Abstract
High protein concentration products for targeted therapeutic use are often freeze-dried to enhance stability. The long-term storage stability of freeze-dried (FD) plasma-derived Immunoglobulin G (IgG) from moderate to high concentrations (10–200 mg/mL) was assessed. Monomer content, binding activity and reconstitution times were evaluated over a 12-month period under accelerated and real-term storage conditions. In the first case study it was shown that FD IgG from 10 to 200 mg/mL had minimal monomer/activity losses at up to ambient temperature after 12 months of storage. However, at 45 °C the sucrose-to-protein ratio played a significant impact on IgG stability above 50 mg/mL. All IgG concentrations witnessed moisture ingress over a 12-month period. The impact of moisture ingress from environmental exposure (between 0.1% and 5% w/w moisture) for IgG 50 mg/mL was assessed, being generated by exposing low moisture batches to an atmospheric environment for fixed time periods. Results showed that at −20 °C and 20 °C there was no significant difference in terms of monomer or antigen-binding activity losses over 6 months. However, at 45 °C, there were losses in monomer content, seemingly worse for higher moisture content samples although model binding activity indicated no losses. Finally, the difference between a low moisture product (0.1–1% w/w) and a moderately high moisture (3% w/w) product generated by alternative freeze-drying cycles, both stoppered under low oxygen headspace conditions, was evaluated. Results showed that at −20 °C and 20 °C there was no difference in terms of binding activity or monomer content. However, at 45 °C, the low moisture samples had greater monomer and binding activity losses than samples from the highest moisture cycle batch, indicating that over-drying can be an issue.
Highlights
There is rising demand in the biopharmaceutical industry for increasingly high concentrations of therapeutic proteins, such as monoclonal antibodies
In this study the Immunoglobulin G (IgG) molar/weight ratio was intentionally varied as the sucrose excipient concentration of 10 mg/mL (1% w/w) was maintained in order to investigate the destabilizing effect of moisture ingress and/or storage temperature conditions during long-term storage
IgG 10 mg/mL with the 1:1 protein:sucrose ratio had greater stability shielding at elevated temperatures compared to concentrations above 50 mg/mL, but this may reflect the need for a higher sucrose content at a higher protein composition
Summary
There is rising demand in the biopharmaceutical industry for increasingly high concentrations of therapeutic proteins, such as monoclonal antibodies (mAbs). With this requirement there comes a host of challenges, including an increased instability, degradation and viscosity, as well as unwanted protein–protein interactions in the liquid state [1,2]. In addition to the continuing development of higher protein concentration formulations in FD formats, there are challenges with regard to selecting the appropriate moisture content for long-term storage stability. During long-term storage, the sugar excipient used in the formulation, moisture content and temperature can all affect the physical and chemical stability of FD cakes. The monomer content, retention of anti-diphtheria/tetanus antibody titres and change in moisture content were evaluated under accelerated and real-time storage conditions (−20 ◦C, ~
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