Study on methods and requirements for quality control of recombinant human tumor necrosis factor receptor Fc fusion protein.
- Author:
Yi ZHANG
1
;
Kai GAO
;
Chun-mei HAN
;
Chun-ming RAO
;
Jun-zhi WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Biotechnology; methods; Cell Division; drug effects; Etanercept; Immunoglobulin G; biosynthesis; chemistry; pharmacology; Mice; Peptide Mapping; Quality Control; Receptors, Tumor Necrosis Factor; biosynthesis; chemistry; Recombinant Fusion Proteins; biosynthesis; chemistry; pharmacology; Technology, Pharmaceutical; standards; Tumor Cells, Cultured
- From: Acta Pharmaceutica Sinica 2003;38(3):165-168
- CountryChina
- Language:Chinese
-
Abstract:
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-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.
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-alpha. The specific activity was up to 8 x 10(4) 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.
