Results of a harmonized endotoxin recovery study protocol evaluation by 14 BioPhorum Operations Group (BPOG) member companies.

ABSTRACT

Significant regulatory and biopharmaceutical manufacturing attention has been recently generated regarding the inability to recover purified lipopolysaccharide in certain biopharmaceutical products, specifically those containing a chelator and surfactant. Disparate data has been generated and submitted during regulatory review. Study design is thought to significantly impact, and cause the confounding data. The BioPhorum Operations Group (BPOG) is an industry consortium of biopharmaceutical manufacturers. A BPOG team developed a collaborative experiment to determine if multiple laboratories could execute a harmonized protocol and reach equivalent conclusions. Twenty-one laboratories across fourteen different companies participated, yielding a robust study and large dataset. The experiment was successful as nearly all of the laboratories reached the same conclusions despite allowable variation in methods, suppliers, reagents, locations and analysts. The results of the study indicate a naturally occurring endotoxin analyte activity was recovered in a chelating buffer containing surfactant (citrate and polysorbate), and a buffer without surfactant (phosphate), while the purified lipopolysaccharide analyte activity was not recovered in the chelating buffer containing surfactant, but was recovered in the buffer without surfactant. Because the harmonized protocol yields consistent results across many different laboratories, the BPOG LER team recommends adoption of this protocol for use in endotoxin recovery studies.