Polycistronic Expression of Human Platelet Factor 4 with Heparin-Neutralizing Activity in <i>Escherichia coli</i>

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  • DUAN Yitao
    Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University
  • WANG Zhe
    Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University
  • WU Wei
    Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University
  • FANG Zhenjiang
    Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University
  • HUANG He
    Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University Key Laboratory of System Bioengineering, Ministry of Education, Tianjin University

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  • Polycistronic Expression of Human Platelet Factor 4 with Heparin-Neutralizing Activity in Escherichia coli

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Abstract

Human platelet factor 4 (hPF4) was evaluated as a clinical alternative to protamine for heparin neutralization, a protector against radiation injury and an anti-neoplastic. To achieve high-level expression of hPF4, expression vectors pET-28a(+)-nf PF4 (n=4, 5, 6) containing n tandem repeats of PF4 were constructed and transformed into the Escherichia coli BL21(DE3) strain. A higher expression level, about 45% of the total proteins (TP), was obtained for E. coli BL21(DE3)/pET28a(+)-nf PF4 (n=4, 5, 6). The purified His-PF4 protein was further identified by cleavage with enterokinase and MS, and its heparin-neutralizing activity was determined by colony formation assay. This study represents a novel approach to large-scale production of PF4 in E. coli, one that might be applied to large-scale production of PF4 protein for possible clinical application. It also provides theoretical points for the expression and purification of other small-molecule peptides.

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