UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA

• Main Author: Merika, Angelia
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/62207
• Institution: Institut Teknologi Bandung

Interferons are a member of cytokines which play important role in immune system especially against virus infection. IFN-?2b belongs to IFN type I that used clinically to treat chronic hepatitis B and C infections. We successfully developed recombinant humanIFN-?2b as well as the mutein forms through site directed mutagenesis technique. The mutein forms were developed by substituting cysteins at position 2 and 99 with aspartic acid. While the in vitro activity of these IFN-?2bs was being performed, in this study we tested for the toxicity to determine LD50 of rIFN-?2b mutein as one requirement for new drug development. Recombinant IFN-?2b mutein and native were overproduced in Eschericia coli BL21 carrying recombinant mutan pET32b IFN?2b plasmid. Protein extract will be collected in supernatant and purified with Nickel column and then characterized using SDS-PAGE. Toxicity study was performed in female mice both for acute and sub chronic. In acute toxicity test, both rIFN-?2bs were subcutaneously administered to the animals at a single dose of 97.5 µg/kg, 975 µg/kg and 9.75 mg/kg and the observation was performed for 14 days. In the sub chronic toxicity test, both rIFN-?2bs were subcutaneously administered to the animals every 3 days with a single dose of 1.95 µg/kg and 19.5 µg/kg for 6 weeks. Standard parameters for toxicity test were observed. Characterization of rIFN-?2b native dan mutein C2D C99D using SDS-PAGE indicates that their molecular weight is arround 37 kDa. At both toxicity studies, we did not observe any death as well as abnormality in body weight, behaviour, main organs, and the value of plasma SGPT and SGOT. Both rIFN-?2b native and mutein C2D C99D are not toxic at the dose up to 9.75 mg/kg (LD50 > 9.75 mg/kg). In addition, administration of these IFN- ?2bs in repeated dose of 1.95 µg/kg bb and 19.5 µg/kg bb every 3 days for 6 weeks do not show toxic effect on the mice. These results suggest that both native and mutein rIFN-?2bs we developed are safe at studied dose. rIFN-?2b native and mutein C2D C99D do not induce the organs damage such as heart, liver and kidney at repetion dose of 1.95 and 19.5 µg/kg bw every 3 days for 6 weeks.