ENHANCING SOLUBLE EXPRESSION AND ONE-STEP PURIFICATION OF AFRICAN SWINE FEVER VIRUS P30 PROTEIN IN E. COLI USING A GST-SNAC DUAL-TAG SYSTEM

Abstract

African Swine Fever (ASF), a highly contagious and deadly disease of swine caused by the African Swine Fever virus (ASFV), leads to significant economic losses due to the lack of effective vaccines or treatments. The ASFV-p30 antigen, an early-expressed and highly immunogenic protein, has significant potential for early detection; however, its structural instability presents challenges for research and recombinant production. In this study, a dual-tag strategy combining Glutathione S-transferase (GST) and sequence-specific nickel-assisted cleavage (SNAC) was used to improve the solubility and purification efficiency of ASFV-p30 in E. coli. The GST tag facilitated proper protein folding, while the SNAC sequence enabled Ni²⁺ chemical cleavage, allowing non-enzymatic GST removal. This approach optimizes purification, reduces processing time, and offers a promising strategy for ASF research and diagnostic applications.