2'3'-cGAMP VacciGrade™

STING-based vaccine adjuvant - Th1/Th2 response

2’3’-cGAMP VacciGrade™ is a preclinical grade preparation of the cyclic dinucleotide 2’3’-cGAMP (cyclic [G(2’,5’)pA(3’,5’)p]), a cyclic dinucleotide (CDN) produced in mammalian cells by cGAS (cGAMP synthase) in response to double-stranded DNA in the cytoplasm.
2’3’-cGAMP is also referred to as “noncanonical” cGAMP due to the presence of the atypical 2’-5’ phosphodiester linkage between the guanosine and the adenosine. Structural and functional studies revealed that noncanonical 2’3’-cGAMP is distinct from the canonical 3’3’-cGAMP produced by bacteria [1, 2].

CDNs have been shown to increase vaccine potency [3]. CDNs activate innate immunity by directly binding the endoplasmic reticulum-resident receptor STING (stimulator of interferon genes), leading to the expression of interferon-β (IFN-β) and nuclear factor-κB (NF-κB) dependent inflammatory cytokines.

2′3′-cGAMP is an effective adjuvant that boosts the production of antigen-specific antibodies and T cell responses in mice [4].

2’3’-cGAMP VacciGrade™ is a high-quality pre-clinical grade.

2’3’-cGAMP VacciGrade™​ is for research use only, and not for human or veterinary use.

Read our review on STING: Deciphering the STING Paradox

References:

1. Diner E. et al., 2013. The Innate Immune DNA Sensor cGAS Produces a Noncanonical Cyclic Dinucleotide that Activates Human STING. Cell Rep. 3(5):1355-61.
2. Gao P. et al., 2013. Cyclic [G(2',5')pA(3',5')p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell. 153(5):1094-107.
3. Dubensky TW. et al., 2013. Rationale, progress, and development of vaccines utilizing STING-activating cyclic dinucleotide adjuvants. Therapeutic Advances in Vaccines 1(4): 131-143.
4. Li XD. et al., 2013. Pivotal roles of cGAS-cGAMP signaling in antiviral defense and immune adjuvant effects. Science. 341(6152):1390-4.

Figures

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). The interferon regulatory factor (IRF) response was assessed by measuring the activity of Lucia luciferase in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia. Data are shown in fold response over non-induced cells (mean ± SEM).

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). The NF-κB-induced SEAP activity was assessed using QUANTI‑Blue™ Solution, a SEAP detection reagent. Data are shown as optical density (OD) at 630 nm (mean ± SEM). Non‑induced cells (NI) have been included as a negative control.

Specifications

Description: STING agonist VacciGrade™

Synonym: Cyclic [G(2’,5’)pA(3’,5’)p], cyclic GMP-AMP; c-GpAp sodium salt

CAS number: 2734858-36-5 / 1441190-66-4 (free acid)

Formula: C₂₀H₂₂N₁₀O₁₃P₂ .2Na

Molecular weight: 718.38 g/ml

Purity: ≥ 95% by LC/MS

Solubility: 50 mg/ml in physiological water

Quality control:

• Sterility guaranteed
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK‑Blue™ TLR4 cells.
• Endotoxin level < 5 EU/mg (measurement by kinetic chromogenic LAL assay)
• The biological activity of  2'3'-cGAMP VacciGrade™ has been assessed by measuring the induction of the interferon pathway in cellular assays.

Contents

• 1 mg (2 x 500 µg) lyophilized 2’3’-cGAMP VacciGrade™
• 10 ml sterile endotoxin-free physiological water (NaCl 0.9%)

2’3’-cGAMP VacciGrade™ is shipped at room temperature

2’3’-cGAMP VacciGrade™should be stored at -20°C.

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

Structure of 2'3'-cGAMP VacciGrade™