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A Novel Clot Lysis Assay for Recombinant Plasminogen Activator Publisher Pubmed



Jamialahmadi O1 ; Fazeli A2 ; Hasheminajafabadi S1 ; Fazeli MR3
Authors

Source: Biotechnology Letters Published:2015


Abstract

Recombinant plasminogen activator (r-PA, reteplase) is an engineered variant of alteplase. When expressed in E. coli, it appears as inclusion bodies that require refolding to recover its biological activity. An important step following refolding is to determine the activity of refolded protein. Current methods for enzymatic activity of thrombolytic drugs are costly and complex. Here a straightforward and low-cost clot lysis assay was developed. It quantitatively measures the activity of the commercial reteplase and is also capable of screening refolding conditions. As evidence for adequate accuracy and sensitivity of the current assay, r-PA activity measurements are shown to be comparable to those obtained from chromogenic substrate assay. © 2014, Springer Science+Business Media Dordrecht.