Enhanced Pharmacokinetic Bioanalysis of Antibody –drug Conjugates using Hybrid Immunoaffinity Capture and Microflow LC-MS/MS

This study aimed to evaluate the feasibility of a hybrid immunoaffinity (IA) capture microflow LC–MS/MS (μLC-MS/MS) method for ADC analysis, utilizin g a minimal sample volume for PK assessments in a preclinical study. A robust workflow was established for the quantitative analysis of ADCs by the implementation of solid-phase extraction (SPE) and semi-automation in µLC-MS/MS. Utilizing the µLC-MS/MS approach in conjunction with 1 µL of ADC-dos ed mouse plasma sample volume, standard curves of two representative surrogate peptides for total antibody (heavy chain, HC) and intact antibody (light chain, LC) ranged from 1.00 ng/mL (LLOQ) to 5000 ng/mL with correlation coefficients (r2) values of  >  0.99. The linear range of the standard curve for payload as a surrogate for the concentration of total ADC was from 0.5 ng/mL (LLOQ) to 2000 ng/mL with high accuracy and precision (<  10% CV at all concentrations). Moreover, a high correlation of concentrations of total antibody between two assay approaches (µLC-MS and ELISA) was achieved with less than 20% difference at all time points, indicating that the two methods are comparable in quantitation of total antibody in plasm a samples. The µLC-MS platform demonstrated a greater dynamic range, sensitivity, robustness, and good reproducibility. These findings demonstrated that the cost-effective µLC-MS method can reduce reagent consumption and minimize the use of mice plasma samples while providing more comprehe...
Source: The AAPS Journal - Category: Drugs & Pharmacology Source Type: research