SELECTION OF CARBOHYDRATE-BINDING CELL PHENOTYPES USING OLIGOSACCHARIDE-COATED MAGNETIC PARTICLES

Neoglycoconjugate coated magnetic beads were assessed for their abilit y to selectively isolate human cells with known anti-carbohydrate reac tivity, Four lung cancer cell lines, NCI-H146, NCI-N417D, SKMES-1, EKV X; two acute lymphoblastic leukemia lines, MOLT-4 and CCRF-CEM; and th e anti- Le(c) (isolactosamine) hybridoma, LU-BCRU-G7, were tested, The neoglycoconjugates (biotinylated pseudopolysaccharides) bound uniform ly to streptavidin coated magnetic beads as demonstrated by FITC label ed lectin, Streptavidin beads alone did not bind to any of the cell ty pes, The anti- Le(c) hybridoma cell line, LU BCRU-G7, demonstrated bin ding only to Le(c) pseudopolysaccharide coated magnetic beads, Subsequ ent incubation in the presence of unlabeled pseudopolysaccharide resul ted in the release of the beads from the cell surface, Although there was some heterogeneity within the individual lung and leukemic cell li nes, positive cells showed strong rosette formation with the coated be ads, The A(di) disaccharide coated beads showed binding in all four lu ng cancer cell lines, with the Le(c) and the H (type1) pseudopolysacch aride-bead conjugates only reactive in the N417 and H146 SCLC lines, T he range of L-selectin ligand-coated beads were all successful in bind ing to the acute lymphoblastic leukemia cell lines MOLT4 and CCRF-CEM, This approach provides a versatile model for the study of cell-surfac e carbohydrate interactions that should find application in many areas of cell biology.