitted through the germline by breeding. These findings will open up unlimited possibilities of modifying milk composition to make it more suitable for human health and also improve the functional properties of milk.

Glycation of the Major Milk Allergen β-Lactoglobulin Changes Its Allergenicity by Alterations in Cellular Uptake and Degradation.

ScopeDuring food processing, the Maillard reaction (МR) may occur, resulting in the formation of glycated proteins. Glycated proteins are of particular importance in food allergies because glycation may influence interactions with the immune system. This study compared native and extensively glycated milk allergen β‐lactoglobulin (BLG), in their interactions with cells crucially involved in allergy.Methods and resultsBLG was glycated in MR and characterized. Native and glycated BLG were tested in experiments of epithelial transport, uptake and degradation by DCs, T‐cell cytokine responses, and basophil cell degranulation using ELISA and flow cytometry. Glycation of BLG induced partial unfolding and reduced its intestinal epithelial transfer over a Caco‐2 monolayer. Uptake of glycated BLG by bone marrow–derived dendritic cells (BMDC) was increased, although both BLG forms entered BMDC via the same mechanism, receptor‐mediated endocytosis. Once inside the BMDC, glycated BLG was degraded faster, which might have led to observed lower cytokine production in BMDC/CD4+ T‐cells coculture. Finally, glycated BLG was less efficient in induction of degranulation of BLG‐specific IgE sensitized basophil cells.ConclusionsThis study suggests that glycation of BLG by MR significantly alters its fate in processes involved in immunogenicity and allergenicity, pointing out the importance of food processing in food allergy.