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建立了依赖色谱保留时间的智能化选择反应监测质谱方法,并与非依赖色谱保留时间的智能化选择反应监测质谱分析方法对不同体系(牛血清白蛋白酶切物、6种标准蛋白质混合物酶切物、腾冲嗜热菌蛋白提取液酶切物)的分析结果进行了系统比较。结果表明,引入色谱保留时间后的智能化选择反应监测质谱方法能够显著提高肽段及蛋白质的鉴定量,并且在复杂体系(如腾冲嗜热菌蛋白提取液酶切物)中效果尤为明显,鉴定到的肽段及蛋白质的覆盖率可分别达到目标肽段和蛋白质数量的89.62%和92.41%,并且灵敏度高、重复性好,能够实现对质荷比相同但保留时间有差异的肽段的准确鉴定。该方法将在复杂生物样本目标蛋白质组高通量、高灵敏度的鉴定、验证和确认中发挥独特作用。
An intelligent selective reaction monitoring mass spectrometry method was established, which was dependent on the retention time of the chromatographic method. The results were compared with those of the other systems (bovine serum albumin digest, enzyme digest of six standard proteins , Tengchong thermophilic protein extract digested) analysis of the results of a systematic comparison. The results showed that the introduction of the chromatographic retention time of intelligent selective reaction monitoring mass spectrometry method can significantly improve the identification of peptide and protein, and in complex systems (such as Tengchong thermophilic protein extract digested) was particularly effective, identification The coverage of the peptides and proteins reached 89.62% and 92.41% respectively of the target peptides and proteins, and the sensitivity and repeatability were good, and the identification of the peptides with the same mass-to-charge ratio but different retention time could be achieved . This method will play a unique role in the high-throughput, high-sensitivity identification, verification and validation of target proteomes in complex biological samples.