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

Lusiana Lusiana, Fransiska Zakaria Rungkat, Endang Prangdimurti, Hendra Wijaya

Abstract


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.

Keywords


Alergen, Monopterus Javanensis La Cepéde, 1800), SDS-PAGE, ELISA, Immunoblotting

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References


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DOI: http://dx.doi.org/10.32765/wartaihp.v38i2.7151

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