UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA
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 thr...
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id-itb.:622072021-12-20T13:44:21ZUJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA Merika, Angelia Indonesia Final Project - INSTITUT TEKNOLOGI BANDUNG https://digilib.itb.ac.id/gdl/view/62207 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. text |
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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.
|
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author |
Merika, Angelia |
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Merika, Angelia UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA |
author_facet |
Merika, Angelia |
author_sort |
Merika, Angelia |
title |
UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA |
title_short |
UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA |
title_full |
UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA |
title_fullStr |
UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA |
title_full_unstemmed |
UJI TOKSISITAS AKUT DAN SUBKRONIK REKOMBINAN INTERFERON ALPHA 2B DAN MUTEIN C2D C99D PADA MECIT SWISS-WEBSTER BETINA |
title_sort |
uji toksisitas akut dan subkronik rekombinan interferon alpha 2b dan mutein c2d c99d pada mecit swiss-webster betina |
url |
https://digilib.itb.ac.id/gdl/view/62207 |
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