High-Throughput Proteomics and Phosphoproteomics of Rat Tissues Using Microflow Zeno SWATH.

High-throughput tissue proteomics has great potential in the advancement of precision medicine. Here, we investigated the combined sensitivity of trap-elute microflow liquid chromatography with a ZenoTOF for DIA proteomics and phosphoproteomics. Method optimization was conducted on HEK293T cell lines to determine the optimal variable window size, MS2 accumulation time and gradient length. The ZenoTOF 7600 was then compared to the previous generation TripleTOF 6600 using eight rat organs, finding up to 23% more proteins using a fifth of the sample load and a third of the instrument time. Spectral reference libraries generated from Zeno SWATH data in FragPipe (MSFragger-DIA/DIA-NN) contained 4 times more fragment ions than the DIA-NN only library and quantified more proteins. Replicate single-shot phosphopeptide enrichments of 50-100 µg of rat tryptic peptide were analyzed by microflow HPLC using Zeno SWATH without fractionation. Using Spectronaut we quantified a shallow phosphoproteome containing 1000-3000 phosphoprecursors per organ. Promisingly, clear hierarchical clustering of organs was observed with high Pearson correlation coefficients >0.95 between replicate enrichments and median CV of 20%. The combined sensitivity of microflow HPLC with Zeno SWATH allows for the high-throughput quantitation of an extensive proteome and shallow phosphoproteome from small tissue samples.

Marine Antifouling Behavior of Lubricant-Infused Nanowrinkled Polymeric Surfaces.

A new family of polymeric, lubricant-infused, nanostructured wrinkled surfaces was designed that effectively retains inert nontoxic silicone oil, after draining by spin-coating and vigorous shear for 2 weeks. The wrinkled surfaces were fabricated using three different polymers (Teflon AF, polystyrene, and poly(4-vinylpyridine)) and two shrinkable substrates (Polyshrink and shrinkwrap), and Teflon on Polyshrink was found to be the most effective system. The volume of trapped lubricant was quantified by adding Nile red to the silicone oil before infusion and then extracting the oil and Nile red from the surfaces in heptane and measuring by fluorimetry. Higher volumes of lubricant induced lower roll-off angles for water droplets, and in turn induced better antifouling performance. The infused surfaces displayed stability in seawater and inhibited growth of Pseudoalteromonas spp. bacteria up to 99%, with as little as 0.9 µL cm-2 of the silicone oil infused. Field tests in the waters of Sydney Harbor over 7 weeks showed that silicone oil infusion inhibited the attachment of algae, but the algal attachment increased as the silicone oil was slowly depleted over time. The infused wrinkled surfaces have high transparency and are moldable, making them suited to protect the windows of underwater sensors and cameras.