• Parasites, Hosts and Diseases
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• v.30 no.4
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• pp.323-328
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• 1992

Analysis of siR isolates of Trichemenes veginalis was carried out with sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunoelectrotransfer blot (EITB). Trichloroacetic acid-treated antigens of the 6 isolates revealed 25 protein profiles ranging 12~170 kDa of molecular weight in SDS-PAGE. In EITB, the specific immunogenic bands were visualized at 51 kDa and 96 kDa when HY-1 antigen was probed with difFerent mice sera immunized with 6 isolates of T. vaginalis. The banding patterns with different sera showed isolate-to-isolate variability. In EITB, homologous antigen (HY-1) did not show any enhanced response in reacting to homologous antiserum(HY-1) when 6 isolates of T. vaginalis were probed with a single serum (HY-1). It is assumed that the different banding patterns of six isolates show isolate-to-isolate variability and immunogenic common bands in 41, 47, 74 and 9:k kDa on EITB may connote the important significance on immune response in T. vaginalis infection.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.9-14
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• 1989

Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

• Journal of Life Science
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• v.5 no.2
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• pp.63-69
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• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.

• Journal of Life Science
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• v.5 no.2
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• pp.7-7
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• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.243-248
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• 1994

Plasma membrane proteins of a Korean isolate of Trichomonus vofinalis HY-1 were fractionated for antigen analysis. Homogenates of T. vaginalis were fractionated by the differential centrifugation using sucrose step-gradient method. The interface layer from the 25%/45% sucrose was collected as a plasma membrane fraction and its purity was examined by transmission electron microscopy. The antigenicity of plasma membrane fraction was analysed by enzyme-linked immunoelectrotransfer blot technique with immune rabbit serum and compared with surface antigen labelled with N. hydroxysuccinimide-biotin. The fluffy fraction of 25%/45% sucrose interface was homogeneous and membrane particles were present as extended sheet and concentric vesicles showing typical trilamellar appearance under transmission electron microscope. Seven fractions at 40, 50, 60, 110, 130, 140 and 150 kDa were identified as the antigenic membrane proteins in EITB with anti HY-1 rabbit serum. The common band at 60 kDa was detected both in antigenic fractions of plasma membrane and surface protein labelled with NHS-biotin. This result indicates that this protein is considered as a major surface antigen of T. vaginalis. The role of this surface antigen at 60 kDa should be studied further.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.1-8
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• 1989

Serodiagnosis of parasitic infections is widely used, since parasites or their eggs are not always detected by ordinary methods. The sensitive tests such as ELISA are highly dependent on the purity of antigens used. To solve this problem. many workers have tried to find species-specific components of antigens, The present study was performed to determine the antigenic profile of crude saline extracts of 3, 5, 8 and 12-week old p. westermani worms, which were collected from experimentally infected cats, based on SDS-PAGE and immunoblot technique. The results were as follows: 1. The SDS-PAGE showed at least 30 Protein bands ranging from 229 kDa to 10 kDa molecular weight. The protein components of p. wsstermani changed chronologically during its developmental period. The 229 kDa band was recognized only in 12-week old worms ($$SEP_{l2}$). 2. Analysis by ELISA showed a significant increase in antibody levels at 3 weeks in infected cats using crude saline extract antigens ($SEP_3,{\;}SEP_5,{\;}SEP_8,{\;}SEP_{l2}$). 3. By EITB using $SEP_3$ and $SEP_5$ infected cats recognisea major protein bands with molecular weight of 60, 35, 28, 25 or 21 kDa at 3~12 weeks of infection, and 3 additional antigens, 19, 13 and 10 kDa, were detected at 8~12 weeks of infections. 4. Using $SEP_8$ 5 antigens, 91, 85, 31, 25 and 21 kDa, were consistently detected by all infected sera tested. In addition, 3 antigens of lg. 13 and 10 kDa were detected at 8~12 weeks of infection. Using $SEP_12$, similar results were obtained with that by using $SEP_8$ and 1 additional antigen of 229 kDa, specifically reacting with the sera from 12 weeks of in(traction, was recognized.

• Parasites, Hosts and Diseases
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• v.26 no.4
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• pp.245-254
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• 1988

We studied the serological reaction between various antigenic components from Cysticercus cellulosae and IgG antibodies in sera of cysticercosis, sparganosis, hydatidosis patients and normal humans by ELISA and EITB. In serological tests by ELISA, we recognized cross reaction of Cysticercus antigenic components with IgG antibodies in heterologous sera such as sparganosis and hydatidosis patients or normal humans. The crude antigenic components of Cysticercus showed lower ELISA sensitivity in homologous sera from cysticercosis patients than heterologous sera from hydatidosis patients. A total of 31 polypeptide bands with 260 KDa~22 KDa molecular weights were detected by SDS-PAGE, and 11 of them showed strong intensity. Total 22 components of them were recognized by IgG antibodies in cysticercosis patients sera. However, 12 of them were recognized also by normal human sera, 11 were by sparganosis sera, and-21 were by hydatidosis patients sera. The crude antigenic components of 104 KDa, 82 KDa, 72 KDa, 59 KDa and 34 KDa molecular weights were nonspecific ones, which cross-reacted with sera of either cysticerco, =is, sparganosis, hydatidosis patients or normal humans.

• The Korean Journal of Zoology
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• v.32 no.4
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• pp.429-440
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• 1989

Spirometru erinocei의 유충인 sparganum에서 추출한 조항원 단백질을 항원으로 하여 sparganosis, cysticercosis, hydatidosis환자의 IgG 항체와 정상인의 IgG 항체를 혈청반응시켜 ELISA와 EITB에 의해 교차반응을 일으키는 비 특이항원 성분과 종특이항원 성분을 추구하였다. 1. sparganum에서 0.01 M PBS (PH 7.4)로 추출한 조항원 단백질을 SDS-PAGE로 전개하여 290 Kd에서 23 Kd범위의 분자량을 가진 25개의 단백질이 분획되었다. 2. sparganum의 조항원을 항원으로하여 ELISA로 sparganosis, cysticercosis, hydatidosis 환자의 190 항체와의 혈청 반응치는 sparganosis환자에게서는 0.44 $\pm$ 0.07에서 1.90 $\pm$ 0.03으로 negative control normal sera의 0.D (0.15 $\pm$ 0.03)를 기준으로 하였을 때 모두 양성이며 민감도(sensitivity)가 100 %이었으며 cysticercosis, hydatidosis환자혈청에서 양성반응이 나타났으며 교차반응도 있었다. 3. EITB에서는 spargancsis환자의 IgG항체에 의해 16개의 항원성분이 인지되었으며 이 중 6개의 항원성분이 정상인의 혈청에서도 인지되어 교차반응을 일으키는 항원성분이었으며 cysticercosis환자혈청에서 인지된 4개의 항원성분 중 2개의 항원성분이 sparganosis 환자혈청에서 인지된 것과 같았으며 hydatidosis환자의 190항체에 의해 인지된 19개의 항원성분 중 12개의 항원성분이 sparganosis환자혈청에서 인지된 항원 성분과 같았다. 4. 290 Kd, 200 Kd, 28 Kd의 항원성분은 sparganosis환자의 196항체에서만 인지되었고 228 Kd, 152 Kd, 66 Kd항원성분은 hydatidosis환자의 19G항체에서만 인지되었으며 66 Kd항원성분은 sparganosis, cysticercosis, hydatidosis, 정상인의 혈청에서 모두 인지되었다. We studied the serological reaction between the specific and nonspecific antigenic components from metacestode (plerocercoid) of spiromeko erimacei and IgG antibodies in sparganosis, cysticercosis. hydatidosis patients and normal human sera by ELISA and EITB. We prepared the crude extracts of sparganlim from snake, Matrix tigrina laterolis and used as antigenic components. By SDS-PAGE, we detected a total 25 peptide bands (fractions) with 290 Kd to 23 Kd molecular weight, and 8 bands of these detected bands developed strongly by silver stain. In serological test, ELISA, we recognized the cross-reaction of antigenic components reacting with IgG antibodies in heterogenous sera, cysticercosis and hydatidosis patients sera. The crude antigenic components of sparganum showed the high sensitivity in sparganosis, hydatidosis patients sera, but showed lower sensitivity in cysticercosis patients sera than the sparasanosis, hydatidosis patients sera. Sixteen antigenic components of these 25 separated bands were recognized by antibodies In sparsanosis patients sera,8 antigenic components in normal human sera, 4 antigenic components in cysticercosis patients sera and 19 antigenic compoenents in hydatidosis patients sera. The crude antigenic compnenets with 290 Kd, 200 Kd, 125 Kd and 28 Kd molecular weight was only recognized in sparganosis patients sera, but 64 Kd antigenic component was nonspecific antigenic components which were also cross-reacted with sparganosis, hydatidosis, cysticercosis patients sera and normal human sera.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.249-258
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• 1994

Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

• Parasites, Hosts and Diseases
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• v.30 no.4
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• pp.289-298
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• 1992

Acanthamoeba sap., free-living amoebae inhabited in moist soil, pond, freshwater, sewage, atmosphere and swimming pool, may be causative protozoa of the fatal primary amoebic meningoence-phalitis in experimental animals and humans. In this study, Acar,thamoeba spry. , including Acan. thamoeba sp. YM-4 (isolated strain from Korea) had been compared by the two-dimensional electrophoresis and hybridoma technique as well as the difference of morphological characteristics. Trophozoite of Acenthamoeba sp. YM-4 is usually uninucleate and show the hyaline filamentous projections (acanthopoda) . No aagellate stage observed. Cysts have two walls, the outer wall is nearly circular, but inner wall is oval or some irregular. As results of SDS-PAGE for Iysate of Acanthamoeba sp. VM-4, 16 major protein fractions are similiar to those of A. cuzbertsoni, but different to A. royreba and A. polyphaga. Findings of two-dimensional electrophoretic patterns of Acanthamceba sp. YM-4 are almost same to those of A. culberssoni, The isotope of monoclonal antibodies produced from McAY 6, McAY 7, McAY 8, McAY 13 and McAY 16 clones were IgGl, and McAY 10 and McAY 11 clones were IsM. As results of the cross-reactivity among various amoebae using ELISA with monoclonal antibodies, McAY 7 monoclonal antibody (molecular weight 43 kDa by EITB) was only reacted with Acanthamoeba sp. YM-4, but McAY 6 and McAY 10 monoclonal antibodies were reacted to A. cuzbertsoni as well as Acanthamoeba sp. YM-4.