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Proteomics technologies are continually advancing, providing opportunities to develop stronger and more robust protein interaction networks (PINs). In part, this is due to the ever‐growing number of high‐throughput proteomics methods that are available. This review discusses how data‐independent acquisition (DIA) and co‐fractionation mass spectrometry (CF‐MS) can be integrated to enhance interactome mapping abilities. Furthermore, integrating these two techniques can improve data quality and network generation through extended protein coverage, less missing data, and reduced noise. CF‐DIA‐MS shows promise in expanding our knowledge of interactomes, notably for non‐model organisms (NMOs). CF‐MS is a valuable technique on its own, but upon the integration of DIA, the potential to develop robust PINs increases, offering a unique approach for researchers to gain an in‐depth understanding into the dynamics of numerous biological processes.

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