Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/81778
Title: Soluble expression and partial purification of recombinant human erythropoietin from E. coli
Authors: Jeong, Taeck-Hyun
Son, Young-Jin
Ryu, Han-Bong
Koo, Bon-Kyung
Jeong, Seung-Mi
Hoang, Phuong
Do, Bich Hang
Song, Jung-A
Chong, Seon-Ha
Robinson, Robert Charles
Choe, Han
Keywords: rhEpo
Escherichia coli expression system
Issue Date: 2014
Source: Jeong, T.-H., Son, Y.-J., Ryu, H.-B., Koo, B.-K., Jeong, S.-M., Hoang, P., et al. (2014). Soluble expression and partial purification of recombinant human erythropoietin from E. coli. Protein Expression and Purification, 95, 211-218.
Series/Report no.: Protein Expression and Purification
Abstract: Human erythropoietin (hEpo) is an essential regulator of erythrocyte production that induces the division and differentiation of erythroid progenitor cells in the bone marrow into mature erythrocytes. It is widely used for the treatment of anemia resulting from chronic kidney disease, chemotherapy, and cancer-related therapies. Active hEpo, and hEpo analogs, have been purified primarily from mammalian cells, which has several disadvantages, including low yields and high production costs. Although an Escherichia coli (E. coli) expression system may provide economic production of therapeutic proteins, it has not been used for the production of recombinant hEpo (rhEpo) because it aggregates in inclusion bodies in the E. coli cytoplasm and is not modified post-translationally. We investigated the soluble overexpression of active rhEpo with various protein tags in E. coli, and found out that several tags increased the solubility of rhEpo. Among them maltose binding protein (MBP)-tagged rhEpo was purified using affinity and gel filtration columns. Non-denaturing electrophoresis and MALDI-TOF MS analysis demonstrated that the purified rhEpo had two intra-disulfide bonds identical to those of the native hEpo. An in vitro proliferation assay showed that rhEpo purified from E. coli had similar biological activity as rhEpo derived from CHO cells. Therefore, we report for the first time that active rhEpo was overexpressed as a soluble form in the cytoplasm of E. coli and purified it in simple purification steps. We hope that our results offer opportunities for progress in rhEpo therapeutics.
URI: https://hdl.handle.net/10356/81778
http://hdl.handle.net/10220/40978
ISSN: 1046-5928
DOI: http://dx.doi.org/10.1016/j.pep.2014.01.001
Rights: © 2014 Elsevier.
Fulltext Permission: none
Fulltext Availability: No Fulltext
Appears in Collections:SBS Journal Articles

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