Extraction of Squid (Photololigo Duvaucelii) Myofibrillar and Sarcoplasmic Proteins to Create A Skin Prick Test Reagent In The Diagnosis of Food Allergy

Wijaya, Hendra; Rungkat-Zakaria, Fransiska; Syah, Dahrul; Prangdimurti, Endang

doi:10.9790/3013-040906014

Cited by 3 publications

(13 citation statements)

References 14 publications

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“…Pereaksi yang digunakan untuk uji tusuk kulit adalah ekstrak protein pangan yang selama ini masih diimpor dari luar negeri sehingga harganya relatif mahal (Wijaya et al 2014). Pada penelitian ini memanfaatkan bahan pangan asli Indonesia salah satunya adalah belut sawah yang jumlahnya cukup melimpah untuk dijadikan ekstrak alergen.…”

Section: Pendahuluanunclassified

“…Standar yang digunakan adalah low molecular weight protein (LMW) Fermentasi® Gel hasil running dipindahkan ke dalam wadah tertutup yang telah berisi pewarna pewarna coomasie brilliant blue G-250. (Wijaya et al 2014, Astuti et al 2016 Sebanyak 10 penderita alergi pangan diambil serumnya oleh tenaga medis dari klinik Alergi dan Asma Indrajana Jakarta (Kode etik penelitian No.168/IT3.KEPMSM-IPB/SK/2019). Subjek penelitian ini berasal dari desa Cigombong dan desa Cijeruk Kab.…”

Section: Ekstraksi Protein Belut Sawah (Ma Et Al 2017)unclassified

“…ELISA (Wijaya et al 2014 2.3.5. Immunoblotting (Wijaya et al 2014) Gel hasil elektroforesis yang tidak diwarnai ditransfer ke membran nitroselulosa (0,45 µm) yang disusun dalam alat transblotting, lalu diisi dengan buffer transfer. Blotting dilakukan pada 90 V selama 90 menit.…”

Section: Analisis Ige Spesifik Serum Dengan Metodeunclassified

“…Larutan yang diperoleh diuji sterilitas dan mikrobiologis sebelum digunakan untuk uji tusuk kulit. (Maleki et al 2010, Jeong et al 2013, Wijaya et al 2014, Wijaya et al 2015 Uji Tusuk Kulit dilakukan oleh dokter alergolog pada Klinik Alergi dan Asma DR. Indrajana, Jakarta. Subyek penelitian untuk uji tusuk kulit ini adalah 10 orang penderita alergi yang dari hasil wawancara alergi terhadap makanan laut dan kacang-kacangan.…”

Section: Persiapan Ekstrak Uji Tusuk Kulitunclassified

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Ekstraksi dan Karakterisasi Alergenisitas Protein Belut Sawah (Monopterus javanensis La Cepéde, 1800) untuk Pembuatan Reagen Uji Tusuk Kulit

Lusiana¹,

Rungkat

Prangdimurti

et al. 2021

J.Wart.Industri.Has.Pertan.

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ABSTRAK: Penelitian ini bertujuan untuk mengisolasi protein alergen belut segar (BS) dan belut rebus (BR) dengan buffer Tris Glisin (TG), Tris Glisin DTT (TGD) dan Phosphat buffer saline (PBS) serta mengkarakterisasi alergenisitas isolat protein dengan uji elektroforesis SDS-PAGE, ELISA dan immunoblotting. Rendemen protein yang dihasilkan pada BSTG, BSTGD dan BSPBS sebesar 2166 mg/L, 2083 mg/L dan 2284 mg/L. Rendemen ekstrak protein BRTG, BRTGD, BRPBS adalah 1445 mg/L, 1805 mg/L dan 1207 mg/L. Hasil SDS-Page menunjukkan profil protein BSTG memiliki 16 pita protein dengan bobot molekul dari 22 kDa-233 kDa dan BSTGD sebanyak 6 pita dengan bobot molekul 22 kDa-112 kDa, BSPBS sebanyak 16 pita dengan bobot molekul 22 kDa-93 kDa, BRTG memiliki 7 pita dengan berat molekul 24 kDa-267 kDa, BRTGD terdeteksi sebanyak 10 pita dengan berat molekul 24 kDa-235 kDa, BRPBS memiliki 10 pita protein dengan berat molekul 24 kDa-297 kDa. Hasil immunobloting dari 10 subjek yang positif alergi menunjukkan ada 3 subjek yang memberikan reaksi positif yaitu subjek 2 pada BSPBS, subjek 4 pada BSTG, BSTGD dan BSPBS serta subjek 8 pada BSTGD, BSPBS, BRTG dan BRTGD. Buffer PBS dapat meningkatkan rendemen ekstrak protein belut dan meningkatkan alergenitas reagen sehingga dapat digunakan sebagai buffer untuk pembuatan reagen uji tusuk kulit.

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Section: Pendahuluanunclassified

Section: Ekstraksi Protein Belut Sawah (Ma Et Al 2017)unclassified

Section: Analisis Ige Spesifik Serum Dengan Metodeunclassified

Section: Persiapan Ekstrak Uji Tusuk Kulitunclassified

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Ekstraksi dan Karakterisasi Alergenisitas Protein Belut Sawah (Monopterus javanensis La Cepéde, 1800) untuk Pembuatan Reagen Uji Tusuk Kulit

Lusiana¹,

Rungkat

Prangdimurti

et al. 2021

J.Wart.Industri.Has.Pertan.

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“…However, specific proteins will trigger their specific IgE antibodies, which can be used to determine the specific allergen-inducing protein (Zakaria et al, 1992b). IgE recognition of its protein allergens is the basis of most allergy analyses and determinations that can identify a single protein allergen in various foods (Wijaya et al, 2014), and it is the principle of the skin prick test (SPT) for allergy diagnosis (Astuti et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Isolation of winged bean (Psophocarpus tetragonolobus L.) allergen protein to create skin prick test reagent

Lusiana¹,

Zakaria²,

Prangdimurti³

et al. 2023

J App Pharm Sc

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Any type of food that contains protein can cause allergies. The winged bean (Psophocarpus tetragonolobus L.) is a type of legume plant that contains high protein, and it is quite abundant in West Java, Indonesia. There are no specific studies discussing allergies to winged bean plants. This study aimed to isolate the protein of the winged bean (P. tetragonolobus L.) and characterize the allergenicity of the protein isolate using ELISA and immunoblotting and test the reactivity by the skin prick testing method. The fresh winged bean (FWB) protein profile had 13 protein bands (22-89 kDa). The boiled winged bean (BWB) contained 10 bands (22-73 kDa). The results of immunoblotting of 10 allergy-positive respondents whose serum was taken showed that the FWB protein can bind to specific IgE antibodies in respondent 1 in the protein band with a molecular weight (MW) of 24 kDa, respondent 5 with a MW of 219 kDa, respondent 7 with MWs of 29 and 23 kDa, and respondent 9 with a MW of 17 kDa. Meanwhile, the BWB protein can bind to specific IgE antibodies in respondents 7 and 8. In the BWB sample, the allergen protein was bound to specific IgE only in respondent 7 with MWs of 31 and 24 kDa and respondent 8 with MWs of 31 and 23 kDa.

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Comparative Seed Proteome Profile Reveals No Alternation of Major Allergens in High-Yielding Mung Bean Cultivars

Shaheen,

Hossen,

Akhter

et al. 2024

J. Agric. Food Chem.

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Mung bean contains up to 32.6% protein and is one of the great sources of plant-based protein. Because many allergens also function as defense-related proteins, it is important to determine their abundance levels in the high-yielding, diseaseresistant cultivars. In this study, for the first time, we compared the seed proteome of high-yielding mung bean cultivars developed by a conventional breeding approach. Using a label-free quantitative proteomic platform, we successfully identified and quantified a total of 1373 proteins. Comparative analysis between the high-yielding disease-resistant cultivar (MC5) and the other three cultivars showed that a total of 69 common proteins were significantly altered in their abundances across all cultivars. Bioinformatic analysis of these altered proteins demonstrated that PDF1 (a defensin-like protein) exhibited high sequence similarity and epitope matching with the established peanut allergens, indicating a potential mung bean allergen that showed a cultivar-specific response. Conversely, known mung bean allergen proteins such as PR-2/PR-10 (Vig r 1), Vig r 2, Vig r 4, LTP1, β-conglycinin, and glycinin G4 showed no alternation in the MC5 compared to other cultivars. Taken together, our findings suggest that the known allergen profiles may not be impacted by the conventional plant breeding method to develop improved mung bean cultivars.

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Extraction of Squid (Photololigo Duvaucelii) Myofibrillar and Sarcoplasmic Proteins to Create A Skin Prick Test Reagent In The Diagnosis of Food Allergy

Cited by 3 publications

References 14 publications

Ekstraksi dan Karakterisasi Alergenisitas Protein Belut Sawah (Monopterus javanensis La Cepéde, 1800) untuk Pembuatan Reagen Uji Tusuk Kulit

Ekstraksi dan Karakterisasi Alergenisitas Protein Belut Sawah (Monopterus javanensis La Cepéde, 1800) untuk Pembuatan Reagen Uji Tusuk Kulit

Isolation of winged bean (Psophocarpus tetragonolobus L.) allergen protein to create skin prick test reagent

Comparative Seed Proteome Profile Reveals No Alternation of Major Allergens in High-Yielding Mung Bean Cultivars

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