Analysis of unconventional approaches for the rapid detection of surface lectin binding ligands on human cell lines
| Autor: | Lily Anne Y. Welty, Steven B. Oppenheimer, Michael L. Summers, Stan Metzenberg, Eileen L. Heinrich, Lisa R. Banner, Larry Baresi, Karina Garcia, Cathy Coyle-Thompson |
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| Rok vydání: | 2006 |
| Předmět: |
Histology
Colon Wheat Germ Agglutinins Plasma protein binding Biology Ligands Article Fluorescence spectroscopy Cell Line Sepharose Cell Line Tumor Lectins Humans Fluorometry Phytohemagglutinins Mass screening Histocytochemistry Ligand Membrane Proteins Reproducibility of Results Cell Biology General Medicine Fluorescence Microspheres Wheat germ agglutinin Biochemistry Cell culture Colonic Neoplasms Plant Lectins Protein Binding |
| Zdroj: | Acta Histochemica. 107:411-420 |
| ISSN: | 0065-1281 |
| Popis: | For over a decade our laboratory has developed and used a novel histochemical assay using derivatized agarose beads to examine the surface properties of various cell types. Most recently, we have used this assay to examine lectin binding ligands on two human cell types, CCL-220, a colon cancer cell line, and CRL-1459, a non-cancer colon cell line. We found that CCL-220 cells bound specific lectins better than CRL-1459, and this information was used to test for possible differential toxicity of these lectins in culture, as a possible approach in the design of more specific anti-cancer drugs. Although we have examined the validity of the bead-binding assay in sea urchin cell systems, we have not previously validated this technique for mammalian cells. Here the binding results of the bead assay are compared with conventional fluorescence assays, using lectins from three species (Triticum vulgaris, Phaseolus vulgaris, and Lens culinaris) on the two colon cell lines. These lectins were chosen because they seemed to interact with the two cell lines differently. Binding results obtained using both assays were compared for frozen, thawed and fixed; cultured and fixed; and live cells. Both qualitative and quantitative fluorescence results generally correlated with those using the bead assay. Similar results were also obtained with all of the three different cell preparation protocols. The fluorescence assay was able to detect lower lectin binding ligand levels than the bead assay, while the bead assay, because it can so rapidly detect cells with large numbers of lectin binding ligands, is ideal for initial screening studies that seek to identify cells that are rich in surface binders for specific molecules. The direct use of frozen, thawed and fixed cells allows rapid mass screening for surface molecules, without the requirement for costly and time consuming cell culture. |
| Databáze: | OpenAIRE |
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