本文選題：糖蛋白　切入點(diǎn)：硼酸　出處：《分析化學(xué)》2017年09期 　論文類(lèi)型：期刊論文

【摘要】：生物樣品中的糖蛋白豐度低,且在檢測(cè)中易受到其它非糖蛋白的抑制和干擾,需在分析檢測(cè)前對(duì)糖蛋白進(jìn)行富集,但常規(guī)的基于固相材料的糖蛋白富集方法不易與生物技術(shù)中最經(jīng)典的酶聯(lián)免疫吸附法(ELISA)兼容。本研究以樹(shù)枝狀聚合物PAMAM 4.0為載體,結(jié)合硼酸親和技術(shù),制備了新型水溶性硼酸親和富集材料(DBC),并將其應(yīng)用于基于ELISA的人肝微粒體中糖蛋白的檢測(cè)。采用標(biāo)準(zhǔn)糖蛋白對(duì)DBC富集條件進(jìn)行優(yōu)化,然后考察其靈敏度和抗干擾能力,將優(yōu)化后的方法應(yīng)用于復(fù)雜樣品人肝微粒體糖蛋白富集。結(jié)果表明,DBC對(duì)糖蛋白的富集選擇性可高達(dá)100000倍,可將糖蛋白的富集信號(hào)提高100倍。以DBC為富集材料,與ELISA分析技術(shù)相結(jié)合,只需一步簡(jiǎn)單的孵育,即可實(shí)現(xiàn)生物樣品中糖蛋白的高靈敏度、高選擇性檢測(cè),為疾病相關(guān)的糖蛋白組學(xué)研究提供了一種有效的檢測(cè)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.
【作者單位】： 大連醫(yī)科大學(xué)藥學(xué)院;大連醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(No.21505015) 遼寧省教育廳科學(xué)研究重點(diǎn)項(xiàng)目(No.LZ2016001) 大連市支持高層次人才創(chuàng)新創(chuàng)業(yè)計(jì)劃(No.2016RQ041) 遼寧省自然科學(xué)基金(No.20170540286) 國(guó)家留學(xué)基金(No.201608210312)資助~~
【分類(lèi)號(hào)】：O647.3;R96

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