ABOUT
CpG ODN, type A (human) - TLR9-based vaccine adjuvant
ODN 2216 VacciGrade™ is a pre-clinical Class A CpG oligonucleotide (ODN) with a preference for the human Toll-like receptor 9 (TLR9). CpG ODNs are short synthetic single-stranded DNA molecules containing unmethylated CpG dinucleotides (CpG motifs). These unmethylated CpG motifs mimic microbial DNA and act as immunostimulants via TLR9.
Class A CpG ODNs, such as ODN 2216, are characterized by a phosphodiester central CpG-containing palindromic motif and a phosphorothioate 3’ poly-G string. ODN 2216 induces robust production of type I interferon by plasmacytoid dendritic cells (pDCs) and highly stimulates natural killer (NK) cells. However, it has little stimulatory effect on B cells [1].
In HEK-Blue™ hTLR9 reporter cells, ODN 2216 efficiently activates hTLR9 compared to the control ODN 2216 (ODN 2243) Moreover, ODN 2216 activates the hTLR9-dependent NF-κB and IRF pathways, as assessed using THP1-Dual™ hTLR9 cells expressing both NF-κB-inducible SEAP and IRF-inducible Lucia luciferase reporters (see figures).
VacciGrade™ is a high-quality pre-clinical grade, suitable for in vivo use.
A standard grade ODN 2216 for in vitro assays is also available.
ODN 2216 VacciGrade™ is for research use only, and not for human or veterinary use. It is not a pharmaceutical preparation fit for vaccine manufacturing.
Get more information about CpG ODNs Classes.
1. Krug A., et al., 2001. Identification of CpG oligonucleotide sequences with high induction of IFN-alpha/beta in plasmacytoid dendritic cells. Eur J Immunol. 31:2154-63.
All products are for research use only, and not for human or veterinary use.
VacciGrade™
VacciGrade™ is a high-quality pre-clinical grade, suitable for in vivo studies. 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.
SPECIFICATIONS
Specifications
TLR9
5’-ggGGGACGATCGTCgggggg-3’ (20 mer)
Note: Regular and capital letters represent phosphorothioate and phosphodiester bonds, respectively, bold letters represent CpG dinucleotides.
20-50 µg/mouse
2 mg/ml in physiological water
Sterility guaranteed
<1 EU/mg (measurement by kinetic chromogenic LAL assay)
Th1 response, TLR9 activation in cellular assays (tested)
Each lot is functionally tested and validated using cellular assays.
CONTENTS
Contents
-
Product:ODN 2216 VacciGrade™
-
Cat code:vac-2216-1
-
Quantity:1 mg
10 ml sterile endotoxin-free physiological water (NaCl 0.9%)
Shipping & Storage
- Shipping method: Room temperature
- -20°C
- Avoid repeated freeze-thaw cycles
Storage:
Caution:
Details
CpG ODNs
Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs), such as ODN 2216, 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 the Toll-like receptor 9 (TLR9), which is expressed 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, as well as 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].
Toll-like receptor 9
The Toll-like Receptor 9 (TLR9) is an endosomal receptor that triggers NF-κB- and interferon regulatory factor (IRF)-mediated pro-inflammatory responses upon the recognition of unmethylated cytosine-phosphorothioate-guanosine (CpG) forms of DNA [6-8]. Unmethylated CpG dinucleotides are a hallmark of microbial (bacterial, viral, fungal, and parasite) DNA, as well as mitochondrial self-DNA [8,9]. These TLR9 agonists can be mimicked by synthetic oligonucleotides containing CpG motifs (CpG ODNs), which have been extensively studied to improve adaptive immune responses in the context of vaccination [6,8].
TLR9 is mainly expressed in subsets of Dendritic Cells and B cells of all mammals. In rodents, but not in humans, TLR9 is also expressed in monocytes and macrophages [8]. The structure of the receptor varies by 24% between human TLR9 (hTLR9) and mouse TLR9 (mTLR9) [8]. They recognize different CpG motifs, the optimal sequences being GTCGTT and GACGTT for hTLR9 and mTLR9, respectively [10].
References:
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. Kumagai Y. et al., 2008. TLR9 as a key receptor of the recognition of DNA. Adv. Drug. Deliv. Rev. 60(7):795-804.
7. Heinz L.X. et al., 2021. TASL is the SLC15A4-associated adaptor for IRF5 activation by TLR7-9. Nature. 581(7808):316-322.
8. Kayraklioglu N. et al., 2021. CpG oligonucleotides as vaccine adjuvants. DNA Vaccines: Methods and Protocols. Methods in Molecular Biology. Vol. 2197. p51-77.
9. Kumar V., 2021. The trinity of cGAS, TLR9, and ALRs: guardians of the cellular galaxy against host-derived self-DNA. Front. Immunol. 11:624597.
10. Bauer S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Natl Acad Sci USA, 98(16):9237-42.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
Need a CoA ?
Powered by Bioz