首页> 外文期刊>Talanta: The International Journal of Pure and Applied Analytical Chemistry >Surface plasmon resonance for real-time study of lectin-carbohydrate interactions for the differentiation and identification of glycoproteins
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Surface plasmon resonance for real-time study of lectin-carbohydrate interactions for the differentiation and identification of glycoproteins

机译:表面等离振子共振用于凝集素-碳水化合物相互作用的实时研究,以区分和鉴定糖蛋白

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A study of specific interactions between lectins and glycoproteins has been carried out using surface plasmon resonance (SPR) in a flow-injection mode. Lectins were covalently immobilised on the surfaces of the microfluidic sensor chip via amine coupling and serum glycoproteins were injected into the flow channels. Specific lectin-glycoprotein interactions caused the shift of refractive index proportional to the mass concentration accumulated on the channel surface. Lectins showed different affinity to the tested glycoproteins and each glycoprotein displayed its own lectin-binding pattern. It is possible to distinguish and identify even glycoproteins with similar sugar structures by simple and quick screening. The working conditions of the assay were optimised. The lectin-based SPR made it possible to carry out the label-free detection of glycoproteins within a broad concentration range with a good linearity. Regeneration conditions for the surface of the sensor chip were found and optimised. Combination of 10 mM HCl and 10 mM glycine-HCl (pH 2.5) removes the bound glycoproteins from the lectin surface without damaging it. The kinetic and affinity parameters of lectin-glycoprotein binding were evaluated. The proposed method was tested on human glycosylated serum. Combination of the lectin panel with SPR is suitable both for specific screening and for sensitive assay of serum glycoproteins.
机译:凝集素和糖蛋白之间的特殊相互作用的研究已经使用表面等离振子共振(SPR)在流动注射模式下进行。通过胺偶联将凝集素共价固定在微流体传感器芯片的表面上,并将血清糖蛋白注射到流动通道中。特定的凝集素-糖蛋白相互作用导致折射率的变化与累积在通道表面的质量浓度成正比。凝集素对测试的糖蛋白表现出不同的亲和力,每种糖蛋白表现出自己的凝集素结合模式。通过简单快速的筛选,甚至可以区分和鉴定具有相似糖结构的糖蛋白。优化了测定的工作条件。基于凝集素的SPR使得可以在宽浓度范围内以良好的线性进行糖蛋白的无标记检测。找到并优化了传感器芯片表面的再生条件。 10 mM HCl和10 mM甘氨酸HCl(pH 2.5)结合使用可从凝集素表面去除结合的糖蛋白,而不会对其造成破坏。评价了凝集素-糖蛋白结合的动力学和亲和力参数。该方法在人糖基化血清上进行了测试。凝集素面板与SPR的组合适用于特异性筛选和血清糖蛋白的敏感测定。

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