發(fā)布時間：2018-05-10 07:32

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

【摘要】：研究背景與目的：針對少量且復雜蛋白質(zhì)組樣品,開發(fā)一種耗時短,操作簡便的預處理方法。研究方法：以人的腦海馬組織蛋白為研究對象,使用反相C18固相萃取小柱進行樣品分離,并與反相高效液相色譜法進行比較分析。根據(jù)四種乙腈梯度設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,兩次重復實驗鑒定蛋白重復率達到70%。研究結(jié)論：該方法操作易行,省時,成本低,是進行蛋白質(zhì)組學分析中少量樣品的一個簡便預處理方法。研究背景與目的：針對尿液樣品有易獲取,處理方法簡便的優(yōu)點,近年來尿蛋白質(zhì)組學引起人們廣泛關注。以往動脈粥樣硬化的尿蛋白質(zhì)組學研究主要應用毛細管電泳-質(zhì)譜聯(lián)用技術(shù),報道多種多肽標記物。本文從尿液蛋白質(zhì)組學入手,目的在于發(fā)現(xiàn)動脈粥樣硬化疾病在蛋白水平的表達差異。研究方法：采用iTRAQ (Isobaric tags for relative and absolute quantitation)定量標記串聯(lián)質(zhì)譜分析技術(shù)從動脈粥樣硬化患者及對照組尿液樣品中發(fā)現(xiàn)差異表達蛋白。研究結(jié)果：經(jīng)過質(zhì)譜分析,Mascot 及 Scaffold數(shù)據(jù)庫檢索從尿液中共鑒定856個蛋白,其中定量蛋白737個。動脈粥樣硬化患者及對照組之間尿液中存在90個差異表達蛋白,倍數(shù)變化設置1.5倍。經(jīng)過IPA功能分析發(fā)現(xiàn)差異表達蛋白與糖類,脂類代謝及炎癥反應密切相關。研究結(jié)論：尿液是動脈粥樣硬化疾病標記物發(fā)現(xiàn)的理想來源。尿液中差異蛋白表達表達情況可以反映動脈粥樣硬化疾病狀態(tài)下細胞內(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.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R541.4

【參考文獻】

相關期刊論文 前1條

1 王璐;周蘭蘭;錢小紅;張養(yǎng)軍;;強陽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：1868414