article published yes Jadranka Miletić Vukajlović author Bojana Ilić author Bella Bruszel author 70abbbb3-88ea-11ec-8e0a-e8c939944834 Tanja Panić-Janković author Goran Mitulović author MassSpec department Background: Low-volume trapping columns are essential for sample enrichment, desalting, and injection profile focusing on nano-LC–MS-based proteomics. They enable higher sample loading, improve chromatographic performance, and protect the analytical column by removing salts and contaminants. Recently, monolithic trap columns with micropillar architecture have emerged as alternatives to conventionally packed traps. This study compares the performance of a packed and a micropillar monolithic trap column for the analysis of tryptic peptides. Methods: A tryptic digest of HeLa cell lysate was analyzed under identical LC–MS conditions using both trap types. Peptides were detected at 214 nm and analyzed by nano-ESI on a Q Exactive Plus Orbitrap. Data were searched against the human UniProt database (February 2023) using FragPipe v20.0, and statistical evaluation of MaxLFQ intensities was performed in Perseus using Welch’s t-test and clustering analysis. Results: Over 2500 proteins were identified with both setups. The packed trap column yielded more total peptides, particularly those with post-translational modifications and higher hydrophilicity, whereas the monolithic column favored peptides of intermediate hydrophobicity. Chromatographic profiles confirmed a slight reduction in the trapping efficiency of hydrophilic peptides by the monolithic trap. Conclusions: Trap column design significantly influences peptide recovery and proteome coverage. https://research-explorer.ista.ac.at/download/21711/21790/2026_Proteomes_Vukajlovic.pdf application/pdfno MDPI2026 eng 2227-7382 4189372510.3390/proteomes14010010 141 Miletić Vukajlović J, Ilić B, Bruszel B, Panić-Janković T, Mitulović G. Comparison of the trapping efficiency for tryptic peptides on particle-packed and micro-pillar trap columns for proteomics analyses. <i>Proteomes</i>. 2026;14(1). doi:<a href="https://doi.org/10.3390/proteomes14010010">10.3390/proteomes14010010</a> Miletić Vukajlović, J., Ilić, B., Bruszel, B., Panić-Janković, T., &#38; Mitulović, G. (2026). Comparison of the trapping efficiency for tryptic peptides on particle-packed and micro-pillar trap columns for proteomics analyses. <i>Proteomes</i>. MDPI. <a href="https://doi.org/10.3390/proteomes14010010">https://doi.org/10.3390/proteomes14010010</a> J. Miletić Vukajlović, B. Ilić, B. Bruszel, T. Panić-Janković, and G. Mitulović, “Comparison of the trapping efficiency for tryptic peptides on particle-packed and micro-pillar trap columns for proteomics analyses,” <i>Proteomes</i>, vol. 14, no. 1. MDPI, 2026. J. Miletić Vukajlović, B. Ilić, B. Bruszel, T. Panić-Janković, G. Mitulović, Proteomes 14 (2026). Miletić Vukajlović, Jadranka, et al. “Comparison of the Trapping Efficiency for Tryptic Peptides on Particle-Packed and Micro-Pillar Trap Columns for Proteomics Analyses.” <i>Proteomes</i>, vol. 14, no. 1, 10, MDPI, 2026, doi:<a href="https://doi.org/10.3390/proteomes14010010">10.3390/proteomes14010010</a>. Miletić Vukajlović J, Ilić B, Bruszel B, Panić-Janković T, Mitulović G. 2026. Comparison of the trapping efficiency for tryptic peptides on particle-packed and micro-pillar trap columns for proteomics analyses. Proteomes. 14(1), 10. Miletić Vukajlović, Jadranka, Bojana Ilić, Bella Bruszel, Tanja Panić-Janković, and Goran Mitulović. “Comparison of the Trapping Efficiency for Tryptic Peptides on Particle-Packed and Micro-Pillar Trap Columns for Proteomics Analyses.” <i>Proteomes</i>. MDPI, 2026. <a href="https://doi.org/10.3390/proteomes14010010">https://doi.org/10.3390/proteomes14010010</a>. 217112026-04-12T22:01:49Z2026-05-04T10:36:21Z