Package: deeprtalign
DeepRTAlign v1.2.2OverviewDeepRTAlign is a deep learning-based retention time alignment tool for large cohort LC-MS data analysis.InstallationInstall Python3.Install Pytorch CPU version, please refer tohttps://pytorch.org/.Usepip install deeprtalignto install deeprtalign OR downloaddeeprtalign-1.2.2-py3-none-any.whlfrom githubreleases, and usepip install deeprtalign-1.2.2-py3-none-any.whlto install deeprtalign. This step will take 3-4 minutes.DeepRTAlign is not dependent on a specific operating system, we have tested it on Windows 10, Ubuntu 18.04 and macOS 12.1.Getting StartedFeature lists and sample list should be prepared before running DeepRTAlign. Feature lists are the output of feature extraction tools (DeepRTAlign supports Dinosaur, OpenMS, MaxQuant, XICFinder and any other text list containing m/z, charge, RT and Intensity information ). The sample list is an excel file recording the correspondences between feature files  and sample names. You can find the test data in the example_files folder.Note that if you use MaxQuant as the feature extraction tool, you should use the allPeptides.txt as the input file or files, and the sample list should correspond to the first column of allPeptides.txt file. DO NOT use "allPeptdes.txt" as the file name in sample list if you use MaxQuant. If you use the old version of MaxQuant (<2.0),please check the 7th column, delete the "Resolution" column if you see it (in MaxQuant v2.0, "Resolution" column is not exists, so we have changed corresponding code).You can get the help information by commanddeeprtalign -h, the basic arguments are as follows:--method {Dinosaur,XICFinder,OpenMS,MaxQuant,TXT,CSV}, -m {Dinosaur,XICFinder,OpenMS,MaxQuant,TXT,CSV}
                        the feature extraction method, support Dinosaur,
                        XICFinder, OpenMS, MaxQuant and any other text list containing m/z, charge, RT and Intensity information
--file_dir FILE_DIR, -f FILE_DIR
                        the data folder
--sample_file SAMPLE_FILE, -s SAMPLE_FILE
                        the sample fileAs an example, to handle the Dinosaur test data in example_files folder, you cancreate a new folderand put the file_dir (containing result files from feature extraction tool ) and sample_file in,switch the working directory to this folder, then use commanddeeprtalign -m Dinosaur -f file_dir -s sample_file.xlsx. On a normal computer, this will take about 30 minutes. We strongly recommend users to use the-pnparameter according to the CPU core numbers. This will make it run much faster.If you choose TXT method (txt file separated by '\t') or CSV method (txt file separated by ','), you must provide optional arguments--mz_col--charge_col--rt_col--intensity_col.optional arguments:--processing_number PROCESSING_NUMBER, -pn PROCESSING_NUMBER
                     processing number, choose according to the number of
                     CPUs, -1 will use all CPUs
--percent PERCENT, -pt PERCENT
                     skip the bins with sample numbers below the percent of
                     total sample numbers
--time_window TIME_WINDOW, -tw TIME_WINDOW
                     min, the time window in the coarse alignment step
--bin_width BIN_WIDTH, -bw BIN_WIDTH
                     the bin width, choose according to the feature
                     extraction step
--bin_precision BIN_PRECISION, -bp BIN_PRECISION
                     the decimal place of bins, choose according to the
                     feature extraction step
--dict_size DICT_SIZE, -ds DICT_SIZE
                     the dict size, choose according to the memory size
--min_time_diff MIN_TIME_DIFF, -mtd MIN_TIME_DIFF
                     min, the time window used to filter the features, only
                     keep the highest feature
--max_mz MAX_MZ, -mm MAX_MZ
                     ppm, the m/z threshold used to filter the features,
                     only align features within the threshold
--max_time MAX_TIME, -mt MAX_TIME
                     min, the time threshold used to filter the features,
                     only align features within the threshold
--max_log_intensity MAX_LOG_INTENSITY, -mli MAX_LOG_INTENSITY
                     log2 intensity value, the intensity threshold used to
                     filter the features, only align features within the
                     threshold
--max_sample_number MAX_SAMPLE_NUMBER, -msn MAX_SAMPLE_NUMBER
                     the max number of candidate feature pairs, if set to
                     -1, all candidate feature pairs will be calculated
--disk_mode DISK_MODE, -dm DISK_MODE
                     if use disk mode,disk mode is slower, but can keep the
                     temp files, 0 do not use disk mode, 1 use disk mode
--keep_temp KEEP_TEMP, -kt KEEP_TEMP
                     if keep the temp files, 0 remove, 1 keep
--begin_step BEGIN_STEP, -bs BEGIN_STEP
                     begin from any step, only work in disk mode
--fdr FDR, -fd FDR    the FDR cutoff
--mz_col MZ_COL, -mz MZ_COL
                     m/z column location, count from 1, for TXT or CSV
                     method
--rt_col RT_COL, -rt RT_COL
                     rt column location, count from 1, for TXT or CSV
                     method
--intensity_col INTENSITY_COL, -int INTENSITY_COL
                     intensity column location, count from 1, for TXT or
                     CSV method
--charge_col CHARGE_COL, -cha CHARGE_COL
                     charge column location, count from 1, for TXT or CSV
                     method
--keep_best_feature KEEP_BEST_FEATURE, -kbf KEEP_BEST_FEATURE
                     0 keep all the candidate features, 1 only keep the
                     best feature for each sample in each group
--keep_best_group KEEP_BEST_GROUP, -kbg KEEP_BEST_GROUP
                     0 keep all the candidate groups, 1 only keep the group
                     with most features if there is conflictprocessing_number (int, default -1) depends on your hardware. percent (float:0-1, default 0) is a threshold, DeepRTAlign will skip the bins with sample numbers below the percent of total sample numbers. time_window (float, default 1) depends on your chromatography, 1 min is the parameter we used in training and is suitable for most situations. bin_width (float, default 0.03) and bin_precision (int, default 2) depends on your feature extraction parameters, bin_width is the m/z window size, and bin_precision is the number of decimal places used in this step. Only the features in the same m/z window will be aligned, default values are suitable for most situations. dict_size (int, default 1024) depends on your memory size, default 1024MB. min_time_diff (float, default 0) is the time window (min) used to used to filter the feature, only keep the highest feature within a time window. max_mz (float, defaut 20),max_time (float, default 5), max_log_intensity (float, default 3) and max_sample_number (int, default 3) are used for reducing calculation. disk_mode will write temp files on your disk, and is not used by default. If you want to keep the temp files, set the keep_temp (int, default 0) to 1. In disk_mode, you can begin from any begin_step (int, default 1). step 1: preprocessing of input files, step 2: coarse alignment, step 3: bining, step 4: filtering, step 5: alignment, step 6: generating result file. fdr (float, 0-1, default 0.01) is the threshold used in quality control part. mz_col, rt_col, intensity_col and charge_col are used in TXT method and CSV method, please note they are counted from 1. Multiple features from a sample may exist in a group after alignment and quality control, keep_best_feature=1 (default) will only keep the feature with the highest score for each sample in each group, or you can set keep_best=0 to keep all candidate features in a group. A feature may exist in multiple groups after alignment and quality control, keep_best_group=1 (default) will only keep the group with the largest number of features, other features of a sample will be divided into new groups, or you can set keep_best=0 to keep all candidate groups.The results will output to the mass_align_all_information folder.If you set kt to 0 (default), there will be only one file in the mass_align_all_information folder: information_target.csv (target results after QC). If you set kt to 1, there will be two files in the mass_align_all_information folder: information_target.csv (target results after QC) and information_all.csv (results before QC, and containing decoy samples).Result DescriptionIn result files, each line represent a feature, the meaning of each column is as follows.Main columnssample: the sample name corresponding to this feature.fraction: the fraction corresponding to this feature.time: the RT (min) corresponding to this feature.mz: the m/z corresponding to this feature.charge: the charge corresponding to this feature.intensity: the intensity corresponding to this feature.score: the score of aligned pairs.group: the group name corresponding to this feature, aligned features share the same group name.In the default workflow, score will be used for quality control (default). This workflow works well in most cases. But when the samples are quite different, or there are too many features with the almost same m/z, the default workflow usually report very few results. In this case, set FDR to 1, and select features with score greater than 0.5 are recomended.Other columnsOther columns are intermediate results and can be ignored.NoteDo not run the different projects under a same folder, the results will be overwritten. That is why we recommendcreate a new folder.DemoThe demo are in the example_files folder. We use part of features in UPS2-Y dataset as examples. On a normal computer, each work will take about 30 minutes. We strongly recommend users to use the-pnparameter according to the CPU core numbers. This will make it run much faster.
We provide Dinosaur, MaxQuant, OpenMS and XICFinder results for user to test DeepRTAlign.To handle the Dinosaur test data in example_files folder, you cancreate a new folderand put the file_dir (containing result files from feature extraction tool ) and sample_file in,switch the working directory to this folder, then use commanddeeprtalign -m Dinosaur -f file_dir -s sample_file.xlsx. The results will output to the mass_align_all_information folder.To handle the MaxQuant test data in example_files folder, you cancreate a new folderand put the file_dir (containing result files from feature extraction tool ) and sample_file in,switch the working directory to this folder, then use commanddeeprtalign -m MaxQuant -f file_dir -s sample_file.xlsx. The results will output to the mass_align_all_information folder.To handle the OpenMS test data in example_files folder, you cancreate a new folderand put the file_dir (containing result files from feature extraction tool ) and sample_file in,switch the working directory to this folder, then use commanddeeprtalign -m OpenMS -f file_dir -s sample_file.xlsx. The results will output to the mass_align_all_information folder.To handle the XICFinder test data in example_files folder, you cancreate a new folderand put the file_dir (containing result files from feature extraction tool ) and sample_file in,switch the working directory to this folder, then use commanddeeprtalign -m XICFinder -f file_dir -s sample_file.xlsx. The results will output to the mass_align_all_information folder.Expected output is in expected_output folder.Code for the experimentsCode for comparing with other alignment tools are in this folder.LicenseGPLv3 (General Public License version 3.0), details in the LICENSE file.ContactsFor any questions involving DeepRTAlign, please contact us by email.Yi Liu,leoicarus@163.comCheng Chang,changchengbio@163.comorchangcheng@ncpsb.org.cn