Selection of carbohydrate-binding cell phenotypes using oligosaccharide-coated magnetic particles

Neoglycoconjugate coated magnetic beads were assessed for theirability to selectively isolate human cells with known anti-carbohydratereactivity. Four lung cancer cell lines, NCI-H146, NCI-N417D,SKMES-1, EKVX; two acute lymphoblastic leukemia lines, MOLT-4and CCRF-CEM; and the anti- Le^c (isolactosamine) hybridoma,LU-BCRU-G7, were tested. The neoglycoconjugates (biotinylatedpseudopolysaccharides) bound uniformly to streptavidin coatedmagnetic beads as demonstrated by FTTC labeled lectin. Streptavidinbeads alone did not bind to any of the cell types. The anti-Le^c hybridoma cell line, LU BCRU-G7, demonstrated binding onlyto Le^c pseudopolysaccharide coated magnetic beads. Subsequentincubation in the presence of unlabeled pseudopolysaccharideresulted in the release of the beads from the cell surface.Although there was some heterogeneity within the individuallung and leukemic cell lines, positive cells showed strong rosetteformation with the coated beads. The A_dl disaccharide coatedbeads showed binding in all four lung cancer cell lines, withthe Le^c and the H (type1) pseudopolysaccharide-bead conjugatesonly reactive in the N417 and H146 SCLC lines. The range ofL-selectin ligand-coated beads were all successful in bindingto the acute lymphoblastic leukemia cell lines MOLT4 and CCRF-CEM.This approach provides a versatile model for the study of cell-surfacecarbohydrate interactions that should find application in manyareas of cell biology. carbohydrate-coated magnetic beads cell selec-tion lectins neoglycoconjugates pseudopolysaccharides