Fatimah Dewi, Akmia (2021) Karakterisasi Antibodi Poliklonal Hasil Induksi Protein Rekombinan CD2vASFV pada Kelinci (Oryctolagus cuniculus) sebagai Kandidat Deteksi ASF. Sarjana thesis, Universitas Brawijaya.
Abstract
Penyebaran African Swine Fever Virus (ASFV) di Indonesia terus berlanjut sampai saat ini, ASF merupakan penyakit infeksius yang menyerang babi dengan mortalitas yang tinggi. Deteksi ASF secara cepat dan efektif sangat dibutuhkan untuk perdagangan internasional maupun lokal agar dapat mencegah penyebaran virus. Tujuan dari penelitian ini adalah untuk mengetahui imunogenesitas protein rekombinan CD2v dengan berat molekul 45 kDa yang dikloning dan diekspresikan pada media kultur sel bakteri E.coli BL21 bersifat imunogenik deteksi dini penyakit ASF. Sampel protein rekombinan CD2v 45 kDa yang digunakan berasal dari PUSVETMA Surabaya. Kemudian dilakukan pemurnian protein target melalui metode elektroelusi gel hasil teknik elektroforesis SDS-PAGE berdasarkan berat molekul 45 kDa. Imunisasi awal dan booster pada kelinci jantan New Zealand (Oryctolagus cuniculus) menggunakan hasil elusi protein rekombinan CD2v diberikan secara subkutan dengan dosis 800µL dengan penambahan adjuvant CFA (Complete Freund’s Adjuvant) dan IFA (Incomplate Freund’s Adjuvant). Uji imunogenisitas protein CD2v menggunakan metode indirect ELISA dan Western blotting. Hasil uji indirect ELISA menunjukkan nilai titer antibodi tertinggi pada pengambian darah terakhir dengan nilai 0,534. Pada uji Western blotting menunjukkan bahwa antibodi antiCD2v dapat mengenali antigen CD2v pada berat molekul 45 kDa.
English Abstract
The spread of African Swine Fever Virus (ASFV) in Indonesia continues to increase until this day. ASF is an infectious disease that attacks pigs with high mortality rate. Fast and effective detection of ASF is needed to prevent the spread of the virus in international and local market trade. The aim of this study was to determine the immunogenicity of the recombinant CD2v protein with a molecular weight of 45 kDa which was cloned and expressed in bacterial cell culture media of E. coli BL21 which is immunogenic for early detection of ASF disease. The recombinant protein sample of CD2v 45 kDa used was obtained from PUSVETMA Surabaya. The target protein then purified using the gel electroelution method of the SDS-PAGE electrophoresis technique based on a molecular weight of 45 kDa. Initial and booster immunization in New Zealand male rabbits (Oryctolagus cuniculus) using the elution of recombinant CD2v protein was administered subcutaneously at a dose of 800µL with the addition of CFA (Complete Freund's Adjuvant) and IFA (Incomplate Freund's Adjuvant) adjuvants. Indirect ELISA and Western blotting methods was used to test the CD2v protein immunogenicity. The results of the indirect ELISA test showed the highest antibody titer value is in the last bleeding with a value of 0.534. The Western blotting test showed that the anti-CD2v antibody could recognize the CD2v antigen at a molecular weight of 45 kDa.
Other obstract
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| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 636.089 |
| Uncontrolled Keywords: | ASF, CD2v, IgG, antibody. |
| Subjects: | 600 Technology (Applied sciences) > 636 Animal husbandry > 636.08 Specific topics in animal husbandry > 636.089 Veterinary medicine |
| Divisions: | Fakultas Kedokteran Hewan > Kedokteran Hewan |
| Depositing User: | Unnamed user with email gaby |
| Date Deposited: | 25 Oct 2021 08:30 |
| Last Modified: | 27 Sep 2024 10:22 |
| URI: | http://repository.ub.ac.id/id/eprint/185989 |
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