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 che- lation 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 success- fully identified the N-terminal peptides of 158 proteins of Thermoanaerobacter tengcongensis incubated at 55 and 75 ℃, 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 55 percent 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 from 0.1 to 10 and both relative standard derivations (RSD) measured at less than 5%. In T. tengcongensis at two incubation tem- peratures, 80 proteins possess quantitative information. In addition, compared with the proteins of T. tengcongensis incubated at 55 ℃, in T. tengcongensis incubated at 75 ℃, 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。