Validation of an immunoassay to selectively quantify the naked antibody of a new Antibody Drug Conjugate--SAR566658--for pharmacokinetic interpretation improvement

Abstract

Given the nature of the ADCs (Antibody Drug Conjugates) as antibodies carrying cytotoxic drugs, two types of immunoassays are usually implemented to perform the analysis of preclinical and clinical study samples during the development phase. The first assay measures the conjugated antibody defined as the ADC carrying at least 1 drug molecule (i.e. drug/antibody ratio greater than or equal to 1). The other measures the total antibody, defined as the ADC irrespective of the drug load (i.e., drug/antibody ratio greater than or equal to 0). One analytical limitation of the total antibody assay is the difficulty to adequately calibrate the assay due to the lack of a representative standard reference for the different circulating entities which change in proportion with time following ADC administration. A new analytical approach that gets round the above highlighted limitation is presented with the development and the validation of a method to quantify selectively naked antibody to support the development of SAR566658 (huDS6-SPDB-DM4). Assessed on 6 separate occasions, the accuracy ranged from -4.3% to 8.9% of nominal values and the precision is 13% at most. The current assay was successfully validated for the quantitation of huDS6 in human LiHe plasma even in the presence of SAR566658 up to 2.00 μg/mL as demonstrated using in vitro spike in quality controls and in actual clinical samples. This innovative assay provides a new tool to document in vivo plasma stability of ADCs and potentially to optimize dose and regimen selection for ADC development.

Keywords: %Diff; ADA; ADC; Anti-Drug Antibodies; Antibody Drug Conjugate; Antibody drug conjugate; CV%; Coefficient of Variation; DM4; Difference from nominal concentrations; EIA; EMA; Enzyme Immunoassay; European Medicines Agency; FDA; Food and Drug Administration; HPAC; Human Pancreatic Adenocarcinoma Cells; Immunobeads; Immunopurification; LC-MS/MS; LLOQ; LiHe; Lithium heparinate; Lower Limit of Quantitation; Maytansin; Naked antibody; PBST; PR; Pharmacokinetics; Phosphate Buffer Saline – Tween; Poor Replicate; Specificity; ULOQ; Upper Limit of Quantitation; VS; Validation Samples; liquid chromatography couple with tandem mass spectrometry; microgram per milliliter; nanogram per milliliter; ng/mL; μg/mL.