• Korean Journal of Veterinary Service
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• v.46 no.4
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• pp.325-334
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• 2023

Edema disease (ED) in pigs is enterotoxemia caused by Shiga toxin type 2e (Stx2e)-producing Escherichia coli (STEC) and frequently occurs in young piglets. Therefore, ED causes enormous economic losses in pig farms. In this study, a modified Stx2e (mStx2e) antigen was expressed and purified using commercial E. coli expression system. Monoclonal antibody was serviced by Ynto Ab Inc., using Phage Display Technique. Anti-Stx2e antibodies in piglets were measured by indirect ELISA using mStx2e antigens. Naive Stx2e in piglets were detected by Sandwich ELISA using Stx2e-monoclonal antibodies and commercial Stx2e-polyclonal antibodies. Among 3,480 piglets, anti-Stx2e antibodies were observed in 2,573 piglets. The 49.4% among 830 piglet serum samples possessed 0.625 ㎍/mL or more of Stx2e proteins. The 18.3% of 830 sera had 0.313 ㎍/mL of Stx2e proteins. The 32.3% of 830 samples held 0.156 ㎍/mL or less of Stx2e proteins. These results show that indirect ELISA using mStx2e antigen and Sandwich ELISA using Stx2e-monoclonal and polyclonal antibodies can be useful to detect ED in piglets.

• Journal of fish pathology
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• v.12 no.1
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• pp.24-31
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• 1999

Optimization and standardization of solid phase enzyme immunoassay were done for the diagnosis of edwardsiellosis in fish. The analyzed degree of immobilized antibody on surface of solid phase with peroxidase saturation method showed the optimized result by using partially purified $50{\mu}g/ml$ of rabbit anti-E. tarda Edk-2 antibody in sodium bicarbonate buffer for overnight incubation to cover the surface of polystyrene beads. Optimized immunoreaction was observed in the treatment of $50{\mu}g/ml$ of biotin conjugated antibody followed extravidin-peroxidase diluted 1 : 2,000 in PBS. The detectable concentrations of the this method were $1{\times}10^5$ cells/ml and $1{\times}10^5$ cells/ml expressed as the source of antigen amount for EDTA extraction and heat extraction, respectively. High cross-reaction of solid phase ELISA with the prepared rabbit and-E. tarda Edk-2 was observed against E. tarda strains isolated from flounder suffering from edwardsiellosis in aquatic farms of Korea. It suggested that the potential of this solid phase of ELISA technique is very powerful for the application to different strains of E. tarda isolated in farms of many different areas.

• Research in Plant Disease
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• v.9 no.2
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• pp.94-98
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• 2003

To produce antibodies against Pseudomonas tolaasii and P. agarici, lyophilized P. tolaasii and P. agarici ($5{\times}10^7$ cfu/ml) and Freund, s adjuvant were immunized into rabbits 4 times. The specificity and sensitivity of the antibodies were evaluated by immunodiffusion test and indirect enzyme-linked immunosorbent assay (id ELISA). The ${\alpha}$-P. tolaasii antibody was very specific only against P. tolaasii, while ${\alpha}$-P. agarici antibody was not specific and showed a high cross reactivity toward P. tolaasii with detection limit concentration of $2{\times}10^3$ cfu/ml. However, the cross reactivities of ${\alpha}$-P. agarici antibody toward the related species including P. reactans were very low. Our results showed that ${\alpha}$-P. tolaasii and ${\alpha}$-P. agarici antibodies against P. tolaasii and P. agarici, respectively, might be useful for rapid and simple detection of the causal agents of bacterial brown and yellow blotches in cultivated oyster mushrooms.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.310-316
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• 1999

BBPI contents in domestic soybean and soybean products were investigated by the measurement of chymotrypsin inhibiting activity(C.I.A) and competitive ELISA method. In order to produce polyclonal antibody, BBPI was purified from soybean trypsin-chymotrypsin inhibitor by ion exchange chromatography and electrophoretic gel slicing. Rabbit anti-BBPI polyclonal antibody was produced with the purified BBPI as immunogen. This antibody showed relatively specific binding to BBPI and then used for the establishment of competitive ELISA method to measure BBPI contents in extracts of soybean and soybean products. The standard curve for the measurement of BBPI in soybean extracts was drawn up within the range 0.03 to $30\;{\mu}g/ml$ of BBPI. The C.I.A. and BBPI contents of 12 soybean cultivars were $8,462{\sim}12,428\;U/g$ and $482{\sim}692\;mg%$, respectively. The C.I.A. and BBPI contents were not detected in most of soybean products except soybean sprouts, which contained $10,695{\sim}13,249\;U/g$ of C.I.A. and $529{\sim}803\;mg%$ of BBPI.

• Korean Journal of Poultry Science
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• v.18 no.3
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• pp.183-196
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• 1991

In order to establish ELISA method to detect antibody against IBV various factors involved were examined. Antigen was prepared from Massachusetts type IBV which is known to be one of serotypes distributed most widely. The virus was grown in embryonated SPF chicken eggs. Allantoic fluid harvested was processed to ultracentrifugation and sucrose density gradient centrifugation to produce a purified antigen The antisera selected from the field samples based on hemagglutination inhibition test were used as the standard positive and negative sera for this study and the results obtained were summarized as follows. 1 , It was found that ELISA test was satisfactory when the purified antigen was coated on the plate in the amount of about 40ng protein per well. In case of the phospholipase treated hemagglutinating antigen it gave satisfactory results when the each well wns coated with 1.2 to 2.5 hemagglutinating unit which was equivalent to 40 to 90ng of protein. 2. There was no significant difference in the ratio of optical density of positive to that of negative serum whether the coated antigen was held for 1 hour at 37$^{\circ}C$ or it was held overnight at 4$^{\circ}C$. The coated antigen could be kept in dried state without change of antigenecity for at least one month of experimental period at 4$^{\circ}C$. 3. There was a big variation in the optical density and P/N values depending on the maker of the plates and on the plate of the same maker. 4. It was found that background optical density was negligible when serum was diluted more than 1：50 and serum dilution of 1：100 appeared to be appropriate as a routine test dilution to screen the antibody. 5. Optical density was fairly constant 15 minutes afterward from the time substrate was treated and during the 4 hours after stopper was treated. 6. There was a low correlation(r＝0.42) between ELISA and HI test. However, when 74serum samples were tested for the IBV antibody, 98.7% were found to be positive by both tests in which titers of 2$^{6}$ or more by HI test and P/N values of 1.4 or more by ELISA were considered to be positive, 7 Day-old IBV vaccinated chickens shows a similar antibody decay and rising pattern until 8 weeks of age by the two tests, ELISA and HI.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.333-342
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• 1989

Effect of raising types and environmental conditions on the infection of Toxoplasma in the swine, the cat and the man were studied in Cheju Island from Sept. 1987 to Aug. 1988. Blood samples were taken from 214 conventionally raised swine in 6 villages and 506 swine raised in swine specialized farms, 122 cats raised under free moving or restraned conditions in 8 locations, 113 butchers, and 210 villagers. Toxoplasma antibody values of the blood sera were determined using the enzymelinked immunosorbent assay (ELISA). The eating type of viscera was also investigated by using questionaires 1. When ELISA method was used, the percentage of Toxoplasma infected swine among the conventionally raised and of those raised in swine specialized farms were 60.7% and 21. 390, respectively. The respective mean of antibody values (${\pm}SD$) were 0.589 (${\pm}0.310$) and 0.385 (${\pm}0.237$) and differed very significantly (p<0.01). A significant difference was also found in antibody values among 6 villages (p<0.05). 2. The mean infection percentage of toxoplasma in the cat was 38.2%. the infection percentage for cats raised under free-moving and re~;trained condition were 37.0% and 38.2% respectively. The respective antibody values(${\pm}SD$) for toxoplasma were 0.600(${\pm}0.614$) and 0.637 (0.645), and did not difference significantly. 3. The infection percentage of toxoplasma in villagers and butchers were 26.2 and 38.3% respectively. The respective antibody values (SD) for toxoplasma were 0.429(${\pm}0.195$) and 0.341 (${\pm}0.236$), and differed very significantly (p<0.01). There were also highly significant differences Pyo-sun and other village (p<0.01). 4. Analysis of the questionaires showed that 26.0% of 392 villages eated liver and some villagers eated other viscera.

• Korean Journal of Veterinary Service
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• v.24 no.4
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• pp.359-367
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• 2001

A serological survey was performed to establish basic data for the prevalence of antibodies to some major diseases of domesticated boar serum samples from January to December 2000. Sera collected in breeding farms in Gyeongbuk province were tested for Aujeszky's disease virus(ADV), Porcine reproductive and respiratory syndrome virus(PRRSV), Porcine parvovirus(PPV), Japanese encephalitis virus (JEV), Bordetella bronchiseptica(B bronchiseptica), Mycoplasma ; APP), Toxoplasma, and Brucella. There was no antibody to ADV in domesticated boars serum samples detected by Anti-ADV-gpI assay kit. Sero-positive samples to PRRS by IFA were 0.9%(3/330) The HI titers to PPV ranged variously from less than 10 to over 1,280. Two hundred ninety-four out of 330 tested sera showed HI titer of less than 10. In HI test to JEV, 90.3% of the sera (298/330) were below 10. The majority of the serum samples had low prevalence of the antibody B bronchiseptica. ELISA titers to M hyopneumoniae ranged variously from $\leq$ 10 to $\geq$ 1,280. Antibody titers to A pleuropneumoniae type 2(APP2) and type 5(APP5) were 58.2% and 52.7%, respectively, and the tested samples showing ELISA antibody titers of less than 20. There was no significant geographical difference between APP2 and APP5 in this study. In the antibody test of Toxoplasma, 11.5%(38/330) were positive and samples were all negative in sera test of Brucella.

• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.93-100
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• 1990

These experiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro maturation of bovine follicular oocytes. Antisera to bovine cumulus cell were produced Japanese Ginat rabbit by repeated immunization of intact or solubilized bovine cumulus cell and purified by ammonium sulfate precipitation and Sepharose CL-4B protein-A affinity chromatography. The bovine cumulus cell-specific antibodies were confirmed by indirect ELISA. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to cumulus cell determined by indirect ELISA using intact or solubilized bovine cumulus cell coated plates was very high in both intact and solubilized cumulus cells. Namely, the optical density at 1:12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. 2. When the follicular oocytes were treated with antibody to intact cumulus cells, the maturation rate of cumulus compacted and removed oocytes was ranged 47.6 to 59.1%. These value is significantly lower(p<0.05) than that(78.8%) of follicular oocytes cultured without the antibody. 3. the maturation rate of cumulus compacted and removed oocytes treated with antibody to solubilized cumulus cells was ranged 46.7 to 59.1%, significantly lower(p<0.05) than that(82.1%) of ooyctes cultured in antibody free medium. From above mentioned results, it could be said that cumulus cells promote nuclear maturation of follicular oocytes and that the beneficial effect of cumulus cells to the oocyte maturation is inhibited by the action of antibody to cumulus cells.

• Journal of Photoscience
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• v.12 no.3
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• pp.123-127
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• 2005

The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

• Journal of Food Hygiene and Safety
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• v.10 no.1
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• pp.1-6
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• 1995

In order to establish the enzyme linked immunosorbent assay(ELISA) of sterigmatocystin produced by Aspergillus versicolor, we experimented and obtained following results. Two of three rabbits which had been immunized with sterigmatocystin-hemiacetal-BSA produced antibodies against sterigmatocystin at 15 weeks. The produced antibodies were specific for sterigmatocystin and sterigmatocystin-hemiacetal but didn't cross react with other sterigmatocystin analogues in a significant degree. DMF : 4% KC1 (18 : 2) mixed solution was most effective to dissolve sterigmatocystin. For the preparation of sample solution to determine sterigmatocystin by ELISA, sample was extracted with CHC13 and dried, than the dried sample was redissolved with 100 ${mu}ell$ DMF + 4% KC1 mixture. 10~1,000 ng/$m\ell$ level of standard sterigmatocystin could be applied to the established ELISA. When artifically contaminated rice were assayed by the ELISA, the average recovery of sterigmatocystin spiked to 25~500 ng/g was 109% (97~116%), and mean interwell coefficient of variation was 21% (11~28%).