Dynamic Data-Independent Acquisition Mass Spectrometry with Real-Time Retrospective Alignment

Data-independentacquisition (DIA) mass spectrometry has grownin popularity in recent years, because of the reproducibility andquantitative rigor of a systematic tandem mass spectrometry (MS/MS)sampling method. However, traditional DIA methods may spend valuableinstrument time acquiring MS/MS spectra with no usable informationin them, affecting sensitivity and quantitative performance. We developeda DIA strategy that dynamically adjusts the MS/MS windows during thechromatographic separation. The method focuses MS/MS acquisition onthe most relevant mass range at each point in time increasingthe quantitative sensitivity by increasing the time spent on eachDIA window. We demonstrate an improved lower limit of quantification,on average, without sacrificing the number of peptides detected.

Automated summary

Data-independent acquisition (DIA) mass spectrometry has become popular due to its reproducibility and quantitative rigor. However, traditional DIA methods may waste instrument time acquiring MS/MS spectra with no usable information. We developed a dynamic DIA strategy that improves quantification sensitivity without sacrificing the number of detected peptides.