Recombinant Human Tumor Necrosis Factor-alpha/TNFSF2, Variant

Synonyms Tumor Necrosis Factor, TNFSF2, Cachectin, Differentiation-inducing factor (DIF), Necrosin, Cytotoxin

Accession P01375

GeneID 7124

Source Escherichia coli.

Molecular Weight Approximately 16.9 kDa, a single non-glycosylated polypeptide chain containing 151 amino acids. Compared with the wild-type, rHuTNF-α Variant has an amino acid sequence (a.a.) deletion from a.a. 1-7, and the following a.a. substitutes Arg8, Lys9, Arg10 and Phe157 which is proven to have more activity and with less inflammatory side effect in vivo.

Quantity 10µg/50µg/1mg

AA Sequence MRKRKPVAHV VANPQAEGQL QWLNRRANAL LANGVELRDN QLVVPSEGLY LIYSQVLFKG QGCPSTHVLL THTISRIAVS YQTKVNLLSA IKSPCQRETP EGAEAKPWYE PIYLGGVFQL EKGDRLSAEI NRPDYLDFAE SGQVYFGIIA F

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

Biological Activity Fully biologically active when compared to standard. The ED50 as determined by a cytotoxicity assay using murine L929 cells is less than 0.01 ng/ml, corresponding to a specific activity of > 1.0 × 107 IU/mg in the presence of actinomycin D.

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

Formulation Lyophilized from a 0.2 μm filtered concentrated solution in PBS, pH 7.0.

Endotoxin Less than 1 EU/μg of rHu TNF-α/TNFSF2, Variant 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 material is offered by Shanghai PrimeGene Bio-Tech for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE.

SDS-PAGE

Reference 1. Davenport C, Kenny H, Ashley DT, et al. 2012. Eur J Clin Invest, 42: 1173-9.

2. Cavalcanti YV, Brelaz MC, Neves JK, et al. 2012. Pulm Med, 2012: 745483.

3. Sheng WS, Hu S, Ni HT, et al. 2005. J Leukoc Biol, 78: 1233-41.

4. Berthold-Losleben MandHimmerich H. 2008. Curr Neuropharmacol, 6: 193-202.

Background The clinical use of the potent antitumor activity of TNF-αhas been limited by the proinflammatory side effects including fever, dose-limiting hypotension, hepatotoxicity, intravascular thrombosis, and hemorrhage. Designing clinically applicable TNF-αmutants with low systemic toxicity has been an intense pharmacological interest. Human TNF-α, which binds to the murine TNF-R55 but not to the murine TNF-R75, exhibits retained antitumor activity and reduced systemic toxicity in mice compared with murine TNF-α, which binds to both murine TNF receptors. Based on these results, many TNF-αmutants that selectively bind to TNF-R55 have been designed. These mutants displayed cytotoxic activities on tumor cell lines in vitro, and exhibited lower systemic toxicity in vivo.