【摘要】： 本文建立一種改進(jìn)的圓盤電泳方法提取人體液中低分子量蛋白質(zhì)的,去除了分子量大于30kD的高豐度高分子量蛋白質(zhì),并結(jié)合HPLC/MS/MS鑒定了提取的低分子量蛋白質(zhì)。 利用改進(jìn)的圓盤電泳方法提取血清中的低分子量蛋白質(zhì),去除了血清中象白蛋白、免疫球蛋白質(zhì)等高豐度、高分子量蛋白質(zhì),并通過膠內(nèi)洗脫時(shí)間、有機(jī)溶劑選擇、沉淀時(shí)間、回收率試驗(yàn)、重復(fù)性試驗(yàn)、電泳分析、MALDI-TOF-MS分析證明了這種方法的可行性。 用建立的圓盤電泳方法,提取血清中的低分子量蛋白質(zhì),結(jié)合熱變性、溶液內(nèi)酶解以及nano-HPLC/MS/MS,通過mascot檢索鑒定了提取的低分子量蛋白質(zhì)。 利用改進(jìn)的圓盤電泳法提取一份癌癥病人胸腔積液中的低分子量蛋白質(zhì),通過熱變性、溶液內(nèi)酶解以及nano-HPLC/MS/MS,鑒定了提取的低分子量蛋白質(zhì),發(fā)現(xiàn)了幾個(gè)可能與疾病相關(guān)的蛋白質(zhì)。 探討非還原圓盤電泳在提取血清中低分子量蛋白質(zhì)的特點(diǎn),以及變徑圓盤電泳潛在提取血清低分子量蛋白質(zhì)的潛在優(yōu)勢(shì)。
[Abstract]:In this paper, an improved disc electrophoresis method was established to extract low molecular weight proteins from human body fluid, and the high abundance high molecular weight proteins with molecular weight greater than 30kD were removed, and the extracted low molecular weight proteins were identified by HPLC/MS/MS. The low molecular weight protein in serum was extracted by the improved disc electrophoresis method. The high abundance of albumin, immunoglobulin and high molecular weight protein were removed, and elution time, organic solvent selection and precipitation time were used. Recovery test, repeatability test, electrophoretic analysis and MALDI-TOF-MS analysis proved the feasibility of this method. The low molecular weight protein was extracted from serum by disc electrophoresis, combined with thermal denaturation, enzymatic hydrolysis in solution and nano-HPLC/MS/MS,. The extracted low molecular weight protein was identified by mascot search. A low molecular weight protein was extracted from pleural effusion of a cancer patient by improved disc electrophoresis. The extracted low molecular weight protein was identified by thermal denaturation, enzymatic hydrolysis in solution and nano-HPLC/MS/MS,. Several proteins have been found that may be associated with disease. The characteristics of low molecular weight protein extracted from serum by non-reducing disc electrophoresis and the potential advantages of variable diameter disc electrophoresis in extracting low molecular weight protein from serum were discussed.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2010
【分類號(hào)】：R341

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