Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein

The enhancement of recombinant therapeutic protein production in mammalian cell culture has been regarded as an important issue in the biopharmaceutical industry. Previous studies have reported that the addition of the recombinant 30Kc19 protein, a silkworm-derived plasma protein with simultaneous c...

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Autores principales: Hyeonjin Cha, Ju-Hyun Park
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Publicado: MDPI AG 2021
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spelling oai:doaj.org-article:24b7c0b6878c402892c2f35cdbf8f5032021-11-25T16:42:53ZRecombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein10.3390/app1122110092076-3417https://doaj.org/article/24b7c0b6878c402892c2f35cdbf8f5032021-11-01T00:00:00Zhttps://www.mdpi.com/2076-3417/11/22/11009https://doaj.org/toc/2076-3417The enhancement of recombinant therapeutic protein production in mammalian cell culture has been regarded as an important issue in the biopharmaceutical industry. Previous studies have reported that the addition of the recombinant 30Kc19 protein, a silkworm-derived plasma protein with simultaneous cell-penetrating and mitochondrial enzyme-stabilizing properties, can enhance the recombinant protein expression in Chinese hamster ovary (CHO) cell culture. Here, we produced an α-helix N-terminal domain of 30Kc19, called (30Kc19α), and investigated its effects on the production of human erythropoietin (EPO), a widely used therapeutic protein for the treatment of anemia, in recombinant CHO cell culture. Similar to the full-length 30Kc19, 30Kc19α was able to be mass-produced in a form of recombinant protein through an <i>Escherichia coli</i> expression system and delivered into EPO-producing CHO (EPO–CHO) cells. Supplementing the medium of EPO–CHO cell culture with 30Kc19α increased the intracellular NADPH/NADP<sup>+</sup> ratio related to the flux of metabolic reducing power for protein biosynthesis, subsequently enhancing EPO production in serum-free culture. 30Kc19α is considered to have certain advantages in the downstream purification process of therapeutic protein production when it is used as a medium supplement due to its small size and low isoelectric point compared to the full-length 30Kc19. These results suggest that 30Kc19α has potential use for manufacturing biopharmaceutical proteins.Hyeonjin ChaJu-Hyun ParkMDPI AGarticle30Kc19α proteinerythropoietin (EPO)Chinese hamster ovary (CHO) cellbiopharmaceuticalsproductivityTechnologyTEngineering (General). Civil engineering (General)TA1-2040Biology (General)QH301-705.5PhysicsQC1-999ChemistryQD1-999ENApplied Sciences, Vol 11, Iss 11009, p 11009 (2021)
institution DOAJ
collection DOAJ
language EN
topic 30Kc19α protein
erythropoietin (EPO)
Chinese hamster ovary (CHO) cell
biopharmaceuticals
productivity
Technology
T
Engineering (General). Civil engineering (General)
TA1-2040
Biology (General)
QH301-705.5
Physics
QC1-999
Chemistry
QD1-999
spellingShingle 30Kc19α protein
erythropoietin (EPO)
Chinese hamster ovary (CHO) cell
biopharmaceuticals
productivity
Technology
T
Engineering (General). Civil engineering (General)
TA1-2040
Biology (General)
QH301-705.5
Physics
QC1-999
Chemistry
QD1-999
Hyeonjin Cha
Ju-Hyun Park
Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein
description The enhancement of recombinant therapeutic protein production in mammalian cell culture has been regarded as an important issue in the biopharmaceutical industry. Previous studies have reported that the addition of the recombinant 30Kc19 protein, a silkworm-derived plasma protein with simultaneous cell-penetrating and mitochondrial enzyme-stabilizing properties, can enhance the recombinant protein expression in Chinese hamster ovary (CHO) cell culture. Here, we produced an α-helix N-terminal domain of 30Kc19, called (30Kc19α), and investigated its effects on the production of human erythropoietin (EPO), a widely used therapeutic protein for the treatment of anemia, in recombinant CHO cell culture. Similar to the full-length 30Kc19, 30Kc19α was able to be mass-produced in a form of recombinant protein through an <i>Escherichia coli</i> expression system and delivered into EPO-producing CHO (EPO–CHO) cells. Supplementing the medium of EPO–CHO cell culture with 30Kc19α increased the intracellular NADPH/NADP<sup>+</sup> ratio related to the flux of metabolic reducing power for protein biosynthesis, subsequently enhancing EPO production in serum-free culture. 30Kc19α is considered to have certain advantages in the downstream purification process of therapeutic protein production when it is used as a medium supplement due to its small size and low isoelectric point compared to the full-length 30Kc19. These results suggest that 30Kc19α has potential use for manufacturing biopharmaceutical proteins.
format article
author Hyeonjin Cha
Ju-Hyun Park
author_facet Hyeonjin Cha
Ju-Hyun Park
author_sort Hyeonjin Cha
title Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein
title_short Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein
title_full Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein
title_fullStr Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein
title_full_unstemmed Recombinant Human Erythropoietin Production in Chinese Hamster Ovary Cells Is Enhanced by Supplementation of α-Helix Domain of 30Kc19 Protein
title_sort recombinant human erythropoietin production in chinese hamster ovary cells is enhanced by supplementation of α-helix domain of 30kc19 protein
publisher MDPI AG
publishDate 2021
url https://doaj.org/article/24b7c0b6878c402892c2f35cdbf8f503
work_keys_str_mv AT hyeonjincha recombinanthumanerythropoietinproductioninchinesehamsterovarycellsisenhancedbysupplementationofahelixdomainof30kc19protein
AT juhyunpark recombinanthumanerythropoietinproductioninchinesehamsterovarycellsisenhancedbysupplementationofahelixdomainof30kc19protein
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