RESUMO
Investigating unidentified weak-acting lectins is important for understanding glycan-related phenomena. We have developed an improved screening method for weak-acting lectins using glycan-conjugated magnetic beads (or glycobeads) involving a partial washing method and named it the mild enrichment procedure. Weak-acting lectins exist in equilibrium between bound lectin and free lectin produced by dissociation, whereas most tight-binding lectin exists in a bound state. The conventional washing step, in which the solution phase is replaced, may remove dissociated lectin from around the glycobeads; therefore, we attempted to leave a buffer space around the glycobeads to maintain the association-dissociation equilibrium of weak-acting lectins. Our results revealed that our mild enrichment procedure for screening for weak interactions, such as maltose-concanavalin A (K(a)â¼10(4)M(-1)) and lactose-peanut agglutinin (K(a)â¼10(3)M(-1)) interactions, was more effective than conventional batch methods.