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システムレベルオミクスのための四重極スキャンを利用した新規DIA分析法の最適化
Introduction
A novel DIA mode of operation, SONAR, which provides both qualitative and quantitative information, has been developed and implemented on a hybrid quadrupole/oa-time-of-flight (Q-TOF) MS. Here we describe the optimization of analytical parameters to obtain the best qualitative and quantitative coverage of complex proteomic samples.
Method
Complex digest samples from E.coli and human cell lines were evaluated. MS data were collected using SONAR acquisition, where a continuously scanning quadrupole, isolates specific m/z regions prior to the oa-TOF. Alternate MS scans comprised precursor and CID product ions respectively. Data were processed and database searched using Symphony, incorporating PLGS, Skyline and Scaffold informatics toolsets, and Progenesis QI providing both qualitative and quantitative results.
Results
SONAR DIA acquisition provides multi dimensional data sets exhibiting improved specificity over other (wide band) DIA methods. SONAR acquisition parameters for omics experiments, i.e. quadrupole isolation window vs. on-column load, scan vs. gradient time, and multi step experiments, can be easily determined for optimal qualitative and quantitative performance. SONAR DIA can be applied in discovery and targeted translational omics studies, including metabolomics, lipidomics and proteomics. Further work will include the targeted extraction of ions to investigate maximum coverage, quantitative precision and throughput (as a function of analysis time).