Fitting bevacizumab aggregation kinetic data with the Finke–Watzky two-step model: Effect of thermal and mechanical stress

摘要: Size exclusion chromatography with light scattering detection (SEC-MALLS) was assessed as a means to characterize the type of bevacizumab aggregates that form under mechanical and thermal stress, quantitatively monitoring aggregation kinetics. The analytical method monitored verified during routine use at two levels: (1) "pre-study" validation shows is specific, linear, accurate, precise, robust stability indicating; (2) "in-study" by inserting quality control samples charts, indicating in statistical stable. kinetics data were interpreted using modified Lumry-Eyring model, but fit can be considered poor (R(2)>0.96), especially higher temperatures. This indicates order reaction could not reliably determined, suggesting different degradation mechanism. kinetic set also minimalistic Finke-Watzky (F-W) 2-step an excellent (R(2)>0.99), yielding first quantitative rate constant for steps nucleation growth aggregation. pharmaceutical preparation contains (initially) dimers, approximately 1.6% total concentration, effect on seeding analyzed F-W model assuming [B]0≠0 (for seeded case). results suggested had no impact Furthermore, Arrhenius equation cannot used extrapolate shelf-life since linear temperature dependence found within range. Although real-time provides basis determining product shelf-life, predictive methodologies such Vogel-Tammann-Fulcher (VFT) or approach misleading result overestimates shelf-life. However, they successfully applied fixing lower upper limits rate, i.e. best worst-case scenarios regarding potential product. In conclusion, present study evaluates application fitting interpretation experimental preparations, SEC-MALLS this context.