Improving bone morphogenetic protein (BMP) production in CHO cells through understanding of BMP synthesis, signaling and endocytosis

Kim, Kyung-Soo; Kim, Mi Gyeom; Lee, Gyun Min

doi:10.1016/j.biotechadv.2022.108080

Cited by 3 publications

(3 citation statements)

References 138 publications

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“…Molecular docking was performed using Discovery Studio 2017 software (Biovia). The lectins (PDB: 3VI4, 1L5G, 3C4M, 4GIQ) were docked to peptides with potential bone-promoting activity [28]. The co-crystal structures of the RGD peptide ligands and the lectins can be downloaded from the Protein Data Bank (PDB: 3VI4, 1L5G, 3C4M, 4GIQ).…”

Section: Molecular Docking Predicts the Analysis Of Osteoactive Pepti...mentioning

confidence: 99%

The Isolation, Purification and Active Mechanism of Sea Cucumber Bone-Strengthening Functional Peptides

Cheng,

Yang,

et al. 2024

Preprint

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Sea cucumber peptides are small molecular peptides with biological function obtained from fresh sea cucumber as feedstock, which were hydrolyzed by protease, separated ,and purified. As the main product of sea cucumber refinement, it more accurately expresses the nutritional effect of sea cucumber and has a wide range of potential applications. It has been demonstrated that sea cucumber has a certain value-added effect on bone mineral density regulation, but the differences in bone mineral density regulation of extracts from different parts of sea cucumber and the techniques contributing to the efficient isolation of bone sea cucumber peptides have not been reported. Firstly, the aim of this study was to reveal the differences in osteogenic activities of enzymatic peptides from three major parts of sea cucumber: the body wall, the egg protein, and the intestinal protein. Then, the bone-enhancing sea cucumber peptides were isolated based on the characteristics of the dif-ferences in molecular weight distribution, hydrophilicity, and ionisation degree of sea cucumber, and to explore the possible mechanisms of their action. The findings indicated that sea cucumber peptides extracted from three distinct sites exhibited a dose-dependent enhancement of osteoblast proliferation and differentiation. Upon evaluating the three methods of sea cucumber peptide separation based on molecular weight distribution, hydrophilicity, and ionization, it was observed that the fraction isolated by hydrophobicity and

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Section: Molecular Docking Predicts the Analysis Of Osteoactive Pepti...mentioning

confidence: 99%

The Isolation, Purification and Active Mechanism of Sea Cucumber Bone-Strengthening Functional Peptides

Cheng,

Yang,

et al. 2024

Preprint

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“…Common mammalian cell lines used for this purpose include HEK293 and Chinese hamster ovary (CHO) cells. [ 22 ] HEK293 cells are easy to grow in suspension culture with serum‐free medium and can be transfected using a variety of methods, making them highly efficient for protein production. [ 23,24 ] However, due to their high metabolic rate, HEK293 cells consume large amounts of glucose under normal culture conditions, leading to the production of growth‐inhibiting byproducts such as ammonia and lactate.…”

Section: Introductionmentioning

confidence: 99%

Recombinant human fibroblast growth factor 7 obtained from stable Chinese hamster ovary cells enhances wound healing

Kim,

Lee,

Jeong

et al. 2024

Biotechnology Journal

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Although fibroblast growth factor 7 (FGF7) is known to promote wound healing, its mass production poses several challenges and very few studies have assessed the feasibility of producing FGF7 in cell lines such as Chinese hamster ovary (CHO) cells. Therefore, this study sought to produce recombinant FGF7 in large quantities and evaluate its wound healing effect. To this end, the FGF7 gene was transfected into CHO cells and FGF7 production was optimized. The wound healing efficacy of N‐glycosylated FGF7 was evaluated in animals on days 7 and 14 post‐treatment using collagen patches (CPs), FGF7‐only, and CP with FGF7 (CP+FGF7), whereas an untreated group was used as the control. Wound healing was most effective in the CP+FGF7 group. Particularly, on day 7 post‐exposure, the CP+FGF7 and FGF7‐only groups exhibited the highest expression of hydroxyproline, fibroblast growth factor, vascular endothelial growth factor, and transforming growth factor. Epidermalization in H&E staining showed the same order of healing as hydroxyproline content. Additionally, the CP+FGF7 and FGF7‐only group exhibited more notable blood vessel formation on days 7 and 14. In conclusion, the prepared FGF7 was effective in promoting wound healing and CHO cells can be a reliable platform for the mass production of FGF7.

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“…Therefore, the properties of the TGF-β superfamily of cytokines are highly variable, and any general strategy to enhance their production in CHO cells remains unavailable. [16] The production of the TGF-β superfamily of cytokines, such as recombinant human bone morphogenetic protein (rhBMP)-2, rhBMP-4, rhBMP-7, and recombinant human growth differentiation factor (rhGDF)-5, has been considerably enhanced in CHO cells by understanding the properties of the individual cytokines with respect to the overall maturation process, signaling pathway and endocytosis. [11][12][13][14][15][16] However, to our knowledge, the properties of rhTGF-β1 in CHO cells have not yet been well characterized.…”

Section: Introductionmentioning

confidence: 99%

Enhancing the production of recombinant human TGF‐β1 through an understanding of TGF‐β1 synthesis, signaling, and endocytosis in CHO cells

Kim,

Jang

et al. 2023

Biotechnology Journal

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To enhance the production of recombinant human transforming growth factor‐beta1 (rhTGF‐β1) in Chinese hamster ovary (CHO) cells, rhTGF‐β1 was first characterized for endocytosis, signaling pathway, and overall maturation process. The mature rhTGF‐β1 used for clinical application was internalized into CHO cells and inhibited the growth of CHO cells in a dose‐dependent manner. However, mature rhTGF‐β1 was mostly produced in the form of latent rhTGF‐β1 in cultures of recombinant CHO (rCHO) cells producing rhTGF‐β1 (CHO‐rhTGF‐β1). The concentration of active mature rhTGF‐β1 in the culture supernatant of CHO‐rhTGF‐β1 cells was not high enough to compromise yield. In addition, a significant amount of unprocessed precursors was produced by CHO‐rhTGF‐β1 cells. Overexpression of PACEsol, a soluble form of furin, in CHO‐rhTGF‐β1 cells was effective for the proteolytic cleavage of unprocessed precursors. The highest mature rhTGF‐β1 concentration (6.4 μg mL−1) was obtained with the PACEsol‐expressing clone, which was approximately 45% higher than that of the parental clone (P < 0.01). Thus, a comprehensive understanding of the intrinsic properties of rhTGF‐β1 with respect to the overall maturation process, signaling pathway, and endocytosis is essential for effectively enhancing the production of mature rhTGF‐β1 in CHO cells.

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Improving bone morphogenetic protein (BMP) production in CHO cells through understanding of BMP synthesis, signaling and endocytosis

Cited by 3 publications

References 138 publications

The Isolation, Purification and Active Mechanism of Sea Cucumber Bone-Strengthening Functional Peptides

The Isolation, Purification and Active Mechanism of Sea Cucumber Bone-Strengthening Functional Peptides

Recombinant human fibroblast growth factor 7 obtained from stable Chinese hamster ovary cells enhances wound healing

Enhancing the production of recombinant human TGF‐β1 through an understanding of TGF‐β1 synthesis, signaling, and endocytosis in CHO cells

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