One of the three basic forms of Salivary Acid is Neu5Gc. Neu5Gc is mostly found in hind-mouth animals. Neu5Ac-CMP is hydroxylated by CMP-Neu5Ac hydroxylase (CMAH) to synthesize Neu5Gc. It has additional oxygen compared to N-Acetylneuraminic Acid (Neu5Ac) and is presented as a hydroxyl group in the N-ethanol acyl group at C-5. Due to the absence of the human CMAH gene, the human body is not able to synthesize Neu5Gc metabolically on its own. It is currently believed that the body obtains Neu5Gc from food. Neu5Gc is a unique antigen that can modify the surface of human cells in a variety of ways. It has been found that some human tumor cell surfaces (e.g., liver, colon, and stomach cancers, etc.) can be found to contain significantly elevated levels of Neu5Gc. Neu5Gc may be linked to the development of cancer in some way. Therefore, the pathway by which high levels of Neu5Gc are generated in cancer patients has become an issue of great interest in the field of cancer research.
Fig.1 Three forms of sialic acid and the synthesis of Neu5Gc and its sialosides. (Kooner, et al., 2019)
CD BioGlyco is equipped with state-of-the-art instrumentation and a dedicated team of testing professionals for salivary acid cell surface glycoengineering service. Neu5Gc and its glycosides are critical tools for understanding Neu5Gc-associated immune responses, analyzing salivary acid-binding proteins and anti-Neu5Gc antibodies, designing potential therapies, etc. In order to provide convenience to our clients who are researching Neu5Gc, we offer Neu5Gc synthesis-related gene editing and analysis services.
Fig.2 Neu5Gc-based cell surface glycoengineering service. (CD BioGlyco)
Neu5Gc synthesis is associated with CMAH and α(1,3) galactosyltransferase (encoded by the GGTA1 gene). We target the introduction and modification of Neu5Gc by Knocking Out GGTA1 or CMAH using gene editing techniques such as CRISPR/Cas9. By knocking out GGTA1 or CMAH, it is possible to detect changes in glycosylation in cells and to investigate the biological role of Neu5Gc.
We modify Neu5Gc by introducing additional acetyl groups, trifluoroacetyl groups, etc., or by converting functional groups to azide groups, etc. Specific modification of cell surface sialic acid is realized through modification of Neu5Gc.
We offer a range of Neu5Gc analysis services including Neu5Gc identification, Neu5Gc quantification, and more. We have specialized kits for salivary acid analysis and quantification. The release of Neu5Gc from glycoproteoglycans is followed by labeling with the fluorescent dye DMB. Finally, DMB-labeled Neu5Gc is separated using reversed-phase liquid chromatography (RPLC) and detected by fluorescence (FLD) or mass spectrometry (MS). Since Neu5Gc is expressed non-humanly, it is potentially immunogenic. Therefore, it is essential to monitor the content of Neu5Gc in therapeutic glycoproteins as well as the levels of different salivary acids. Our assays cover a wide range of biological products. Samples do not need to be dried before the assay. Enzymatic release greatly reduces the degradation of Neu5Gc.
Fig.3 Process of Neu5Gc analysis. (CD BioGlyco)
CD BioGlyco aims to provide efficient services for research on biotherapeutic glycoproteins and more. We will provide a detailed Neu5Gc glycoengineering report. In addition to Neu5Gc, we also provide Neu5Ac and 2-Keto-3-deoxy-D-glycero-D-galacto-Nononic Acid glycoengineering services. If you have any need about our glycoengineering process, technology, or collaboration, please feel free to contact us. We are always ready to answer your questions.
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