Tumor-Associated 9NHAc-GD2 Ganglioside Antigen Production Service at CD BioGlyco

CD BioGlyco specializes in Carbohydrate-based Vaccine Development. We possess an advanced Glyco™ Vaccine Development Service Platform, offering tumor-associated 9NHAc-GD2 ganglioside antigen production in the field of glycobiology. CD BioGlyco produces a 9NHAc-GD2 ganglioside to represent O-acetyl-GD2 to generate anti-cancer vaccines.

Why DO We Produce This Ganglioside?

9-O-acetylated GD2 (9O-Ac-GD2) is highly expressed on tumor cells that also express GD2. Notably, it is not present on human peripheral nerve fibers, indicating that anti-9OAc-GD2 antibodies may have lower toxicity compared to anti-GD2 antibodies. As a result, 9OAc-GD2 can be a promising antigenic target for developing anti-cancer vaccines.

Fig.1 The primary route for the biosynthesis of O-Ac-GD2 ganglioside. (Fleurence, et al., 2017)

It is difficult to synthesize O-Ac-GD2. Therefore, we design a new GD2 derivative (9NHAc-GD2) bearing an N-acetamide at its non-reducing end neuraminic acid to mimic the O-Ac-GD2.

• Firstly, we synthesize Neu5Ac9NAcA by an efficient chemoenzymatic synthetic strategy. In this strategy, a simplified one-step reduction and simultaneous acetylation process are utilized to synthesize ManNAc6NAc from ManNAc6N₃. This process involves the addition of thioacetic acid to a pyridine solution containing ManNAc6N₃. After that, we utilize a reaction catalyzed by a sialic acid aldolase from Pasteurella multocida (PmAldolase) to obtain Neu5Ac9NAc from ManNAc6NAc.
• CJCost II shows excellent ability to accept modifications at the C-9 position. Therefore, we utilize CjCstII to convert 9NHAc-Neu5Ac and GM3-ProN₃ into 9NHAc-GD3. Subsequently, we produce 9NHAc-GD2 through the transfer of GalNAc to 9NHAc-GD3 by CjCgtA, followed by catalytic hydrogenolysis. It remains stable after prolonged storage in buffers, retaining its acetyl moiety.
• We employ molecular dynamics simulations to explore the conformational space, followed by quantum mechanical calculations for optimizing geometry and calculating energy. It is suggested that the lowest energy conformations identified for 9OAc-GD2 and 9NHAc-GD2 exhibit a high degree of similarity.
• We also explore anti-cancer vaccines based on this antigen. In order to induce robust and enduring antibody responses, the 9NHAc-GD2 antigen is coupled to an immunogenic carrier. The bacteriophage Qβ virus-like particle, a potent carrier for tumor-associated carbohydrate antigens in vaccine design, is used to conjugate with 9NHAc-GD2 antigen at CD BioGlyco. The amine groups of 9NHAc-GD2 are converted to thiocyanate 9NHAc-GD2 with thiophosgene, followed by their addition to bacteriophage Qβ virus-like particles to produce Qβ-9NHAc-GD2 conjugate. We utilize mass spectrometry for the analysis of the resulting particles. With Qβ-9NHAc-GD2 conjugates in hand, immunological studies are performed.

Fig.2 Diagram of Qβ-9NHAc-GD2 conjugate for eliciting robust IgG antibodies against cancer. (CD BioGlyco)

Frequently Asked Questions

• In which animals are the immunization experiments with Qβ-9NHAc-GD2 carried out?

We conduct immunization experiments in mice and canines.

• What is the immune effect of Qβ-9NHAc-GD2 in the mouse model?

Mice immunized with Qβ-9NHAc-GD2 produce significantly higher levels of IgG antibodies compared to those immunized with Qβ-GD2, indicating that the NHAc substitution can enhance its antigenicity.

• What is the immune effect of Qβ-9NHAc-GD2 in the canine model?

The conjugate leads to the production of IgG antibodies, indicating the immunogenicity of Qβ-9NHAc-GD2 in canines. Moreover, there are no notable alterations in pain tolerance, serum chemistry, or hematology, demonstrating the safety of Qβ-9NHAc-GD2 immunization.

Advantages

• We have experience in the utilization of molecular dynamics simulations and quantum mechanical calculations for conformational and energy analysis, ensuring accurate understanding and design of antigen structures.
• Our service evaluates a Qβ glycan conjugate in a canine model, suggesting its promising prospects for translation to human patients.
• We are proficient in chemoenzymatic synthesis strategies, enabling efficient and tailored production of complex glycans.
• We are able to conduct comprehensive immunological studies to evaluate the efficacy and potential applications of the produced antigens and vaccine candidates.

With a deep understanding of chemoenzymatic synthesis and molecular dynamics simulations, CD BioGlyco excels in the tailored production of complex antigens and vaccines. Our proficiency in mass spectrometry analysis and immunological studies ensures precision and efficacy in vaccine development. We devote ourselves to delivering innovative and potent solutions for advancing carbohydrate-based vaccine strategies. Should our services pique your interest, please do not hesitate to contact us.

References

1. Fleurence, J.; et al. Targeting O-acetyl-GD2 ganglioside for cancer immunotherapy. Journal of immunology research. 2017, 1-16.
2. From Wikipedia: https://en.wikipedia.org/wiki/Antibody#/media/File:Antibody.svg