Magnetic beads-assisted mild enrichment procedure for weak-binding lectins |
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Authors: | Kiichiro Totani Haruka Miyazawa Shino Kurata Yukishige Ito |
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Institution: | aDepartment of Materials and Life Science, Seikei University, 3-3-1 Kichijoji-kitamachi, Musashino-shi, Tokyo 180-8633, Japan;bRIKEN Advanced Science Institute and ERATO Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan |
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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 (Ka ∼ 104 M−1) and lactose–peanut agglutinin (Ka ∼ 103 M−1) interactions, was more effective than conventional batch methods. |
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Keywords: | Lectin Weak binding Magnetic beads Enrichment Concanavalin A (ConA) Peanut agglutinin (PNA) |
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