The human blood plasma proteome contains an immense amount of information for determining a patient’s phenotype on the molecular level. The changes in protein levels as an evidence for a trait have to meet strict requirements regarding accuracy and reproducibility to become a clinical assay fulfilling personalized and precision medicine endeavors. Here, we present an assay repertoire of 474 isotopically labelled protein standards mapping towards 351 proteins that can be used for multiplexed absolute quantification of proteins by mass spectrometry (MS) operated in data independent acquisition (DIA) mode. Each of these standards is overlapping each target protein sequence by shorter 50-150 amino acid stretches and they are added directly to the plasma sample prior to trypsin digestion. This enables robust quantification while each protein standard spike-in is individually tuned to its endogenous level, thereby allowing for precise one-point calibration.