• 한국환경농학회지
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• 제13권1호
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• pp.76-82
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• 1994

Metalaxyl을 ELISA기법을 이용하여 분석하기 위하여 metalaxyl-HSA를 합성하고 이에 대한 항체를 생산하여 competitive indirect ELISA를 위한 최적조건을 조사하였다. Metalaxyl-protein conjugate는 metalaxyl을 가수분해시켜 생성된 metalaxyl acid에 단백질(HSA, OA)을 mixed anhydride방법이나 EDC 첨가방법으로 조제하였고 생성된 항체의 역가는 1:16,000으로 나타났다. Coating Ag의 최적농도는 $8\;{\mu}g/ml$, 배양시간은 $4\;^{\circ}C$에서 1시간 이상, $20\;^{\circ}C$와 $37\;^{\circ}C$에서는 4시간으로 나타났으며 항체의 희석배수는 1:2,000이 최적으로 나타났다. 항원과 항체의 혼합비율은 0.5 ml의 항원에 대해 1 ml의 항체가 최적이었으며 이 혼합액의 배양시간은 1시간, antirabbit Ig G-peroxidase conjugate의 배양시간은 30분이 최적으로 나타났다.

• 약학회지
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• 제45권6호
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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.

• 대한미생물학회지
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• 제22권3호
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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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• 제30권2호
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• pp.83-90
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• 1992

Toxoplasma증의 혈청학적 진단에 있어서 민감도를 증가시키기 위해 간접 latex 응집반응의 결과와 비교하면서 ELISA를 개발하였으며, 뇌척수액의 검사 시료로서의 가능성을 검토하였다. 아울러 중추신경계 질환환자로부 터기생충질환을 감별하기 위하여 1986년부터 1991년까지 전국 카 병원에서 채취한 혈청과 뇌척수액에 대하여 간접 latex 응집반응(ILA)과 ELISA를 실시하여 Toxoplasma 항체 보유 양상을 비교 검토하였다. 전체 2,016 건의 혈청에 대해 ILA를 실시하여 76건(3.8%)의 양성 (1:32이상의 titer)을 얻었다. 그러나 양성 혈청환자에서 채취한 뇌척수액에서는 낮은 titer의 반응은 있었으나 양성은 나타나지 않았다. 이들 양성 혈청의 양성 혈청 및 음성 혈청에 대하여 ELISA로 항체검사를 실시한바 ILA의 titer가 1 : 32인 군에서 통계적으로 유의한 차이를 나타내는 항체값을 얻었으며, 그 흡반도는 0.40이었다. 뇌척수액에 대한 ELISA로는 ILA의 1 : 64 titer군에서 통계 적으로 유의한 차이가 나타났고 그때의 흡광도 0.27을 양성 판단의 기준으로 사용하였다. ELISA에 의한 항체 검사상 전체 혈청에서 7.0%의 양성을 검출하여 ILA보다 약 2배 정도의 높은 민감도를 보였으며, 뇌척수액에서 는 5.6%의 양성률을 보여 ELISA는 뇌척수액에서의 항체 검출시 유용한 방법이라고 판단하였다. ILA에 비하여 ELISA는 약 2배 정도 높은 양성률을 내었고 양성률은 나이에 마라 40대 이후 급격한 증가를 보였으며, 여성보 다는 남성에서 약 2배 정도 양성률이 높게 나타났다. ELISA에 의한 뇌척수액의 항체 검사에서는 양성률의 성별 차이를 나타내지 않았다. 이상의 결과로 판단할 때, ELISA가 ILA보다 Toxoplasma 항체 검출의 민감도가 높았으며, 뇌척수액은 ELISA의 좋은 검사시료가 되며, 특히 중추신경계 Tocxoplnsma증의 진단에 있어 뇌척수액에 대한 항체 검사에서 ELISA가 유용하다고 판단하였다.

• 한국미생물·생명공학회지
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• 제18권1호
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• pp.61-65
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• 1990

Murine hybridoma 세포배양시 lectin 계통의 생산 증진제가 무혈청 배지의 단계적 유입에 의해 약 40mg/l의 단일항체를 생산함으로서 단일항체의 생산성을 증가시킬 수 있음이 확인됐으며, 또한 이 배양공법으로 무혈청 배지에서 연속배양이 불가능한 세포의 배양이 가능했다. 이같은 무혈청 배지의 단계적 유입공법으로 약 28 x $10^{-10}$mg/cells/h의 속도로 단일항체가 생산된 것에 비해, 10 혈청이 포함된 배지로는 연속 배양시 11.1 x $10^{-10}$mg/cells/h 의 속도로 생산됐으며 회분배양의 경우는 4.0 x $10^{-11}$mg/cells/h의 속도로 생산됐다.

• 한국동물위생학회지
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• 제46권4호
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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.

• 대한수의학회지
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• 제37권4호
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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.

• 농촌의학ㆍ지역보건
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• 제16권1호
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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.

• 대한미생물학회지
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• 제21권4호
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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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• 제10권1호
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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.