Purpose: Introduction: When solid protein dosage form is desirable, lyophilization (freeze drying) is one of the most common processing method for the formulation of biopharmaceuticals with the increased stability and shelf life. Objective of the study is to understand the stabilization effect of HPβCD, PVP, and NaCMC on protein formulation during lyophilization using BSA as a model protein with sucrose used as protectant control. BSA is a structurally well characterized protein which contains 583 amino acid. Isolated freeze-dried BSA is typically stored at refrigerated temperature. It was reported that formulated injectable BSA solution forms disulfide-linked BSA aggregates and gelation. Thermal treatment also alters the secondary and tertiary structures, denatures the protein and causes aggregation.
Purpose: To investigate the stabilization effect of HPβCD, PVP, and NaCMC in BSA lyophilization formulation.
Methods: Size exclusion HPLC (SEC) is used to monitor the aggregation of BSA. Stock solution was prepared by weighing 800 mg of BSA into 20 mL volumetric flask followed by the addition 10 mL of phosphate buffer solution (1XPBS), gently vortex until dissolve followed by q.s to 20 mL. Each sample was prepared by pipette 4 mL of stock into 20 mL vials followed by the addition of the Stabilizers in Table 1
Results: HPβCD (Cavitron W7) and PVP (C-12) are comparable as sucrose in the lyophilization process. Lyophilization formulations using HPβCD, PVP at accelerated storage conditions at 4 weeks (Figure 2) are comparable or better than sucrose in the monomer purity. Reconstitution time of lyophilization formulation using HPβCD, PVP, and sucrose is 105, 56, and 68 seconds using three replicates.
Conclusion: HPβCD and PVP are comparable with Sucrose in the Lyophilization process and better stabilizing the BSA formulation at accelerated storage condition at 25 °C for 4 weeks. Reconstitution time of lyophilization formulation using PVP gave comparable results as sucrose lyophilization formulation. Reconstitution time of lyophilization formulation using HPβCD is 37 seconds longer than sucrose lyophilization product.
Wucheng Wen– Ashland Specialty Ingredients, Delaware
Kamaru Lawal– Ashland Specialty Ingredients
Yunxia Bi– Technical Director, Ashland Specialty Ingredients
Thomas Dürig– Senior Director, Ashland Specialty Ingredients, Wilmington, Delaware