Abstract
Poloxamer 188, also known as Pluronic F-68, is an excipient added to the biotherapeutic protein-manufacturing process. Poloxamer 188 (P188) is a nonionic triblock copolymer surfactant that can be used as a shear protective excipient in bioreactors. In the current study, a method for the process clearance monitoring of poloxamer 188 during downstream processing of biotherapeutics using liquid chromatography-triple-quadrupole mass spectrometry was developed and validated. Chromatographic separation of P188 was achieved using a Phenomenex, Luna 3 μm phenyl-hexyl, 150 × 2 mm column, and quantitation was achieved using a triple-quadrupole mass spectrometer operated in selected reaction monitoring mode. Linearity was assessed over a working range of 250-10,000 ng/mL. Precision and accuracy were within 15% of the theoretical spike levels assessed across the three different concentration levels. For this study, two different IgG1 antibodies were used for the method validation activities. Analyte specificity and selectivity were deemed acceptable based on no extraneous peaks present. System suitability was evaluated throughout this study in anticipation of the introduction of this method into the quality control environment. This method was successfully validated and used to monitor the clearance of poloxamer 188 in a tangential flow filtration purification step during biotherapeutic downstream processing. In addition, the capability of the method to successfully support poloxamer 188 mixing studies is presented in this work.