• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.459-465
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• 2015

We tested biomarker systems [enzyme-linked immunosorbent assay (ELISA) and immunochromatography assay (ICG) kits] for the screening of endocrine-disrupting chemicals in contaminated environments using antibodies resulting from $17{\beta}$-Estradiol-induced vitellogenin (Vtg) in the wild scorpion fish Sebastiscus marmoratus. Monoclonal antibodies of two clones (S28 and S15) were used as capture and tracer antibodies for ELISA and ICG assays. ELISA detected Vtg at levels greater than $0.1{\mu}g/mL$, while ICG detected Vtg at levels greater than $1{\mu}g/mL$. However, the ICG system was able to detect antibodies from $17{\beta}$-Estradiol-induced Vtg serum that had been diluted 1,000 times. Our results suggest that previously developed biomarker assays can be used as detection systems to detect known endocrine-disrupting chemicals in contaminated environments, and to measure their activity.

• Korean Journal of Veterinary Service
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• v.15 no.1
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• pp.32-45
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• 1992

In order to establish and enzyme-linked immunosorbent assay to ILTV, field virus strain of ILTV was propagated in chorioallantoic membrane of the embryonated eggs. purified and used as antigen. The antisera selected from the field samples and immunized chickens based on serum neutralization test were used as the standard positive and negative sera in all tests. It was found that optimal antigen concentration was $2{\mu}g$ of protein per well and a 1 : 100 dilution of standard serum showed low background optical density with negative serum and high P/N values of positive sera. A 1 : 500 dilution of the rabbit anti-chicken IgG peroxidase conjugate produced a high P/N values and thirty minutes was chosen as suitable time to read the optical density of the enzyme substrate reaction and optical density was consistent during the 16 hours after stopper was treated. When coated antigen was kept on microplate for varying time up to 16 hours at $4^{\circ}C$ or $37^{\circ}C,$ no significant difference was observed between the treatment. The coated antigen could be kept without change of antigenicity for at least one month at $-70^{\circ}C,\; -20^{\circ}C,\; 4^{\circ}C$ and room temperature. When blocking buffer contanining bovine serum albumin was mixed directly with conjugate and serum at 10% level induced higher P/N values compared to blocking antigen coated microplate with the same blocking buffer. The coefficience of correlation between ELISA and SN test was 0.577. When antibody response of chickens, vaccinated with ILTV, was examined by ELISA and SN test, antibody rising and decay pattern between the two test was similar until 11 weeks of age. However 12 weeks onward antibody titer checked on by SN test was slightly lower than that tested by ELISA.

• BMB Reports
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• v.29 no.3
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• Microbiology and Biotechnology Letters
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• v.10 no.3
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• pp.205-209
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• 1982

The enzyme-linked immunosorbent assay using the so-called "double-sandwich"technique was applied to determine Clostridium botulinum type F toxin. Polystyrene tubes were coated with horse anti-type F toxin serum and then toxin sample was added. The tubes were subsequently treated with rabbit anti-type F toxin IgG and sheep anti-rabbit serum IgG-horseradish peroxidase conjugate. By this technique, about 10 mouse intraperitoneal 50% lethal doses (ip LD/50/) of type F toxin could be detected. Low back-ground reading was achieved with the use of phosphate-buffered saline containing 0.05% Tween 20 and 1% bovine serum albumin as diluents of rabbit IgG and conjugate. Addition of EDTA in the diluents of toxin increased ELISA extinction value significantly. No cross-reaction was observed with botulinum type A and B toxin, but type E toxin gave sleight cross-reaction.

• Korean Journal of Veterinary Service
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• v.22 no.1
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• pp.9-13
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• 1999

Mycoplasmal pneumonia of swine(MPS) cause by Mycoplasma hyopneumoniae has been recognized as a serious impediment to swine production due to chronic respiratory disorder which result in the weight loss and decreased feed conversion. The disease causes a great economic losses in pig industry by characterizing with high morbidity, low mortality, growth retardation and low feed efficiency. The present study was conducted to investigate the titers of antibody against M hyopneumoniae from the regional and seasonal groups of the slaughtered pigs by enzyme-linked immunosorbent assay(ELISA). The result have shown that the average seropositive rate of M hyopneumoniae infection was 84.6% . The regional seropositive rate in Korea showed 87.4% in Kyonggj, 83.4n in Kangwon, 89.2% in Chungnam and 77.6% in Chungbuk area, respectively. Also the seasonal seropositive rate was appeared as 78.6% in spring,90.1% in summer, 76.9% in autumn and 83.8% in winter, respectively.

• Bulletin of the Korean Chemical Society
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• v.23 no.8
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• pp.1116-1119
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• 2002

Monoclonal antibodies have been generated against a generic hapten, ο,ο-diethyl ο-(5-carboxy-2-fluorophenyl) phosphorothioate, for the determination of organophosphorus (OP) pesticides in a class-specific manner. In an indirect competitive enzyme-linked immunosorbent assay (ELISA) format, employing a heterologous coating antigen, these monoclonal antibodies showed desirable properties for use in the class-specific determination, i.e., broad specificity and high sensitivity. The IC50 values of four commonly used ο,ο-diethyl OP pesticides were fairly uniform ranging from 0.1 to 0.3 ㎛/mL. The IC50 values of three ο,ο-dimethyl derivatives were between 0.3 and 1.4 ㎛/mL. These values, together with the limits of detection (LOD), were better, in terms of the specificity and sensitivity, compared with the values obtained previously with polyclonal antibodies.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.150-153
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• 2007

The aim of this study was to investigate the prevalence of bovine viral diarrhea virus (BVDV) infection in Chonbuk province. Blood samples were taken from 92 korean calves to determined their serological status against BVDV, Capture enzyme-linked immunosorbent assay (ELISA) was used to test for antigen. The number of seropositive calves ranged from 3.3% to 12.9%. Antigens against BVDV were detected in 3.3% of healthy calves, 6.4% of digestive symptom calves, 12.9% of respiratory symptom calves, respectively. Sex and age of calves had no significant differences on the prevalence of BVDV. The results indicate that transmission of BVDV may have become exposed as a result of contact with acute infected or persistently infected cattle.

• Parasites, Hosts and Diseases
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• v.39 no.3
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• pp.261-264
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• 2001

Trichinellosis is a parasitic zoonosis of public health importance. It is caused by Trichinella spiralis which has a wide host range including humans. In the present communication, the ELISA technique was employed on a total of 803 blood samples from 7 selected pig breeding farms in 1996 for diagnosis and surveillance of trichinellosis. Out of the entire 803 samples, nine were found to be suspected while one was positive by ELISA. But western blot analyses employed for further confirmation have shown that all of 10 samples did not react to larval excretory-secretory product antigens. These results indicate that pig breeding farms included in the present study are free from trichinellosis . However, it does not mean Korea is free from trichinellosis since human trichinellosis has recently been reported. The necessity of continued surveillance for trichinellosis in both pigs and wild animals was discussed.

• Journal of Korean Society of Environmental Engineers
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• v.29 no.7
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• pp.771-777
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• 2007

Several endocrine disrupting chemicals(EDCs) were monitored to evaluate the estrogenic activities and the concentrations by yeast two-hybrid assay and enzyme-linked immunosorbent assay(ELISA) in sewage treatment plant(STP) which consist of industrial and domestic line. In the influent of domestic line, estrone, 17$\beta$-estradiol, 17$\alpha$-ethinylestradiol and alkylphenolethoxylate(APE) were detected up to 167.1, 39.7, 7.3 and 145.4 ng/L, respectively. The average removal efficiency of 17$\beta$-estradiol after the activated sludge process was 77.5% and further removed to 80.8% after the sand filtration-ozonation step. These results suggests that the activated sludge process has limited potential to remove the estrogenic activity effectively. The contributions of the estrogenic chemicals to the estrogenic activities were 70.7, 23.3, 3.7 and 2.3% for estrone, 17$\beta$-estradiol 17$\alpha$-ethinylestradiol and APE, respectively, in the domestic line effluents. Therefore, 17$\beta$-estradiol and estrone contributed most of the estrogenic activity in the domestic line effluents.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.226-230
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• 1988

Antibody against cellobiohydrolase purified from Trichoderma viride had been obtained by injection to rabbit. The antibody had a high specificity against the cellobiohydroase evidienced by absence of immunological reaction to other isozymes from Trichoderma viride. Assay limit of cellobiohydrolase was 1-10 $\mu\textrm{g}$ by column single immunodiffusion and by enzyme linked immunosorbent assay, it was 10-140 ng and 100-1200 pg when the dilution of antibody was 10$^{-6}$ and 10$^{-5}$, respectively.