Recombinant Human Pegylated Granulocyte-Colony Stimulating Factor - rHuPEG-G-CSF



Data Sheet
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Recombinant Human Pegylated Granulocyte-Colony Stimulating Factor - rHuPEG-G-CSF

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. rHuPEG-G-CSF 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 dlaton.. G-CSF is purified by proprietary chromatographic techniques.
Source: Escherichia Coli.
Physical Appearance: Sterile filtered liquid formulation 0.85 mg/ml
Formulation: The protein was extensive dialyzed against 10mM sodium acetate buffer PH= 4 and 5% mannitol was added.
Solubility: It is recommended to reconstitute the lyophilized rHuPEG-G-CSF in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability: rHuPEG-G-CSF although stable at 15°C for 1 week, should be stored between 2°C-8°C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Purity: Greater than 95.0% as determined by: (a) Analysis by RP-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.
Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity: ProSpec's rHuPEG-G-CSF is fully biologically active when compared to standard. The ED50, calculated by the dose-dependant proliferation of murine NFS-60 indicator cells (measured by 3H-thymidine uptake) isless then0.1 ng/ml, corresponding to a Specific Activity of 1 x 108 IU/mg.
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuPEG-G-CSF.
Protein content: Protein quantitation was carried out by two independent methods: UV spectroscopy at 280 nm using the absorbency value of 0.815 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). Analysis by RP-HPLC, using a calibrated solution of PEG-G-CSF as a Reference Standard.
Usage: This material is offered for research, laboratory or further manufacturing purposes.