Development and Validation of Analytical Methods to Determine the Prolonged in-use Stability of Anticancer Monoclonal Antibodies in Clinical Practice

Abstract

Background/purpose: Personalized dosing of monoclonal antibodies (mAbs) based on body weight and surface area may result in leftovers that need to be discarded due to the absence of stability data. After manufacturing, these mAbs undergo extensive quality control with a panel of methods. With some of these similar methods, these mAbs could be tested to see if stability is maintained after the opening of the pharmaceutical product vial and dilution for administration in a daily clinical setting. Prolonged in-use stability may reduce unnecessary waste and minimize the financial loss of these expensive drugs. Methods: Previously, based on extensive literature research, a complementary panel of methods was selected including visual inspection colour, apparency and, clarity (CAC), size exclusion chromatography (SEC), enzyme-linked immunosorbent assay (ELISA), dynamic light scattering (DLS), thermal denaturation (Tm), and aggregation (Tagg) determination, pH and dye ingress method. We evaluated, developed, and validated these methods with respect to the accuracy, precision, reproducibility, stability-indicating capability, carry-over, limit of quantification (LOQ) and limit of detection (LOD) for nivolumab and pembrolizumab as prototype anticancer mAbs. Results: Analytical results show that stability can be determined from a physiochemical, biological, and microbiological point of view with the selected methods. Forced degradation studies showed that even small instabilities can be detected using the panel of methods together, therefore the stability-indicating capability has been established. CAC shows mainly aggregation and discolouration. SEC shows aggregation and fragmentation/degradation, DLS gives information about turbidity and soluble aggregates, Tm and Tagg give information changes intra- and inter conformational changes, respectively, in the mAb molecule. ELISA shows potential loss in bioactivity and the pH of the sample is essential to the chemical composition of the mAb molecules in the solution. Conclusion: A panel of analytical methods was selected, enabling in-use stability determination of anticancer mAbs. This panel will be used in future studies to determine the prolonged use of leftover anticancer mAb products and infusion solutions, starting with nivolumab and pembrolizumab.