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

Hee Seon Lee; Mi Na Kim; Jung Yeon Hong; Won Il Heo; Kyung Won Kim; Myung Hyun Sohn; Kyu-Earn Kim; Kyung Eun Lee; Jung-Won Park

doi:10.4168/aard.2014.2.4.259

Journal List > Allergy Asthma Respir Dis > v.2(4) > 1059037

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Lee, Kim, Hong, Heo, Kim, Sohn, Kim, Lee, and Park: The effect of heat treatment or hydrolysis on cow's milk protein distributions and antigenicities

Original Article

Allergy, Asthma & Respiratory Disease 2014; 2(4): 259-265.

Published online: 30 September 2014

DOI: https://doi.org/10.4168/aard.2014.2.4.259

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

Hee Seon Lee^1,^2,^*, Mi Na Kim^1,^2,^3,^*, Jung Yeon Hong^1,^2,³, Won Il Heo^1,^2,³, Kyung Won Kim^1,², Myung Hyun Sohn^1,^2,^3,⁴, Kyu-Earn Kim^1,^2,³, Kyung Eun Lee^1,^2,^3,⁴, Jung-Won Park^2,^3,⁵

¹Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea.

²Institute of Allergy, Yonsei University College of Medicine, Seoul, Korea.

³Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.

⁴Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea.

⁵Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea.

Correspondence to: Kyung Eun Lee. Department of Pediatrics and Institute of Allergy, Biomedical Science Institute, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul 120-752, Korea. Tel: +82-2-2228-0750, Fax: +82-2-2227-8129, KELEE616@yuhs.ac

Equal contributor:

*These authors contributed equally to this study and should be considered co-first authors.

Received 8 November 2013 Revised 8 April 2014 Accepted 14 April 2014

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/).

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 α-casen, β-lactoglobulin (BLG), α-lactalbumin (ALA), and crude extract of cow's milk in 100℃ 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.

Keywords: Milk protein, Heating, Hydrolysis, Antigenicity

Figures and Tables

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.

Table 1

Clinical characteristics of 11 subjects with milk sensitization

Notes

This study was supported by a grant from the Korea Healthcare Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (A092076).

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