Pustaka Ilmiah Universitas Padjadjaran

Abstrak

Faktor Suhu Dan Lioprotektan Sukrosa Terhadap Stabilisasi Fibronektin Dalam Sediaan Serbuk Serum Otologus

Serum otologus mengandung faktor pertumbuhan seperti EGF, TGF-?, vitamin A, dan fibronektin, namun sediaan dalam bentuk cairan mudah rusak dan hanya stabil selama 1 bulan pada penyimpanan suhu 40C. Serum otologus dibuat dalam bentuk sediaan kering melalui proses liofilisasi. Dalam tahapan liofilisasi protein mengalami stress denaturation, karena itu dilakukan penambahan lioprotektan sukrosa sebagai stabilisator protein. Tujuan penelitian ini adalah untuk mengetahui kestabilan kadar dan karakter fibronektin pada serum otologus serbuk beku-kering yang telah ditambahkan sukrosa dan disimpan pada suhu 40C. Penelitian ini dilakukan studi komparatif dan uji non klinis pada darah 3 orang sampel, dilakukan pengambilan darah 100 mL, dibekukan, sentrifugasi pada 5000 g (10 menit), dan diperoleh serum sebanyak ~20 mL. Sampel serum diencerkan dan sebagian sampel serum ditambahkan sukrosa 60 mM. Pengukuran kestabilan kadar fibronektin menggunakan metode ELISA dan karakter fibronektin menggunakan HPLC. Hasil penelitian menunjukan bahwa kadar fibronektin yang disimpan pada suhu 40C lebih stabil dibandingkan dengan yang disimpan pada suhu ruang selama lima bulan dengan penurunan 80%, dengan kadar fibronektin masih ? 21 ?g/mL (kadar Fibronektin dalam serum yang masih dapat digunakan), dan karakter fibronektin sebelum dan sesudah liofilisasi tidak terjadi perbedaan dimana pola penurunan luas area mengikuti penurunan persen kadar fibronektin.

Autologous serum contains growth factors such as EGF, TGF-?, and fibronectin, however serum in liquid form can be easly damaged and stable just 1 month at 40C storage. The autologous serum is made in dry forms through lyophilization process. In lyophilization stage, protein through stress denaturation, therefore an addition of sucrose was used as a protein stabilizer or lyoprotectant. The purpose of this research is to determine the stability of fibronectin concentration and character in solid proteins of serum with lyoprotectant sucrose and stored in 40C. This research was conducted comparative studies and clinical trials on three blood samples from humans, 100 mL for each samples, and than freeze the sample, sample was centrifuged at 5000 g (10 min), to obtained ~20 mL of serum. Serum was diluted, and partially of sampel serum was added with 60 mM sucrose. The measurement of fibronectin concentration stability using ELISA method, and the character of fibronectin using HPLC. The results showed that the fibronectin concentration in 40C more stable than in room temperature for five months, with decrease until 80%, with the concentration of fibronectin still ? 21 ?g/mL (fibronectin in the serum that are still usable), and the character of fibronectin before and after lyophilization is no differences in the pattern of decline in the area following the percent decrease in fibronectin concentration.