Sample preparation increasingly sets the throughput and reproducibility of mass spectrometry (MS)-based proteomics. StageTips (stop-and-go extraction tips) and variants thereof have long been common implements to purify samples, and we recently extended the concept to solid-phase extraction capture (SPEC) tips, in which the entire digestion takes place in sub-microliter volumes. Here we replace the hand-packed bed with a strong anion-exchange (SAX) monolith photopolymerized directly inside the pipette tip from a defined recipe (pSPEC). A liquid-handling robot casts 384 tunable tips in minutes, at low cost and in any format, adding negligibly to the workflow's variance. Across biofluids, pSPEC added ~20% more identifications than in-solution plasma and reached 3,500 protein groups from a single injection of healthy urine and 4,800 from saliva at 100 samples per day, depths usually requiring depletion or fractionation. The same light-cast chemistry should extend to single cells, affinity capture, and population-scale studies.
Korff, K., Henneberg, L., Oliinyk, D., Eikmeier, N., Schuele, A. M., Heymann, T., Schebesta, A.-S., Albrecht, V., Kaiser, C. J. O., Mann, M., Mueller-Reif, J. B.
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