ODN 2006 VacciGrade™

CpG ODN, type B (human) - TLR9-based adjuvant

CpG ODN 2006 (ODN 7909) is a synthetic immunostimulatory oligonucleotide (ODN) that contains unmethylated CpG dinucleotides.

ODN 2006 is a class B CpG ODN and is a ligand of choice for human TLR9. It strongly activates B cells but weakly stimulates IFN-α secretion in pDCs. CpG ODN 2006 generates Th1-dominant immune responses.

ODN 2006 VacciGrade™ is a high-quality pre-clinical grade.
ODN 2006 VacciGrade™​ is for research use only, and not for human or veterinary use.

Specifications

ODN 2006 sequence: 5’-tcgtcgttttgtcgttttgtcgtt-3’ (24 mer)

Description: TLR9 agonist VacciGrade™

Synonyms: ODN 7909, PF_3512676

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 < 2 EU/mg (determined using the HEK-Blue™ LPS Detection Kit 2)

Contents

• 1 mg (129.8 nmol) of sterile lyophilized ODN 2006 VacciGrade™.
• 10 ml sterile endotoxin-free physiological water (NaCl 0.9%).

ODN 2006 VacciGrade™ is shipped at room temperature.

ODN 2006 VacciGrade™ should be stored at -20°C for up to 1 year.
Upon resuspension, prepare aliquots of ODN 2006 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), such as ODN 2006, 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 2006 is a type B CpG ODN specific for mouse TLR9.

Type B CpG ODNs contain a full phosphorothioate backbone with one or more CpG dinucleotides and can strongly activate B cells  [4]. ODN 2006 has been tested as an adjuvant in numerous animal models  [7-9].

Research has demonstrated that ODN 2006 is an effective adjuvant for stimulating both antibody [8-9] and interferon-mediated cellular immune responses [9].

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. Hartmann G. et al., 2000. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primate immune responses in vitro and in vivo. J. Immunol. 164(3): 1617 - 1624.
8. conforti VA. et al., 2007. The effectiveness of a CpG motif-based adjuvant (CpG ODN 2006) for LHRH immunization. Vaccine 25: 6537-43.
9. Zhang Y. et al., 2003. CpG ODN 2006 and IL-12 are comparable for priming Th1 lymphocyte and IgG responses in cattle immunized with a rickettsial outer membrane protein in alum. Vaccine 21:3307-18.