Characterization and Identification of Allergen Protein in Shrimp Before and After Heated with SDS-PAGE Method

Emma Emawati (1) , Idar Idar (2) , Resta Ramadiyanti (3)
(1) Sekolah Tinggi Farmasi Bandung , Indonesia
(2) Sekolah Tinggi Farmasi Bandung , Indonesia
(3) Sekolah Tinggi Farmasi Bandung , Indonesia

Abstract

Food allergies are one of the most common allergies in Indonesian society. Generally, when children aged 5-6 years food allergies will disappear, except peanut allergies and allergies to seafood, such as fish, shellfish and crustaceans. This study aims to determine the pattern of separation of allergen proteins in shrimp using anion exchange column chromatography method and identify allergen proteins in shrimp using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. Protein extraction from shrimp using Phosphate buffer saline (PBS) pH 7.2 and centrifuged at 10,000 rpm for 10 min at 4°C. Protein separation was carried out by anion-exchange column chromatography method, and the fraction obtained was measured at 280nm wavelength. The highest yield at absorbance was identified by using SDS-PAGE. Polyacrylamide gel electrophoresis was used to determine the protein profile and molecular weight of shrimp extract. Coloring of protein bands using silver staining. Data were analyzed descriptively based on the migration value of the sample protein bands compared to the marker protein band (Rf). The results of protein allergen profile analysis on shrimp using SDS-PAGE showed that the shrimp contained a protein band with a molecular weight of 37.77 kDa for cooked shrimp and 37.03 kDa for fresh shrimp.

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References

Bonner, P. 2018. Protein Purification. 2nd Edition. Boca Raton: CRC Press, Taylor & Francis.
Ellenbogen, Y., Jimenez-Saiz, R., Spill, P., Chu, D.K., Waserman, S., Jordana, M. 2018. The Initiation of Th2 Immunity Towards Food Allergens. International Journal of Molecular Sciences. 19(5):1447. https://dx.doi.org/10.3390/ijms19051447
Gomaa, A., Boye, J. 2015. Simultaneous detection of multi-allergens in an incurred food matrix using ELISA, multiplex flow cytometry and liquid chromatography mass spectrometry (LC-MS). Food Chemistry. 175:585-592. https://doi.org/10.1016/j.foodchem.2014.12.017
Hafiz, A. 2004. Principles and Reactions of Protein Extraction, Purification, and Characterization. Boca Raton: CRC Press, Taylor & Francis. https://doi.org/10.1201/9780203507438
Lopata, A.L., Kleine-Tebbe, J., Kamath, S.D. 2016. Allergens and molecular diagnostics of shellfish allergy: Part 22 of the Series Molecular Allergology. Allergo Journal International. 25(7):210-218. https://dx.doi.org/10.1007/s40629-016-0124-2
Musmand, J.J., Dul, C.B., Lehrer, S.B. 1993. Crustacea Allergy. Clinical & Experimental Allergy. 23(9):722-732. https://doi.org/10.1111/j.1365-2222.1993.tb00359.x
Nowakowski, A.B., Wobig, W.J., Petering, D.H. 2014. Native SDS-PAGE: High Resolution Electrophoretic Separation of Proteins with Retention of Native Properties Including Bound Metal Ions. Metallomics. 6(5):1068-1078. https://dx.doi.org/10.1039/c4mt00033a
Oh-Ishi, M., Maeda, T. 2002. Separation techniques for high-molecular-mass proteins. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 771(1-2):49-66. https://doi.org/10.1016/S1570-0232(02)00112-5
Rabilloud, T., Vaezzadeh, A.R., Potier, N., Lelong, C., Leize-Wagner, E., Chevallet, M. 2009. Power and limitations of electrophoretic separations in proteomics strategies. Mass Spectrometry Reviews. 28(5):816-843. https://doi.org/10.1002/mas.20204
Rath, A., Cunningham, F., Deber, C.M. 2013. Acrylamide concentration determines the direction and magnitude of helical membrane protein gel shifts. Proceedings of the National Academy of Sciences of the United States of America. 110(39):15668-15673. https://dx.doi.org/10.1073/pnas.1311305110
Rosenberg, I.M. 1996. Protein Analysis and Purification. Boston: Birkhäuser, Springer. https://doi.org/10.1007/978-1-4612-2056-5
Rudge, S.R., Monnig, C.A. 2000. Electrophoresis Techniques. Separation and Purification Methods. 29(1):129-148. https://doi.org/10.1081/SPM-100100006
Tan, S.C., Yiap, B.C. 2009. DNA, RNA, and Protein Extraction: The Past and The Present. Journal of Biomedicine and Biotechnology. 2009:574398. https://dx.doi.org/10.1155/2009/574398
Thalayasingam, M., Gerez, I.F., Yap, G.C., Llanora, G.V., Chia, I.P., Chua, L., Lee, C.J., Ta, L.D., Cheng, Y.K., Thong, B.Y., Tang, C.Y., Yan Bever, H.P., Shek, L.P., Curotto de Lafaille, M.A., Lee, B.W. 2015. Clinical and immunochemical profiles of food challenge proven or anaphylactic shrimp allergy in tropical Singapore. Clinical & Experimental Allergy. 45(3):687-697. https://doi.org/10.1111/cea.12416
Tong, W.S., Yuen, A.W.T., Wai, C.Y.Y., Leung, N.Y.H., Chu, K.H., Leung, P.S.C. 2018. Diagnosis of fish and shellfish allergies. Journal of Asthma and Allergy. 11:247-260. https://dx.doi.org/10.2147/JAA.S142476

Authors

Emma Emawati
emma.emawati@stfb.ac.id (Primary Contact)
Idar Idar
Resta Ramadiyanti
1.
Emawati E, Idar I, Ramadiyanti R. Characterization and Identification of Allergen Protein in Shrimp Before and After Heated with SDS-PAGE Method. Borneo J Pharm [Internet]. 2019Nov.30 [cited 2024Dec.21];2(2):87-93. Available from: https://journal.umpr.ac.id/index.php/bjop/article/view/901

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