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Large-scale preparation of the Δ10 form of staphylokinase by in vitro processing of recombinant staphylokinase with purified human plasminogen

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Abstract

The authors have developed a rapid and convenient method for purification of a low molecular weight form (Δ10) of the bacterial plasminogen activator, staphylokinase. Recombinant staphylokinase is expressed inEscherichia coli, with an amino terminal extension that facilitated purification by immobilized metal-affinity chromatography. Purified staphylokinase is treated with human plasminogen, and the resulting truncated form is purified using a combination of immobilized metal affinity chromatography and hydrophobic interaction chromatography. Purified protein is characterized by amino terminal sequencing and in vitro plasminogen activation assay.

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Authors and Affiliations

  1. Center for Macromolecular Crystallography, University of Alabama at Birmingham, 276 Basic Health Science Building, 1918 University Boulevard, 35294-0005, Birmingham, AL

    Debasish Chattopadhyay, Jerry E. Stewart & Lawrence J. DeLucas

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  1. Debasish Chattopadhyay
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  2. Jerry E. Stewart
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  3. Lawrence J. DeLucas
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Chattopadhyay, D., Stewart, J.E. & DeLucas, L.J. Large-scale preparation of the Δ10 form of staphylokinase by in vitro processing of recombinant staphylokinase with purified human plasminogen. Appl Biochem Biotechnol 69, 147–156 (1998). https://doi.org/10.1007/BF02788810

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  • DOI: https://doi.org/10.1007/BF02788810

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