Ramadhani, AnggiaNoor (2017) Produksi Antibodi Poliklonal Human Lp-PLA2 Dengan Induksi Protein Lp-PLA2 Wild Type Dan V279F Mutan. Sarjana thesis, Universitas Brawijaya.
Abstract
Penyakit jantung koroner merupakan salah satu penyebab kematian tertinggi di Indonesia. Penyakit jantung terjadi, karena adanya disfungsi endotel pada arteri serta inflamasi yang berlebih. Lipoprotein-associated phospholipase A2 (Lp-PLA2) atau dikenal juga sebagai platelet-activating factor acetylhydrolase (PAF-AH) merupakan enzim phospholipase A2 yang dikode oleh gen PLA2G7. Protein Lp-PLA2 berperan dalam inflamasi pada tubuh sehingga protein ini dapat digunakan sebagai penanda awal terjadinya penyakit jantung koroner. Tujuan penelitian ini adalah untuk mengetahui respon produksi antibodi human Lp-PLA2 dari mencit (Mus musculus) terhadap antigen protein Lp-PLA2 wild type dan mutan. Serum darah pasien penyakit jantung koroner dari RSSA (Rumah Sakit Saiful Anwar) diisolasi protein Lp-PLA2 dengan imunopresipitasi. Kemudian dilakukan SDS-PAGE untuk konfirmasi protein yang berhasil diisolasi. Terdapat 5 perlakuan injeksi antigen yaitu injeksi protein Lp-PLA2 mutan, wild-type, lipoprotein (Lee Biosolution (No. Katalog 400-42 W128511), adjuvant (Lee Biosolution (400-42 W128511) , dan kontrol negatif. Lp-PLA2 mutan dan wild type didapatkan dari hasil imunopresipitasi serum pasien, dan adjuvan yang digunakan adalah CFA dan IFA. Setelah 5 minggu injeksi dengan booster CFA pada minggu 1 dan IFA pada mingu berikutnya, mencit dilakukan dislokasi dan darah dari jantung mencit diambil. Screening antibodi yang memiliki high-sensitivity tertinggi dilakukan dengan menggunakan Indirect ELISA. Kelompok mencit yang diinjeksi dengan protein Lp-PLA2 mutan memiliki sifat imunogen dan life span lebih tinggi dibandingkan dengan kelompok injeksi wild type.
English Abstract
Coronary artery disease (CAD) has emerged as a leading cause of death in Indonesia nowadays. CAD occured because of endothelial dysfunction in the arteries and over inflammation. Lipoprotein-associated phospholipase A2 (Lp-PLA2) also known as platelet-activating factor acetylhydrolase (PAF-AH) is a phospholipase A2 enzyme,encoded by the PLA2G7 gene. This protein can be used as a biomarker for CAD in early phase, cause this protein plays a key role in inflamatory of phospholipids. Phospholipase driven inflammation is associated with endothelial dysfunction and plaque formation. Thus the purpose of this research is to know the response of human Lp-PLA2 antibody production from mouse (Mus musculus) against wildvtype and mutant Lp-PLA2 protein antigen. Protein Lp-PLA2 from blood serum’s heart disease positive patients at Saiful Anwar State Hospital (RSSA) is isolated by immunoprecipitation, then followed by SDS-PAGE. After that, immunoprecipitated protein administered to mice (Mus musculus) by intraperitoneal injection. There are 5types of treatment: (1) mutant protein; (2) wild-type protein; (3) lipoprotein; (4) adjuvants; and (5) control treatment. Mutant and wild-type Lp-PLA2 protein were obtained from patient serum immunoprecipitation results, and the adjuvants used were CFA and IFA. After 5 weeks of injection with a CFA booster at frist week and IFA on the next week, the mice performed dislocations and blood from the heart of the mice are collected. Screening of antibodies that have the highest sensitivity is Indirect ELISA is used to find antibodies that have the highest sensitivity. The group of mice injected with the mutant Lp-PLA2 protein had higher concentrations than the other groups. This is presumably because Lp-PLA2 has higher antigenicity than other proteins.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/MIPA/2017/222/051704399 |
| Subjects: | 500 Natural sciences and mathematics > 570 Biology |
| Divisions: | Fakultas Matematika dan Ilmu Pengetahuan Alam > Biologi |
| Depositing User: | Kustati |
| Date Deposited: | 14 Jun 2017 13:02 |
| Last Modified: | 14 Jun 2017 13:02 |
| URI: | http://repository.ub.ac.id/id/eprint/155264 |
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