人体液中低分子量蛋白质的圆盘凝胶电泳提取
发布时间：2018-11-21 19:11
【摘要】： 本文建立一种改进的圆盘电泳方法提取人体液中低分子量蛋白质的,去除了分子量大于30kD的高丰度高分子量蛋白质,并结合HPLC/MS/MS鉴定了提取的低分子量蛋白质。 利用改进的圆盘电泳方法提取血清中的低分子量蛋白质,去除了血清中象白蛋白、免疫球蛋白质等高丰度、高分子量蛋白质,并通过胶内洗脱时间、有机溶剂选择、沉淀时间、回收率试验、重复性试验、电泳分析、MALDI-TOF-MS分析证明了这种方法的可行性。 用建立的圆盘电泳方法,提取血清中的低分子量蛋白质,结合热变性、溶液内酶解以及nano-HPLC/MS/MS,通过mascot检索鉴定了提取的低分子量蛋白质。 利用改进的圆盘电泳法提取一份癌症病人胸腔积液中的低分子量蛋白质,通过热变性、溶液内酶解以及nano-HPLC/MS/MS,鉴定了提取的低分子量蛋白质,发现了几个可能与疾病相关的蛋白质。 探讨非还原圆盘电泳在提取血清中低分子量蛋白质的特点,以及变径圆盘电泳潜在提取血清低分子量蛋白质的潜在优势。
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R341
本文编号：2348023
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R341
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