An automated, sheathless capillary electrophoresis‐mass spectrometry platform for discovery of biomarkers in human serum

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Abstract
A capillary electrophoresis‐mass spectrometry (CE‐MS) method has been developed to perform routine, automated analysis of low‐molecular‐weight peptides in human serum. The method incorporates transient isotachophoresis for in‐line preconcentration and a sheathless electrospray interface. To evaluate the performance of the method and demonstrate the utility of the approach, an experiment was designed in which peptides were added to sera from individuals at each of two different concentrations, artificially creating two groups of samples. The CE‐MS data from the serum samples were divided into separate training and test sets. A pattern‐recognition/feature‐selection algorithm based on support vector machines was used to select the mass‐to‐charge (m/z) values from the training set data that distinguished the two groups of samples from each other. The added peptides were identified correctly as the distinguishing features, and pattern recognition based on these peptides was used to assign each sample in the independent test set to its respective group. A twofold difference in peptide concentration could be detected with statistical significance (p‐value < 0.0001). The accuracy of the assignment was 95%, demonstrating the utility of this technique for the discovery of patterns of biomarkers in serum.