Construction of IL-1Ra-HSA fusion protein and analysis of its bioactivity and pharmacokinetics.
- Author:
Yi HUANG
1
;
Lei HU
;
Yan-Qun YANG
;
Xue-Ping HU
;
Yong-Su ZHEN
;
Meng-Yuan LIU
Author Information
1. Center for Infection and Immunity Research, Life Sciences College of Hubei University, Wuhan 430062, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Apoptosis;
drug effects;
Arthritis, Experimental;
metabolism;
Cell Line, Tumor;
Forelimb;
metabolism;
Half-Life;
Humans;
Interleukin 1 Receptor Antagonist Protein;
genetics;
metabolism;
pharmacokinetics;
pharmacology;
Interleukin-1beta;
toxicity;
Male;
Melanoma;
pathology;
Mice;
Mice, Inbred DBA;
Pichia;
genetics;
metabolism;
Plasmids;
Recombinant Fusion Proteins;
genetics;
metabolism;
pharmacokinetics;
pharmacology;
Serum Albumin;
genetics;
metabolism;
pharmacokinetics;
pharmacology;
Tissue Distribution
- From:
Acta Pharmaceutica Sinica
2012;47(9):1210-1218
- CountryChina
- Language:Chinese
-
Abstract:
In order to increase the plasma half-life and tissue specificity of IL-1 receptor antagonist, a recombinant fusion protein IL-1Ra-HSA, linked by a rigid peptide linker PAPAP, was engineered and expressed by the Pichia pastoris host cells. The fusion protein was secreted to the host cells culture, identified by Western blot, and purified by affinity chromatography. This was followed by a further examination of its bioactivity and pharmacokinetics. Our results demonstrated that the fusion protein retained the antagonist activity of IL-1Ra, capable of binding specifically to the IL-1 receptor on human melanoma A375.S2 cells, and inhibits the cytolytic effect of IL-1beta to A375.S2 cells. Albumin fusion dramatically extended the half-life of IL-1Ra and resulted in a specific accumulation of IL-1Ra in the arthritic paws and a lower distribution of IL-1Ra in other organs such as liver, kidney, spleen and lung in mice with collagen-induced arthritis. The findings reported herein indicate that the fusion protein is likely to have greater clinical applications in areas such as the treatment of rheumatoid arthritis.
