Department of Materials and Life Science, Seikei University, Musashino-shi, Tokyo, Japan.
Anal Biochem. 2011 Apr 1;411(1):50-7. doi: 10.1016/j.ab.2010.12.023. Epub 2010 Dec 22.
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.
研究弱活性凝集素对于了解糖相关现象非常重要。我们开发了一种使用糖缀合磁珠（或糖珠）的改进的弱活性凝集素筛选方法，涉及部分洗涤方法，并将其命名为温和富集程序。弱活性凝集素存在于结合的凝集素和由解离产生的游离凝集素之间的平衡中，而大多数强结合的凝集素存在于结合状态。传统的洗涤步骤，即用溶液相替代，可能会从糖珠周围去除解离的凝集素；因此，我们试图在糖珠周围留下缓冲空间，以维持弱活性凝集素的缔合-解离平衡。我们的结果表明，我们用于筛选弱相互作用的温和富集程序，例如麦芽糖结合刀豆球蛋白 A（K(a)∼10(4)M(-1))和乳糖-花生凝集素（K(a)∼10(3)M(-1))相互作用，比传统的批量方法更有效。
