本文關(guān)鍵詞：微量生物樣品中糖蛋白N-糖鏈電噴霧電離質(zhì)譜分析前處理方法的建立　出處：《分析化學(xué)》2016年02期 　論文類型：期刊論文

  更多相關(guān)文章： 微量分析 糖蛋白 N-糖鏈 電噴霧質(zhì)譜 前處理

【摘要】：基于電噴霧電離質(zhì)譜檢測技術(shù),建立了一種可靠、高效、簡單,適合于微量糖蛋白N-糖鏈解離、富集純化的方法。以糖蛋白牛胰核糖核酸酶(Rib B)和卵清白蛋白(OVA)為模型蛋白直接酶解,比較了4種方法純化酶解樣品的效果。比較了直接酶解和經(jīng)過聚偏氟乙烯(PVDF)膜富集后酶解微量復(fù)雜生物來源樣品胎牛血清的酶解效果,最終建立了微量生物樣品中糖蛋白N-糖鏈的質(zhì)譜分析前處理方法。采用PVDF膜吸附復(fù)雜生物樣品中的糖蛋白,N-糖苷酶F(PNGase F)酶直接在膜上完成糖鏈釋放(37℃,24 h),采用微晶纖維素柱結(jié)合石墨碳柱對(duì)糖鏈進(jìn)行富集純化,用于微克級(jí)胎牛血清和健康人血清中N-糖鏈質(zhì)譜分析的前處理。本方法通用性好,在微量生物樣品糖鏈質(zhì)譜分析檢測的前處理方面具有一定應(yīng)用價(jià)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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