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Optimised Sample Handling Minimises Peptide Adsorption to Plastics to Enable High Sensitivity Evosep Based Chemical Proteomics

ABSTRACT: Chemical proteomics approaches often yield low peptide recovery because they aim to enrich low-abundance proteins. Poor sample handling further reduces recovery through peptide adsorption to plastic surfaces and losses due to vacuum evaporation. We systematically mapped these losses across buffers, volumes, plastics and pH, then developed an Evotip-compatible handling protocol that minimises adsorption and removes the need for vacuum evaporation. Losses to plastic adsorption and vacuum evaporation were most apparent at low inputs (< 200 ng) and with larger volumes (> 20 µL) but were also dependent on acidification as well as the buffer and plastic used. The optimised workflow: direct acidification of peptides, frozen storage if needed, and direct loading onto Evotips, resulted in up to ~100-fold gains versus common drying practices. This optimised sample handling method enables high-throughput chemical proteomics by reducing manual handling steps and enables the characterisation of low abundance proteins previously lost during sample preparation.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Jack Houghton

LAB HEAD: Edward Tate

PROVIDER: PXD069322 | Pride | 2026-06-23

REPOSITORIES: Pride

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			Action	DRS
	1-combined_peptide.tsv	Tabular
	1-psm.xlsx	Xlsx
	2-combined_peptide.tsv	Tabular
	3-combined_peptide.tsv	Tabular
	4-combined_peptide.tsv	Tabular

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