A screen for peptide agonists of the G-CSF receptor

Conzelmann, Nadine; Schneider, Armin

doi:10.1186/1756-0500-4-194

Cited by 4 publications

(3 citation statements)

References 31 publications

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“…The EC 50 values for hGCSF purified from MBP-hGCSF (2.83 pM) and PDIb'a'-hGCSF (3.38 pM) were consistent with a previous study that reported an EC 50 value in the range of 0.8–6 pM for hGCSF [25], [54], [55]. At high concentrations, the purified hGCSF proteins induced mild inhibition of cell proliferation, resulting in a bell-shaped biphasic dose-response curve (Figure 5E).…”

Section: Discussionsupporting

confidence: 91%

Soluble Prokaryotic Overexpression and Purification of Bioactive Human Granulocyte Colony-Stimulating Factor by Maltose Binding Protein and Protein Disulfide Isomerase

et al. 2014

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Human granulocyte colony-stimulating factor (hGCSF), a neutrophil-promoting cytokine, is an effective therapeutic agent for neutropenia patients who have undergone several cancer treatments. Efficient production of hGCSF using E. coli is challenging because the hormone tends to aggregate and forms inclusion bodies. This study examined the ability of seven different N-terminal fusion tags to increase expression of soluble hGCSF in E. coli. Four tag proteins, namely maltose-binding protein (MBP), N-utilization substance protein A, protein disulfide isomerase (PDI), and the b'a' domain of PDI (PDIb'a'), increased the solubility of hGCSF under normal conditions. Lowering the expression temperature from 30°C to 18°C also increased the solubility of thioredoxin-tagged and glutathione S-transferase-tagged hGCSF. By contrast, hexahistidine-tagged hGCSF was insoluble at both temperatures. Simple conventional chromatographic methods were used to purify hGCSF from the overexpressed PDIb'a'-hGCSF and MBP-hGCSF proteins. In total, 11.3 mg or 10.2 mg of pure hGCSF were obtained from 500 mL cultures of E. coli expressing PDIb'a'-hGCSF or MBP-hGCSF, respectively. SDS-PAGE analysis and silver staining confirmed high purity of the isolated hGCSF proteins, and the endotoxin levels were less than 0.05 EU/µg of protein. Subsequently, the bioactivity of the purified hGCSF proteins similar to that of the commercially available hGCSF was confirmed using the mouse M-NFS-60 myelogenous leukemia cell line. The EC50s of the cell proliferation dose-response curves for hGCSF proteins purified from MBP-hGCSF and PDIb'a'-hGCSF were 2.83±0.31 pM, and 3.38±0.41 pM, respectively. In summary, this study describes an efficient method for the soluble overexpression and purification of bioactive hGCSF in E. coli.

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Section: Discussionsupporting

confidence: 91%

Soluble Prokaryotic Overexpression and Purification of Bioactive Human Granulocyte Colony-Stimulating Factor by Maltose Binding Protein and Protein Disulfide Isomerase

et al. 2014

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“…201 So far, the search for peptide agonists of the G-CSF receptor has been unsuccessful. 202 Biomimetic Self-Assembling Peptides. Self-assembling peptides are short (∼5−50 aa) polypeptides that can be rationally designed to undergo hierarchical assembly into nanostructures.…”

Section: ■ Mechanism Of Angiogenesismentioning

confidence: 99%

“…Clinical trials of G-CSF have been associated with an inconsistent pattern of vascularization . So far, the search for peptide agonists of the G-CSF receptor has been unsuccessful …”

Section: Angiogenic Factors and Functional Mimicsmentioning

confidence: 99%

Angiogenic Self-Assembling Peptide Scaffolds for Functional Tissue Regeneration

et al. 2018

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Implantation of acellular biomimetic scaffolds with proangiogenic motifs may have exciting clinical utility for the treatment of ischemic pathologies such as myocardial infarction. Although direct delivery of angiogenic proteins is a possible treatment option, smaller synthetic peptide-based nanostructured alternatives are being investigated due to favorable factors, such as sustained efficacy and high-density epitope presentation of functional moieties. These peptides may be implanted in vivo at the site of ischemia, bypassing the first-pass metabolism and enabling long-term retention and sustained efficacy. Mimics of angiogenic proteins show tremendous potential for clinical use. We discuss possible approaches to integrate the functionality of such angiogenic peptide mimics into self-assembled peptide scaffolds for application in functional tissue regeneration.

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