title: A novel clot lysis assay for recombinant plasminogen activator.
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.
authors.affiliation: Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, P.O. Box: 14115-114, Tehran, Iran, oveis.jamialahmadi@gmail.com.
authors.forename: Oveis
authors.initials: O
authors.lastname: Jamialahmadi
authors.forename: Ahmad
authors.initials: A
authors.lastname: Fazeli
authors.forename: Sameereh
authors.initials: S
authors.lastname: Hashemi-Najafabadi
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Fazeli
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005342:Fibrinolysis; D005343:Fibrinolytic Agents / Q000032:analysis*; D010960:Plasminogen Activators / Q000032:analysis* / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D010960:Plasminogen Activators