Biotin-based Cell Surface Glycoengineering Service at CD BioGlyco

Cell surface glycans are involved in cell migration, immune regulation, cell aging and death, and the occurrence and development of diseases in organisms. Cell surface glycoengineering provides a powerful means to modulate and redesign cell surface glycans. CD BioGlyco is a professional glycobiology company dedicated to helping clients solve their difficulties and problems in the field of glycobiology. To better help clients analyze cell surface glycan-mediated signaling events and their downstream cellular molecular mechanisms, we have established a first-class Cell Surface Glycoengineering platform. We provide clients with glycoengineering services for cell surface glycans based on Metabolic Pathways, Chemical Conjugation, Liposome Fusion, and Genetic methods to achieve new biological insights and new cellular functions of cell surface glycans.

At CD BioGlyco, we covalently install many reactive groups (such as azides, thiols, isonitriles, etc.) and chemical groups carrying reporters such as biotin, fluorescent dyes, cross-linkers, etc. onto cell surface glycans through bioorthogonal chemistry, thus imparting new chemical functions to the glycan structure. Common examples are the fluorescent labeling or imaging of cell surface glycans through glycoengineering techniques based on chemical conjugation.

Biotin is ideal for labeling proteins and macromolecules and is commonly used in enzyme-linked immunosorbent assays (ELISA), western blots, and many other applications. Biotin labeling has three advantages:

• Biotin has a strong affinity.
• Biotin is relatively smaller than globular proteins, minimizing interference in the protein.
• Biotin has a valeric acid side chain, easy to derivatize and conjugate with chemical structures.

We introduce biotin onto cell surface glycan as follows:

1) Introduction of aldehydes onto cell surface sialic acid by mild periodate oxidation.

2) Under the nucleophilic catalysis of aniline, biotin-tagged molecules are attached to cell surface glycans through oximes and hydrazones.

3) Evaluation of biotin labeling efficiency by fluorescence imaging of sialylated proteins and gel electrophoresis.

Fig.1 Biotin-based cell surface glycoengineering service. (CD BioGlyco)

Publication

Paper Title: High-efficiency labeling of glycoproteins on living cells

Technology: Biotin-labeled cell surface glycoproteins

Journal: Scientific Reports

IF: 48.0

Published: 2009

Results: The authors used two methods to introduce aminooxy-biotin onto cell surface sialic acid: mild periodate oxidation and 9-deoxy-9-N-carboxy- benzaldehyde-NeuAc (^BANeuAc) metabolic labeling. BJA-B subclone K88 (K88) with wild-type levels of sialic acid was used as a control. The results showed that K88 cells showed high levels of biotinylation after periodate oxidation. K88 cells cultured with ^BANeuAc were not biotinylated, possibly due to competition with endogenous NeuAc pools that were not incorporated.

Fig.2 The extent of aminooxy-biotin ligation to K88 and K20 cells by flow cytometry. (Zeng, et al., 2009)

Highlights

• Glycan engineering allows reporters such as biotin, fluorescent dyes, and cross-linking agents to be installed on cell surface glycans, providing an effective method for molecular imaging of glycans.
• Glycoengineering controls the glycan group displayed on the cell surface, providing a unique opportunity to deeply dissect the molecular interactions and biological functions of glycans.
• Biotin has the advantages of strong affinity, low interference, and easy conjugation making it widely used in various detections such as ELISA and western blotting.

CD BioGlyco has rich experience in glycoengineering and has continuously improved our glycoengineering technology and proposal. We would like to share our knowledge in this area to help you achieve modification and analysis of cell surface glycans. Please feel free to contact us for more information.