ExSTA: External Standard Addition Method for Accurate High‐Throughput Quantitation in Targeted Proteomics Experiments

摘要

PurposeTargeted proteomics using MRM with stable‐isotope‐labeled internal‐standard (SIS) peptides is the current method of choice for protein quantitation in complex biological matrices. Better quantitation can be achieved with the internal standard‐addition method, where successive increments of synthesized natural form (NAT) of the endogenous analyte are added to each sample, a response curve is generated, and the endogenous concentration is determined at the x‐intercept. Internal NAT‐addition, however, requires multiple analyses of each sample, resulting in increased sample consumption and analysis time.