Recombinant Human Platelet-derived Growth Factor BB

(rHuPDGF-BB)

Certificate of Analysis and Data Sheet

 

×                  Description:

Recombinant Human PDGF-BB produced in E.Coli is a double, non-glycosylated,

polypeptide chain containing 109 amino acids and having a molecular mass of 24525.18 Dalton.                  

          rHuPDGF-BB 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 2mM sodium phosphate buffer pH= 7.4±0.1.

 

×                  Solubility:

The lyophilized rHuPDGF-BB is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=11.85).

 

×                  Stability:

Lyophilized rHuPDGF-BB although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuPDGF-BB is best stored refrigerated at 4°C.

 

×                  Purity:

Greater than 98.0% as determined by:

 (a) Analysis by RP-HPLC.

           (b) Anion-exchange FPLC.

         (c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained.

(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 PDGF-BB.

 

×                  Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Ser-Leu-Gly-Ser-Leu, conforming the sequence of native human PDGF-BB.

 

×                  Dimers and aggregates:

Less than 1% as determined by silver stained SDS-PAGE gel analysis.

 

×                  Biological Activity:

Our rHuPDGF-BB is fully biologically active when compared to standard.

The ED50,  calculated by the dose-dependant proliferation of murine BALB/c 3T3 indicator cells (measured by ³H-thymidine uptake) is < 0.1 ng/ml, corresponding to a Specific Activity of 1 MIU/mg.

 

×                  Endotoxin:

Less than 0.1 ng/µg (IEU/µg) of rHuPDGF-BB.

 

×                  Protein content:

          Protein quantitation was carried out by two independent methods:

 

UV spectroscopy at 280 nm.

 

2.  Analysis by RP-HPLC, using a standard solution of PDGF-BB as a Reference Standard.

 

×                  Usage:

This material is offered for research, laboratory or further manufacturing purposes.