Recombinant Human Bone Morphogenetic Protein-2

Description

　　Human bone morphogenetic protein-2 (BMP-2) is involved in the development of many organs and tissues as well as in the estab-lishment of the basic embryonic body plan. This implies its potential use in the regeneration of many types of tissues following injuries and dis-eases. It has the unique capacity of inducing bone regeneration and ectopic bone formation in adult vertebrates. In fact, clinical trials performed recently have shown that, when delivered properly, recombinant BMP-2 is effective and safe alterna-tives to autologous bone grafting. Further therapeutic applications, including its application for osteointegration of metallic implants, tendon and ligaments reunion with bones, cartilage repair and even in non-skeletal diseases, are foreseen.

　　Biologically active BMP-2 is a homodimer, linked by a single disulfide bond. The monomers have three intrachain disulfide bridges, forming a so-called cystine knot. In vivo, the monomer is synthesized as a large precursor, which after dimerization, is prote-olytically cleaved releasing the smaller C-terminal fragment forming the mature dimer

　　Our product produced in genetically engineered Eschericha coli is a non-active aggregated form. After refolding and purification, the recombinant human bone morphogenetic protein-2 is active and with high homogeneity.

Source

　　Escherichia Coli.

Physical Appearance

　　Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation

　　Lyophilized from a concentrated (1mg/ml) solution in water containing 20mM Sodium Acetic buffer, pH4.0.

Solubility

　　It is recommended to reconstitute the lyophilized BMP-2 in distilled H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability

　　Lyophilized BMP-2 although stable at room temperature for 3 weeks, should be stored desiccated below -18℃. Upon reconstitution BMP-2 should be stored at 4℃ between 2-7 days and for future use below -18℃ . For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
　　Please avoid freeze-thaw cycles.

Quality Standard

DETECTION ITEM	ANALYSIS METHOD	SPECIFICATION
PROTEIN CONTENT	Lowry Method	1.0mg/vial±10%
PURITY	NON-REDUCED SDS-PAGE	≥95%
	RP-HPLC	≥95%
MOLECULAR WEIGHT	REDUCED SDS-PAGE	12kD±10%
	MALDI-Tof MS	12110Da
UV ABSORBANCE	UV SCAN	278±3nm
ZYMOGRAM	Trypsin Digest, RP-HPLC	Same as Control
N-TERMINAL SEQUENCE	Edman Degradation	MQAKHKQRKRLKSS
IMMUNOLOGIC SPECIFICITY	Western-blotting	Positive
ISOELECTRIC POINT	Isoelectric focusing(IEF)	7.0-8.0
DAN RESIDUE	Dot blotting	≤10ng/3mg
HOST CELL PROTEIN RESIDUE	ELISA	≤0.02%
BIOLOGICAL ACTIVITY	Mice Muscle Implantation	≥1000BU/mg

Amino-Acid Sequence

　　The sequence of the first five N-terminal amino acids of monomer determined and was found to be Met-Gln-Ala-Lys-His.

Monomers

　　Less than 1% as determined by non-reduced SDS-PAGE gel analysis.

Biological Activity

　　Recombinant human BMP-2 is fully biologically active when compared to standards. When implanted in the mouse thighbone muscle and grow for 14 days, Muscle tissue was harvested and examined biochemically for calcium uptake. Our unit defined as 1 microgram calcium absorbed. The specific activity is more than 1000 Units/mg.