Data independent acquisition (DIA) strategies have emerged as a powerful alternative to classic data dependent acquisition (DDA) in shotgun proteomics. DIA offers systematic measurement of all peptide ions regardless of their intensity by co-fragmenting all co-eluting peptides in broader precursor isolation windows. This provides a wider dynamic range of the proteomes analyzed, improved reproducibility for identification and enables better sensitivity and accuracy for quantification.

While modern mass spectrometers have developed towards faster scanning speeds and enhanced selectivity by ion mobility, it is indeed the elaborate processing algorithms for identification, that has made the most significant contributions to the field over the last couple of years. The analysis of DIA data has historically been difficult and driven by specialists in the field. One of the most important requirements to perform a DIA analysis has been to generate a project specific library, which is made from extensive offline fractionation with the same LC gradient and column as used in the actual DIA experiment.

It is now also possible to search DIA data without the need for DDA based spectral libraries, which is a game-changer, facilitating simpler and more powerful workflows. Further implementation of deep-learning based models has significantly contributed to increased proteome coverage. These major developments over the last years have positioned DIA as an important acquisition strategy and the preferred method in many labs.

The trend continues

This is a especially positive, as DIA is the ideal match for short and standardized gradients on the Evosep One. It has been demonstrated that more than 5000 proteins can be identified in just 5 min using dia-PASEF with the fast 200 samples per day method. This proteome coverage is increased to nearly 7000 proteins when extending the gradient time using the 60 samples per day method. With such a coverage obtained in short time, it is possible to gain relevant biological insight for discovery proteomics with large cohorts of clinical samples.

Webinar: Unleash the power of DIA

The steep development over the last years, have positioned DIA as an important acquisition strategy to generate record high identifications with short gradients on the Evosep One. Join this webinar to explore how our customers have used Evosep One to generate record high IDs per minute for large-scale sample cohorts.

You can also read more about the research published using DIA methodologies from our users here.

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MORE RESEARCH ON DIA

Here you can see publications available using DIA featuring Evosep One. For a full overview of publications published using the Evosep One Technology visit our Literature room here