Schistosoma japonicum: Efficient and rapid purification of the tetraspanin extracellular loop 2, a potential protective antigen against schistosomiasis in mammalian

• Published in: Experimental parasitology 2010-12, Vol.126 (4), p.456-461
• Main Authors: Yuan, Chuang, Fu, Yi-jie, Li, Jie, Yue, Yu-fei, Cai, Lu-lu, Xiao, Wen-jing, Chen, Jian-ping, Yang, Li
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Amino Acid Sequence; Animals; Antibodies, Helminth - blood; Antigens, Helminth - chemistry; Antigens, Helminth - immunology; Antigens, Helminth - isolation & purification; Biological and medical sciences; Child; Co-affinity; Enterokinase (Enzyme Commission Number: 232-761-1); Enteropeptidase - metabolism; Female; Fluke; Fundamental and applied biological sciences. Psychology; Gene Expression; Humans; Life cycle. Host-agent relationship. Pathogenesis; Male; Mammalia; Membrane Proteins - chemistry; Membrane Proteins - immunology; Membrane Proteins - isolation & purification; Mice; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - immunology; Nerve Tissue Proteins - isolation & purification; pET32; Plasmids; Protozoa; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; Schistosoma japonicum; Schistosoma japonicum - immunology; Schistosoma mansoni; Schistosomiasis japonica - immunology; Schistosomiasis japonica - prevention & control; Sequence Alignment; Small peptide; Tetraspanin; Tetraspanins; Trx-Tag; Vaccination; Vertebrates: general zoology, morphology, phylogeny, systematics, cytogenetics, geographical distribution; Young Adult
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2010.05.018

The extracellular loop 2 of a tetraspanin from Schistosoma japonicum (Sj-TSP-2) is homologous to Schistosoma mansoni TSP-2. In our initial study, Sj-TSP-2 is an identical antigen against schistosomiasis caused by S. japonicum. Through the pET32 vector system and nickel (Ni)-absorbed chelating Sepharose, Sj-TSP-2 was expressed and purified as a soluble fusion constructed with an N-terminal thioredoxin-His 6-EK protease site tag (Trx-TSP-2). In phosphate buffer (PB) with a low concentration of imidazole, the Trx-TSP-2 fusion protein was efficiently cleaved by enterokinase (EK). Sj-TSP-2 was isolated and enriched using cobalt (Co)-absorbed chelating Sepharose and HiTrap SP column. Character of the protein was analyzed via animal experiments and then clinical trials. The purification approach yielded pure Sj-TSP-2, which will provide feasible advices for discovering vaccines against schistosomiasis.