In situ polymerized monolith tips for reproducible, format-flexible proteomic sample preparation applied to biofluids
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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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 workflows 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.
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