High yield secretion of human erythropoietin from tobacco cells for ex vivo differentiation of hematopoietic stem cells towards red blood cells
Document Type
Article
Publication Title
Journal of biotechnology
PubMed ID
35777457
MeSH Headings (Medical Subject Headings)
Cell Differentiation; Erythroid Precursor Cells (metabolism); Erythropoiesis (physiology); Erythropoietin (genetics, metabolism); Hematopoietic Stem Cells (metabolism); Humans; Nicotiana (genetics)
Abstract
Human erythropoietin (EPO) is a key cytokine in erythropoiesis by regulating differentiation of erythroid progenitor cells into red blood cells (RBCs). Plant cell cultures are considered as promising alternative bioproduction platforms for EPO. To overcome the bottlenecks of low protein productivity and secretion, EPO was expressed in tobacco BY-2 cells with a designer peptide tag, termed (SP) that consists of 20 tandem repeats of a "Ser-Pro" motif. This de novo designed tag directed extensive O-glycosylation on each Pro residue in plant cells and acted as a molecular carrier to promote the extracellular secretion of EPO. To facilitate the establishment of stable and high-expression BY-2 cell lines, EPO molecules were co-expressed with a reporter protein GFP, which could be used as a visual marker to monitor the protein expression during the subculture. The engineered (SP) glycomodule substantially increased the secreted yields of EPO up to 4.31 μg/mL. The (SP)-tagged EPOs exhibited the expected activity in promoting the proliferation of TF-1 cells, though their EC50 was 12-fold higher than that of EPO standard. The (SP)-tagged EPOs could also stimulate the ex vivo expansion and differentiation of hematopoietic stem cell (CD34 cells) towards RBCs.
First Page
10
Last Page
20
DOI
10.1016/j.jbiotec.2022.06.010
Publication Date
8-20-2022
Recommended Citation
Karki, Uddhab; Wright, Tristen; and Xu, Jianfeng, "High yield secretion of human erythropoietin from tobacco cells for ex vivo differentiation of hematopoietic stem cells towards red blood cells" (2022). Arkansas Biosciences Institute. 28.
https://arch.astate.edu/abi/28