Magnetic beads-assisted mild enrichment procedure for weak-binding lectins
| Autor: | Shino Kurata, Haruka Miyazawa, Yukishige Ito, Kiichiro Totani |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Biophysics
Buffers Biochemistry Dissociation (chemistry) Weak binding Magnetics Mice Peanut Agglutinin Agglutinin Polysaccharides Lectins Screening method Concanavalin A Animals Maltose Molecular Biology Chromatography biology Chemistry Lectin Cell Biology Solution phase Microspheres biology.protein Cattle Protein Binding |
| Zdroj: | Analytical biochemistry. 411(1) |
| ISSN: | 1096-0309 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect