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Kevin Yueju Wang

Kevin Yueju Wang

Northeastern State University, USA

Title: Seed-derived plasminogen activators (t-PA and DSPAa1) dissolve fibrin and blood clot

Biography

Biography: Kevin Yueju Wang

Abstract

Stroke has remained the top major killer worldwide. It is also the leading cause of adult disability. Recombinant human tissue plasminogen (rt-PA) is the only FDA-approved treatment for acute ischemic stroke. The yields of rt-PA protein produced from CHO cells, however, are, at present, relatively low while the cost of production is high. It often causes side effects which may lead to disability and death. Vampire bat (Desmodus rotundus) salivary plasminogen activators (DSPA1) have been found to be both more active than t-PA and to have fewer side effects. In this study, both DSPA1 and t-PA were targeted in tobacco seed under seed-specific promoter (phas) to minimize protein degradation. Both t-PA and DSPA1 were codon-optimized for tobacco plant usage to increase foreign protein expression level. 6xHis tag was fused to C-terminal for recombinant protein purification. The ER signal peptide sequence, KDEL, will be used to accumulate recombinant proteins to the ER. The geminivirus-based single DNA replicon system (pBY was used to rapidly produce t-PA and DSPA1, respectively in tobacco leaf. Studies showed that recombinant proteins caused leaf tissue death. Seed-derived proteins can dissolve fibrin and human blood clots. Our study demonstrated that seed-based system can rapidly produce functional recombinant t-PA and DSPA1 for stroke patient treatment. This work was supported by the National Institute of General Medical Sciences of the National Institutes of Health (8P20GM103447) and National Institute of Neurological Disorders and Strokes (1R03NS095246-01).