Soluble expression and partial purification of recombinant human erythropoietin from E. coli

Taeck Hyun Jeong, Young Jin Son, Han Bong Ryu, Bon Kyung Koo, Seung Mi Jeong, Phuong Hoang, Bich Hang Do, Jung A. Song, Seon Ha Chong, Robert Charles Robinson, Han Choe

Research output: Contribution to journalArticle

17 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)211-218
Number of pages8
JournalProtein Expression and Purification
Volume95
DOIs
Publication statusPublished - Mar 1 2014
Externally publishedYes

Fingerprint

Erythropoietin
Escherichia coli
Cytoplasm
Erythrocytes
Maltose-Binding Proteins
Erythroid Precursor Cells
CHO Cells
Kidney Neoplasms
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Inclusion Bodies
Therapeutics
Chronic Renal Insufficiency
Disulfides
Solubility
Gel Chromatography
Electrophoresis
Anemia
Proteins
Bone Marrow
Economics

Keywords

  • ErhEpo
  • Escherichia coli expression system
  • Maltose binding protein (MBP)
  • Therapeutic protein

ASJC Scopus subject areas

  • Biotechnology

Cite this

Jeong, T. H., Son, Y. J., Ryu, H. B., Koo, B. K., Jeong, S. M., Hoang, P., ... Choe, H. (2014). Soluble expression and partial purification of recombinant human erythropoietin from E. coli. Protein Expression and Purification, 95, 211-218. https://doi.org/10.1016/j.pep.2014.01.001

Soluble expression and partial purification of recombinant human erythropoietin from E. coli. / 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.

In: Protein Expression and Purification, Vol. 95, 01.03.2014, p. 211-218.

Research output: Contribution to journalArticle

Jeong, TH, Son, YJ, Ryu, HB, Koo, BK, Jeong, SM, Hoang, P, Do, BH, Song, JA, Chong, SH, Robinson, RC & Choe, H 2014, 'Soluble expression and partial purification of recombinant human erythropoietin from E. coli', Protein Expression and Purification, vol. 95, pp. 211-218. https://doi.org/10.1016/j.pep.2014.01.001
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. / Soluble expression and partial purification of recombinant human erythropoietin from E. coli. In: Protein Expression and Purification. 2014 ; Vol. 95. pp. 211-218.
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