on 09-26-201603:56 AM - edited on 10-15-202111:50 AM by AnalyteGuru
Alexander Boychenko1, Mike Baynham2, Stephan Meding1; Martin Ruehl1; Martin Samonig1; Remco Swart1 International Hupo 2016 We systematically investigated the effect of flow rate and column ID on MS sensitivity using samples with different complexity. The sensitivity of capillary flow LC-MS (capLC-MS) was around 20 times higher at 5 μL/min in comparison with analytical flow LC-MS analysis at 450 μL/min. The optimized capLC-MS conditions resulted in high-throughput separations with ~ 10 min total analysis time for targeted quantification of peptides at amole level. CapLC-MS can also be used for routine and robust proteome profiling of complex bio-samples. Around 70% of proteins were identified with capLC-MS in comparison with nanoLC-MS using the same sample amount and column length. The increase in loading amount allowed to identify around 80% of protein groups found with nanoLC-MS. Capillary flow LC together with plug-and-spray EASY-Spray emitters for electrospray ionization unites the advantages of high-throughput analytical flow LC separations with high sensitivity of nano flow LC-MS analysis. It is a valuable tool for targeted peptide quantification
as well as bottom-up proteomics profiling of large sample cohorts for translational medicine studies.