Considering The Advantages and Pitfalls of The Use of Isotopically Labeled Protein Standards for Accurate Protein Quantification

Abstract

To ensure comparability of results in clinical proteomics, methods for accurate and traceable quantification of proteins are required. Typically this is done for recombinant proteins using isotopically labeled peptides as internal standards (IS). However, in order to perform quantification in complex matrices such as human serum, isotopically labeled protein standards have been suggested for use as IS to account for losses in sample preparation. The isotopic diluent must be chemically and physically identical to the analyte of interest, having the same amino acid sequence, post-translational modifications, secondary and tertiary structure. It must not be assumed but rather proven that the isotopic diluent is a true mimic, and here we consider both the advantages and potential pitfalls encountered when using isotopically labeled protein IS.