本文为2015年毛细管电泳(CE)技术的年度回顾。归纳了ISI Web of Science中检索的2015年度CE技术相关的论文,从毛细管电泳-质谱(CE-MS)联用技术、方法学研究、检测与富集、手性分子拆分及CE技术应用5个方面进行了分类说明;简要介绍了2015年涉及CE技术的国际会议6个,国内会议2个以及各会议的研究报道情况。最后,介绍了目前国内外的单克隆抗体、水质、酒类和食品分析中涉及的毛细管电泳的方法标准。
By means of the reaction between a DOTA-NHS-ester bifunctional reagent and N-terminal peptides of proteins,and then chelation of lanthanide metal ions as tags,we established a novel method for the identification of N-terminal peptides of proteins and their relative quantification using metal-element-chelated tags coupled with mass spectrometry.The experimental results indicate that metal elements are able to completely label N-terminal peptides at the protein level.The N-terminal peptides are enriched as the peptides digested with trypsin are selectively eliminated by isothiocyanate-coupled silica beads.We successfully identified the N-terminal peptides of 158 proteins of Thermoanaerobacter tengcongensis incubated at 55 and 75°C,among which N-terminal peptides of 24 proteins are partially acetylated.Moreover,metal-element tags with high molecule weights make it convenient for N-terminal peptides consisting of less than 6 amino acids to be identified;these make up 55percent of the identified proteins.Finally,we developed a general approach for the relative quantification of proteins based on N-terminal peptides.We adopted lysozyme and ribonuclease B as model proteins;the correlation coefficients(R2)of the standard curves for the quantitative method were 0.9994 and 0.9997,respectively,with each concentration ratio ranging from0.1 to 10 and both relative standard derivations(RSD)measured at less than 5%.In T.tengcongensis at two incubation temperatures,80 proteins possess quantitative information.In addition,compared with the proteins of T.tengcongensis incubated at 55°C,in T.tengcongensis incubated at 75°C,7 proteins upregulate whereas 16 proteins downregulate,and most differential proteins are related to protein synthesis.
本文为2014年毛细管电泳(CE)技术的年度回顾。介绍了2014年涉及CE技术的国际会议5个,国内会议2个,总结了各会议的研究报道情况;归纳了在ISI Web of Science中检索到的2014年度发表的与CE技术相关的论文,并对以上论文在生物医药研究、检测器使用以及重要分析化学杂志发表的情况进行了分类说明。最后,回顾和比较了2012-2014年的CE进展。
