Soper Anastasiya S, Aird Steven D
Department of Chemistry, Norfolk State University, 700 Park Avenue, Norfolk, VA 23504, USA.
J Chromatogr A. 2007 Jun 22;1154(1-2):308-18. doi: 10.1016/j.chroma.2007.03.126. Epub 2007 Apr 6.
Some glycoproteins bind so tightly to concanavalin A Sepharose that common desorption techniques are ineffective, so a systematic exploration of factors affecting desorption of cottonmouth venom glycoproteins was undertaken. Glycoprotein desorption is greatly improved by introducing up to four pauses of 5-10 min duration into the elution step. Eluent concentrations above 250 mM methylglucoside or 500 mM methyl-mannoside reduced glycoprotein desorption. Eluent NaCl diminished glycoprotein desorption. Most venom glycoproteins desorb more readily as pH diminishes from 6.0 to 4.0, but phosphodiesterase shows the opposite pattern. Eluents recommended by the supplier for desorbing solutes or for column cleaning were ineffectual.
一些糖蛋白与伴刀豆球蛋白A琼脂糖结合紧密，以至于常用的解吸技术无效，因此对影响棉口蛇毒糖蛋白解吸的因素进行了系统探索。通过在洗脱步骤中引入多达四个持续时间为5 - 10分钟的停顿，糖蛋白的解吸得到了极大改善。洗脱液中甲基葡萄糖苷浓度高于250 mM或甲基甘露糖苷浓度高于500 mM会降低糖蛋白解吸。洗脱液中的氯化钠会减少糖蛋白解吸。随着pH从6.0降至4.0，大多数毒液糖蛋白更容易解吸，但磷酸二酯酶表现出相反的模式。供应商推荐的用于解吸溶质或柱清洗的洗脱液无效。
