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ProtPipe2

ProtPipe2 provides reproducible workflows for downstream proteomics analysis through the ProtPipe R package and a Shiny interface. The package supports data import into SummarizedExperiment, quality control, filtering, normalization, imputation, batch correction, dimensionality reduction, clustering, differential intensity analysis, pathway analysis, and abundance profiling. For a no-code interface, use our web application which you can launch from here

Installation

if (!requireNamespace("pak", quietly = TRUE)) {
  install.packages("pak", repos = "https://cran.rstudio.com")
}
pak::pak("NIH-CARD/ProtPipe2", dependencies = TRUE)
library(ProtPipe)

If the installation step fails on a Bioconductor dependency, install BiocManager and retry after installing the missing package:

install.packages("BiocManager", repos = "https://cran.rstudio.com")
BiocManager::install("<missing-package>")

Quick start with bundled example data

library(ProtPipe)
library(SummarizedExperiment)

data("neuron_differentiation_intensities")
data("neuron_differentiation_metadata")
data("ipsc_stem_cell_genes")

# 1. Create a SummarizedExperiment from the bundled tables

se <- ProtPipe::create_se(
  data = neuron_differentiation_intensities,
  sample_metadata = neuron_differentiation_metadata,
  creation_method = "bundled neuron differentiation example"
)

# 2. Plot protein group counts
ProtPipe::plot_pg_counts(se, condition = "differentiation_day")

# Prepare the object for PCA, limma, and heatmaps
se_ready <- se |>
  ProtPipe::median_normalize() |>
  ProtPipe::impute_left_dist()

# 3. Plot PCA
ProtPipe::plot_PCs(se_ready, condition = "differentiation_day")

# 4. Run limma differential expression (day 28 vs day 0)
de_day0_vs_day28 <- ProtPipe::do_limma_binary(
  se_ready,
  condition = "differentiation_day",
  control_group = "day_0",
  treatment_group = "day_28"
)
ProtPipe::plot_volcano(de_day0_vs_day28, label_col = "PG.Genes")

# 5. Plot a heatmap using bundled iPSC stem cell markers
p<-ProtPipe::plot_proteomics_heatmap(
  se_ready,
  protmeta_col = "PG.Genes",
  genes = ipsc_stem_cell_genes$Gene,
  condition = "differentiation_day",
  cluster_rows = TRUE,
  cluster_cols = FALSE,
  title = "Stem cell markers across differentiation"
)

About

ProtPipe is an object-oriented R package for analyzing proteomics data. The included shiny app offers a web interface for visualization and analysis. Functionality includes data-preprocessing, quality control, clustering, differential expression and more!

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