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Korean J Parasitol > Volume 29(1):1991 > Article

Original Article
Korean J Parasitol. 1991 Mar;29(1):1-7. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1991.29.1.1
Copyright © 1991 by The Korean Society for Parasitology
Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography
Y Kong,S Y Kang and S Y Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract

Out of many component proteins in crude saline extract of Spirometra mansoni plerocercoid (sparganum), 36 kDa and 29 kDa proteins were found to be the most antigenic and were already purified by immunoaffinity chromatography using monoclonal antibody as a ligand. In this study, a single step purification of these potent antigenic proteins of sparganum extract was investigated. When the crude saline extract was charged to gelatin-Sepharose 4B affinity column, 36 kDa and 29 kDa protein fractions were bound. SDS-polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE/immunoblot confirmed that the bound protein to gelatin was serologically pure. When evaluated by ELISA with patients sera, the purified protein of 36 and 29 kDa also showed improved antigenicity.

Figures


Fig. 1
Findings on reducing SDS-PAGE of sparganum extract. For protein separation, 10~15% linear gradient gel was used.

Mr: Molecular weight in kDa, C: Crude extract U: Unbound protein, B: Bound protein



Fig. 2
Western blot analysis of antigenicity of the crude extract (C), unbound protein(U) and bound protein(B) suing a patient serum of surgery confirmed sparganosis.

Mr: Molecular weight in kDa, A: Amido black B: stained nitrocellulose strip of crude extract.


Tables


Table 1
Mean absorbance and standard deviations of sparganum-specific IgG antibody by ELISA in human sparganosis and other conditions against crude and purified sparganum antigens


Table 2
Comparison of sensitivity and specificity of the purified antigen in 27 sparganosis and 91 other conditions

References
1. Anders K, Foley K, Stern E, Brown WJ. Intracranial sparganosis: an uncommon infection. Case report. J Neurosurg 1984;60(6):1282–1286.
  
2. Anegawa S, Hayashi T, Ozuru K, Kuramoto S, Nishimura K, Shimizu T, Hirata M. Sparganosis of the brain. Case report. J Neurosurg 1989;71(2):287–289.
  
3. Chan ST, Tse CH, Chan YS, Fong D. Sparganosis of the brain. Report of two cases. J Neurosurg 1987;67(6):931–934.
  
4. Chang KH, Cho SY, Chi JG, Kim WS, Han MC, Kim CW, Myung H, Choi KS. Cerebral sparganosis: CT characteristics. Radiology 1987;165(2):505–510.
 
5. Cho SY, Bae JH, Seo BS. Some Aspects Of Human Sparganosis In Korea. Korean J Parasitol 1975;13(1):60–77.
 
6. Cho SY, Kang SY, Kong Y. Purification of antigenic protein of sparganum by immunoaffinity chromatography using a monoclonal antibody. Korean J Parasitol 1990;28(3):135–142.
 
7. Choi SH, Kang SY, Kong Y, Cho SY. Antigenic protein fractions reacting with sera of sparganosis patients. Korean J Parasitol 1988;26(3):163–167.
 
8. Dresden MH, Rege AA, Murrell KD. Strongyloides ransomi: proteolytic enzymes from larvae. Exp Parasitol 1985;59(2):257–263.
  
9. Duve C. Exploring cells with a centrifuge. Science 1975;189(4198):186–194.
  
10. Engvall E, Ruoslahti E. Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int J Cancer 1977;20(1):1–5.
  
11. Fan KJ, Pezeshkpour GH. Cerebral sparganosis. Neurology 1986;36(9):1249–1251.
  
12. Fukase T, et al. Jpn J Parasitol 1986;34(5):351–360.
13. Gelder FB, Brown ST 3rd. Fibronectin gelatin-binding activity: effects of platelet thrombospondin, serum, and plasma. J Lab Clin Med 1987;110(5):548–557.
 
14. Katzman M, Lederman MM, Spagnuolo PJ. Fibronectin and cellular cytotoxicity: evidence against a role for fibronectin in natural killer activity. J Lab Clin Med 1987;110(1):75–82.
 
15. Kim CH, Yang J. Imunological characterization of antigens from cysticercus and sparganum and their application to immunodiagnosis 1. Immunological characteristics of crude antigenic components from Cysticercus cellulosae. Korean J Parasitol 1988;26(4):245–254.
 
16. Kim H, Kim SI, Cho SY. Serological Diagnosis Of Human Sparganosis By Means Of Micro-ELISA. Korean J Parasitol 1984;22(2):222–228.
 
17. Laclette JP, Alagon A, Willms K, Torre-Blanco A. Purification of antigen B from Taenia solium cysticerci by affinity to mammalian collagen. J Parasitol 1990;76(2):273–275.
  
18. Laemmli UK. Nature 1970;227:681–685.
 
19. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
 
20. McDonald JA, Kelley DG, Broekelmann TJ. Role of fibronectin in collagen deposition: Fab' to the gelatin-binding domain of fibronectin inhibits both fibronectin and collagen organization in fibroblast extracellular matrix. J Cell Biol 1982;92(2):485–492.
  
21. McKerrow JH. Parasite proteases. Exp Parasitol 1989;68(1):111–115.
  
22. McKerrow JH, Jones P, Sage H, Pino-Heiss S. Proteinases from invasive larvae of the trematode parasite Schistosoma mansoni degrade connective-tissue and basement-membrane macromolecules. Biochem J 1985;231(1):47–51.
 
23. McLaren M, Draper CC, Roberts JM, Minter-Goedbloed E, Ligthart GS, Teesdale CH, Amin MA, Omer AH, Bartlett A, Voller A. Studies on the enzyme linked immunosorbent assay (ELISA) test for Schistosoma mansoni infections. Ann Trop Med Parasitol 1978;72(3):243–253.
 
24. Mineura K, Mori T. Sparganosis of the brain. Case report. J Neurosurg 1980;52(4):588–590.
  
25. Ruoslahti E, Hayman EG, Pierschbacher M, Engvall E. Fibronectin: purification, immunochemical properties, and biological activities. Methods Enzymol 1982;82(Pt A):803–831.
  
26. Song CY, et al. Proc Mol Biol Genet 1990;5:241–246.
27. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 1979;76(9):4350–4354.
  
28. Tsang VC, Peralta JM, Simons AR. Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods Enzymol 1983;92:377–391.
  
29. Velge P, Ouaissi MA, Cornette J, Afchain D, Capron A. Identification and isolation of Trypanosoma cruzi trypomastigote collagen-binding proteins: possible role in cell-parasite interaction. Parasitology 1988;97(Pt 2):255–268.
  
30. Yamada KM. Lymphokine Reports 1980;1:231–254.
31. Yamada KM, Olden K. Fibronectins--adhesive glycoproteins of cell surface and blood. Nature 1978;275(5677):179–184.
  
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