Structural Characterization of Antibody Drug Conjugates in Animal Plasma by High Resolution Mass Spectrometry

摘要

In this study, methodologies have been established for the analysis of ADCs by measuring DARs. Extraction with IgSelect column offers good results but only applicable for endogen IgG very different from ADCs. Two extraction methods utilizing antigen-antibody specificity, immobilization of antigen on sepharose or on magnetic beads, have been explored. The former method consume too much antigen. Magnetic beads coupled to biotinylated antigen gave satisfying results with better sensitivity and less quantity of antigen. Results obtained for stability and PK studies show the potential of the methods to analyze ADCs. In the future, other liquid chromatography such as capillary RP-HPLC might be used for achieving better sensitivity and separation of the ADCs from the impurity (aggregates, salts, etc.). Ultra high resolution mass spectrometry might be employed for a further understand of drug release mechanism.