# Pho-Tip: One-Pot Dephosphorylation for Rapid and Sensitive Analysis of DIA Phosphoproteomics Data

## TL;DR

Pho-Tip is a new method that improves the analysis of phosphoproteomics data by simplifying the detection of phosphorylated peptides in DIA experiments.

## Contribution

Pho-Tip introduces a lossless one-pot dephosphorylation strategy for more efficient and sensitive DIA phosphoproteomics analysis.

## Key findings

- Pho-Tip enables comprehensive mapping of phosphorylated peptide sequences.
- The method facilitates the creation of experiment-focused in silico predicted spectral libraries.
- Pho-Tip improves detection of low-abundant phosphorylated peptides in low sample amounts.

## Abstract

Recent advances in instrumentation and data processing
have transformed
data-independent acquisition (DIA) proteomics into a reliable technology
for quantitative profiling of post-translational modifications. However,
analysis of DIA phosphoproteomics data is challenging due to the large
search space, wherein all combinations of phosphosites on a peptide
need to be considered. Current approaches therefore face significant
hurdles in detecting low-abundant phosphorylated peptides, in particular
when working with low sample amounts. Here we introduce Pho-Tip, a
lossless one-pot dephosphorylation strategy. We show that Pho-Tip
enables comprehensive mapping of phosphorylated peptide sequences,
facilitating streamlined creation of experiment-focused in
silico predicted spectral libraries and thus rapid and sensitive
analysis of DIA phosphoproteomics experiments.