||CD40 Ligand, CD154, TRAP, TNFSF5, gp39
||CD40 Ligand (CD40L/CD154/TRAP) is a membrane glycoprotein and differentiation antigen expressed on the surface of T-cells. The CD40 ligand stimulates B-cell proliferation and secretion of all immunoglobulin isotypes in the presence of cytokines. It also costimulates proliferation of activated T-cell and this is accompanied by the production of IFN-γ, TNF-α, and IL2. CD40 ligand has been shown to induce cytokine production and tumoricidal activity in peripheral blood monocytes.
Recombinant Human CD40 Ligand (CD40L/CD154/TRAP) produced in E. coli is a non-glycosylated, polypeptide chain containing 149 amino acids and having a molecular mass of 16,308 Da.
||Recombinant human CD40 Ligand (CD40L/CD154/TRAP) is fully biologically active when compared to standard. The ED50 as determined by the dose-dependant stimulation of IL-12 & IL-8 induction by PMB (Peripheral Mononuclear) cells was found to be 5-10 ng/ml, corresponding to a Specific Activity of 1.0 x 105 IU/mg.
||The sequence of the first five N-terminal amino acids was determined and was found to be Met-Gln-Lys-Gly-Asp.
||The protein was lyophilized after extensive dialysis against 50mM Tris-HCl, pH8.0, 250mM NaCl buffer.
||It is recommended to reconstitute the lyophilized recombinant human CD40 Ligand (CD40L/CD154/TRAP) in sterile 18 MΩ-cm H2O not less than 100 µg/ml, which can then be further diluted to other aqueous solutions.
||Greater than 95.0% as determined by:
1. Analysis by RP-HPLC.
2. Anion-exchange FPLC.
3. Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
||Less than 0.1 ng/µg (1 EU/µg) of recombinant human CD40 Ligand (CD40L) as determined by LAL test.
||Lyophilized recombinant human CD40 Ligand (CD40L/CD154/TRAP) although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution CD40L should be stored at 4°C between 2-7 days and for future use below -18°C. For long-term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Oertelt S, et al. Soluble CD40L in plasma of patients with primary biliary cirrhosis. Ann. N. Y. Acad. Sci. Jun 2005; 1051: 205-210.
Danese S, et al. TNF-alpha blockade down-regulates the CD40/CD40L pathway in the mucosal microcirculation: a novel anti-inflammatory mechanism of infliximab in Crohn's disease. J. Immunol. Feb 2006; 176(4): 2617-2624.