Lectin Staining for Detection of Sialic Acid Residues on Cell Surface of Wild Type Chinese Hamster Ovary (CHO) and HeLa Cells

Sialic acid plays important roles in human physiology particularly in cell-cell interaction and communication, cell-cell signalling, carbohydrate-protein interactions, cellular aggregation, development processes, immune reactions, reproduction, neurobiology, and human diseases. In this study, we inv...

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Published inAsia-Pacific journal of molecular biology and biotechnology Vol. 31; p. 96
Main Authors Rahman, Salina Abdul, Omar, Affandi, Nordin, Fatimah Diana Amin, Kamarudin, Balqis, Muhamad, Nur Jannaim, Jalil, Julaina Abdul
Format Journal Article
LanguageEnglish
Published Kuala Lumpur Malaysian Society for Molecular Biology and Biotechnology 01.06.2023
Subjects
Acids
Carbohydrates
Cell interactions
Cell surface
Exo-a-sialidase
Fluorescence
Fluorescence microscopy
Galactose
Glycolipids
Glycoproteins
Labeling
Lectins
Nervous system
Neurosciences
Peptide nucleic acids
Polysaccharides
Protein interaction
Residues
Rhodamine
Staining
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Summary:Sialic acid plays important roles in human physiology particularly in cell-cell interaction and communication, cell-cell signalling, carbohydrate-protein interactions, cellular aggregation, development processes, immune reactions, reproduction, neurobiology, and human diseases. In this study, we investigated the presence of sialic acid on the surface of CHO cells by using rhodamine-linked peanut agglutinin (PNA-Rho) lectin, which binds to Galꞵ1-3GlcNAc only if the terminal sialic acid is absent. Wild type CHO and HeLa cells were seeded on cover slips in a 24 well plate. Wild type HeLa cells were chosen as a positive control. After overnight incubation, one set of wild type CHO and HeLa cells were treated with neuraminidase to cause release of sialic acid residues from surface glycoproteins and glycolipids. Subsequently PNA-Rho was used to stain all cells and the samples were visualised using fluorescence microscopy. Lectin staining without neuraminidase treatment showed no fluorescent labelling. In contrast, following neuraminidase treatment, fluorescent detection of PNA-Rho was detected in both wild type CHO and HeLa cells. This indicating neuraminidase treatment successfully removed sialic acid normally present on cell surface glycans, exposing more galactose residues for PNA-Rh binding. In summary, sialic acid residues are found on the surface of both wild type CHO and HeLa cells. Future work will be done to see if these lectins can be used to monitor changes in sialic acid levels on cell surfaces especially when its production is impaired due to mutations.
ISSN:0128-7451