Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography

, Kong, Y; , Kang, S Y; , Cho, S Y

doi:1991.29.1.1

Korean J Parasito > 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. M_r: 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. M_r: 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

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