Recombinant Human Interleukin-15

( rHuIL-15)

Certificate of Analysis and Data Sheet

Description:

Recombinant Human IL-15 produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 114 amino acids and having a molecular mass of 13007 Dalton. rHuIL-15 is purified by proprietary chromatographic techniques

Source: Escherichia Coli.

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

Formulation & Packaging: The protein was lyophilized from a concentrated (1mg/ml) solution with 5mM Tris pH 8.0. 

Solubility:: The lyophilized rHuIL-15 is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=5.7).

Stability: Lyophilized rHuIL-15 although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuIL-15  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 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 IL-15.

Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Asn-Trp-Val-Asn, which agrees with the sequence of native Human IL-15.

 Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis. 

Biological Activity: Our rHuIL-15 is fully biologically active when compared to standard. The ED50 as determined by the dose-dependant stimulation of the proliferation of CTLL-2 was found to be < 0.45 ng/ml, corresponding to a Specific Activity of 3 x 10⁶ IU/mg.

Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuIL-15.

Protein content: Protein quantitation was carried out by two independent methods:

1.     UV spectroscopy at 280 nm using the absorbency value of 0.55 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 IL-15 as a Reference Standard.

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