Ex-vivo Clot Lysis Evaluation of Fibrinolytic Enzyme 'Actinokinase'

Cardiovascular diseases, including cerebral stroke and myocardial infarction, are increasing in prevalence worldwide. Current fibrinolytic therapies, such as urokinase, can cause side effects like hemolysis, hemorrhage, and immunogenicity, prompting the search for novel thrombolytic agents. Actinokinase, a fibrinolytic enzyme isolated from thermophilic Streptomyces sp. MCMB 379, presents a promising alternative. This study investigates the enzyme's specificity and its potential as a thrombolytic agent. The enzyme was purified using DEAE ion exchange chromatography, and its purity was assessed via SDS-PAGE analysis. Ex-vivo fibrin clot lysis was conducted at 37°C, comparing Actinokinase's efficacy to that of urokinase at equivalent protein concentrations. The lysate was evaluated for fibrin degradation products (FDP) and D-dimer using immunochemical methods. Actinokinase demonstrated fibrin specificity and was non-hemolytic and non-allergenic. In vitro clot lysis was performed using fibrinogen and thrombin purified from human blood, with varying incubation times and urokinase concentrations. The fibrinolytic activity, assessed by the release of D-dimer and FDP, was comparable between Actinokinase and urokinase, indicating Actinokinase's potential as an effective fibrinolytic enzyme.