Schistosoma japonicum: efficient and rapid purification of the tetraspanin extracellular loop 2, a potential protective antigen against schistosomiasis in mammalian.
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.
Yuan
Chuang
C
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Gaoxin District, Chengdu, Sichuan, PR China.
Fu
Yi-jie
YJ
Li
Jie
J
Yue
Yu-fei
YF
Cai
Lu-lu
LL
Xiao
Wen-jing
WJ
Chen
Jian-ping
JP
Yang
Li
L
D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002648:Child; D004765:Enteropeptidase / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012549:Schistosoma japonicum / Q000276:immunology*; D012554:Schistosomiasis japonica / Q000276:immunology / Q000517:prevention & control*; D016415:Sequence Alignment; D059470:Tetraspanins; D014611:Vaccination; D055815:Young Adult
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; C512650:TSPAN2 protein, human; D059470:Tetraspanins; D004765:Enteropeptidase