水溶性硼酸亲和富集材料的制备及基于酶联免疫吸附法检测人肝微粒体糖蛋白的应用

发布时间：2018-03-14 14:51

本文选题：糖蛋白　切入点：硼酸　出处：《分析化学》2017年09期 　论文类型：期刊论文

【摘要】：生物样品中的糖蛋白丰度低,且在检测中易受到其它非糖蛋白的抑制和干扰,需在分析检测前对糖蛋白进行富集,但常规的基于固相材料的糖蛋白富集方法不易与生物技术中最经典的酶联免疫吸附法(ELISA)兼容。本研究以树枝状聚合物PAMAM 4.0为载体,结合硼酸亲和技术,制备了新型水溶性硼酸亲和富集材料(DBC),并将其应用于基于ELISA的人肝微粒体中糖蛋白的检测。采用标准糖蛋白对DBC富集条件进行优化,然后考察其灵敏度和抗干扰能力,将优化后的方法应用于复杂样品人肝微粒体糖蛋白富集。结果表明,DBC对糖蛋白的富集选择性可高达100000倍,可将糖蛋白的富集信号提高100倍。以DBC为富集材料,与ELISA分析技术相结合,只需一步简单的孵育,即可实现生物样品中糖蛋白的高灵敏度、高选择性检测,为疾病相关的糖蛋白组学研究提供了一种有效的检测手段。
[Abstract]:The abundance of glycoprotein in biological samples is low, and it is easy to be inhibited and interfered by other non-glycoproteins in the detection, so it is necessary to enrich the glycoproteins before analysis and detection. However, the conventional method based on solid phase material for glycoprotein enrichment is not easy to be compatible with the most classical enzyme linked immunosorbent assay (Elisa) in biotechnology. In this study, dendritic polymer PAMAM 4.0 was used as carrier and boric acid affinity technique was used. A new water-soluble boric acid affinity enrichment material was prepared and applied to the detection of glycoproteins in human liver microsomes based on ELISA. The enrichment conditions of DBC were optimized by standard glycoproteins, and the sensitivity and anti-interference ability were investigated. The optimized method was applied to the enrichment of human liver microsomal glycoproteins in complex samples. The results showed that the enrichment selectivity of DBC to glycoprotein was up to 100000 times, and the enrichment signal of glycoprotein could be increased by 100 times. DBC was used as enrichment material. In combination with ELISA analysis technique, the high sensitivity and selectivity detection of glycoprotein in biological samples can be realized by one simple incubation, which provides an effective method for the study of glycoproteomics related to disease.
【作者单位】：大连医科大学药学院;大连医科大学基础医学院;
【基金】：国家自然科学基金(No.21505015) 辽宁省教育厅科学研究重点项目(No.LZ2016001) 大连市支持高层次人才创新创业计划(No.2016RQ041) 辽宁省自然科学基金(No.20170540286) 国家留学基金(No.201608210312)资助~~
【分类号】：O647.3;R96

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