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Title: A human expression system based on HEK293 for the stable production of recombinant erythropoietin
Authors: Chin, Christine Lin
Goh, Justin Bryan
Srinivasan, Harini
Liu, Kaiwen Ivy
Gowher, Ali
Shanmugam, Raghuvaran
Lim, Hsueh Lee
Choo, Matthew
Tang, Wen Qin
Tan, Andy Hee-Meng
Nguyen-Khuong, Terry
Tan, Meng How
Ng, Say Kong
Keywords: Engineering::Chemical engineering
Issue Date: 2019
Source: Chin, C. L., Goh, J. B., Srinivasan, H., Liu, K. I., Gowher, A., Shanmugam, R., . . . Ng, S. K. (2019). A human expression system based on HEK293 for the stable production of recombinant erythropoietin. Scientific Reports, 9(1), 16768-. doi:10.1038/s41598-019-53391-z
Journal: Scientific Reports
Abstract: Mammalian host cell lines are the preferred expression systems for the manufacture of complex therapeutics and recombinant proteins. However, the most utilized mammalian host systems, namely Chinese hamster ovary (CHO), Sp2/0 and NS0 mouse myeloma cells, can produce glycoproteins with non-human glycans that may potentially illicit immunogenic responses. Hence, we developed a fully human expression system based on HEK293 cells for the stable and high titer production of recombinant proteins by first knocking out GLUL (encoding glutamine synthetase) using CRISPR-Cas9 system. Expression vectors using human GLUL as selection marker were then generated, with recombinant human erythropoietin (EPO) as our model protein. Selection was performed using methionine sulfoximine (MSX) to select for high EPO expression cells. EPO production of up to 92700 U/mL of EPO as analyzed by ELISA or 696 mg/L by densitometry was demonstrated in a 2 L stirred-tank fed batch bioreactor. Mass spectrometry analysis revealed that N-glycosylation of the produced EPO was similar to endogenous human proteins and non-human glycan epitopes were not detected. Collectively, our results highlight the use of a human cellular expression system for the high titer and xenogeneic-free production of EPO and possibly other complex recombinant proteins.
ISSN: 2045-2322
DOI: 10.1038/s41598-019-53391-z
Rights: © 2019 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Te images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit
Fulltext Permission: open
Fulltext Availability: With Fulltext
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