nf-core/mhcquant

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Introduction

This document describes the output produced by the pipeline.

nextflow run mhcquant -profile test,<docker/singularity/institute>

The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory.

Pipeline overview

The pipeline is built using Nextflow and processes data using the following steps:

• General
  • Quantification
  • Intermediate results
• VCF
• Class prediction
  • Class (1|2) bindings
• Rotation time prediction
• Workflow reporting and documentation
  • MultiQC
  • Pipeline information

General

Quantification

The CSV output file is a table containing all information extracted from a database search throughout the pipeline. See the OpenMS or PSI documentation for more information about annotated scores and format.

MAP contains information about the different mzML files that were provided initially

#MAP    id      filename        label   size

RUN contains information about the search that was performed on each run

#RUN    run_id  score_type      score_direction date_time       search_engine_version   parameters

PROTEIN contains information about the protein ids corresponding to the peptides that were detected (No protein inference was performed)

#PROTEIN        score   rank    accession       protein_description     coverage        sequence

UNASSIGNEDPEPTIDE contains information about PSMs that were identified but couldn’t be quantified to a precursor feature on MS Level 1

#UNASSIGNEDPEPTIDE      rt      mz      score   rank    sequence        charge  aa_before       aa_after        score_type      search_identifier       accessions      FFId_category   feature_id      file_origin     map_index       spectrum_reference      COMET:IonFrac   COMET:deltCn    COMET:deltLCn   COMET:lnExpect  COMET:lnNumSP   COMET:lnRankSP  MS:1001491      MS:1001492      MS:1001493      MS:1002252      MS:1002253      MS:1002254      MS:1002255      MS:1002256      MS:1002257      MS:1002258      MS:1002259      num_matched_peptides    protein_references      target_decoy

CONSENSUS contains information about precursor features that were identified in multiple runs (eg. run 1-3 in this case)

#CONSENSUS      rt_cf   mz_cf   intensity_cf    charge_cf       width_cf        quality_cf      rt_0    mz_0    intensity_0     charge_0        width_0 rt_1    mz_1    intensity_1     charge_1        width_1 rt_2    mz_2    intensity_2     charge_2        width_2 rt_3    mz_3    intensity_3     charge_3        width_3

PEPTIDE contains information about peptide hits that were identified and correspond to the consensus features described below

#PEPTIDE        rt      mz      score   rank    sequence        charge  aa_before       aa_after        score_type      search_identifier       accessions      FFId_category   fea

Intermediate results

This folder contains the intermediate results from various steps of the MHCquant pipeline (e.g. (un)filtered PSMs, aligned mzMLs, features)

The output mzTab contains many columns annotating the most important information - here are a few outpointed:

PEP   sequence   accession   best_search_engine_score[1]   retention_time   charge   mass_to_charge   peptide_abundance_study_variable[1]

Most important to know that in this format we annotated the q-value of each peptide identification in the best_seach_engine_score[1] column and peptide quantities in the peptide_abundance_study_variable` columns.

mzTab is a light-weight format to report mass spectrometry search results. It provides all important information about identified peptide hits and is compatible with the PRIDE Archive - proteomics data repository.

VCF

Reference fasta

Neoepitopes

These CSV files list all of the theoretically possible neoepitope sequences from the variants specified in the vcf and neoepitopes that are found during the mass spectrometry search, independant of binding predictions, respectively

found_neoepitopes

This CSV which lists all neoepitopes that are found during the mass spectrometry search, independant of binding predictions The format is as follows:

peptide sequence   geneID

vcf_neoepitopes

This CSV fils contains all theoretically possible neoepitope sequences from the variants that were specified in the vcf The format is shown below

Sequence        Antigen ID       Variants

Class prediction

Class (1|2) bindings

This folder containing the binding predictions of all detected class 1 or 2 peptides and all theoretically possible neoepitope sequences The prediction outputs are comma-separated table (CSV) for each allele, listing each peptide sequence and its corresponding predicted affinity scores:

peptide   allele   prediction   prediction_low   prediction_high   prediction_percentile

Rotation time prediction

Workflow reporting and documentation

MultiQC

MultiQC is a visualization tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in the report data directory.

The pipeline has special steps which allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see http://multiqc.info.

Pipeline information

Nextflow provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to troubleshoot errors with the running of the pipeline and provide you with other information such as launch commands, run times and resource usage.