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Korean J Parasitol > Volume 28(3):1990 > Article

Original Article
Korean J Parasitol. 1990 Sep;28(3):135-142. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1990.28.3.135
Copyright © 1990 by The Korean Society for Parasitology
Purification of antigenic protein of sparganum by immunoaffinity chromatography using a monoclonal antibody
S Y Cho,S Y Kang and Y Kong
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.

The quality improvement of antigen (crude saline extract) of Spirometra mansoni pleroceroid (sparganum) was investigated by protein purification. The crude extract was fractionated by gel filtration through Sephacryl S-300 Superfine. Its third fraction was purified by affinity chromatography using a monoclonal antibody as ligand. When observed by SDS-PAGE, the purified protein was composed of 2 bands of 36 kDa and 29 kDa which were found already as the most sensitive components in the crude extract by immunoblots with patients sera. The quality of the purified antigen was evaluated in comparison with the crude extract by enzyme-linked immunosorbent assay (ELISA) for the specific (IgG) antibody in sera of human sparganosis, other parasitic and neurologic diseases, and normal control. When the purified antigen was used, the sensitivity was not altered but remained high (96.4%) while the specificity was increased from 86.8% to 96.9%.


Fig. 1
Fractionation of crude saline extract of sparganum through Sephacryl S-300 Super-fine. 1.6(ϕ)×40cm sized column was used. Eluent: 0.15 M PBS containing 0.01% merthiolate. Flow rate: 30 ml·cm-2·hour. Collection volume: 35 drops/tube.

Fig. 2
Disc-PAGE findings of the crude and fractions at 10% gel concentration.

C: Crude extract, 1: Fraction 1, 2: Fraction 2, 3: Fraction 3, 4: Fraction 4, 5: Fraction 5.

Fig. 3
SDS-PAGE findings of the crude and fractions. Ten-15% linear gradient gel was used for protein separation. Mr: Molecular weight in kDa, C, 1-5: Same as described in Fig. 2.

Fig. 4
SDS-PAGE/immunoblot finding of SP-53-2 monoclonal antibody to sparganum antigen.

Fig. 5
Elution profile of immunoaffinity chromatography.

UB: Unbound protein, B: Bound protein.

Fig. 6
Findings of SDS-PAGE of the crude and purified proteins.

Mr: Molecular weight in kDa, C: Crude extract, 1: UB of crude extract, 2: Fraction 3, 3: UB of crude Fraction 3, 4: B of crude extract, 5: B of Fraction 3.


Table 1
Mean absorbance for sparganum-specific IgG antibody in human sparganosis and other conditions against the crude and processed antigens of sparganum

Table 2
Comparison of sensitivity and specificity in 28 sparganosis and 99 other conditions in different antigens

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. Beaver PC, Rolon FA. Proliferating larval cestode in a man in Paraguay. A case report and review. Am J Trop Med Hyg 1981;30(3):625–637.
4. Cha CY, et al. J Korean Med Ass 1984;27:253–263.
5. Chan ST, Tse CH, Chan YS, Fong D. Sparganosis of the brain. Report of two cases. J Neurosurg 1987;67(6):931–934.
6. 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.
7. Cho SY, Kim SI, Kang SY, Choi DY, Suk JS, Choi KS, Ha YS, Chung CS, Myung HJ. Evaluation of enzyme-linked immunosorbent assay in serological diagnosis of human neurocysticercosis using paired samples of serum and cerebrospinal fluid. Korean J Parasitol 1986;24(1):25–41.
8. 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.
9. Davis BJ. Disc Electrophoresis. II. Method and Application to Human Serum Proteins. Ann N Y Acad Sci 1964;121:404–427.
10. Fan KJ, Pezeshkpour GH. Cerebral sparganosis. Neurology 1986;36(9):1249–1251.
11. 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.
12. Kim H, Kim SI, Cho SY. Serological Diagnosis Of Human Sparganosis By Means Of Micro-ELISA. Korean J Parasitol 1984;22(2):222–228.
13. Kim SI, Kang SY, Cho SY, Hwang ES, Cha CY. Purification of cystic fluid antigen of Taenia solium metacestodes by affinity chromatography using monoclonal antibody and its antigenic characterization. Korean J Parasitol 1986;24(2):145–148.
14. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680–685.
15. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
16. 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.
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