Generation of a fusion protein of the extracellular domain of BR3 with the Fc fragment of human IgG1 (sBR3-Fc) in Pichia pastoris as an antagonist for BLyS

Applied Microbiology and Biotechnology
Peng CaoChangliang Luo

Abstract

Elevated levels of B cell-activating factor of the TNF family (BlyS) have been implicated in the pathogenesis of autoimmune diseases in human. Removal of pathogenic B lymphocytes by decoy receptors has demonstrated clinical benefit in both oncological and immunological diseases. In this report, we have constructed vectors for the convenient and rapid expression of the extracellular domain of BR3(sBR3) fused to the Fc fragment (hinge, CH2, CH3) of human IgG1 in the methylotrophic yeast, Pichia pastoris. SDS-PAGE assays of culture broth from a methanol-induced expression strain demonstrated that the recombinant sBR3-Fc fusion protein is secreted and recovered from the culture medium as a disulfide-linked, glycosylated homodimer. The recombinant protein was purified to >95% using protein A affinity chromatography and size exclusion chromatography steps. Bioactivity of the recombinant sBR3-Fc was confirmed by the ability of the protein to inhibit mouse B lymphocyte proliferation induced by BLyS in vitro. Our results suggest that the P. pastoris expression system can be used to produce large quantities of fully functional sBR3-Fc fusion protein for both research and industrial purposes.

References

Nov 1, 1989·Analytical Biochemistry·S C Gill, P H von Hippel
Jun 8, 1999·The Journal of Experimental Medicine·P SchneiderJ Tschopp
Jan 7, 2004·The Journal of Experimental Medicine·Brian P O'ConnorRandolph J Noelle

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Related Concepts

Related Feeds

B cell Activation

B cell activation is initiated by the ligation of the B cell receptor with antigen and ultimately results in the production of protective antibodies against potentially pathogenic invaders. Here is the latest research.

Autoimmune Diseases

Autoimmune diseases occur as a result of an attack by the immune system on the body’s own tissues resulting in damage and dysfunction. There are different types of autoimmune diseases, in which there is a complex and unknown interaction between genetics and the environment. Discover the latest research on autoimmune diseases here.