#TITLE_ALTERNATIVE#

• Main Author: DAMANHURI (NIM 10704126), FERRY
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/10437
• Institution: Institut Teknologi Bandung

Streptokinase is a therapeutic protein used as a potent thrombolytic agent. However, its use as a therapy is limited by short biological half-life and imunogenity. One of the effort to overcome these problems are pegylation. Pegylation is a conjugation process of a polyethylene glycol (PEG) into a molecule. The aims of this research are to produce pure mutant K59Q-K386Q streptokinase and to pegylate this mutant streptokinase to increase its fibrinolytic activity. Mutant streptokinase was overproduced in E. coli BL21. Protein was isolated and purified with nikel afinity coloumn chromatography. PEG was activated with 1,1 carbonyl diimidazol and identified with Dragendorff test, ultraviolet (UV) and infra red (IR) spectrophotometry and melting point test. Various factors of pegylation were incubation time and molarity ratio of protein and activated PEG. Fibrinolytic activity of pegylated mutant streptokinase (PEG-streptokinase) was tested in vitro for blood clot lysis. Mutant K59Q-K386Q streptokinase was successfully overproduced, purified and pegylated. Optimum condition for pegylation process is incubation for 48 hours with ratio streptokinase:activated PEG 1:400. In vitro result of fibrinolytic activity assay showed that PEG-streptokinase has better fibrinolytic activity than non-pegylated form. It can be concluded that mutant K59Q-K386Q streptokinase was successfully pegylated with increased in vitro fibrinolytic activity as compared to non-pegylated form.