Study on methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein

AimTo establish methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein (rhTNFR-Fc). MethodsBiological potency of rhTNFR-Fc was determined by neutralizing the bioactivity of TNF-α. rhTNFR-Fc samples were reduced by β-mercaptoethanol and the peptide map was performed by tryptic digestion. Residual protein A and the host cell protein content were detected by ELISA. Anti-TNFR and anti-Fc antibodies were used in ELISA for detection of the rhTNFR-Fc content. ResultsThe quality control methods, such as bioassay, peptide map, residual protein A detection, were established and used for quality control of rhTNFR-Fc. The unit of rhTNFR-Fc (AU) was defined according to the international unit of TNF-α. The specific activity was up to 8×10⁴ AU·mg^-1. The requirements for quality control of rhTNFR-Fc were established. ConclusionThe methods and requirement were used for quality control of rhTNFR-Fc products.