Construction, expression and preliminary pharmacokinetic analysis of IL-1ra mutants / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 472-476, 2006.
Article
Dans Chinois
| WPRIM
| ID: wpr-286265
ABSTRACT
Interleukin-1 receptor antagonist (IL-1ra), a member of IL-1 family, is a naturally occurring IL-1 inhibitor as "receptor antagonist", which blocks biological responses mediated by IL-1. Recombinant human IL-1ra (rhIL-1ra, Kineret) was introduced in clinical trials involving patients with RA. Between 2001 to approximately 2002, rhIL-1 ra was approved by the US Food and Drug Administration and the European Agency for the Evaluation of Medicine Procedure. Unfortunately, 10,000 to 100,000-fold excess amounts of IL-1ra are needed to relieve disease because minimal IL-1 can induce complete biological responses, and the dosage of 100 to approximately 150mg/day in a RA patient is so big that it greatly influence patients' physical, psychological and economical situation. In this study, IL-1ra mutants were established by site-specific mutagenesis to improve its stability. The sites of mutagenesis included R6 K7-AA,R93 K94-AA and K97 R98-AA. IL-1ra and its mutants were expressed in E. coli BL21 (DE3) using pTIG-Trx expressing system with the induction of IPTG. The recombinant proteins were purified by Ni2+ chelate chromatography and Sephadex G75 gel filtration chromatography. The activity of mutants is as high as IL-1ra. We characterized the pharmacokinetic profile of IL-1ra and its mutants. The third mutant's half life is 2.26 times than wt IL-1ra. The study has provided some approaches and experience for further research to improve the metabolism stability of IL-1ra.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Protéines recombinantes
/
Pharmacocinétique
/
Mutagenèse dirigée
/
Escherichia coli
/
Protéines mutantes
/
Antagoniste du récepteur à l'interleukine-1
/
Génétique
/
Métabolisme
/
Méthodes
Limites du sujet:
Animaux
/
Femelle
/
Humains
langue:
Chinois
Texte intégral:
Chinese Journal of Biotechnology
Année:
2006
Type:
Article
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