Vaccine Grade

TDB-HS15

  • Product Name
  • TDB-HS15 / Trehalose-6,6-dibehenate formulated with Kolliphor HS 15
  • Cat. No.
  • ADJU-021CL
  • Content
  • 2 x 1 mg Trehalose-6,6-dibehenate (TDB) formulated with Kolliphor HS 15.
    10 ml sterile endotoxin-free physiological water (NaCl 0.9%)
  • Storage and Stability
  • TDB-HS15 is provided as a sterile powder and shipped at room temperature. Store at -20°C. Lyophilized product is stable for 1 year at -20°C.
  • Description

  • Trehalose-6,6-dibehenate (TDB) is a non-toxic synthetic analogue of the mycobacterial cell wall component trehalose 6,6’ dimycolate (TDM, also known as cord factor). 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, 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. In order to produce a homogenous suspension, TDB was formulated using Kolliphor HS 15 (CAS no. 70142-34-6), a low toxicity non-ionic surfactant.
  • Preparation of Stock Suspension (1 mg/ml)
  • Add 100 µl DMSO to 1 mg TDB-HS15, heat at 60°C (approx. 15 - 30 seconds) and vortex.
    - Once resuspended, immediately add 900 µl sterile physiological water (provided) or sterile phosphate buffered saline, heat for 10 - 15 minutes at 60°C and homogenize by vortexing for 30 seconds.
    Note: Following the addition of PBS or physiological water, the suspension may appear slightly cloudy containing floating fine particles.
    - Store at 4°C or prepare dilutions using a buffered solution for immediate use. Prior to each use, bring suspension to room temperature and homogenize by vortexing for 30 seconds. 
    OR
  • Preparation of DDA: TDB Complex
  • The protocol described below is based on the method described in Henriksen-Lacey M. et al., 2010. Liposomes based on dimethyldioctadecylammonium promote a depot effect and enhance immunogenicity of soluble antigen. J Control Release. 142(2):180-6.
    - 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-HS15 (dissolve 1 mg TDB-HS15 using 0.2 ml chloroform/methanol (ratio 9:1 v/v)).
    - Add 1 ml DDA solution (5 mg/ml) to 0.2 ml TDB-HS15 solution (5 mg/ml) providing a ratio of 5:1 w/w (DDA:TBD-HS15).
    - 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.
  • Chemical Properties

  • CAS number: 66758-35-8
    formula: C56H106O13
    Molecular weight: 987.43
    Structure of TDB:

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