MALDI-ToF mass spectrometry coupled with multivariate pattern recognition analysis for the rapid biomarker profiling of Escherichia coli in different growth phases.
MALDI-ToF mass spectrometry coupled with multivariate pattern recognition analysis for the rapid biomarker profiling of Escherichia coli in different growth phases.
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MALDI-ToF 质谱与多变量模式识别分析相结合，可快速分析不同生长阶段大肠杆菌的生物标志物。
DOI：
10.1007/s00216-013-7245-y
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发表时间：
2013
影响因子：
4.3
通讯作者：
Momo RA
中科院分区：
文献类型：
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作者：
Momo RA
基质辅助激光解吸/电离飞行时间质谱（MALDI-ToF MS）已被广泛应用于微生物学领域，用于细菌物种的表征，用于早期疾病诊断和细菌鉴定的生物标志物的检测。在这里，偏最小二乘判别分析（PLS-DA）的多变量数据分析技术应用于从大肠杆菌样品获得的“完整细胞”MALDI-ToF MS数据，以确定这种方法是否可以用来区分，和区分，不同的生长阶段。PLS-DA是一种通过识别包含潜在信息的低维子空间来从大型和噪声数据集中提取系统变化的技术。PLS-DA应用于从不同生长阶段的细胞获得的MALDI-ToF数据导致根据细菌培养物的生长阶段成功地对样品进行分类。分析的另一个结果是，可以识别有助于样品分类的质荷比（m/z）峰或离子信号。然后使用Swiss-Prot/TrEMBL数据库和主要文献将少量的thesem/zion信号暂时分配给蛋白质，这些初步分配显示，来自指数期的主要贡献者是核糖体蛋白。对于稳定期和衰退期培养物，可能进行其他分配，其中鉴定的蛋白质与先前观察到的生物学解释一致。总之，结果显示MALDI-ToF MS、PLS-DA和蛋白质数据库搜索可以组合使用以区分“完整细胞”E. colicell样品在不同的生长阶段，因此可以潜在地用作生物加工工业中的工艺开发的工具，以增强细胞生长和细胞工程策略。
Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF MS) has been exploited extensively in the field of microbiology for the characterisation of bacterial species, the detection of biomarkers for early disease diagnosis and bacterial identification. Here, the multivariate data analysis technique of partial least squares-discriminant analysis (PLS-DA) was applied to ‘intact cell’ MALDI-ToF MS data obtained fromEscherichia colicell samples to determine if such an approach could be used to distinguish between, and characterise, different growth phases. PLS-DA is a technique that has the potential to extract systematic variation from large and noisy data sets by identifying a lower-dimensional subspace that contains latent information. The application of PLS-DA to the MALDI-ToF data obtained from cells at different stages of growth resulted in the successful classification of the samples according to the growth phase of the bacteria cultures. A further outcome of the analysis was that it was possible to identify the mass-to-charge (m/z) ratio peaks or ion signals that contributed to the classification of the samples. The Swiss-Prot/TrEMBL database and primary literature were then used to provisionally assign a small number of thesem/zion signals to proteins, and these tentative assignments revealed that the major contributors from the exponential phase were ribosomal proteins. Additional assignments were possible for the stationary phase and the decline phase cultures where the proteins identified were consistent with previously observed biological interpretation. In summary, the results show that MALDI-ToF MS, PLS-DA and a protein database search can be used in combination to discriminate between ‘intact cell’E. colicell samples in different growth phases and thus could potentially be used as a tool in process development in the bioprocessing industry to enhance cell growth and cell engineering strategies.