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A novel clot lysis assay for recombinant plasminogen activator

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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.

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Acknowledgments

We are grateful to Zistdaru Danesh Company for their generous gift of r-PA inclusion bodies.

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Authors and Affiliations

  1. Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, P.O. Box: 14115-114, Tehran, Iran

    Oveis Jamialahmadi & Sameereh Hashemi-Najafabadi

  2. Department of Research & Development, Zistdaru Danesh Company, Tehran, Iran

    Ahmad Fazeli

  3. Department of Drug & Food Control and Pharmaceutical Quality Assurance Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

    Mohammad Reza Fazeli

Authors
  1. Oveis Jamialahmadi
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  2. Ahmad Fazeli
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  3. Sameereh Hashemi-Najafabadi
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  4. Mohammad Reza Fazeli
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Corresponding author

Correspondence to Sameereh Hashemi-Najafabadi.

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Jamialahmadi, O., Fazeli, A., Hashemi-Najafabadi, S. et al. A novel clot lysis assay for recombinant plasminogen activator. Biotechnol Lett 37, 593–600 (2015). https://doi.org/10.1007/s10529-014-1711-2

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  • DOI: https://doi.org/10.1007/s10529-014-1711-2

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