Pegylated Recombinant Human Granulocyte-Colony Stimulating Factor (PEG-GCSF)
(Cat. No.: C028)
Background:
A glycoprotein of MW 20 kDa containing internal disulfide bonds. It induces the survival, proliferation, and differentiation of neutrophilic granulocyte precursor cells and functionally activates mature blood neutrophils. Among the family of colony-stimulating factors, G-CSF is the most potent inducer of terminal differentiation to granulocytes and macrophages of leukemic myeloid cell lines.
The synthesis of G-CSF can be induced by bacterial endotoxins, TNF, IL-1 and GM-CSF. Prostaglandin E2 inhibits the synthesis of G-CSF. In epithelial, endothelial, and fibroblastic cells secretion of G-CSF is induced by IL-17.
Description:
Recombinant Human G-CSF produced in E. coli is a single, non-glycosylated, polypeptide chain containing 175 amino acids and having a molecular mass of 18,800 Dalton. PEG-GCSF is manufactured by attaching a 20,000 Dalton methoxypolyethylene glycol propionaldehyde (mPEG-ALD) to the N-terminal amino acid of G-CSF having a total molecular mass of 38,800 Dalton.
Quality Control:
Biological Activity: PEG-GCSF is fully biologically active when compared to standard. The ED50, calculated by the dose-dependant proliferation of murine NFS-60 indicator cells is less then 0.1 ng/ml, corresponding to a Specific Activity of 1.0 x 107 IU/mg.
Purity: Greater than 95% as determined by:
(a) Analysis by HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Thr-Pro-Leu-Gly.
Endotoxin: Less than 0.1 ng/µg (1IEU/µg) of PEG-GCSF.
Formulation: The protein was extensively dialyzed against 10mM sodium acetate buffer PH= 4 and 5% mannitol was added.
Storage: PEG-GCSF although stable at 15oC for 1 week, should be stored between 2oC -8oC. For long-term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.
Reconstitution: It is recommended to reconstitute the lyophilized PEG-GCSF in sterile 18M¦¸-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
