Karakterisasi Imunogenesitas Protein Pilli Salmonella Typhi: Analisis Absorban Antibodi dan Spesifisitas dengan Western Blot pada Mencit Balb/C
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Abstract
Protein pili berperan dalam adhesi dan bersifat imunogenik, berpotensi sebagai kandidat vaksin subunit. Penelitian ini menganalisis imunogenesitas protein pili S. typhi isolat BA07.4 melalui pengukuran absorbansi antibodi dan karakterisasi spesifisitasnya. Protein pili diisolasi dengan metode biphasic, dimurnikan, dan digunakan untuk imunisasi 30 ekor mencit Balb/C (6 kelompok: kontrol dan dosis 2, 3, 4, 5, 6 µg) secara intraperitoneal tiga kali. Kadar antibodi diukur dengan inhouse ELISA (λ 405 nm) dan spesifisitasnya dianalisis dengan Western blot. Rerata absorbansi tertinggi pada dosis 5 µg (2,827), diikuti dosis 6 µg (2,678), 4 µg (2,456), 3 µg (2,134), 2 µg (1,856), dan kontrol (0,423). Analisis One Way ANOVA menunjukkan perbedaan signifikan antar kelompok (p=0,000) dengan semua kelompok perlakuan berbeda signifikan terhadap kontrol (p<0,05). Western blot menunjukkan pita spesifik pada berat molekul 87, 58, dan 42 kDa pada kelompok dosis 5 µg, sementara kontrol tidak menunjukkan pita. Protein pili S. typhi bersifat imunogenik dengan dosis optimum 5 µg, menghasilkan antibodi yang secara spesifik mengenali sub unit protein pili 87, 58, dan 42 kDa, mendukung potensinya sebagai kandidat vaksin subunit.
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