Allergy Asthma Respir Dis.  2014 Sep;2(4):259-265. 10.4168/aard.2014.2.4.259.

The effect of heat treatment or hydrolysis on cow's milk protein distributions and antigenicities

• Lee HS
• Kim MN
• Hong JY
• Heo WI
• Kim KW
• Sohn MH
• Kim KE
• Lee KE
• Park JW

• KMID: 2262393
• DOI: http://doi.org/10.4168/aard.2014.2.4.259

Abstract

PURPOSE
Cow's milk protein is one of the most common and strongest food allergen. We investigated the effects of heat treatment on the distribution and antigenicities of major allergens from cow's milk. We also compared the protein distribution and antigenicities among cow's milk formula and its substitutes.
METHODS
We heated alpha-casen, beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), and crude extract of cow's milk in 100degrees C boiling water for 1 hour. We prepared crude extracts from cow's milk formula, partially hydrolyzed milk formula (pHF) and extensively hydrolyzed milk formula (eHF). The protein compositions of all the samples were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigenicities were determined by IgE immunoblotting with pooled serum collected from 11 patients with milk allergy.
RESULTS
After heating, no significant alteration was found in casein, and the aggregates of ALA and BLG were detected with molecular weights of about 30 and 45 kDa, respectively. The antigenicities of newly detected aggregates were increased. The new aggregates of BLG with increased antigenicities were also found in heated milk total protein. Major milk allergens were not found in pHF, and residual components with a molecular weight below 10 KDa did not show IgE-binding activity. We failed to observe the residual components and antigenicities of eHF.
CONCLUSION
Changes in protein distribution and antigenicity of milk total protein induced by heat treatment may not be significantly different from those of each major allergen. The residual components of pHF could have little IgE-binding capacity, and there may be few or no antigenic components in eHF.

Keyword

Milk protein; Heating; Hydrolysis; Antigenicity

MeSH Terms

Allergens
Caseins
Complex Mixtures
Electrophoresis
Heating
Hot Temperature*
Humans
Hydrolysis*
Immunoblotting
Immunoglobulin E
Lactalbumin
Lactoglobulins
Milk
Milk Hypersensitivity
Milk Proteins*
Molecular Weight
Sodium
Water
Allergens
Caseins
Complex Mixtures
Immunoglobulin E
Lactalbumin
Lactoglobulins
Milk Proteins
Sodium
Water

Figure

• 

  Fig. 1 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis of milk proteins. M, molecular weight marker. Lane 1: milk total protein; Lane 2: purified α-lactalbumin; Lane 3: purified β-lactoglobulin; Lane 4: purified α-casein; Lane 5: IgE immunoblot of milk total protein with patients serum; Lane 6: IgE immunoblot of milk total protein with control buffer.

• 

  Fig. 2 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of heat treated milk proteins. After heating, The 30 KDa band of ALA and the 45 KDa band of BLG were increased in intensity (arrows). M, molecular weight marker. Lane 1: α-lactalbumin; Lane 2: heat treated α-lactalbumin; Lane 3: β-lactoglobulin; Lane 4: heat treated β-lactoglobulin; Lane 5: α-casein; Lane 6: heat treated α-casein; Lane 7: milk total protein; Lane 8: heat treated milk total protein; Lane 9: milk total protein after boiling.

• 

  Fig. 3 IgE immunoblot analysis of heat treated milk proteins. M, molecular weight marker. After heating, the 30 KDa band of ALA and the 45 KDa band of BLG were increased in intensity (arrows). Lane 1: α-lactalbumin; Lane 2: heat treated α-lactalbumin; Lane 3: β-lactoglobulin; Lane 4: heat treated β-lactoglobulin; Lane 5: α-casein; Lane 6: heat treated α-casein; Lane 7: milk total protein; Lane 8: heat treated milk total protein; Lane 9: milk total protein after boiling.

• 

  Fig. 4 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of milk proteins. M, molecular weight marker. Lane 1: ultra-high-temperature pasteurized milk; Lane 2: low-temperature pasteurized milk; Lane 3: cow's milk formula; Lane 4: partially hydrolyzed cow's milk formula; Lane 5: extensively hydrolyzed cow's milk formula.

• 

  Fig. 5 Immunoblot analysis of milk proteins. M, molecular weight marker. Lane 1: ultra-high-temperature pasteurized milk; Lane 2: low-temperature pasteurized milk; Lane 3: cow's milk formula; Lane 4: partially hydrolyzed cow's milk formula; Lane 5: extensively hydrolyzed cow's milk formula.

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