Zinc supplementation protects human endostatin Fc fusion against proteolytic degradation during cell culture

Hao, Gang; Dawson, Jessica; Lee, Gene; Longley, Robert L.

doi:10.1016/j.pep.2013.10.005

Cited by 4 publications

(2 citation statements)

References 30 publications

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“…Another modification to endostatin is the addition of an iRGD (internalization RGD) sequence that allows endostatin to penetrate further into tumor tissue and increase its distribution volume in the tumor [257]. Fusing endostatin to the Fc region of IgG also increases its half-life from 2 hours to more than 2 weeks, and the addition of zinc can even further decrease the Fc-endostatin degradation [182, 258]. The use of nanoparticles for endostatin delivery allows for controlled drug release.…”

Section: Discussionmentioning

confidence: 99%

Endostatin's emerging roles in angiogenesis, lymphangiogenesis, disease, and clinical applications

Walia

Yang

Huang

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

154

126

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Background Angiogenesis is the process of neovascularization from pre-existing vasculature and is involved in various physiological and pathological processes. Inhibitors of angiogenesis, administered either as individual drugs or in combination with other chemotherapy, have been shown to benefit patients with various cancers. Endostatin, a 20-kDa C-terminal fragment of type XVIII collagen, is one of the most potent inhibitors of angiogenesis. Scope of review We discuss the biology behind endostatin in the context of its endogenous production, the various receptors to which it binds, and the mechanisms by which it acts. We focus on its inhibitory role in angiogenesis, lymphangiogenesis, and cancer metastasis. We also present emerging clinical applications for endostatin and its potential as a therapeutic agent in the form a short peptide. Major conclusions The delicate balance between pro- and anti-angiogenic factors can be modulated to result in physiological wound healing or pathological tumor metastasis. Research in the last decade has emphasized an emerging clinical potential for endostatin as a biomarker and as a therapeutic short peptide. Moreover, elevated or depressed endostatin levels in diseased states may help explain the pathophysiological mechanisms of the particular disease. General significance Endostatin was once sought after as the ‘be all and end all’ for cancer treatment; however, research throughout the last decade has made it apparent that endostatin’s effects are complex and involve multiple mechanisms. A better understanding of newly discovered mechanisms and clinical applications still has the potential to lead to future advances in the use of endostatin in the clinic.

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Section: Discussionmentioning

confidence: 99%

Endostatin's emerging roles in angiogenesis, lymphangiogenesis, disease, and clinical applications

Walia

Yang

Huang

et al. 2015

Biochimica et Biophysica Acta (BBA) - General Subjects

154

126

View full text Add to dashboard Cite

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“…Thus, a combination of factors such as the action of proteases and glycosidases released from dead cells and instability under the culture conditions could be responsible for the reduced molecular weight and decreased amount of mature BMP‐7 at the end of cultures. Although little is known about the proteases from CHO cells, a number of strategies have been employed to minimize the degradation of glycoproteins including optimization of process and culture conditions and supplementation of protease inhibitors (Arden et al, ; Hao et al, ; Yang and Butler, ; Yoon et al, ). Specific protease inhibitors may be assayed to cultured cells with the expectation of maximizing BMP‐7 production.…”

Section: Discussionmentioning

confidence: 99%

Characterization and expression of proprotein convertases in CHO cells: Efficient proteolytic maturation of human bone morphogenetic protein‐7

Sathyamurthy

Kim

Bang

et al. 2014

Biotech & Bioengineering

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Bone morphogenetic protein-7 (BMP-7) is synthesized as a precursor that requires proteolytic cleavage of the propeptide by proprotein convertases (PCs) for its functional activity. A high-level expression of BMP-7 in CHO cells (CHO-BMP-7) resulted in secretion of a mixture of inactive precursor and active BMP-7. In an effort to achieve efficient processing of BMP-7 in CHO cells, PCs responsible for cleavage of the precursors in CHO cells were characterized. Analysis of the mRNA expression levels of four PCs (furin, PACE4, PC5/6, and PC7) revealed that only furin and PC7 genes are expressed in CHO-BMP-7 cells. Specific inhibition of the PCs by hexa-D-arginine (D6R) or decanoyl-RVKR-chloromethyl ketone (RVKR-CMK) further revealed that furin is mainly responsible for the proteolytic processing of BMP-7. To identify a more efficient PC for BMP-7 processing, the four PC genes were transiently expressed in CHO-BMP-7 cells, respectively. Among these, PC5/6 was found to be the most efficient in BMP-7 processing. Stable overexpression of PC5/6ΔC, a secreted form of PC5/6, significantly improved mature BMP-7 production in CHO-BMP-7 cells. When the maximum BMP-7 concentration was obtained in the culture of CHO-BMP-7 cells, approximately 88% of BMP-7 was unprocessed. In contrast, no precursor was found in the culture of PC5/6ΔC-overexpressing cells (clone #97). Furthermore, the in vitro biological activity of the mature BMP-7 from PC5/6ΔC-overexpressing cells was comparable to that from CHO-BMP-7 cells. Taken together, the present results indicate that overexpression of PC5/6ΔC in CHO-BMP-7 cells is an efficient means of increasing the yield of BMP-7.

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Zinc-containing Mohs’ paste affects blood flow and angiogenesis suppression

et al. 2021

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Zinc supplementation protects human endostatin Fc fusion against proteolytic degradation during cell culture

Cited by 4 publications

References 30 publications

Endostatin's emerging roles in angiogenesis, lymphangiogenesis, disease, and clinical applications

Endostatin's emerging roles in angiogenesis, lymphangiogenesis, disease, and clinical applications

Characterization and expression of proprotein convertases in CHO cells: Efficient proteolytic maturation of human bone morphogenetic protein‐7

Zinc-containing Mohs’ paste affects blood flow and angiogenesis suppression

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