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For activity measurements, samples were thawed at room temperature and diluted with 3% bovine serum albumin in tris buffered saline to achieve a tPA concentration of 0.36, 0.73, 1.45, 2.9 or 3.6 (IU/ml). Solution samples were then stored at -80 C as 0.5–2 mL aliquots for 0.5 to 7.2 years in cryogenic vials.
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The tPA was obtained as a lyophilized powder and mixed with sterile water to a concentration of 1 mg/mL (580,000 IU/ml), as per the manufacturer's instructions. In this work, the enzymatic activity of tPA stored at -80 C over time was measured. Therefore, methods to store reconstituted tPA for long durations while maintaining activity would be of assistance to laboratories using this medication. According to the manufacturer, tPA can be used for 8 hours after reconstitution in sterile water at room temperature this is a short period of time for laboratory use. Therefore the preservation of tPA for laboratory use in such studies is of substantial interest to laboratories engaged in such research.Ĭurrently, tPA is available as a lyophilized powder from the manufacturer (Genentech, San Francisco, CA). Such optimization requires substantial in- vitro and in- vivo work. As a result, there is much interest in optimizing lytic therapy using tPA in the treatment of such disease. Recombinant tissue plasminogen activator (tPA) is widely to treat thrombotic disorders such as acute ischemic stroke, deep venous thrombosis. Such cryogenic storage is a viable method for the preservation of tPA solution for laboratory investigations of tPA-based lytic therapies. There was no decrease in tPA activity for samples stored up to 7 years. Results are reported as means with standard errors, and 12 measurements were performed for each sample age. For a given sample, aliquots were assayed for tPA activity, and compared with a tPA standard to determine relative enzymatic activity. Sample of tPA solution mixed to a concentration of 1 (mg/ml) were stored in cryogenic vials at -80 C for up to 7 years. In this work, the enzymatic activity of tPA stored at -80 C over time was measured, using an ELISA technique that measured the amount of active tPA bound to plasminogen activator inhibitor 1 (PAI-1) in a given sample. Therefore, methods to store reconstituted tPA for long durations while maintaining activity would be of assistance to laboratories using this enzyme.
#Tissue plasminogen activator (tpa) full
However, tPA reconstituted in solution exhibits full activity for only 6–8 hours, according to the manufacturer. This has led to much interest in tPA based lytic therapies leading to laboratory based in-vitro and in-vivo investigations using this drug. Recombinant tissue plasminogen activator (tPA) is a thrombolytic widely used clinically in the treatment of acute thrombotic disease such as ischemic stroke, myocardial infarction, and deep venous thrombosis.