Abstract
Avian reovirus (ARV) causes a number of severe chicken diseases and death. One of the outer structural proteins, sC, is linked to cell fusion and neutralizing antibody induction, and therefore becomes the prime candidate for vaccine development and diagnosis tool. In this study, we expressed sC using baculovirus/insect cell system and characterized the production profiles. The yield reached 96 mg/106 cells, which is high compared to other viral proteins expressed in this system. Besides, the protein was fused with additional histidine tag and an EK cleavage site to facilitate the downstream purification and subsequent removal of the histidine tag. Successful and efficient one-step purification using immobilized metal affinity chromatography (IMAC) was demonstrated and the recovery was roughly 58%. The purity by a single step reached 94%. Additionally, the histidine tag was successfully removed by incubating the purified protein with EK. Overall, the fusion strategy resulted in high level expression of sC properly fused to a histidine tag and an EK cleavage site, the recombinant protein conformation conferred reasonable accessibility of the histidine tag to Ni2+ for IMAC purification and the enterokinase cleavage, and enabled successful, simple downstream purification.