A hydrophobic interaction chromatography method suitable for quantitating individual monoclonal antibodies contained in co-formulated drug products

摘要

A co-formulated monoclonal antibody (mAb) product containing two or more antibodies offers several therapeutic advantages. However, quantitating the individual antibodies in a co-formulated product is challenging due to the similar biochemical and biophysical properties of mAbs. To identify a method suitable to support the development of a co-formulated drug product with three mAbs, a hydrophobic interaction chromatography method was developed, utilizing a Dionex ProPac HIC-10 column, 100 mM phosphate buffer (pH 7.0), and an ammonium sulfate gradient. Compared to other methods that were evaluated, the HIC method showed the best separation, as well as accurate quantitation of the three mAbs in the co-formulated drug product. The calibration curves were linear over column loads of 225 mu g to 900 mu g (R-2 > 0.99) and the accuracy was between 91% and 106%. Intra-day and inter-day precisions (RSD) were less than or equal to 0.6 % and 1.7%, respectively. The method was used to quantitate individual mAb concentrations in the co-formulated drug product and to monitor any changes in concentration during stability studies. (C) 2020 Regeneron Pharmaceuticals, Inc. Published by Elsevier B.V.