ODN 2395 VacciGrade™

CpG ODN, class C (human / mouse) - TLR9-based adjuvant

ODN 2395 is a synthetic oligodeoxynucleotide containing unmethylated CpG motifs (CpG ODNs). CpG ODNs are recognized by TLR9, which is expressed exclusively on human B cells and plasmacytoid dendritic cells (pDCs), thereby inducing Th1-dominated immune responses.

ODN 2395 is a class C CpG ODN specific for mouse TLR9. In vivo studies have demonstrated that C-class ODNs, such as ODN 2395, are very potent Th1 adjuvants.

ODN 2395 VacciGrade™ is a high-quality pre-clinical grade.
ODN 2395 VacciGrade™​ is for research use only, and not for human or veterinary use.

Specifications

ODN 2395 sequence: 5’-tcgtcgttttcggcgc:gcgccg-3’ (22 mer)

Description: TLR9 agonist VacciGrade™

Polarization of adaptive immune response: Th1 response

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 (141.85 nmol) of sterile lyophilized ODN 2395 VacciGrade™
• 10 ml sterile endotoxin-free physiological water (NaCl 0.9%)

ODN 2395 VacciGrade™ is shipped at room temperature

Should be stored at -20°C for up to 1 year.
Upon resuspension, prepare aliquots of ODN 2395  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

Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs) have been extensively studied as adjuvants [1]. These CpG motifs are present at a 20-fold greater frequency in bacterial DNA compared to mammalian DNA [2].
CpG ODNs are recognized by TLR9, which is expressed exclusively on human B cells and plasmacytoid dendritic cells (pDCs), thereby inducing Th1-dominated immune responses [3].
Pre-clinical studies, conducted in rodents and non-human primates, and human clinical trials have demonstrated that CpG ODNs can significantly improve vaccine-specific antibody responses [1].

Three types of stimulatory CpG ODNs have been identified, types A, B and C, which differ in their immune-stimulatory activities [4-5]. Furthermore, CpG ODNs activate TLR9 in a species-specific manner [6].

ODN 2395 is a type C CpG ODN specific for mouse TLR9 [7]. Type C CpG ODNs contain a full phosphorothioate backbone with one or more CpG dinucleotides. Type C CpG ODNs are potent inducers of IFN-α from pDC and strong B cell activators [8, 9]. In vivo studies have demonstrated that type C ODNs which combine the effects of types A and B ODNs, such ODN 2395, are very potent Th1 adjuvants [10].

1. Steinhagen F. et al., 2011. TLR-based immune adjuvants. Vaccine 29(17):3341-55.
2. Hemmi H. et al., 2000. A Toll-like receptor recognizes bacterial DNA. Nature 408: 740-5.
3. Coffman RL. et al., 2010. Vaccine adjuvants: Putting innate immunity to work. Immunity 33(4):492-503.
4. 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.
5. 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.
6. Bauer S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. PNAS, 98(16):9237-42.
7. Moseman EA. et al., 2004. Human plasmacytoid dendritic cells activated by CpG oligodeoxynucleotides induce the generation of CD4+CD25+ regulatory T cells. J Immunol. 173:4433-4442.
8. Jurk M. et al., 2004. C-Class CpG ODN: sequence requirements and characterization of immunostimulatory activities on mRNA level. Immunobiology. 209(1-2):141-54.
9. Abel K. et al., 2005. Deoxycytidyl-deoxyguanosine oligonucleotide classes A, B, and C induce distinct cytokine gene expression patterns in rhesus monkey peripheral blood mononuclear cells and distinct alpha interferon responses in TLR9-expressing rhesus monkey plasmacytoid dendritic cells. Clin Diagn Lab Immunol. 12(5):606-21.
10. Vollmer J. et al., 2004. Characterization of three CpG oligodeoxynucleotide classes with distinct immunostimulatory activities. Eur J Immunol. 34(1):251-62.