2025/11/04 by Ndungu, Teresia, Jana Zecha, Zecha, Jana +4
Biochemistry, Genetics and Molecular Biology · Chemistry · #Advanced Biosensing Techniques and Applications #Advanced Proteomics Techniques and Applications #Cancer and biochemical research #FOS: Biological sciences #Genomics (q-bio.GN) #Quantitative Methods (q-bio.QM)
paper · pdf · doi:10.48550/arxiv.2511.02932
openalex publication_date 2025/11/04 · openalex created_date 2025/11/07 · openalex updated_date 2026/07/28
In clinical proteomics, available input is often limited. In addition, phospho-proteomics is of particular interest since the dysregulation of these post-translational modifications (PTMs) has been implicated in various diseases such as cancer. We therefore assessed the feasibility of low input phospho-proteomics via phospho-bulk titration and low-input starting material. We found that there was identification of more phospho-peptides through phospho-bulk titration because of sample loss during preparation of low input starting material. Additionally, we explored various lysis buffers and boiling times for efficiency of decrosslinking formalin-fixed cells since cells and tissues are often fixed for preservation and sorting via FACS. We found that boiling in 0.05M Tris pH 7.6 with 5% SDS for 60 min yielded the highest number of phospho-peptides. Lastly, we applied Evotips Pure and phospho-bulk titration to treated Jurkat cells and identified 7 phospho-sites involved in T-cell stimulation.