Protocol for simultaneous isolation of three important banana allergens
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2014
Authors
Nikolić, JasnaMrkić, Ivan
Grozdanović, Milica
Popović, Milica M.
Petersen, Arnd
Jappe, Uta
Gavrović-Jankulović, Marija
Article (Published version)
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Banana fruit (Musa acuminata) has become an important food allergen source in recent years. So far, 5 IgE reactive banana proteins have been identified, and the major allergens are: Mus a 2 (a class I chitinase, 31 kDa),Mus a4 (thaumatin-like protein, 21 kDa), and Mus a 5 (beta-1,3-glucanase,33 kDa). Due to variations in allergen expression levels, diagnostic reagents for food allergy can be improved by using individual allergen components instead of banana allergen extracts. The purpose of this study was to optimize the purification protocol of the three major allergens present in banana fruit: Mus a 2, Mus a 4 and Mus a 5. By employing a three-step purification protocol (a combination of anion-exchange, cation-exchange and reversed-phase chromatography) three important banana allergens were obtained in sufficient yield and high purity. Characterization of the purified proteins was performed by both biochemical (2-D PAGE, mass fingerprint and N-terminal sequencing) and immunochemical ...(immunoblot) methods. IgE reactivity to the purified allergens was tested by employing sera of five allergic patients. The purified allergens displayed higher sensitivity in IgE detection than the routinely used extracts. The three purified allergens are good candidates for reagents in component-based diagnosis of banana allergy. (C) 2014 Elsevier B.V. All rights reserved.
Keywords:
Allergens / Purification / Banana / Chitinase / TLP / GlucanaseSource:
Journal of Chromatography B: Analytical Technologies in the Biomedical and L, 2014, 962, 30-36Publisher:
- Elsevier Science Bv, Amsterdam
Funding / projects:
- Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-MESTD-Basic Research (BR or ON)-172049)
DOI: 10.1016/j.jchromb.2014.05.020
ISSN: 1570-0232
PubMed: 24887593
WoS: 000338406800005
Scopus: 2-s2.0-84901634866
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Institution/Community
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Nikolić, Jasna AU - Mrkić, Ivan AU - Grozdanović, Milica AU - Popović, Milica M. AU - Petersen, Arnd AU - Jappe, Uta AU - Gavrović-Jankulović, Marija PY - 2014 UR - https://cherry.chem.bg.ac.rs/handle/123456789/1804 AB - Banana fruit (Musa acuminata) has become an important food allergen source in recent years. So far, 5 IgE reactive banana proteins have been identified, and the major allergens are: Mus a 2 (a class I chitinase, 31 kDa),Mus a4 (thaumatin-like protein, 21 kDa), and Mus a 5 (beta-1,3-glucanase,33 kDa). Due to variations in allergen expression levels, diagnostic reagents for food allergy can be improved by using individual allergen components instead of banana allergen extracts. The purpose of this study was to optimize the purification protocol of the three major allergens present in banana fruit: Mus a 2, Mus a 4 and Mus a 5. By employing a three-step purification protocol (a combination of anion-exchange, cation-exchange and reversed-phase chromatography) three important banana allergens were obtained in sufficient yield and high purity. Characterization of the purified proteins was performed by both biochemical (2-D PAGE, mass fingerprint and N-terminal sequencing) and immunochemical (immunoblot) methods. IgE reactivity to the purified allergens was tested by employing sera of five allergic patients. The purified allergens displayed higher sensitivity in IgE detection than the routinely used extracts. The three purified allergens are good candidates for reagents in component-based diagnosis of banana allergy. (C) 2014 Elsevier B.V. All rights reserved. PB - Elsevier Science Bv, Amsterdam T2 - Journal of Chromatography B: Analytical Technologies in the Biomedical and L T1 - Protocol for simultaneous isolation of three important banana allergens VL - 962 SP - 30 EP - 36 DO - 10.1016/j.jchromb.2014.05.020 ER -
@article{ author = "Nikolić, Jasna and Mrkić, Ivan and Grozdanović, Milica and Popović, Milica M. and Petersen, Arnd and Jappe, Uta and Gavrović-Jankulović, Marija", year = "2014", abstract = "Banana fruit (Musa acuminata) has become an important food allergen source in recent years. So far, 5 IgE reactive banana proteins have been identified, and the major allergens are: Mus a 2 (a class I chitinase, 31 kDa),Mus a4 (thaumatin-like protein, 21 kDa), and Mus a 5 (beta-1,3-glucanase,33 kDa). Due to variations in allergen expression levels, diagnostic reagents for food allergy can be improved by using individual allergen components instead of banana allergen extracts. The purpose of this study was to optimize the purification protocol of the three major allergens present in banana fruit: Mus a 2, Mus a 4 and Mus a 5. By employing a three-step purification protocol (a combination of anion-exchange, cation-exchange and reversed-phase chromatography) three important banana allergens were obtained in sufficient yield and high purity. Characterization of the purified proteins was performed by both biochemical (2-D PAGE, mass fingerprint and N-terminal sequencing) and immunochemical (immunoblot) methods. IgE reactivity to the purified allergens was tested by employing sera of five allergic patients. The purified allergens displayed higher sensitivity in IgE detection than the routinely used extracts. The three purified allergens are good candidates for reagents in component-based diagnosis of banana allergy. (C) 2014 Elsevier B.V. All rights reserved.", publisher = "Elsevier Science Bv, Amsterdam", journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and L", title = "Protocol for simultaneous isolation of three important banana allergens", volume = "962", pages = "30-36", doi = "10.1016/j.jchromb.2014.05.020" }
Nikolić, J., Mrkić, I., Grozdanović, M., Popović, M. M., Petersen, A., Jappe, U.,& Gavrović-Jankulović, M.. (2014). Protocol for simultaneous isolation of three important banana allergens. in Journal of Chromatography B: Analytical Technologies in the Biomedical and L Elsevier Science Bv, Amsterdam., 962, 30-36. https://doi.org/10.1016/j.jchromb.2014.05.020
Nikolić J, Mrkić I, Grozdanović M, Popović MM, Petersen A, Jappe U, Gavrović-Jankulović M. Protocol for simultaneous isolation of three important banana allergens. in Journal of Chromatography B: Analytical Technologies in the Biomedical and L. 2014;962:30-36. doi:10.1016/j.jchromb.2014.05.020 .
Nikolić, Jasna, Mrkić, Ivan, Grozdanović, Milica, Popović, Milica M., Petersen, Arnd, Jappe, Uta, Gavrović-Jankulović, Marija, "Protocol for simultaneous isolation of three important banana allergens" in Journal of Chromatography B: Analytical Technologies in the Biomedical and L, 962 (2014):30-36, https://doi.org/10.1016/j.jchromb.2014.05.020 . .