G CSF Human, HEK
Description
Manufacturing Process
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Expression System: HEK293 cells enable high-yield, protein-free production with mammalian glycosylation patterns .
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Purification: Proprietary chromatographic techniques achieve >95% purity .
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Activity: Potent myelopoietic activity, with an ED50 <0.01 ng/mL in murine M-NFS-60 cell proliferation assays .
| Parameter | Specification |
|---|---|
| Molecular Weight | 21–25 kDa (glycosylated) |
| Purity | >95% (SDS-PAGE) |
| Endotoxin Level | <1 EU/µg |
| Biological Activity | ED50 <0.01 ng/mL (proliferation assays) |
Biological Functions and Mechanisms
G-CSF activates the JAK/STAT3 pathway upon binding to GCSFR, driving neutrophil maturation and release . Key roles include:
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Neutrophil Regulation: Promotes survival, proliferation, and chemotaxis of neutrophils .
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Immunomodulation: Attenuates pro-inflammatory cytokines (e.g., IL-1β, TNF-α) and enhances anti-inflammatory mediators (e.g., IL-1ra) .
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Hematopoietic Stem Cell (HSC) Mobilization: Critical for peripheral blood stem cell (PBSC) collection in clinical transplantation .
In Vitro and Preclinical Studies
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G-CSF Detection: HEK-Blue™ G-CSF cells (engineered HEK293) enable colorimetric detection of bioactive G-CSF via SEAP reporter assays .
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Drug Development: Used to screen anti-G-CSF antibodies and small-molecule inhibitors targeting the G-CSF pathway .
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Fusion Proteins: G-CSF-transferrin (G-CSF-Tf) fusion proteins produced in HEK293 cells show oral bioavailability in mice, enabling GI absorption studies .
Clinical Findings
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Immune Modulation: Transient suppression of T-cell proliferation post-G-CSF administration, reversible within 14 days .
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GVHD Outcomes: Murine studies show reduced graft-versus-host disease (GVHD) severity with donor G-CSF pretreatment, though human trials report mixed results .
Comparative Analysis with Other Production Systems
| Feature | HEK293-Produced G-CSF | CHO-Produced G-CSF |
|---|---|---|
| Glycosylation Pattern | Human-like, O-linked sialic acid | Non-human sialylation patterns |
| Yield | High (protein-free medium) | Moderate |
| Cost | Higher | Lower |
Therapeutic Implications
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Neutropenia Management: Reduces infection risk in chemotherapy patients .
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Stem Cell Mobilization: Enhances PBSC yield for transplantation .
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Oral Drug Delivery: G-CSF-Tf fusion proteins demonstrate potential for non-invasive administration .
Key Research Challenges
Product Specs
Product Science Overview
Granulocyte-Colony Stimulating Factor (G-CSF) is a glycoprotein that plays a crucial role in hematopoiesis, the process of forming blood cellular components. It specifically stimulates the bone marrow to produce granulocytes and stem cells and release them into the bloodstream . The human recombinant form of G-CSF, expressed in Human Embryonic Kidney (HEK) cells, is a significant advancement in biotechnology and medicine.
Human recombinant G-CSF is produced using HEK 293 cells, a specific cell line derived from human embryonic kidney cells. These cells are widely used in biotechnology for their ability to produce high yields of recombinant proteins. The recombinant G-CSF expressed in HEK cells is biologically active and structurally similar to the naturally occurring protein .
G-CSF is a proliferation, differentiation, survival, and activation factor for hematopoietic cells of the neutrophilic granulocyte lineage . It is produced by various cells, including macrophages, monocytes, fibroblasts, and endothelial cells, in response to specific stimuli such as endotoxins and cytokines . The primary function of G-CSF is to stimulate the bone marrow to produce and release neutrophils, a type of white blood cell essential for fighting infections .
G-CSF binds to its specific receptor on the surface of target cells, primarily neutrophils and their precursors . This binding activates intracellular signaling pathways that promote the proliferation, differentiation, and survival of these cells. Additionally, G-CSF enhances the functional activity of mature neutrophils, including their ability to combat infections .
Recombinant human G-CSF is used clinically to treat neutropenia, a condition characterized by low levels of neutrophils . It is commonly administered to patients undergoing chemotherapy, which can reduce white blood cell counts and increase the risk of infections . G-CSF helps to restore neutrophil levels, reducing the risk of infections and allowing patients to continue their chemotherapy treatments on schedule . It is also used in stem cell transplantation to mobilize stem cells from the bone marrow into the bloodstream for collection and subsequent transplantation .
