The molecular masses from m/z 0–2 k were excluded from analysis because they were mainly the signal noises of the energy absorbing molecule (EAM). The Biomarker Wizard (Ciphergen Biosystems) was subsequently used to make peak detection and clustering across all spectra in the training set with the following settings: signal/noise (first pass): 5; minimum peak threshold: 15% of all; mass error: 0.3%; and signal/noise (second pass): 2 for the m/z 2–20 k mass Akt inhibitor range. MEK162 price Corresponding peaks in the spectra from the test set were likewise identified using the clustering data from the training set by the same software. The spectral data of the training

set were then exported as spreadsheet files and then further analyzed by the see more Biomarker Pattern Software (BPS) (version 4.0; Ciphergen Biosystems) to develop a classification tree. Decision Tree Classification One of the objectives of SELDI-TOF MS data analysis is to build a Decision Tree that is able to determine the target condition (case or control, cancer or non-cancer) for a given patient’s profile. Peak mass and intensity were exported to an excel file, then transferred to BPS. The classification model was built up with BPS. A Decision Tree was set up to divide the training dataset into either the

cancer group or the control group through multiple rounds of decision-making. When the dataset was first transferred to BPS, the dataset formed a “”root node”". The software tried to find the best peak to separate this dataset into two “”child

nodes”" based on peak ID-8 intensity. To achieve this, the software would identify the best peak and set a peak intensity threshold. If the peak intensity of a blind sample was lower than or equal to the threshold, this peak would go to the left-side child node. Otherwise, the peak would go to the right-side child node. The process would go on for each child node until a blind sample entered a terminal node, either labeled as cancer or control. Peaks selected by the process to form the model were the ones that yielded the least classification error when these peaks were combined to be used. The double-blind sample dataset was used to challenge the model. Peaks from the blind dataset were selected with Biomarker Wizard feature of the Software, following the exact conditions under which peaks from the training dataset were selected. The peak intensities were then transferred to BPS, and each sample was identified as either control or cancer based on the model. The results were compared to clinical data for model evaluation. To characterize the protein peaks of potential interest, serum profiling of patients with NPC and normal control was compared. Mean peak intensity of each protein was calculated and compared (nonparametric test) in each group of serum samples [11]. Statistical analysis Sensitivity was calculated as the ratio of the number of correctly classified diseased samples to the total number of diseased samples.