ODN 1585 VacciGrade™

CpG ODN, type A (mouse) - TLR9-based adjuvant

CpG ODN 1585 is a synthetic immunostimulatory oligonucleotide (ODN) that contains unmethylated CpG dinucleotides.

ODN 1585 is a class A CpG ODN specific for mouse Toll-like receptor 9 (TLR9).

In plasmacytoid dendritic cells (pDCs) it induces high IFN-α production but is a weak stimulator of TLR9-dependent NF-κB signaling (IL-6). It generates Th1-dominant immune responses.

ODN 1585 VacciGrade™ is a high-quality pre-clinical grade.
ODN 1585 VacciGrade™​ is for research use only, and not for human or veterinary use.

Specifications

ODN1585 sequence: 5’- ggGGTCAACGTTGAgggggg -3’ (20 mer)

Description: TLR9 agonist VacciGrade™

Polarization of adaptive immune response: Th1 response

Solubility: 2 mg/ml in physiological water

Working concentration: 20 - 50 μg/mouse

Quality control:

• Sterility guaranteed
• The absence of bacterial contamination (lipoproteins & endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells
• Endotoxin level < 1 EU/mg (measurement by kinetic chromogenic LAL assay)

Contents

• 1 mg (155.5 nmol) sterile lyophilized ODN 1585 VacciGrade™
• 10 ml sterile endotoxin-free physiological water (NaCl 0.9%)

ODN 1585 VacciGrade™ is shipped at room temperature

Should be stored at -20°C for up to 1 year.
Upon resuspension, prepare aliquots of ODN 1826 VacciGrade™ and store at -20°C.

Product is stable 6 months at -20°C when properly stored.

Avoid repeated freeze-thaw cycles.

VacciGrade™

VacciGrade™ is a high-quality pre-clinical grade. VacciGrade™ products are filter-sterilized (0.2 µm) and filled under strict aseptic conditions in a clean room*. The absence of bacterial contamination is assessed by a sterility test using a pharmacopeia-derived assay. The level of bacterial contaminants (endotoxins and lipoproteins) in each lot is verified using a LAL assay and/or a TLR2 and TLR4 reporter assay.
*Except for LPS VacciGrade™, which is prepared in a laminar flow hood dedicated to LPS.

Details

CpG ODNs are synthetic oligonucleotides that contain unmethylated CpG dinucleotides in particular sequence contexts (CpG motifs) [1]. These CpG motifs are present at a 20-fold greater frequency in bacterial DNA compared to mammalian DNA. CpG ODNs are recognized by Toll-like receptor 9 (TLR9) leading to strong immunostimulatory effects [2].

Three types of stimulatory CpG ODNs have been identified, types A, B and C, which differ in their immune-stimulatory activities [3-4]. Type A CpG ODNs are characterized by a phosphodiester central CpG-containing palindromic motif and a phosphorothioate 3’ poly-G string. They induce high IFN-α production from plasmacytoid dendritic cells (pDC) but are weak stimulators of TLR9-dependent NF-κB signaling. Type B CpG ODNs contain a full phosphorothioate backbone with one or more CpG dinucleotides. They strongly activate B cells but stimulate weakly IFN-α secretion. Type C CpG ODNs combine features of both types A and B. They contain a complete phosphorothioate backbone and a CpG-containing palindromic motif. Type C CpG ODNs induce strong IFN-α production from pDC and B cell stimulation.

ODN 1585 is a CpG ODN type A specific for mouse TLR9 [5].

1. Krieg, A.M. et al., 1995. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature, 374(6522):546-9.
2. Bauer, S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. PNAS. 98(16):9237-42.
3. Krug A. et al., 2001. Identification of CpG oligonucleotide sequences with high induction of IFN- alpha/beta in plasmacytoid dendritic cells. Eur J Immunol, 31(7): 2154-63.
4. Marshall JD. et al., 2005. Superior activity of the type C class of ISS in vitro and in vivo across multiple species. DNA Cell Biol. 24(2):63-72.
5. Ballas ZK. et al., 2001. Divergent therapeutic and immunologic effects of oligodeoxynucleotides with distinct CpG motifs. J Immunol. 167(9):4878-86.