• Korean Journal of Environmental Agriculture
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• v.13 no.1
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• pp.76-82
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• 1994

A competitive indirect enzyme-linked immunosorbent assay(ELISA) was developed to detect and quantify levels of the fungicide metalaxyl in crops. Antiserum against metalaxyl was demonstrated in rabbits immunized with metalaxyl-human serum albumin(HSA) conjugate. Metalaxyl-protein conjugate was prepared by mixed anhydride and peptide coupling method with EDC. In this assay, metalaxyl-ovalbumin(OA) was $coated(8{\mu}g/ml)$ on the microtiter plate, which was incubated for 1 hr at $4^{\circ}C$ or 4 hr at $37^{\circ}C$ with diluted antiserum(1:2,000). The optimum volume ratio of antigen and antibody mixture was 0.5: 1, which was incubated for 1 hr at $20^{\circ}C$. The detection of metalaxyl bound on the surface of wells was determined by the reaction(30 min) of antirabbit Ig G-peroxidase conjugate with its substrate.

• YAKHAK HOEJI
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• v.45 no.6
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• pp.670-676
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• 2001

Effect of the butanol fraction of Astragali Radix (BFAR) on the humoral immune response were investigated in ICR mice. Mice were divided into 4 groups and BFAR at doses of 5,25 and 125 mg/kg were administered orally to mice daily for 3 weeks, and the normal animals were given vehicle. The results of this study are summarized as follows; the relative weight of spleen was markedly increased by BFAR treatment, compared with that in normal mice. However, the body weight gain and the relative weight of liver were not affected. Splenic plaque forming cells and hemagglutination titers to sheep red blood cells, and the secondary IgG antibody response to bovine serum albumin were also dose-dependently enhanced by BFAR treatment. In these mice, BFAR did not increase serum alanine aminotransferase total protein, sect albumin and albumin/globulin ratio when compared with those in normal mice. Thus, these findings indicate that BFAR significantly enhances humoral immune response to antigen in concentrations that do not affected liver function.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.309-321
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• 1987

For the investigation of microbiological and immunological specificity of Bacteroides gingivalis, Bacteroides gingivalis were isolated, enumerated and characterized from 13 Korean rapidly progressive periodontitis and 7 healthy control by anaerobic culture technique. The total proportion of black-pigmented Bacteroides from Korean R.P.P. patients and healthy control were 8.78% and 0.92%, respectively, among total isolated black-pigmented Bacteroides. In antibiotic susceptibility test, Bacteroides gingivalis isolated from R.P.P. patients were sensitive to Ampicillin and Tetracycline, and resistant to Gentamicin and Erythromycin in disc diffusion method. In antibiotic broth dilution method, the minimum inhibitory concentration(MIC) to Bacteroides gingivalis was 2 unit/ml of Penicillin and $0.25{\sim}1{\mu}g/ml$ of Tetracycline, respectively. The concentration of serum IgG in rapidly progressive periodontitis patients were sigificantly higher than that of healthy control, and concentration of diluted gingival crevicular IgG has not any significant differences between two groups. Serum and gingival crevicular IgG antibody to Bacteroides gingivalis were significantly higher titer in rapidly progressive periodontitis patients to compare with healthy control. The lipopolysaccharide profiles of 2 Korean B. gingivalis in silver stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis were similar to type strains of B. gingivalis and typical LPS band were appeared around the 24-Kd molecular weight. Immunodiffusion test and immunoelectrophoresis of the L.P.S. extracted from 2 Korean B. gingivalis and 2 kinds of type strains of B. gingivalis showed that B. gingivalis Korean-1 was reacted identically to B. gingivalis ATCC 33277. In trypsin and ${\alpha}$-glucosidase activity test of 2 Korean B. gingivalis, both of them revealed positive trypsin and negative ${\alpha}$-glucosidase activity, respectively. These investigation suggested that B. gingivalis is important pathogenic plaque bacteria for the pathogenesis of periodontitis and further study is needed to purify and characterize of the species-specific antigens of this organisms to develop monoclonal antibody and potential diagnostic reagents.

• Parasites, Hosts and Diseases
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• v.30 no.2
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• pp.83-90
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• 1992

Sensitivity of anti-Texoplasma antibody (IgG) test by enzyme·linked immunosorbent assay (ELISA) was evaluated in comparison with indirect laten agglutination (ILA) using 2,016 paired human samples of serum and cerebrospinal quid (CSF) . The samples were collected from neurologic patients in Korea with mass lesions in central nervous system(CNS) as revealed by imaging diagnosis(CTIMRI). When the sera were screened for anti-Toxoplasma antibody by ILA, 76 cases (3.8%) were positive (1:32 or higher titers). In the pairs samples of CSF, no positive reactions were observed. When ELISA was performed using PBS extract of Percoll purified tachysoites as antigen, cut-off absorbance was determined as 0.40 for serum and 0.27 for CSF tests. The antibody positive rates by ELISA were 7.0% in serum and 5.6% in CSF Of them, 40 cases(2.0%) showed positive reactions in both serum and CSF, The antibody positive rates were higher in groups older than 40 years, The rates were higher in male(4.7% by ILA, 8.3% by ELISA) than in female(2.2% by ILA, 5.0% by ELISA). The rates in CSF showed no such sex difference. ELISA showed twice higher positive rates when serum was tested, and was sensitive enough to detect specific antibodies in CSF. Etiologic relations between positive antibody tests and CNS lesions remained unknown.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.61-65
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• 1990

Lectin related inducer can enhance IgG$_1$ production rate from murine hybridoma cells by employing step-feeding of serum free media with producing about 40 mg/$\ell$ of monoclonal antibodies. This step-feeding perfusion process also proves to be able to cutivate animal cells when serum free media can not support the growth of these cells in perfusion process, as well as to improve production rate. This process yields about 28 x 10$^{-10}$ mg of MAb/cells/h compared to 11.1 x 10$^{-10}$ and 4.0 x 10$^{-11}$ mg/cells/h for perfusion process and batch cultivation with 10% serum containing media, respectively.

• Korean Journal of Veterinary Service
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• v.46 no.4
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• pp.263-268
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• 2023

Despite regular vaccinations, equine influenza virus (EIV) and Streptococcus equi subsp. equi (strangles) are the cause of highly contagious respiratory infections in horses. Many recent studies have reported that the concurrent administration of two vaccines could simplify horse management and minimize veterinary expenses. However, there is little information available regarding the efficacy of concurrent vaccinations against EIV and strangles. In this study, we evaluated EIV-specific antibody responses following the single EIV vaccination with the recombinant viral-vectored EIV vaccine or concurrent vaccination with the EIV and inactivated strangles vaccines. Blood samples were collected at 1-, 2-, 4-, and 8 weeks post-immunization (wpi) from each group. EIV-specific antibodies were evaluated by enzyme-linked immunosorbent assay (ELISA) and hemagglutination inhibition (HAI) assay. Both single and concurrent vaccination showed similar levels of EIV-specific serum immunoglobulin g (IgG) at 1 and 2 wpi. However, at 4 to 8 wpi, the EIV-only vaccination group showed significantly higher serum IgG levels than those from the concurrently vaccinated group. The HAI titers showed similar trends as the ELISA data, except at 8 wpi when both groups presented HAI titers with no significant differences. These data demonstrate that the concurrent vaccination against EIV and strangles could compromise the humoral immune response to equine influenza between vaccination intervals, which suggests the use of the consecutive vaccination protocol for EIV and strangles rather than concurrent vaccination.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.925-934
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• 1997

This study was directed at inducing the production of antibodies by immunizing heifers with bovine sperm antigen and on measuring the serum antibodies using indirect immunofluorescence assay(IFA) and agglutination test. The effect of antisperm antibodies on fertilizing capacity of bovine spermatozoa was evaluated. 1. Three heifers between 12- and 15- month old were immunized with bovine spermatozoa or phosphate-buffered saline. In heifers immunized with bovine spermatozoa serum IgG level was highest between 3 weeks and 5 weeks postimmunization detected by IFA. The antibody levels persisted through week 7 and slowly declined until week 20 and then antisperm antibodies were localized on spermatozoa. The fluorescent antisperm antibodies were detected at 2~20 weeks and at 6~9 weeks postinoculation on acrosome and tail, respectively. Among 21 sera from repeat breeder cows, only one cow has shown positive antisperm antibody response detected by IFA. 2. In spite of vital rate of bovine sperm after swim-up was not significantly affected by different concentration of antisperm antibodies in sera, the numbers of bovine sperm after swim-up were significantly reduced in proportion to the increased concentration of antibodies. Above 1/512 dilution of antibody neither influence on vital rate and numbers of bovine sperm nor sperm agglutination after swim-up. The study has also shown that the vital rate and number of sperm after swim-up and capacitation were also significantly reduced by the addition of antisperm antibodies. Although antisperm antibodies did not influence on the acrosome reaction rate of sperm during swim-up, did significantly reduce the sperm acrosome reaction rate after capacitation. The studies have resulted that the bovine antisperm antibodies can prevent the sperm motility by agglutination and block the capacitation and acrosome reaction of bovine sperm.

• Journal of agricultural medicine and community health
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• v.16 no.1
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• pp.70-78
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• 1991

Antibody changes in experimental anisakiasis were observed by ELlSA and SDS-PAGE/EITB using various antigens : whole worm extract antigen(WWE), somatic antigen(SOM), excretory-secretory antign(ES), and hemoglobin antigen(HB) of Anisakis Type 1. The results obtained were as follows. l) Serum levels of IgG antibody by ELISA increased from 1st week of infection and reached their maximum titer at 5th week after infection, and decreased gradually thereafter. 2) The best result expressed as positive/negative ratio could be obtained when ES antigen was used. 3) Silver stained SDS-PAGE of each antigen showed at least 20 protein bands : In WWE, 286, 278, 262, 38, 18 Kd bands ; In SOM, 38 Kd band : In ES, 286, 65, 13 Kd bands ; In Hb, 61, 55, 38, 28, 26, 22, 20, 16, 15 Kd bands iepntibied as were major bands. 4) By EITB using WWE, Serum antibody recognized major protein with molecular weight of 86 Kd and 16 Kd. Using ES, 69, 59, 16 Kd bands were observed and using Hb, 28 Kd band was observed as specific band. In conclusion, excretory-secretory antigen(ES) of Anisakis larvae was most usable for ELISA.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.447-453
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• 1986

To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.95-98
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• 2000

An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.