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Fluorescent carbohydrate probes for cell lectins.
O Galanina1, A Feofanov, A B Tuzikov
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
|October 18, 2001
Summary
Fluorescein-labeled carbohydrate probes specifically detect cellular lectins in cell cultures and tissue sections. These advanced probes enable precise visualization and analysis of lectin binding sites, even at low expression levels.
Area of Science:
- Biochemistry
- Cell Biology
- Analytical Chemistry
Background:
- Cellular lectins play crucial roles in biological processes.
- Developing specific tools to study lectin-carbohydrate interactions is essential.
- Existing methods may lack specificity or sensitivity for low lectin expression.
Purpose of the Study:
- To synthesize and validate fluorescein-labeled carbohydrate probes for lectin detection.
- To assess the specificity and utility of these probes in cellular and tissue contexts.
- To demonstrate the application of confocal spectral imaging for high-resolution lectin analysis.
Main Methods:
- Synthesis of fluorescein-labeled carbohydrate-polyacrylamide (Glyc-PAA-flu) probes.
- Flow cytometry assays with lectin-transfected cells (CHO-E-selectin, COS-siglec-9).
- Tumor xenograft models in nude mice and analysis of tumor sections.
- Confocal spectral imaging for high-resolution probe localization and quantification.
Main Results:
- Fluorescent Glyc-PAA-flu probes showed specific binding to lectin-transfected cells.
- Binding was inhibited by unlabeled carbohydrate ligands, confirming specificity.
- Probes detected E-selectin on tumor sections from transfected cells.
- Confocal spectral imaging allowed accurate separation of signals and 3D submicron resolution analysis.
Conclusions:
- Fluorescein-labeled carbohydrate probes are effective tools for specific lectin detection.
- These probes are applicable to both cultured cells and tissue sections.
- Confocal spectral imaging enhances lectin visualization and quantification, especially for low expression levels.