Tumor-Associated GT3 Ganglioside Antigen Production Service
GT3 Ganglioside Is an Attractive Target
The existence of GT3, a trisialosyl lactosylceramide, in the central nervous system, was first unveiled in 1979. GT3 gangliosides are distributed in mammalian tissue. They act as precursors for the synthesis of more intricate gangliosides. Besides, it is suggested that ganglioside GT3 plays a distinct role in the organization and functionality of the tissue. It is highly expressed due to altered ganglioside metabolism in melanoma. Therefore, targeting GT3 ganglioside antibodies has become an appealing approach for immunotherapeutic strategies.
Fig 1. The ganglioside biosynthetic pathway and related pathologies. (Sipione, et al., 2020)
Tumor-Associated GD3 Ganglioside Antigen Production Service at CD BioGlyco
CD BioGlyco provides GT3 ganglioside synthesis by chemoenzymatic strategy.
- Cst-II, a glycosyltransferase gene from Campylobacter jejuni, is responsible for coding bifunctional α-(2→3/8)-sialyltransferase (α3/8SiaT). We clone the genes encoding cst-II into the expression vector pCWori+ and subsequently express them in the AD202 E. coli cell line. We obtain high enzymes with high activities from crude cell lysates during large-scale fermentation after subjecting the cells to a microfluidizer and removing cellular debris. The activity of Cst-II is determined by measuring α-(2→3)-activity using lactose as the acceptor substrate and cytidine-5’monophospho-N-acetylneuraminic acid (CMP-Neu5Ac) as the donor substrate.
- To address the issue of the higher cost of UDP-GalNAc compared to UDP-GlcNAc, we develop a rat UDP-N-acetylglucosamine-4′epimerase (GalNAcE) enzyme. UDP-GlcNAc is transformed into UDP-GalNAc by this enzyme.
- We also clone the rat epimerase gene into the pCWori+ vector and express it in E. coli. The cell lysates are used directly in the enzymatic reaction mixtures.
- In order to synthesize ganglio-oligosaccharide structures for conjugation purposes, we introduce the 2-azidoethyl-functionality to the reducing end of lactose before enzymatic modifications. These modified derivatives can be directly used in competitive inhibition experiments, conjugated to various supports, or converted to other functional groups as needed. We employ carefully controlled conditions with limited amounts of CMP-Neu5Ac, as the Cst-II enzyme exhibits extensive α-(2→8) multi-sialylation activities under excess CMP-Neu5Ac conditions. By elongating 2-azidoethyllactoside using Cst-II and CMP-Neu5Ac, we successfully obtain GT3.
We also provide antigen conjugation service. GT3 ganglioside is conjugated to keyhole limpet hemocyanin or a derivative of keyhole limpet hemocyanin. This conjugate has the potential to stimulate and enhance in a subject to which the vaccine is administered, the production of an antibody that recognizes a ganglioside. This conjugate has the potential as a vaccine candidate.
Fig. 2 The production process of GT3 conjugate. (CD BioGlyco)
Applications
- GT3 ganglioside can be used for therapeutic development of neurodegenerative disorders.
- GT3 ganglioside has the potential to be a biomarker for melanoma.
- GT3 ganglioside antibodies serve as an attractive target for immunotherapeutic approaches.
Advantages
- Our production process is efficient synthesis by using microbial glycosyltransferases we produce.
- The substrate specificities of bacterial glycosyltransferases we produce are generally more flexible than those of mammalian enzymes, which can be beneficial for various synthetic applications.
- We utilize an E. coli-based production system on a large scale to achieve the gram-scale synthesis of ganglioside oligosaccharides.
CD BioGlyco, a leading company in the field of glycobiology, provides specialized Tumor-associated Ganglioside Antigen Production Service. We have a professional research team with expertise and state-of-the-art technologies. Our research team offers customized solutions for the isolation, purification, and characterization of these antigens. If our proposition piques your interest, we encourage you to contact us for further information and discussions.
Reference
- Sipione, S.; et al. Gangliosides in the brain: physiology, pathophysiology and therapeutic applications. Frontiers in neuroscience. 2020, 14: 572965.
