Vicia villosa B(4) (VVL-B(4)) is an important lectin for detecting exposed Tn (GalNAcalpha1-Ser/Thr) determinants on cancer cells. In order to elucidate the binding factors involved in VVL-B(4) and glycotope interaction, the binding properties of this lectin were analyzed by enzyme-linked lectinosorbent and inhibition assays. From the results, it is concluded that the most critical factor affecting VVL-B(4) binding is polyvalency at the alpha anomer of Gal with -NH CH(3)CO at carbon-2 (Tn epitope), which enhances the reactivity by 3.3x10(5) times over monovalent Gal. The reactivities of glycotopes can be ranked as follows: high density Tn cluster >Tn glycopeptides (MW<3.0x10(3) > monomeric Tn to tri- Tn glycopeptides > other GalNAcalpha/beta-related structural units>Gal and Galalpha- or beta-linked ligands, demonstrating the essential role of the polyvalency of Tn glycotopes in the enhancement of the binding.
