Liquid chromatography – tandem mass spectrometry (LC-MS/MS) is an indispensable tool in the bioanalysis of small molecules and biologics, offering sensitivity, selectivity, and flexibility to multiplex. This presentation will focus on an application of LC-MS/MS for indirect measurement of anti-drug antibody (ADA) responses. Three-month, repeat dosing studies of Therapeutic A in rat and NHPs led to significant drug accumulation as determined by PK assessment. In order to investigate the reason for the observed unexpected pharmacokinetic profiles in both species, a workflow was established which employed immunoprecipitation (IP) of immune complexes from plasma followed by LC-MS/MS measurement of the Therapeutic A. This indirect, semi-quantitative assay allowed detection of immunoglobulin associated Therapeutic A. These results were further corroborated by an orthogonal method, thus strengthening the evidence of ADA formation.
Learning Objectives:
Upon completion, participants will have developed an in-depth understanding of hybrid LC-MS/MS workflow employing immunoprecipitation followed by LC-MS/MS analysis of the analytes of interest and its applications in ADA semi-quantitation.
Upon completion, participant have gained insights on the challenges and future directions of LC-MS/MS as a supplementary and orthogonal tool to the traditional LBA approach for measuring ADA responses.
Upon completion, participant will have deep understanding of a case study of ADA measurement by LC-MS/MS and have learned the design of direct and indirect assay formats and key considerations in method development.
