Use of a Design of Experiments (DoE) Approach to Optimize Large-Scale Freeze-Thaw Process of Biologics

Large volumes of protein solutions are commonly stored in a frozen state before further drug product fill and finish. This study aimed to establish design space perform large-scale freeze-thaw (F/T) processes biotherapeutics without inducing destabilization. A response surface model was designed evaluate the following main factors interactions: volume solution 1-L containers, distance among nine containers during both F/T, freezer set temperature, novel forced air flow methodology thawing. The analysis from 46 experimental runs indicated over 4-fold increase freezing rate by lowering temperature -20 -80°C, at 98 fpm doubled thawing rate. Furthermore, multivariate linear regression modeling revealed significant impact all investigated on lactate dehydrogenase (LDH) quality attributes. factor that most strongly affected retention LDH activity loading distance: ≥ 5 cm positively 50.6%. retained tetramers towards lower range -80°C (2.2% higher tetramer compared -20°C freezing, due faster rate). In summary, this DoE-based systematic increased F/T process understanding large scale, identified critical parameters, confirmed feasibility applying procedures maintain stability subject F/T.