微量生物样品中糖蛋白N-糖链电喷雾电离质谱分析前处理方法的建立

发布时间：2018-01-18 14:22

  本文关键词：微量生物样品中糖蛋白N-糖链电喷雾电离质谱分析前处理方法的建立　出处：《分析化学》2016年02期 　论文类型：期刊论文

  更多相关文章： 微量分析 糖蛋白 N-糖链 电喷雾质谱 前处理

【摘要】：基于电喷雾电离质谱检测技术,建立了一种可靠、高效、简单,适合于微量糖蛋白N-糖链解离、富集纯化的方法。以糖蛋白牛胰核糖核酸酶(Rib B)和卵清白蛋白(OVA)为模型蛋白直接酶解,比较了4种方法纯化酶解样品的效果。比较了直接酶解和经过聚偏氟乙烯(PVDF)膜富集后酶解微量复杂生物来源样品胎牛血清的酶解效果,最终建立了微量生物样品中糖蛋白N-糖链的质谱分析前处理方法。采用PVDF膜吸附复杂生物样品中的糖蛋白,N-糖苷酶F(PNGase F)酶直接在膜上完成糖链释放(37℃,24 h),采用微晶纤维素柱结合石墨碳柱对糖链进行富集纯化,用于微克级胎牛血清和健康人血清中N-糖链质谱分析的前处理。本方法通用性好,在微量生物样品糖链质谱分析检测的前处理方面具有一定应用价值。
[Abstract]:Based on the electrospray ionization mass spectrometry (EI-MS), a reliable, efficient, simple and suitable for the dissociation of N-sugar chain of trace glycoprotein was established. The method of enrichment and purification. Rib B) and ovalbumin ovalbumin (OVA) were directly hydrolyzed with glycoprotein bovine pancreatic ribonuclease (Rib B) and ovalbumin (OVA) as model proteins. The effects of four methods on the purification of samples were compared. The enzymatic hydrolysis of fetal bovine serum was compared after direct enzymatic hydrolysis and polyvinylidene fluoride PVDF (polyvinylidene fluoride PVDF) membrane enrichment. Finally, a pre-treatment method for the analysis of glycoprotein N-sugar chain in trace biological samples by mass spectrometry was established. PVDF membrane was used to adsorb glycoprotein from complex biological samples. N-glucosidase (FN) PNGase Fase was released directly on the membrane at 37 鈩，

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