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Immunoelectrophoretic analysis of major component proteins in cystic fluid of Taenia solium metacestodes
Y Kong,S Y Cho,S I Kim,** and S Y Kang
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract
When cystic fluid of Taenia solium metacestodes (CF) was filtrated through Sephacryl S-300 Superfine, major proteins were in fractions III and IV. Major protein in fraction III was Band C protein of 150 kDa and that in fraction IV was Band N protein (Choi et al., 1990). When CF was electrophoresed in 0.9% agarose gel and reacted with anti-CF rabbit serum (RACF), two main bands, a long outer and a short inner band, were precipitated, together with 8 minor bands. RACF reacted with fraction III forming the long outer band whereas RACF formed the short inner band with fraction IV in immunoelectrophoresis (IEP). The long outer precipitin band of CF fraction III was similar to antigen B in hydatid fluid (HF) of Oriol et al. (1971), while the short inner band of CF fraction IV was similar to HF antigen 5 of Capron et al. (1967). When HF was reacted with RACF, the short inner band was immunoprecipitated without forming the long outer band. Common antigenicity between CF and HF seemed to exist in fraction IV rather than in fraction III of CF. Patient sera of neurocysticercosis reacted more frequently with fraction III than with fraction IV.
Figures
Fig. 1 Elution profile of CF through Sephacryl S-300 Superfine. The major fractions were fraction III and fraction IV.
Fig. 2 Disc-PAGE findings of CF and its fractions in 8% gel. CF: Crude cystic fluid of T. solium metacestodes, III: fraction III, IV: fraction IV of CF. Letters at left border stand for the designated band names.
Fig. 3 SDS-PAGE/immunoblot finding of CF and its fractions using RACF as antibody. A: Molecular mass markers (Mr) and CF transfer blotted to nitrocellulose paper by electrophoresis and stained by amido black B (Cr). R: proteins in cystic fluid (Cr), Fraction III (III) and Fraction IV (IV) was separated by reducing SDS-PAGE and immunoblotted with RACF. N: CF, fraction III and IV were separated by non-reducing SDS-PAGE and immunoblotted with RACF.
Fig. 4 Immunoelectrophoresis finding of CF and its fractions using RACF before absorption (A) or after absorption test (B). Troughs were filled with RACF. Antigens in the wells were shown as: CF, crude cystic fluid; Fr III, fraction III; Fr IV, fraction IV; CP, parenchymal extract of T. solium metacestodes; HF, hydatid fluid. ◂, Long outer precipitin bands; ★, short inner bands.
Fig. 5 Comparative finding of immunoelectrophoresis using CF and CP as antigens and reacted with patient sera of neurocysticercosis. Numbers at troughs mean different patient sera.
Fig. 6 Comparative finding of immunoelectrophoresis using Fraction III and Fraction IV as antigen and reacted with patient sera of neurocysticercosis. Numbers at troughs mean different patient sera.
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