Recombinant Human Granulocyte Colony Stimulating Factor
Certificate of Analysis and Data Sheet
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 18802.65 Dalton. rHuG-CSF is purified by proprietary chromatographic techniques.
Source: Escherichia Coli.
Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation & Packaging: The protein was lyophilized after extensive dialysis against 10mM sodium acetate buffer pH = 4.
Solubility: The lyophilized rHuG-CSF is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=5.59).
Stability: Lyophilized rHuG-CSF, although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuG-CSF is best stored refrigerated at 4°C.
Purity: Greater than 99.0% as determined by:
(a) Analysis by RP-HPLC
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing silver-stained SDS-PAGE
(Limit of acceptance:³98.0%. No more than 2% total impurities; no single impurity greater than 1%)
Amino Acid Composition:
In total agreement with the expected amino acid
composition of native
human G-CSF.
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, conforming to the sequence of native human G-CSF. N-terminal methionine has been completely removed enzymatically.
Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel.
Biological Activity: Our rHuG-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) is < 0.1 ng/ml, corresponding to a Specific Activity of 1 x 108 IU/mg.
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuG-CSF.
Protein content: Protein quantitation was carried out by two independent methods:
1.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).
2. Analysis by RP-HPLC, using a calibrated solution of G-CSF as a Reference Standard.
Usage: This material is offered for research, laboratory or further manufacturing purposes.
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