Starburst PAMAM dendrimers ending with mannopyranoside residues were readily synthesized in large scale and good yields from commercially available dendrimers bearing high-density amine functionality on their surface and p-isothiocyanatophenyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranoside. The first four generations of this novel class of monodispersed neoglycoconjugates having up to 32 mannoside units were evaluated as ligands for the phytohemagglutinins from concanavalin A (Con A) and Pisum sativum (pea lectin) using enzyme-linked lectin assay (ELLA) and turbidimetric analyses. The binding properties of these glycodendrimers, together with reference monosaccharides, were determined using yeast mannan as a coating antigen and peroxidase-labeled lectins. These mannosylated dendrimers were demonstrated to be potent inhibitors with IC50 values 400 times better than those of monomeric methyl alpha-D-mannopyranoside taken as a standard. Their lipophilic character was shown to be sufficient for their direct use as coating antigens in microtiter plate assays. Moreover, their ability to bind and form insoluble carbohydrate-lectin complexes was also demonstrated by radial double immunodiffusion and turbidimetric analyses. Furthermore, the ability of these ligands to selectively precipitate a mannose-binding protein (Con A) from a crude lectin mixture was also demonstrated using polyacrylamide gel electrophoresis (SDS-PAGE). These multivalent neoglycoconjugates were shown to constitute novel biochromatography materials of high affinity for the easy isolation of carbohydrate-binding proteins.