title: Magnetic beads-assisted mild enrichment procedure for weak-binding lectins.
abstract: 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)∼10(4)M(-1)) and lactose-peanut agglutinin (K(a)∼10(3)M(-1)) interactions, was more effective than conventional batch methods.
authors.affiliation: Department of Materials and Life Science, Seikei University, Musashino-shi, Tokyo, Japan. ktotani@st.seikei.ac.jp
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Totani
authors.forename: Haruka
authors.initials: H
authors.lastname: Miyazawa
authors.forename: Shino
authors.initials: S
authors.lastname: Kurata
authors.forename: Yukishige
authors.initials: Y
authors.lastname: Ito
mesh_terms: D000818:Animals; D002021:Buffers; D002417:Cattle; D003208:Concanavalin A / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D008280:Magnetics / Q000379:methods*; D008320:Maltose / Q000378:metabolism; D051379:Mice; D008863:Microspheres; D019887:Peanut Agglutinin / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding
chemical_list: D002021:Buffers; D037102:Lectins; D019887:Peanut Agglutinin; D011134:Polysaccharides; D003208:Concanavalin A; D008320:Maltose