Mass Fabrication of Capillary Columns Based on Centrifugal Packing

Packed
capillary columns have become the standard front-end separation
device for mass spectrometry-based proteomics. The development of
simple, fast, and robust capillary column technology, especially that
with mass-fabrication capacity, can greatly improve analytical throughput
and reproducibility in omics research. In this technical note, we
report a centrifugal packing technology, which has the capability
to mass fabricate high quality capillary columns with a 2886 columns/day
fabrication throughput. The centrifugally packed columns presented
significantly improved efficiency (reduced plate height hmin = 1.6, 37%–40% improvement compared with slurry
packed columns), advanced kinetic performance limit, and excellent
column-to-column reproducibility (2.0% RSD for retention time, 50
columns). Such columns enabled ∼5300 HeLa proteins identified
in single-shot proteomic analysis, displaying both intercolumn and
inter-run retention time stability (retention time RSD = 0.94% between
nine replicates on three columns for probing peptide sequence). The
mass-fabrication technology reported in this technical note may support
disposable use of high quality chromatographic columns in large-scale
bioanalysis.