Shipped at room temperature and it should be stored at room temp. Store at -20 °C. Lyophilized product is stable for 1 year at -20 °C.
Preparation of stock suspension (1 mg/ml)
Add 100 µl DMSO to 1 mg TDB, heat at 60°C (approx. 15 -30 seconds) and vortex.
Once resuspended, immediately add 900 µl sterile phosphate buffered saline (PBS), heat for 10 - 15 minutes at 60°C and homogenize by vortexing for 30 seconds
Preparation of DDA:TDB complex
Prepare a 5 mg/ml solution of dimethyldioctadecylammonium bromide (DDA; not provided) in chloroform/methanol (ratio 9:1 v/v). - Prepare a 5 mg/ml solution of TDB VacciGrade™ (dissolve 1 mg TDB VacciGrade™ using 0.2 ml chloroform/methanol (ratio 9:1 v/v)). - Add 1 ml DDA solution (5 mg/ml) to 0.2 ml TDB VacciGrade™ solution (5 mg/ml) providing a ratio of 5:1 w/w (DDA:TBD). - Use a roto-evaporator to remove the chloroform/methanol followed by flushing with N2, to produce a thin lipidic film containing the DDA:TDB complex. - Store DDA:TDB complex at -20 °C for 3 months or use immediately. - Hydrate the lipidic film with 2 ml TRIS base buffer (10 mM; pH 7.4). Heat the suspension for 20 minutes at 60 °C. Allow to cool before use. This suspension should not be stored, use within 1 day.
Trehalose-6,6-dibehenate (TDB) is a synthetic analog of rehalose6,6-dimycolate (TDM, also known as cord factor), which is the most studied mmunostimulatory component of Mycobacterium tuberculosis.
TDB was recently shown to rely on the C-type lectin Mincle and the signaling molecules Syk and Card9 to trigger a Th1/Th17 innate immunity . Incorporation of TDB combined with the synthetic amphiphilic cationic lipid compound dimethyldioctadecylammonium (DDA) into liposomes, known as CAF01, has been shown to strongly enhance cellular and humoral responses against a protein antigen . Adjuvanticity of the cationic DDA:TDB liposomes and sustained protection against disease challenge has been demonstrated in particular with a tuberculosis vaccine candidate , and has good potential for application in a range of other diseases .
Supplied as ready to use, lyophilized, sterile powder,
2 x 1 mg TDB VacciGrade™
10 ml sterile endotoxin-free physiological water (NaCl 0.9%)