Effect of crosslinking degree and thickness of thermosensitive imprinted layers on recognition and elution efficiency of protein imprinted magnetic microspheres
Effect of crosslinking degree and thickness of thermosensitive imprinted layers on recognition and elution efficiency of protein imprinted magnetic microspheres
复制标题
热敏印迹层交联度和厚度对蛋白质印迹磁性微球识别和洗脱效率的影响
DOI：
10.1016/j.snb.2015.11.082
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发表时间：
2016-03
期刊：
影响因子：
--
通讯作者：
Zhang Qiuyu
中科院分区：
文献类型：
--
作者：
Li Xiangjie;Zhou Jingjing;Tian Lei;Li Wei;Ali Zafar;Ali Nisar;Zhang Baoliang;Zhang Hepeng;Zhang Qiuyu
由于热敏印迹材料的响应性能和识别能力是相互矛盾的关系，在同时考虑二者的前提下，很难确定其交联度。此外，热敏印迹层厚度对目标蛋白识别的影响也难以定量评估。针对这两个问题，制备了一系列具有不同交联度和印迹层厚度的热敏磁性印迹微球。吸附实验结果表明，印迹层厚度为17 nm时，BSA印迹效果最佳。通过同时考虑响应性能和识别能力，确定交联度为20%作为平衡点。在此条件下，一次洗涤后的洗脱效率可达78.60%，保证了良好的再生性能。吸附容量和印迹因子分别可达42.01 mg/g和3.41，与以往报道的印迹材料相比有较大提高。并将该印迹微球应用于蛋白质混合物样品中BSA的分离，得到了令人振奋的结果。这些结果表明，该印迹微球有望应用于BSA的快速分离纯化。
Crosslinking degree is difficult to determine on the premise of simultaneously considering response performance and recognition capability in the thermosensitive imprinted materials due to their mutually contradictory relationship. In addition, the effect of thickness of thermosensitive imprinted layers on the recognition of target protein is also difficult to evaluate quantitatively. To solve these two problems, a series of thermosensitive magnetic imprinted microspheres with different crosslinking degree and imprinted layer thickness were fabricated. The adsorption results showed that imprinted layer thickness of 17 nm was the most appropriate for BSA imprinting. Crosslinking degree of 20% was determined as the balance point by simultaneously considering response performance and recognition capability. Under such conditions, the elution efficiency after once washing process was up to 78.60%, which guaranteed the satisfactory regeneration capability. The adsorption capacity and the imprinting factor could reach 42.01 mg/g and 3.41, respectively, which were greatly increased compared to those imprinted materials reported previously. Furthermore, the practical separation performance of the imprinted microspheres was investigated by separating BSA from the protein mixture sample, and the results were exciting. All these results indicated that these imprinted microspheres were expected to be applied to the rapid isolation and purification of BSA.
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