Surface Plasmon Resonance (SPR) method development and characterization studies of a monoclonal antibody (AVT04)

Abstract

Introduction: Ligand-binding activity studies using Surface Plasmon Resonance (SPR) are key for measuring functional performances of monoclonal antibody (mAb) medicines, both for characterization, biosimilarity assessment and confirmation of biological activity.Aim: The aim of this project was to set up and optimize a ligand-binding assay for a mAb using SPR (Biacore technology). Specific aims were to set up and optimize calibration-free concentration analysis (CFCA) and single-cycle kinetics (SCK) method, prepare and analyze forced degraded samples of a mAb.Methods: The fully formulated mAb drug substance (DS) was subjected to forced degradation under oxidative, thermal, alkaline and acidic conditions. The cytokine (CK01) binds specifically to the captured mAb on the sensor chip surface. The binding and the dissociation are detected and quantified by SPR in the SCK method and for evaluation of relative binding the KD value of the sample is normalized with the KD value of the reference. CFCA method was used prior to SCK method to determine the active concentration of CK01.Results: For degraded samples, kinetic parameters ka, kd and KD were determined and relative binding of the samples was normalized with the reference sample (100%). Sample exposed to 0.008% H2O2 showed less relative binding (KD was 83%). The binding was also reduced for samples stored at 50 °C for 7 and 14 days, 93% and 87% relative binding, respectively. For the sample subjected to pH 2.5, the binding increased to 144% relative binding. Discussion/conclusions: The results from this project are useful for further analysis of specific modifications on the degraded samples and in determining critical quality attributes (CQA) of the particular mAb used in this study.