Development of a multidose formulation for a humanized monoclonal antibody using experimental design techniques

Area of Science:

• Pharmaceutical Sciences
• Biotechnology
• Protein Chemistry

Background:

• Multidose protein formulations require effective preservatives to maintain stability and prevent microbial contamination.
• Selecting appropriate preservatives is crucial to avoid compromising protein integrity and therapeutic efficacy.
• Traditional screening methods can be time-consuming and costly.

Purpose of the Study:

• To identify optimal antimicrobial preservatives for a multidose humanized monoclonal antibody formulation.
• To assess the impact of various preservatives on protein stability using multiple analytical techniques.
• To develop a cost-effective screening method and utilize experimental design for preservative selection.

Main Methods:

• Utilized size-exclusion chromatography, differential scanning calorimetry, light scattering, and UV spectroscopy to evaluate protein stability.
• Employed cell-based fluorescence-activated cell sorting for potency testing.
• Implemented a quick preservative screening test and an I-optimal experimental design to examine preservative combinations.

Main Results:

• Methylparaben and propylparaben demonstrated the highest protein stability.
• Benzyl alcohol and chlorobutanol were compatible at low concentrations; phenol and m-cresol were incompatible.
• An I-optimal design identified benzyl alcohol as a promising individual preservative and several effective combinations.

Conclusions:

• The developed screening test and experimental design approach successfully identified optimal preservative concentrations for multidose protein formulations.
• Benzyl alcohol and a combination of methylparaben and chlorobutanol were identified as leading candidates meeting regulatory requirements for preservative efficacy and protein stability.