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Korean J Parasitol > Volume 29(4):1991 > Article

Original Article
Korean J Parasitol. 1991 Dec;29(4):363-369. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1991.29.4.363
Copyright © 1991 by The Korean Society for Parasitology
Component proteins in crude extract of adult Paragonimus westermani purified by immunoaffinity chromatography using monoclonal antibodies
S Y Kang,Y Kong and S Y Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract

The nature of 2 component proteins in crude saline extract of adult Paragonimus westermani was investigated. By immunoaffinity chromatography using monoclonal antibodies (MAb) as ligands, the proteins were purified from the crude extract. Band 1 protein in disc-polyacrylamide gel electrophoresis (PAGE) was purified by PFCK-136 MAb. The protein, known to have molecular mass of 440 kDa, was composed of 23, 46 and 92 kDa subunits when observed by reducing SDS-PAGE and SDS-PAGE/immunoblot. This protein was originated from eggs of the worm as revealed by immunohistochemical staining with PFCK-136 Mab. Another affinity purified protein utilizing PFCK-44 MAb was the band 4 protein of 17 kDa in disc-PAGE. This was a monomer protein in reducing SDS-PAGE and SDS-PAGE/immunoblot. The protein was produced at intestinal epithelium of the worm.

Figures


Fig. 1
Findings of SDS-PAGE/immunoblot of MAbs reacted to crude saline extract of adult P. westermani.

A: PFCK-21, B: PFCK-44, C: PFCK-136, D: PFCK-189



Fig. 2
Findings in disc-PAGE of the component proteins of adult P. westermani obtained in affinity chromatography.

C: Crude extract, 1: Unbound protein(UP) from affinity chromatography using PFCK-44, 2: Bound protein(BP) of PFCK-44, 3: UP of PFCK-136, 4: BP of PFCK-136, 5: UP of PFCK-44 and PFCK-136



Fig. 3
SDS-PAGE findings of the component and purified proteins of P. westermani in 10~15% gradient gel.

Mr: Molecular mass in kDa, C: Crude extract, 1: UP of PFCK-44, 2: BP of PFCK-44, 3: UP of PFCK-136, 4: BP of PFCK-136, 5: UP of PFCK-44 and PFCK-136



Figs. 4-9
Immunohistochemical stainings of P. westermani and its surrounding granuloma with MAbs and paragonimiasis patient serum.

Fig. 4. Negative control (×100). Normal human serum was reacted as a primary antibody. No organs were reacted with AEC chromogen.

Fig. 5. Negative control reacted with normal human serum on granuloma wall with eggs (×100).

Fig. 6. A patient serum of paragonimiasis was used as a primary antibody. Strong reactions were shown (see text).

Fig. 7. Section of eggs in granuloma wall reacted with a patient serum (×100).

Fig. 8. Reactivity of PFCK-136 MAb to eggs in granuloma wall (×100).

Fig. 9. Reactivity of PFCK-44 MAb to intestinal wall of the worm (×200).


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