Expression and purification of recombinant mTLNE. (A) Schematic representation of the mTLNE constructs. Three linear neutralizing epitopes (VP1-SP55, VP1-SP70 and VP2-SP28) was sequentially linked with (Gly4Ser)3 sequence. Thioredoxin (Trx) was fused at the N-terminal, and His-Patch at the C-terminal. (B) SDS-PAGE results of recombinant proteins. The induced cells were harvested by centrifugation, and the pellet was re-suspended completely by mixing in PBS. Following sonication, the supernatant and the precipitate were harvested and assessed on by SDS-PAGE. Prominent protein bands of about 30 kDa and 20.8 kDa were visible in induced fractions. Lanes 1 and 2: the precipitate and supernatant from E. coli receiving pET32a-mTLNE. Lanes 4 and 5: the precipitate and supernatant from E. coli receiving pET32a plasmid. Lanes 3 and 6: the purified recombinant mTLNE and Trx protein.