[Study on methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein].
OBJECTIVE
To establish methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein (rhTNFR-Fc).


METHODS
Biological potency of rhTNFR-Fc was determined by neutralizing the bioactivity of TNF-alpha. rhTNFR-Fc samples were reduced by beta-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.


RESULTS
The 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-alpha. The specific activity was up to 8 x 10(4) AU.mg-1. The requirements for quality control of rhTNFR-Fc were established.


CONCLUSIONS
The methods and requirement were used for quality control of rhTNFR-Fc products.
Zhang
Yi
Y
Department of Biochemistry, National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China.
Gao
Kai
K
Han
Chun-mei
CM
Rao
Chun-ming
CM
Wang
Jun-zhi
JZ
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D002455:Cell Division / Q000187:drug effects; D000068800:Etanercept; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D051379:Mice; D010449:Peptide Mapping; D011786:Quality Control; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis* / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D013678:Technology, Pharmaceutical / Q000592:standards; D014407:Tumor Cells, Cultured
D007074:Immunoglobulin G; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; D000068800:Etanercept