Analysis of unconventional approaches for the rapid detection of surface lectin binding ligands on human cell lines.
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.
Welty
Lily Anne Y
LA
Department of Biology and The Center for Cancer and Developmental Biology, California State University Northridge, 18111 Nordhoff Street, Northridge, CA 91330-8303, USA.
Heinrich
Eileen L
EL
Garcia
Karina
K
Banner
Lisa R
LR
Summers
Michael L
ML
Baresi
Larry
L
Metzenberg
Stan
S
Coyle-Thompson
Cathy
C
Oppenheimer
Steven B
SB
D002460:Cell Line; D045744:Cell Line, Tumor; D003106:Colon / Q000737:chemistry* / Q000473:pathology; D003110:Colonic Neoplasms / Q000737:chemistry* / Q000473:pathology; D005470:Fluorometry / Q000379:methods*; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D037102:Lectins / Q000032:analysis / Q000378:metabolism*; D008024:Ligands; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D008863:Microspheres; D010835:Phytohemagglutinins / Q000032:analysis / Q000378:metabolism; D037121:Plant Lectins / Q000032:analysis / Q000378:metabolism; D011485:Protein Binding; D015203:Reproducibility of Results; D012685:Sepharose; D014909:Wheat Germ Agglutinins / Q000032:analysis / Q000378:metabolism
D037102:Lectins; D008024:Ligands; D008565:Membrane Proteins; D010835:Phytohemagglutinins; D037121:Plant Lectins; D014909:Wheat Germ Agglutinins; C033619:lentil lectin; D012685:Sepharose