表面辅助激光解吸附/离子化质谱(Surface-assisted laser desorption/ionization mass spectrometry,SALDI-MS)是一种利用无机纳米粒子或纳米结构表面作为基质,辅助待测分子的解吸附和离子化的质谱技术。由于其具有灵敏度高、耐盐性好...表面辅助激光解吸附/离子化质谱(Surface-assisted laser desorption/ionization mass spectrometry,SALDI-MS)是一种利用无机纳米粒子或纳米结构表面作为基质,辅助待测分子的解吸附和离子化的质谱技术。由于其具有灵敏度高、耐盐性好、操作简便、重现性好、检测通量高等优势,已经被广泛应用于食品安全、环境监测、生命科学等诸多领域。该文总结了近5年来,SALDI基质材料(金属及金属氧化物材料、碳材料、硅材料、金属有机骨架化合物材料等)的最新研究进展及其在生物检测领域中的应用,并对SALDI-MS基质材料的发展及应用进行了展望。展开更多
Erythropoietin (EPO) is a glycoprotein with three N-glycosylation sites and one O-glycosylation sites. According to the reference, all of the N-glyco-chains were acid complex type. The amount of sialic acid in rhEPO w...Erythropoietin (EPO) is a glycoprotein with three N-glycosylation sites and one O-glycosylation sites. According to the reference, all of the N-glyco-chains were acid complex type. The amount of sialic acid in rhEPO was determined and two N-glycosylation sites on it were confirmed by matrix-assisted laser desorption time of flight-time of flight mass spectrometry (MALDI-TOF-TOFMS) in this paper. The total amount of sialic acid in rhEPO accounts for 12.97%. Two N-glycosylation sites (Asn38,Asn83) were identified successfully.展开更多
The protein profiles of the drug-resistant cell line K562/A02 and its sensitive parental cell line K562 were compared by using high resolution two-dimensional gel electrophoresis with IPG strip of pH 4\^7\5\^9. One pr...The protein profiles of the drug-resistant cell line K562/A02 and its sensitive parental cell line K562 were compared by using high resolution two-dimensional gel electrophoresis with IPG strip of pH 4\^7\5\^9. One protein spot was found to be differentially expressed, which was identified as the soluble resistance-related calcium binding protein（sorcin）. Tandem mass spectrometric analysis of one tryptic peptide by a TOF-TOF analyzer as well as the western blot analysis substantiates the result derived from peptide mapping fingerprint data.展开更多
目的:探索胶上蛋白原位酶切、肽质量指纹图(PMF)分析以及数据库检索方法的优化条件,建立适合于亚细胞比较蛋白质组学研究的质谱鉴定策略。方法:从考马斯亮蓝染色的2-DE胶上取下牛血清白蛋白(BSA)标准蛋白进行胶内原位酶切,与基质(CHCA)...目的:探索胶上蛋白原位酶切、肽质量指纹图(PMF)分析以及数据库检索方法的优化条件,建立适合于亚细胞比较蛋白质组学研究的质谱鉴定策略。方法:从考马斯亮蓝染色的2-DE胶上取下牛血清白蛋白(BSA)标准蛋白进行胶内原位酶切,与基质(CHCA)混匀后进行MALDI-TOF MS分析;对来源于人肝细胞癌细胞株QGY-7703线粒体的差异表达的候选蛋白点L9,按照通过BSA标准蛋白优化的条件进行MALDI-TOF质谱鉴定;其肽质量指纹图(PMF)经MS-Fit检索。结果:经数据库检索,BSA的肽质量指纹图(PMF)匹配肽质量数为10/11,序列覆盖率为19%,说明实验条件完全可信。候选蛋白点L9的PMF数据经数据库检索后,排名前两位的蛋白均为OXCT(3-oxoacid CoA transferase 1 precursor),其匹配肽段为9/13,序列覆盖率为24%;且OXCT的理论分子量(56kda),理论等电点(7.1),与胶上L9蛋白的位置相符。故候选蛋白点L9确认为OXCT(3-oxoacid CoA transferase 1 precursor)。结论:本文所确定的鉴定策略适用于线粒体比较蛋白质组学的研究。展开更多
文摘表面辅助激光解吸附/离子化质谱(Surface-assisted laser desorption/ionization mass spectrometry,SALDI-MS)是一种利用无机纳米粒子或纳米结构表面作为基质,辅助待测分子的解吸附和离子化的质谱技术。由于其具有灵敏度高、耐盐性好、操作简便、重现性好、检测通量高等优势,已经被广泛应用于食品安全、环境监测、生命科学等诸多领域。该文总结了近5年来,SALDI基质材料(金属及金属氧化物材料、碳材料、硅材料、金属有机骨架化合物材料等)的最新研究进展及其在生物检测领域中的应用,并对SALDI-MS基质材料的发展及应用进行了展望。
文摘Erythropoietin (EPO) is a glycoprotein with three N-glycosylation sites and one O-glycosylation sites. According to the reference, all of the N-glyco-chains were acid complex type. The amount of sialic acid in rhEPO was determined and two N-glycosylation sites on it were confirmed by matrix-assisted laser desorption time of flight-time of flight mass spectrometry (MALDI-TOF-TOFMS) in this paper. The total amount of sialic acid in rhEPO accounts for 12.97%. Two N-glycosylation sites (Asn38,Asn83) were identified successfully.
文摘The protein profiles of the drug-resistant cell line K562/A02 and its sensitive parental cell line K562 were compared by using high resolution two-dimensional gel electrophoresis with IPG strip of pH 4\^7\5\^9. One protein spot was found to be differentially expressed, which was identified as the soluble resistance-related calcium binding protein（sorcin）. Tandem mass spectrometric analysis of one tryptic peptide by a TOF-TOF analyzer as well as the western blot analysis substantiates the result derived from peptide mapping fingerprint data.
文摘目的:探索胶上蛋白原位酶切、肽质量指纹图(PMF)分析以及数据库检索方法的优化条件,建立适合于亚细胞比较蛋白质组学研究的质谱鉴定策略。方法:从考马斯亮蓝染色的2-DE胶上取下牛血清白蛋白(BSA)标准蛋白进行胶内原位酶切,与基质(CHCA)混匀后进行MALDI-TOF MS分析;对来源于人肝细胞癌细胞株QGY-7703线粒体的差异表达的候选蛋白点L9,按照通过BSA标准蛋白优化的条件进行MALDI-TOF质谱鉴定;其肽质量指纹图(PMF)经MS-Fit检索。结果:经数据库检索,BSA的肽质量指纹图(PMF)匹配肽质量数为10/11,序列覆盖率为19%,说明实验条件完全可信。候选蛋白点L9的PMF数据经数据库检索后,排名前两位的蛋白均为OXCT(3-oxoacid CoA transferase 1 precursor),其匹配肽段为9/13,序列覆盖率为24%;且OXCT的理论分子量(56kda),理论等电点(7.1),与胶上L9蛋白的位置相符。故候选蛋白点L9确认为OXCT(3-oxoacid CoA transferase 1 precursor)。结论:本文所确定的鉴定策略适用于线粒体比较蛋白质组学的研究。
