Recombinant Human Granulocyte-Macrophage Colony Stimulating Factor GMP

Synonyms Granulocyte/Macrophage Colony-Stimulating Factor, CSF-2, MGI-1GM, Pluripoietin-α

Accession P04141

GeneID 1437

Source Escherichia coli.

Molecular Weight Approximately 14.5 kDa, a single non-glycosylated polypeptide chain containing 127 amino acids.

Quantity 5µg/100µg/1mg

AA Sequence APARSPSPST QPWEHVNAIQ EARRLLNLSR DTAAEMNETV EVISEMFDLQ EPTCLQTRLE LYKQGLRGSL TKLKGPLTMM ASHYKQHCPP TPETSCATQI ITFESFKENL KDFLLVIPFD CWEPVQE

Purity > 98 % by SDS-PAGE and HPLC analyses.

Biological Activity Fully biologically active when compared to standard. The ED50 as determined by a cell proliferation assay using human TF-1 cells is less than 0.1 ng/ml, corresponding to a specific activity of > 1.0 × 107 IU/mg.

Physical Appearance Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation Lyophilized from a 0.2 μm filtered concentrated solution in PBS, pH 7.4.

Endotoxin Less than 0.01 EU/μg of rHuGM-CSF GMP as determined by LAL method.

Reconstitution We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1 % BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions.

Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

- 12 months from date of receipt, -20 to -70 °C as supplied.

- 1 month, 2 to 8 °C under sterile conditions after reconstitution.

- 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Usage This GMP product can be for research use or further manufacturing use.

Quality statement The manufacture and testing of this product is in compliance with ICH Q7a guidelines.

Reference 1. Wang JM, Chen ZG, Colotta F, et al. 1988. Behring Inst Mitt: 270-3.

2. 1989. N Engl J Med, 320: 253-4.

3. Nissen-Druey C. 1989. Nouv Rev Fr Hematol, 31: 99-101.

4. Eager RandNemunaitis J. 2005. Mol Ther, 12: 18-27.

5. Tran T, Fernandes DJ, Schuliga M, et al. 2005. Br J Pharmacol, 145: 123-31.

Background Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) is secreted by a number of different cell types (including activated T cells, B cells, macrophages, mast cells, endothelial cells and fibroblasts) in response to cytokine or immune and inflammatory stimulation. It was initially characterized as a growth factor that can support the in vitro colony formation of granulocyte-macrophage progenitors and has functions of stimulates the growth and differentiation of hematopoietic precursor cells from various lineages. GM-CSF has also been reported to have a functional role on non-hematopoietic cells and can induce human endothelial cells to migrate and proliferate. Additionally, it can stimulate the proliferation of a number of tumor cell lines, including osteogenic sarcoma, carcinoma and adenocarcinoma cell lines. Human GM-CSF shares 54 % sequences identity with mouse GM-CSF, but has no biological effects across species. GM-CSF is used as a medication to stimulate the production of white blood cells following chemotherapy and has also recently been evaluated in clinical trials for its potential as a vaccine adjuvant in HIV-infected patients.