Liquid high concentration IgG1 antibody formulations by precipitation

Area of Science:

• Biopharmaceutical Manufacturing
• Protein Chemistry
• Formulation Science

Background:

• High-concentration antibody formulations are crucial for therapeutic efficacy and patient convenience.
• Traditional methods like ultrafiltration can be time-consuming and may impact protein stability.
• Developing efficient and scalable methods for antibody concentration is an ongoing challenge.

Purpose of the Study:

• To investigate an alternative manufacturing approach for liquid high-concentration antibody formulations.
• To evaluate the feasibility of using precipitation and re-dissolution for antibody concentration.
• To assess the impact of this method on antibody recovery, structure, activity, and stability.

Main Methods:

• Investigated antibody precipitation using ammonium sulphate, sodium citrate, and PEG 4000.
• Concentrated IgG1 antibody solutions from 20 to 100 mg/mL.
• Assessed recovery, secondary structure, binding activity, and stability post-re-dissolution.
• Evaluated scalability from 1 to 100 mL and the effect of residual precipitants.

Main Results:

• Achieved antibody recovery exceeding 95% with retention of native secondary structure and binding activity.
• Quantitative precipitation was reproducible with tested agents at specified conditions.
• Scalability was confirmed, and final protein concentration was influenced by precipitant removal and scale.
• Similar protein quality was observed post-re-dissolution, irrespective of the precipitant, but short-term storage led to increased aggregate formation.

Conclusions:

• Intermediate precipitation followed by re-dissolution is a viable method for producing high-concentration antibody formulations (up to 100 mg/mL).
• The method demonstrates high recovery and preserves critical quality attributes, comparable to standard ultrafiltration.
• Further optimization is needed to mitigate aggregate formation during short-term storage of precipitated particles.