Effect of molecular weight and ratio of poly ethylene glycols’ derivatives in combination with trehalose on stability of freeze-dried IgG

Abstract

The influence of poly ethylene glycol (PEG) at different molecular weights (MWs) and ratios was studied on the stability of freeze-dried immune globulin G (IgG). PEGs (600-4000 Dalton) at concentrations of 0.5 and 5% W/V were applied in the presence of 40 and 60% W/W of trehalose to prepare freeze-dried IgG formulations. Size-exclusion chromatography, infra-red spectroscopy, differential scanning calorimeter, and gel electrophoresis were performed to characterize lyophilized samples. Pure IgG demonstrated the highest aggregation of 5.77 ± 0.10% after process and 12.66 ± 0.50% as well as 44.69 ± 0.50% upon 1 and 2 months of storage at 45 °C, respectively. 5% W/V of PEGs 4000 in combination with 40% W/W trehalose, significantly suppressed aggregation, 0.05 ± 0.01%, with minimum aggregation rate constant of 0.32 (1/month). The integrity of IgG molecules and secondary conformation were properly preserved in all formulations comparing native IgG. It could be concluded that appropriate concentration and MW of PEGs, prominently augmented stabilizing effect of trehalose on freeze-dried antibody through inserting additional supportive mechanisms of actions.

Stability of Monoclonal Antibodies as Solid Formulation for Auto-Injectors: A Pilot Study

“…Different PEGs have been successfully used as cryoprotectants of solid formulations, proteins, and even cells [ 31 , 32 , 33 , 34 ]. In most cases, high molecular weight PEGs (from 2000 to 10,000) are preferred for cryoprotection.…”

“…Sugars are the most widely known excipients for cryoprotection and protein stability, among which Tre and Man can be highlighted [ 31 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ]. These excipients, due to their low MW, were added to C-ADA after the concentration process.…”

“…The need to include other excipients in the concentrated formulation is therefore clear, the primary objective being the improvement of the monoclonal antibody stability after the concentration and freeze-drying step. Different PEGs have been successfully used as cryoprotectants of solid formulations, proteins, and even cells [31][32][33][34]. In most cases, high molecular weight PEGs (from 2000 to 10,000) are preferred for cryoprotection.…”