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ODN 1018 VacciGrade™

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ODN 1018 VacciGrade™

ODN, Class B (human/mouse) - TLR9 agonist

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1 mg

vac-1018-1
+-
$950

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

TLR9 activation with ODN 1018
TLR9 activation with ODN 1018

ODN 1018 VacciGrade™ is for research use only, and not for human or veterinary use. It is not a pharmaceutical preparation fit for vaccine manufacturing.

CpG ODN 1018 Vaccigrade™ is a synthetic immunostimulatory oligonucleotide (ODN) that contains unmethylated CpG dinucleotides. This product is a class B CpG ODN containing CpG motifs that are able to bind and activate both the rodent and human Toll-like receptor 9 (TLR9) [1]. ODN 1018 VacciGrade™ has an identical sequence to 1018 ISS

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InvivoGen's ODN 1018 VacciGrade™ successfully activates mouse (m)TLR9, but not human (h)TLR9 in HEK-derived cells [in-house data]. In monocytic THP1-Dual™ hTLR9 cells, however, ODN 1018 VacciGrade™ shows a strong hTLR9-mediated NF-κB and IRF induction (see figures). ODN 1018 VacciGrade™ is a high-quality pre-clinical grade, suitable for in vivo use.

 

 

1. Campbell JD, 2017. Development of the CpG Adjuvant 1018: A Case Study. Methods Mol Biol. 1494:15-27.

Figures

Murine and human TLR9 activation in HEK-Blue™-derived cells
Murine and human TLR9 activation in HEK-Blue™-derived cells

ODN 1018 VacciGrade™ is a potent activator of murine TLR9 in HEK-derived cells. HEK-Blue™ murine (m)TLR9 and HEK-Blue™ human (h)TLR9 cells were incubated in HEK-Blue™ Detection reagent with increasing concentrations of ODN 1018 VacciGrade™. After 24 hours, SEAP activity in the cell culture supernatant was assessed. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Human TLR9 activation in THP1-Dual™-derived cells
Human TLR9 activation in THP1-Dual™-derived cells

ODN 1018 VacciGrade™ is a potent activator of human TLR9 in THP-1-derived cells. THP1-Dual™ and THP1-Dual™ human (h)TLR9 cells were incubated with increasing concentrations of ODN 1018 VacciGrade™. After 24h, the hTLR9-induced (A) NF-κB and (B) IRF responses were assessed by measuring SEAP and Lucia activity using QUANTI-Blue™ and QUANTI-Luc™, respectively. Data are shown as optical density (OD) at 650 nm or in fold response over non-induced cells (mean + SEM).

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Specifications

ODN 1018 sequence: 5′-TGACTGTGAACGTTCGAGATGA-3′ (22-mer)

Description: TLR9 agonist VacciGrade™

Synonyms: 1018 ISS

CAS number: 937402-51-2

Polarization of adaptive immune response: Th1 response

Solubility: 2 mg/ml in physiological water

Working concentration: 1 ng/ml - 10 µg/ml

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)
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Contents

  • 1 mg (139.8 nmol) of sterile lyophilized ODN 1018 VacciGrade™
  • 10 ml sterile endotoxin-free physiological water (NaCl 0.9%)

ODN 1018 VacciGrade™ is shipped at room temperature.

ODN 1018 VacciGrade™ should be stored at -20°C.

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

Avoid repeated freeze-thaw cycles.

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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.

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Details

CpG ODNs

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 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]. 

1018 ISS

1018 immunostimulatory sequence (ISS) is a type B CpG ODN with the sequence 5′-TGACTGTGAACGTTCGAGATGA-3′. It mimics bacterial single-stranded (ss) DNA to activate innate immunity through binding to TLR9. The first CpG motif (underlined) is a sequence active on mouse TLR9, whereas the second CpG motif (bold) is active on human and non-human primate TLR9 [6]. Type B CpG ODNs contain a full phosphorothioate backbone with one or more CpG dinucleotides and were described to strongly activate the Th1 cell and B cell populations [4,6]. CpG motifs present in the 1018 sequence stimulate the innate immune system, which results in efficacious immunological effects, such as the production of IL-12, IL-18, and IFN-γ from macrophages and natural killer (NK) cells, promoting Th1 responses, and its involvement in antibody production and B cell proliferation [7]. 

The GMP-produced CpG 1018® produced by Dynavax Technologies is currently the only FDA-approved CpG adjuvant for human vaccine use. It is a component of several approved vaccines, including the licensed hepatitis B virus vaccine HEPLISAV-B® [8].

 

Note: CpG 1018® is a registered trademark of Dynavax Technologies and is used for comparative purposes only. ODN 1018 VacciGrade™​ is not made by, affiliated with, sponsored by, or endorsed by Dynavax Technologies.

 

 

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. Campbell JD, 2017. Development of the CpG Adjuvant 1018: A Case Study. Methods Mol Biol. 1494:15-27.
7. Eng NF, et al., 2013. The potential of 1018 ISS adjuvant in hepatitis B vaccines: HEPLISAV™ review. Hum Vaccin Immunother.(8):1661-72.
8. Strohmeier S, et al., 2022. A CpG 1018 adjuvanted neuraminidase vaccine provides robust protection from influenza virus challenge in mice. NPJ Vaccines. 2022 Jul 22;7(1):81.

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