• Parasites, Hosts and Diseases
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• v.33 no.2
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• pp.101-106
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• 1995

Soluble protein of purified Pneumocvstis cnrinii was prepared from experimentally infected rats. SDS-PAGE of the crude antigen resolved about 20 protein bands from 20 to 200 kDa. Out of thenl, 116 kDa band strongly reacted and 45-55 and 100 kDa bands reacted weakly to the positive reference human serum from U.S.A. Western blot analysis with sera of 130 normal children and 15 newborns in Korea revealed specific IgG antibody reaction to 40-55 and 116 kDa protein bands. Forty percent (40.0%) of the 145 sera were positive with any of the antigenic protein bands of R corinii. The positive rate was 56% in 50 males and 33.3% in 48 females. The protein bands 40-55 and 116 kDa from rat P. carinii were confirmed to cross-react with human sera in Korea.

• Parasites, Hosts and Diseases
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• v.31 no.1
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• pp.37-42
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• 1993

Surface proteins of Trichomonqs unginnlis (T vqsinalis) were analyzed to study the antigenic variation. The surface proteins of protozoa were labelled by N- hydrokysuccinimide-biotin (NHS-biotins, the NHS-biotin-labelled proteins were immunoprecipitated with rabbit antiserum to purifjr the antigenic fractions and analysed by SDS-PAGE plus electroblotting. The results obtained in this study were as follows; Biotinylated T. uaginalis-proteins obtained from intact cell and cells disrupted prior to labelling were detected by antibiotin-peroxidase in Western blots. Labelled proteins were immunoprecipitated by T. vcqinalis-immunized rabbit serum and the six bands with, the molecular weights of 46, 60, 68, 90, 130 and 220 kDa were identified as having antigenicity. T unginalis HY-1,HY-15 and ATCC 50148 were immunoprecipitated by immune rabbit serum after biotinylation and there were no difference from antigenic bands among these strains by this tehchnique. In conclusion with the results obtained in the present study, it was assumed that surface proteins of T vaqinclis were labelled by biotinylation and the six labelled bands at 46, 60, 68, 90, 130 and 220 kDa in their molecular weight were identified as having antigenicity by immunoprecipitation (IPI and this biotinylation-IP technique may be used for further study of surface antigen of T vaginalis.

• Parasites, Hosts and Diseases
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• v.35 no.1
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• pp.31-38
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• 1997

The skin ulcer in Leishmcnio mcior infection is known to be variable according to the inoculation sites even in a susceptible host. The present study traced the immunoblot patterns by the site of inoculation and duration of infection in BALB/c mice. L. mqior were subcutaneously inoculated on the nose, footpad, and back of the mice, in a dose of 3 × 106 promastigotes. Sera of the mice were collected every 10 days after inoculation. SDS-PAGE separated soluble protein bands of the promastigotes and immunoblot was carried out with the infection sera. The skin ulcer first appeared on the nose at 15 days, and on the footpad at 17 days after inoculation. The ulcer on the back appeared after 90 days. In the mice with ulcer on the nose or footpad, serum IgG antibody reacted to 202, 139, 98, 83, 81, 67, 65, 62, 59, 54, 52, 42, 26, and 23 kDa bands at 20 days after inoculation. In mice inoculated on the back, however, the immunoblot showed visible reactions with 202, 83, 81 74, 67, 65, 62, 59, 54, 52,20 and 17 kDa bands at 90 days after inoculation. The present result showed that the antigenic protein bands of L. mqior promastigotes were differed by the inoculation site and duration of infection. Since the skin ulcer and the serum antibodies to antigenic bands between 67-52 kDa appeared simultaneously, it is suggested that the serum IgG antibodies may play a role in formation of the skin ulcer in BALB/c mice.

• BMB Reports
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• v.29 no.6
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• pp.540-544
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• 1996

Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.

• Journal of Microbiology and Biotechnology
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• v.5 no.6
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• pp.359-363
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• 1995

The antigenic variation of B. thuringiensis subsp. satto have been investigated for 120 hours during sporulation and crystallization by using SDS-PAGE and Western blot. Most antigens of a vegetative cell were found to disappear as it was in sporulation and crystallization, but protein antigens of 46, 29, 27, and 21 kDa continued to be expressed. The new protein bands of 293, 138, 119, 75, and 68 kDa appeared on days 2 through 5 in modified GYS medium. They were thought to be involved in sporulation and crystallization. The protein of 138 kDa was found to be a major protein of both crystal and spore. The expression patterns were immunologically analyzed by Western blot. The polyclonal antisera against the intact crystal showed strong immunoreactivity to proteins with molecular masses of 293, 138, 68, and 46 kDa. The polyclonal antisera against the spore recognized proteins of 293, 138, 68, and 46 kDa. Both crystals and spores appeared to express the common protein antigens.

• Parasites, Hosts and Diseases
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• v.41 no.3
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• pp.155-163
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• 2003

The antigenic characterizations and serological reactions of human liver flukes, Clonorchis sinensis and Opisthorchis viverrini, were analyzed by immunoblot. The antigenic profiles of the crude extract of Clonorchis contained major proteins of 8, 26-28, 34-37, 43, and 70 kDa, and those of Opisthorchis 34-37, 43, 70, and 100 kDa. Of these, the 8, 26-28 and 34-37 kDa bands of Clonorchis and the 100 kDa of Opisthorchis were major components of each excretory-secretory antigen. The 8 and 26-28 kDa bands were specific to Clonorchis but the 100 kDa of Opisthorchis cross-reacted with the sera of clonorchiasis, and the 34-37, 70 and 100 kDa bands cross-reacted with sera of other helminthiases. The frequency and intensity of the immunoblot reactions were positively correlated with the intensity of the liver fluke infection.

• Parasites, Hosts and Diseases
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• v.37 no.2
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• pp.109-115
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• 1999

The present study observed the variation of antigenicity of Pneumocystis carinii and serum IgG antibody reastion to the antigens from different localtities in Korea. Antigens of rat P,carinii and sera of inhabitants were collected at Chunchon, Chungju, Kwangju and Seoul during 1995-1996. Enzyme-Linked Immunosorbent Assay and immunoblot were used for immune reaction. Absorbance of 1,294 human sera ranged between 0.01and 0.93. Sera from Chunchon showed higher absorbances than those from other areas. Immunoblotting revealed IgG antibody reactions to 116,100, and 45-55 kDa antigenic bands of rat P.carinii, but the frequencies of positive reaction to individual bands were variable by localities. Total 62.6% reacted to 100kDa band and 32.0% to 45-55 kDa bands. For the reaction to 116kda, the reaction rate was 60.0% of the sera showed the reaction to 116kda band while 37.7% reacted to 100kda, the reaction rate was 60.0% to 82.6% by localities. It is found that the reaction rates of the human sera to rat P.carinii antigen are variable according to the localities. Also, the high molecular antigen of 116kDa of rat P.carinii is the most frequent antigenic band reaction to human sera.

• Food Science and Biotechnology
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• v.15 no.3
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• pp.441-446
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• 2006

The biological effects of heating and chemical treatment on castor meal were investigated in order to develop a procedure to inactivate its antigenic activity in a way that is suitable for industrial applications. A 1% solution of purified castor bean allergen (CB-1A) was heat-treated with or without exposure to NaOH and NaOCI (250 ppm each). CB-1A exhibited extreme stability when heat-treated alone. In the presence of NaOH and NaOCl, CB-1A showed a drastic decrease in antigenic activity as the temperature surpassed the critical level of $70^{\circ}C$. The gradual disappearance of disc gel electrophoresis bands presumably responsible for the allergenicity of CB-1A, along with the significant losses of the amino acids phenylalanine, methionine, arginine, histidine, and cysteine correlated with the loss of CB-1A activity. CB-1A showed a single symmetrical band in SDS acrylamide gel electrophoresis with an estimated molecular weight of 6,000 daltons. The chemical and heat treatments reduced the disulfide bond content of CB-1A by 9.1% with a coincident increase in sulfhydryl bonds.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.197-202
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• 1997

Diagnosis of early paragonimiasis is difficult because parasitological evidence is not easily obtained. Antibody tests have been proposed as a good substitute for classical diagnostic techniques. Using the crude extracts of Parnsonimus westermnni eggs, metacercariae. 4- and 7-week Juveniles, and 16-week adults as antigens, we observed the early antibody responses. Sera were obtained from 4 experimental cats, fed 50 metacercariae each, at intervals until 13 weeks post-infection. Antibody (IgG) responses were identified by ELISA using extracts of 4-week juveniles, followed by those of 7- and 16-week worms. Antibody responses were minimal against the metacercarial extracts. Antibodies to p. westemoni egg extracts were elevated after 10 weeks post-infection. In immunoblot analysis, more than nine protein bands in 4-week juveniles reacted with the early infection sera. Antigenic proteins in adult worms were different from those of juveniles. After four weeks of infection, 32 and 35 kDa bands in the adult extracts were increasingly reactive. Egg specific proteins at 28, 46 and 94 kDa were reactive only after 10 weeks. Antigenic components reactin료 to the early infection sera changed during the maturation stages of P. westermani; almost all juvenile antigens were replaced by adult antigen components.

• Journal of agricultural medicine and community health
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• v.16 no.1
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• pp.61-69
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• 1991

Serodiagnosis of Clonorchis sinensis infections will probably be a first choice tool for screening of clonorchiasis in a future because of increasing difficulties in collection and examination of stools. The sensitive test such as ELISA can he used effectively. However there are some limitations in serological diagnosis for the detection of serum antibody. One of the major problems is the non-specificity of the antigens which produce cross reaction with other helminthic infection sera. To solve this problem. many investigators have tried to purify the antigens used. In this study, we determined the antigenic profile of the crude saline extract antigen of C. sinensis at early developmental stage based on SDS-PAGE and immunoblotting techniques for the purpose of understanding the nature of C. sinensis worm antigen The following results were obtained : 1) The SDS-PAGE showed many protein hands ranging from 10Kd to 91Kd relative molecular weight. Among them, 66, 46, 40, 33, 27, 24, 16, 14 and 10Kd bands were observed as a principle bands. The protein components of C. sinensis changed chronologically during their early developmental period. 44Kd band was stained unclearly in antigen of 2 weeks worm, but changed to concentrated state in antigen of 5 weeks worm. 35Kd band was found in antigen of 2 weeks worm, however this band was disappeared in antigen of 5 weeks worm. 22Kd band also lost its staining property gradually. 2) In spite of differences in antigenic profile, there was no differences in the data obtained by microplate ELISA using each antigen preparation. Absorbance value began to rise in between 2 to 3 weeks after infection. 3) By EITB. serum antibody recognized major protein bands with molecular weight of 91, 85, 63, 46, 40, 33, 24, 14 and 10Kd hand respectively. Among them 66, 33, 17 and 14Kd bands were observed as non-specific band because they reacted even in normal control sera. Generally, gradual increase of positive reactions were observed as the infection period of C. sinensis was prolonged. In other hand, the reaction of 10Kd hand did not occurred when 26th week sera was tested. 4) The positive reactions using antigens of 2 weeks worm, especially on 40 and 24Kd bands, were most strong and sharply demarcated compared to those of 3~5 weeks worm antigen.