Evaluating the Robustness of Micro-Pillar Array Columns for High-Throughput Proteomics

Nanoflow liquid chromatography capable of delivering consistent and reliable results across extensive sample sets is essential for the advancement of mass spectrometry-based proteomics. Micro-Pillar Array Columns (μPACs) represent a significant breakthrough, offering durability and performance stability. Here, we evaluate the robustness of μPACs by comparing a column used continuously for over 16 months with more than 7,000 injections (μPAC1) to a nearly new column (μPAC2). Analysis of two TMT-labeled yeast TKO standards (TKOpro10u, a 10-plex unit-resolved standard; and TKOpro12, a 12-plex isotopolog-inclusive standard) showed that μPAC1 and μPAC2 yielded comparable numbers of unique peptides and proteins, exhibited similar reproducibility, and delivered equivalent chromatographic and spectral quality. Notably, these data showed that μPAC1 maintained high performance with minimal degradation of data quality, highlighting the exceptional durability of μPAC technology. These findings underscore that μPAC could contribute to reduced workflow disruptions in high-throughput analytical workflows, notably in proteomics workflows using mass spectrometry.