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Update OSD-514 proteomics analysis#2

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Update OSD-514 proteomics analysis#2
jelardaquino wants to merge 6 commits into
OpenScienceDataRepo:mainfrom
jelardaquino:main

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Copilot AI review requested due to automatic review settings July 7, 2026 18:32

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Pull request overview

This PR updates the OSD-514 proteomics analysis for an “ms3fix” processing path by adding refreshed sample metadata, regenerated differential expression/GSEA result tables, and a reproducible FragPipe TMT runner + export tooling, while removing older local-path R scripts.

Changes:

  • Added ms3fix sample/assay metadata and regenerated model summary tables (condition/batch/sex and sex-stratified).
  • Added a reproducible FragPipe TMT (MS3) headless runner plus Python helpers to extract a protein matrix and export PD-like TMT_all-style files.
  • Removed prior workstation-specific R scripts (CombineTMT.r, Proteomics_QC.r).

Reviewed changes

Copilot reviewed 108 out of 198 changed files in this pull request and generated 9 comments.

Show a summary per file
File Description
Mitochondria/OSD-514/Proteomics/sample_metadata_ms3fix.csv New per-sample metadata (sample/run/condition/sex) for ms3fix workflow
Mitochondria/OSD-514/Proteomics/a_OSD-514_protein-expression-profiling_mass-spectrometry_Orbitrap Fusion.txt Added assay labeling metadata used to build TMT annotations/mappings
Mitochondria/OSD-514/Proteomics/fragpipe_dmel_tmt/run_fragpipe_dmel_tmt.sh End-to-end FragPipe TMT (MS3) runner script
Mitochondria/OSD-514/Proteomics/fragpipe_dmel_tmt/README.md Documentation for running FragPipe TMT pipeline and expected outputs
Mitochondria/OSD-514/Proteomics/fragpipe_dmel_tmt/build_tmt_annotation.py Builds per-plex FragPipe annotation files from assay metadata / TMT_all headers
Mitochondria/OSD-514/Proteomics/fragpipe_dmel_tmt/extract_tmt_protein_matrix.py Extracts protein × sample matrix from FragPipe TMT outputs
Mitochondria/OSD-514/Proteomics/fragpipe_dmel_tmt/export_tmt_all_like.py Legacy exporter to PD-like per-sample files from tmt-report tables
Mitochondria/OSD-514/Proteomics/fragpipe_dmel_tmt/export_tmt_all_like_from_psm.py Recommended PD-like exporter aggregating from psm.tsv
Mitochondria/OSD-514/Proteomics/Proteomics_QC.r Removed old QC script with workstation-specific paths
Mitochondria/OSD-514/Proteomics/CombineTMT.r Removed old combine script with workstation-specific paths
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/sample_metadata_ms3fix.csv Result-bundled copy of ms3fix sample metadata
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/sex_stratified_significant_counts_summary.csv Sex-stratified DE significant-count summary
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/sex_stratified_significant_counts_direction_summary.csv Sex-stratified DE counts with up/down directionality
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/all_model_significant_counts_summary.csv Combined significant-count summary across models
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_significant_counts_summary.csv Model output: sex-stratified significant-count summary
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/significant_counts_summary.csv Model output: sex-stratified condition+batch significant-count summary
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/significant_counts_direction_summary.csv Model output: sex-stratified condition+batch directional counts
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/model_metadata_SFug_males_vs_Earth_males.csv Model metadata for SFug males vs Earth males
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/model_metadata_SFug_females_vs_Earth_females.csv Model metadata for SFug females vs Earth females
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/model_metadata_SF1g_males_vs_Earth_males.csv Model metadata for SF1g males vs Earth males
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/model_metadata_SF1g_females_vs_Earth_females.csv Model metadata for SF1g females vs Earth females
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/design_matrix_SFug_males_vs_Earth_males.csv Design matrix for sex-stratified condition+batch model (SFug males vs Earth males)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/design_matrix_SFug_females_vs_Earth_females.csv Design matrix for sex-stratified condition+batch model (SFug females vs Earth females)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/design_matrix_SF1g_males_vs_Earth_males.csv Design matrix for sex-stratified condition+batch model (SF1g males vs Earth males)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/design_matrix_SF1g_females_vs_Earth_females.csv Design matrix for sex-stratified condition+batch model (SF1g females vs Earth females)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/sex_stratified_condition_batch/Limma_SF1g_males_vs_Earth_males_significant.csv Significant proteins for SF1g males vs Earth males
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/significant_counts_summary.csv Condition-only model: significant-count summary
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/model_metadata_SFug_vs_Earth.csv Condition-only model metadata (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/model_metadata_SF1g_vs_SFug.csv Condition-only model metadata (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/model_metadata_SF1g_vs_Earth.csv Condition-only model metadata (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/Limma_SFug_vs_Earth_significant.csv Condition-only: significant results table (empty header-only export)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/Limma_SF1g_vs_SFug_significant.csv Condition-only: significant results table (empty header-only export)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/Limma_SF1g_vs_Earth_significant.csv Condition-only: significant results table (empty header-only export)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/design_matrix_SFug_vs_Earth.csv Condition-only design matrix (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/design_matrix_SF1g_vs_SFug.csv Condition-only design matrix (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_only/design_matrix_SF1g_vs_Earth.csv Condition-only design matrix (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/significant_counts_summary.csv Condition+batch model: significant-count summary
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/model_metadata_SFug_vs_Earth.csv Condition+batch model metadata (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/model_metadata_SF1g_vs_SFug.csv Condition+batch model metadata (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/model_metadata_SF1g_vs_Earth.csv Condition+batch model metadata (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/Limma_SF1g_vs_SFug_significant.csv Condition+batch: significant proteins (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/Limma_SF1g_vs_Earth_significant.csv Condition+batch: significant proteins (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/design_matrix_SFug_vs_Earth.csv Condition+batch design matrix (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/design_matrix_SF1g_vs_SFug.csv Condition+batch design matrix (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch/design_matrix_SF1g_vs_Earth.csv Condition+batch design matrix (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/significant_counts_summary.csv Condition+batch+sex model: significant-count summary
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/model_metadata_SFug_vs_Earth.csv Condition+batch+sex model metadata (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/model_metadata_SF1g_vs_SFug.csv Condition+batch+sex model metadata (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/model_metadata_SF1g_vs_Earth.csv Condition+batch+sex model metadata (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/Limma_SF1g_vs_SFug_significant.csv Condition+batch+sex: significant proteins (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/Limma_SF1g_vs_Earth_significant.csv Condition+batch+sex: significant proteins (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/design_matrix_SFug_vs_Earth.csv Condition+batch+sex design matrix (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/design_matrix_SF1g_vs_SFug.csv Condition+batch+sex design matrix (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/model_outputs/condition_batch_sex/design_matrix_SF1g_vs_Earth.csv Condition+batch+sex design matrix (SF1g vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_significant_counts_summary_ms3fix.csv Overall limma significant-count summary for ms3fix
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_SFug_vs_Earth_significant.csv Significant proteins table (non-ms3fix naming)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_SFug_vs_Earth_significant_ms3fix.csv Significant proteins table for ms3fix (SFug vs Earth)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_SF1g_vs_SFug_significant.csv Significant proteins table (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_SF1g_vs_SFug_significant_ms3fix.csv Significant proteins table for ms3fix (SF1g vs SFug)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_SF1g_vs_Earth_significant.csv Significant proteins table (empty header-only export)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/Limma_SF1g_vs_Earth_significant_ms3fix.csv Significant proteins table for ms3fix (empty header-only export)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/tables/design_matrix_condition_batch_sex_ms3fix.csv Design matrix used for ms3fix condition+batch+sex model
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Molecular_Function_GeneRIF.csv GSEA enrichment results (SFug vs Earth, MF, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Molecular_Function_GeneRIF_Predicted_zscore.csv GSEA enrichment results (SFug vs Earth, MF, predicted z-score, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Molecular_Function_AutoRIF.csv GSEA enrichment results (SFug vs Earth, MF, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Molecular_Function_AutoRIF_Predicted_zscore.csv GSEA enrichment results (SFug vs Earth, MF, predicted z-score, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Molecular_Function_2018.csv GSEA enrichment results (SFug vs Earth, MF, 2018 gene sets)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Cellular_Component_GeneRIF.csv GSEA enrichment results (SFug vs Earth, CC, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Cellular_Component_GeneRIF_Predicted_zscore.csv GSEA enrichment results (SFug vs Earth, CC, predicted z-score, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Cellular_Component_AutoRIF.csv GSEA enrichment results (SFug vs Earth, CC, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Cellular_Component_AutoRIF_Predicted_zscore.csv GSEA enrichment results (SFug vs Earth, CC, predicted z-score, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Cellular_Component_2018.csv GSEA enrichment results (SFug vs Earth, CC, 2018 gene sets)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Biological_Process_GeneRIF.csv GSEA enrichment results (SFug vs Earth, BP, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Biological_Process_GeneRIF_Predicted_zscore.csv GSEA enrichment results (SFug vs Earth, BP, predicted z-score, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Biological_Process_AutoRIF.csv GSEA enrichment results (SFug vs Earth, BP, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SFug_vs_Earth_GO_Biological_Process_2018.csv GSEA enrichment results (SFug vs Earth, BP, 2018 gene sets)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Molecular_Function_GeneRIF.csv GSEA enrichment results (SF1g vs SFug, MF, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Molecular_Function_GeneRIF_Predicted_zscore.csv GSEA enrichment results (SF1g vs SFug, MF, predicted z-score, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Molecular_Function_AutoRIF_Predicted_zscore.csv GSEA enrichment results (SF1g vs SFug, MF, predicted z-score, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Molecular_Function_2018.csv GSEA enrichment results (SF1g vs SFug, MF, 2018 gene sets)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Cellular_Component_GeneRIF.csv GSEA enrichment results (SF1g vs SFug, CC, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Cellular_Component_GeneRIF_Predicted_zscore.csv GSEA enrichment results (SF1g vs SFug, CC, predicted z-score, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Cellular_Component_AutoRIF.csv GSEA enrichment results (SF1g vs SFug, CC, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Cellular_Component_AutoRIF_Predicted_zscore.csv GSEA enrichment results (SF1g vs SFug, CC, predicted z-score, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Cellular_Component_2018.csv GSEA enrichment results (SF1g vs SFug, CC, 2018 gene sets)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Biological_Process_GeneRIF_Predicted_zscore.csv GSEA enrichment results (SF1g vs SFug, BP, predicted z-score, GeneRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Biological_Process_AutoRIF_Predicted_zscore.csv GSEA enrichment results (SF1g vs SFug, BP, predicted z-score, AutoRIF)
Mitochondria/OSD-514/Proteomics/RESULTS_OSD514_ms3fix/GSEA/tables/SF1g_vs_SFug_GO_Biological_Process_2018.csv GSEA enrichment results (SF1g vs SFug, BP, 2018 gene sets)

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Comment on lines +44 to +48
if [[ -s "${TARGET_DECOY_FASTA}" ]]; then
echo "[1/6] Using existing FASTA: ${TARGET_DECOY_FASTA}"
elif [[ -s "${FALLBACK_DB}" ]]; then
echo "[1/6] Copying FASTA from LFQ setup"
cp -f "${FALLBACK_DB}" "${TARGET_DECOY_FASTA}"
Comment on lines +57 to +63
echo "[2/6] Generating TMT manifest..."
python3 - << 'PY'
from pathlib import Path
import re

mzml_dir = Path('/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/mzML')
out = Path('/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/fragpipe_dmel_tmt/manifests/brain_awg_tmt.fp-manifest')
Comment on lines +89 to +96
echo "[3/6] Generating TMT annotation file..."
# Clear old annotation files to avoid parser ambiguity
find "${MZML_DIR}" -maxdepth 1 -type f -name '*annotation.txt' -delete

python3 "/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/fragpipe_dmel_tmt/build_tmt_annotation.py" \
--tmt-all-dir "${TMT_ALL_DIR}" \
--assay-file "${ASSAY_LABEL_FILE}" \
--mzml-dir "${MZML_DIR}"
Comment on lines +122 to +126
python3 - << 'PY'
from pathlib import Path
wf = Path('/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/fragpipe_dmel_tmt/manifests/brain_awg_dmel_tmt10.workflow')
db = '/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/fragpipe_dmel_tmt/db/dmel_UP000000803_uniprot_target_decoy.fasta'
lines = wf.read_text().splitlines()
Comment on lines +190 to +193
echo "[6/7] Extracting protein matrix from TMT output..."
python3 "/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/fragpipe_dmel_tmt/extract_tmt_protein_matrix.py" \
--workdir "${OUT_DIR}" \
--out "${WORK_DIR}/protein_abundance_tmt_matrix.tsv"
Comment on lines +196 to +201
echo "[7/7] Exporting TMT_all-like per-sample txt files..."
TMT_ALL_EXPORT_DIR="/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/TMT_all_from_psm_pdlike_${RUN_SUBDIR}"
python3 "/home/AD.UNLV.EDU/Shared_Data/AlternativeSplicing/brain_awg/proteomics/fragpipe_dmel_tmt/export_tmt_all_like_from_psm.py" \
--results-dir "${OUT_DIR}" \
--assay-file "${ASSAY_LABEL_FILE}" \
--out-dir "${TMT_ALL_EXPORT_DIR}"
Comment on lines +98 to +100
if any(k in cl for k in ["abundance", "intensity", "reporter", "ratio"]):
sample_cols.append(c)
continue
Comment on lines +70 to +78
reporter_cols = [f"Intensity {plex}_{c}" for c in ["126", "127N", "127C", "128N", "128C", "129N", "129C", "130N", "130C", "131N"]]
existing_reporters = [c for c in reporter_cols if c in psm.columns]

keep_cols = ["Protein", "Qvalue", "Probability"] + existing_reporters
psm = psm[keep_cols].copy()

# Sum reporter intensities per protein.
agg_sum = psm.groupby("Protein", as_index=False)[existing_reporters].sum(min_count=1)

Comment on lines +23 to +27
1. Reuses your D. melanogaster target-decoy FASTA.
2. Builds a TMT manifest grouping files by plex (`TMTA`, `TMTB`, `TMTC`).
3. Auto-generates TMT experiment annotation from assay metadata (`a_OSD-514_protein-expression-profiling_mass-spectrometry_Orbitrap Fusion.txt`).
4. Runs FragPipe headless using built-in `TMT10` workflow template.
5. Extracts a clean protein abundance matrix from the TMT output.
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