This dataset was generated to validate the real-time quality control and automated data analysis capabilities of Apus, an end-to-end solution for large-cohort mass spectrometry-based proteomics. The dataset consists of 23 longitudinally acquired data-dependent acquisition (DDA) runs from HEK 293T cell digest samples, prepared and analyzed under routine laboratory operating conditions on an Orbitrap Astral mass spectrometer (Thermo Scientific) coupled to a Vanquish Neo nanoflow LC system. The dataset was processed through the complete Apus automated workflow, including real-time data aggregation (ApuPickup), parallelized protein identification (ApuPioneer), high-throughput label-free quantification (ApuHorizon), and comprehensive quality control (ApuMonitor), to demonstrate the system's capability for proactive quality management in a live laboratory setting.