本文選題：反相色譜 + C18固相萃取小柱��； 參考：《北京協(xié)和醫(yī)學(xué)院》2015年碩士論文

【摘要】：研究背景與目的：針對(duì)少量且復(fù)雜蛋白質(zhì)組樣品,開(kāi)發(fā)一種耗時(shí)短,操作簡(jiǎn)便的預(yù)處理方法。研究方法：以人的腦海馬組織蛋白為研究對(duì)象,使用反相C18固相萃取小柱進(jìn)行樣品分離,并與反相高效液相色譜法進(jìn)行比較分析。根據(jù)四種乙腈梯度設(shè)置方案的分離效果以確定最佳的洗脫條件。研究結(jié)果：八種乙腈濃度洗脫僅耗時(shí)10 min,為反相高效液相色譜法的1/4,但鑒定蛋白總數(shù)占高效液相色譜法的85.5%。乙腈洗脫濃度為5%,15%,20%,90%時(shí)分離30μg人的海馬多肽混合物可以得到較優(yōu)的分離效果,兩次重復(fù)實(shí)驗(yàn)鑒定蛋白重復(fù)率達(dá)到70%。研究結(jié)論：該方法操作易行,省時(shí),成本低,是進(jìn)行蛋白質(zhì)組學(xué)分析中少量樣品的一個(gè)簡(jiǎn)便預(yù)處理方法。研究背景與目的：針對(duì)尿液樣品有易獲取,處理方法簡(jiǎn)便的優(yōu)點(diǎn),近年來(lái)尿蛋白質(zhì)組學(xué)引起人們廣泛關(guān)注。以往動(dòng)脈粥樣硬化的尿蛋白質(zhì)組學(xué)研究主要應(yīng)用毛細(xì)管電泳-質(zhì)譜聯(lián)用技術(shù),報(bào)道多種多肽標(biāo)記物。本文從尿液蛋白質(zhì)組學(xué)入手,目的在于發(fā)現(xiàn)動(dòng)脈粥樣硬化疾病在蛋白水平的表達(dá)差異。研究方法：采用iTRAQ (Isobaric tags for relative and absolute quantitation)定量標(biāo)記串聯(lián)質(zhì)譜分析技術(shù)從動(dòng)脈粥樣硬化患者及對(duì)照組尿液樣品中發(fā)現(xiàn)差異表達(dá)蛋白。研究結(jié)果：經(jīng)過(guò)質(zhì)譜分析,Mascot 及 Scaffold數(shù)據(jù)庫(kù)檢索從尿液中共鑒定856個(gè)蛋白,其中定量蛋白737個(gè)。動(dòng)脈粥樣硬化患者及對(duì)照組之間尿液中存在90個(gè)差異表達(dá)蛋白,倍數(shù)變化設(shè)置1.5倍。經(jīng)過(guò)IPA功能分析發(fā)現(xiàn)差異表達(dá)蛋白與糖類(lèi),脂類(lèi)代謝及炎癥反應(yīng)密切相關(guān)。研究結(jié)論：尿液是動(dòng)脈粥樣硬化疾病標(biāo)記物發(fā)現(xiàn)的理想來(lái)源。尿液中差異蛋白表達(dá)表達(dá)情況可以反映動(dòng)脈粥樣硬化疾病狀態(tài)下細(xì)胞內(nèi)外的病理生理變化。
[Abstract]:Background & AIM: to develop a simple and time consuming pretreatment method for a small amount of complex proteome samples. Methods: the human brain tissue protein was separated by reverse phase C 18 solid phase extraction column, and compared with that of the reverse phase high performance liquid chromatography (RP-HPLC). According to the separation effect of four acetonitrile gradient setting schemes, the best elution conditions were determined. The results showed that the elution time of eight acetonitrile concentrations was only 10 min, which was 1 / 4 of that by RP-HPLC, but the total number of identified proteins was 85.5% of that by HPLC. The separation of 30 渭 g human hippocampal polypeptide mixture with acetonitrile elution concentration was 20% and 90 渭 g, and the protein repetition rate reached 70%. Conclusion: this method is easy to operate, time saving and low cost. It is a simple pretreatment method for a small amount of samples in proteomics analysis. Background & AIM: because urine samples are easy to be obtained and processed, proteomics of urine has attracted more and more attention in recent years. In the past, urinary proteomics of atherosclerosis was mainly reported by capillary electrophoresis-mass spectrometry. The aim of this paper is to find out the difference of protein expression in atherosclerotic diseases. Methods: the differentially expressed proteins were found in urine samples from patients with atherosclerosis and control group by iTRAQ tags for relative and absolute quantitative tandem mass spectrometry. Results: a total of 856 proteins, including 737 quantitative proteins, were identified from urine by Mascot and Scaffold database. There were 90 differentially expressed proteins in urine between atherosclerotic patients and controls. IPA functional analysis showed that differentially expressed proteins were closely related to carbohydrate, lipid metabolism and inflammation. Conclusion: urine is an ideal source for the discovery of markers for atherosclerosis. The expression of differential protein in urine can reflect the pathophysiological changes of cells in atherosclerotic state.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R541.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王璐;周蘭蘭;錢(qián)小紅;張養(yǎng)軍;;強(qiáng)陽(yáng)離子交換色譜分離多肽混合物的條件優(yōu)化[J];色譜;2010年04期

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本文編號(hào)：1868414