Abstract
Objective
Thrombopoietin mimetic peptide (TMP), an analog of natural thrombopoietin, can be used to treat primary immune thrombocytopenia. However, the short half-life of TMP limits its application in clinics. The present study aimed to improve the stability and biological activity of TMP in vivo via genetic fusion to the albumin-binding protein domain (ABD).
Results
TMP dimer was genetically fused to the N-terminal or C-terminal of ABD, denoted as TMP-TMP-ABD and ABD-TMP-TMP. A Trx-tag was used to improve the fusion proteins’ expression levels effectively. ABD-fusion TMP proteins were produced in Escherichia coli and purified by Ni2+-NTA and SP ion exchange column. Albumin binding studies in vitro showed that the fusion proteins could effectively bind to serum albumin to extend their half-lives. The fusion proteins effectively induced platelet proliferation in healthy mice, and the platelet count was increased by more than 2.3-fold compared with the control group. The increased platelet count induced by the fusion proteins lasted 12 days compared with the control group. The increasing trend was maintained for 6 days before a decline occurred after the last injection in the fusion-protein-treated mice group.
Conclusions
ABD can effectively improve the stability and pharmacological activity of TMP by binding to serum albumin, and the ABD-fusion TMP protein can promote platelet formation in vivo.
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Acknowledgements
We would like to thank Editage (www.editage.cn) for English language editing.
Funding
This work was supported by Key Research Foundation of Hwa Mei Hospital, University of Chinese Academy of Sciences, China (Grant No. 2019HMKY17); Key Medical Subjects of Joint Construction Between Provinces and Cites (Infectious Diseases), China (Grant No. 2016017); Key Laboratory of Diagnosis and Treatment of Digestive System Tumors of Zhejiang Province (Grant No. 2019E10020).
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Li, D., Gao, G., Zhu, B. et al. Improving pharmacological activities of thrombopoietin mimetic peptide by genetic fusion to albumin-binding domain. Biotechnol Lett 45, 439–448 (2023). https://doi.org/10.1007/s10529-023-03345-3
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DOI: https://doi.org/10.1007/s10529-023-03345-3