• Title/Summary/Keyword: IgG-ELISA

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The Comparative Study of IgG, IgM, and IgA in Laboratory Animal Administrated Red-ginseng, Using Immunoglobulin Productivity Assay (Immunoglobulin productivity assay를 이용(利用)한 홍삼투여(紅蔘投與) 실험동물(實驗動物)의 IgG, IgM, IgA 비교(比較) 연구(硏究))

  • Lee, Beom-Jun;So, Hyung-Jin;Kim, Jae-Wan;Lew, Jae-Hwan
    • The Journal of Internal Korean Medicine
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    • v.28 no.4
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    • pp.886-895
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    • 2007
  • Objective : The immune system is a complex of systems, all of which work together to clear infection from the body. In Korea, red ginsenghas been one of the herbs most widely used to enhance the immune system for thousand of years. More recently, red ginseng has been reported to have many positive effects on the immune system. The purpose of this study was evaluate the effects of Korean red ginseng and Chinese red ginseng on IgG, IgM, and IgA, using immunoglobulin productivity assay. Methods : Male SD rats were separated into 3 groups. We administered Korean red ginseng (KRG) to one group and Chinese red ginseng (CRG) to another, with normal saline for the Control group consecutively and orally for 3 months. The dose of red ginseng was 500mg per day, as a powder with soluble water. Immunoglobulin levels from spleen cell were estimated by ELISA kit. Results : In immunoglobulin productivity assay (cell), the IgG level of the KRG group significantly increased but there was no significant difference in the IgG of the CRG group. The IgM level of the KRG group significantly increased stimulated with PWM. When it was unstimulated, the level of IgM in KRG and CRG increased together. The IgA level of the KRG group significantly increased when it was stimulated with PWM and unstimulated. Conclusion : According to the above results, oral administration of red ginseng for 3 months is considered useful for immunomodulatory effect, and Korean red ginseng may be superior to Chinese red ginseng in that effect.

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A STUDY ON SYSTEMIC AND LOCAL PRODUCTION OF IMMUNOGLOBULINS IN HUMAN PERIAPICAL CYSTS (치근단 낭종환자의 혈청 및 낭종내의 항체수준에 관한 연구)

  • Park, Kye-Yang;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.17 no.1
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    • pp.222-234
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    • 1992
  • The purpose of this study was to evaluate the systemic and local production of immunoglobulins and their levels in patients with periapical cysts using Enzyme - Linked Immunosorbent Assay. Streptococcus sanguis, Bacteroides gingivalis, and Bacteroides intermedius were grown for use as antigen and they were harvested by centrifugation. The patients were divided into two groups: patients of periapical cysts and normal control. 5 patients of each group were selected and their blood were obtained via intravenous puncture prior to surgical operation. Sera were prepared by centrifugation of each blood samples. Cyst fluid were aspirated from cystic cavity and cyst wall were excised at operation. Control tissue were also excised at extraction site of impacted wisdom teeth from normal control. Each tissue was prepared by homogenization and centrifugation. Then antibodies of each sample were measured by modified ELISA. The following results were obtained: 1. Serum IgG and IgM levels were not significantly different between patients with periapical cyst and normal control. 2. IgG and IgM levels of cyst fluid to Bacteroides gingivalis and Bacteroides intermedius were significantly higher than those of serum of patients with periapical cyst, but there was no significant difference to Streptococcus sanguis. 3. IgG and IgM levels of cyst wall to Bacteroides gingivalis and Bacteroides intermedius were significantly higher than those of control tissue, but there was no significant difference to Streptococcus sanguis. 4. IgG and IgM levels in cyst fluid and IgG levels in cyst wall were highest to Bacteroides gingivalis, and IgM levels in cyst wall were highest to Bacteroides intermedius.

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Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A (보툴리눔 신경독소 A를 중화하는 재조합 항체의 제조와 특성 분석)

  • Park, Hong-Gyu;Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.1
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    • pp.295-301
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    • 2017
  • Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

Use of enzyme-linked immunosorbent assay (ELISA) for detection of toxoplasmosis in dogs (ELISA 법을 이용한 개 톡소플라즈마병의 조기진단에 관한 연구)

  • Suh, Myung-deuk;Joo, Hoo-don;Lee, Byung-hoon
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.491-500
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    • 1991
  • This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate. Tachyzoites of T gondii harvested from mouse peritoneal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows; 1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of $l{\mu}g/ml$ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate. 2. Cut-off value in this IgG ELISA was 0.375 that was determined by mean absorbance (at 492nm) of IFA negative serum added with the dauble value of the standard deviation $(mean{\pm}2S.D.)$. 3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation. 4. Stability of the antigen absorbed in the microplates that were preserved at $4^{\circ}C$ and $-25^{\circ}C$ separately were prolonged up to 3 weeks and 10 weeks at $4^{\circ}C$ and $-25^{\circ}C$, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer. 5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively. 6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.

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Monitoring the Rate of Frozen Denaturation of Bovine Myosin by Competitive Indirect ELISA Method (Competitive Indirect ELISA를 이용한 Bovine Myosin의 동결 변성도 측정)

  • Kim, Seong-Bae;Lee, Ju-Woon;Park, Jong-Heum;Do, Hyung-Ki;Hyun, Chang-Kee;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.862-870
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    • 1998
  • This study shows the application of Ci-ELISA method for monitoring the denaturation of myosin by the frozen treatment in order to differentiate thawed beef from chilled. Hanwoo M.Semitendinosus (n=25) was treated under the two different frozen process as follows; simple frozen treatment (Exp-1) at 4 different temperatures, -10, -20, -50 and $-80^{\circ}C$, respectively, and repeated thawing-refreezing treatment (Exp-2) stored at 4 different temperatures, -10, -20, -50 and $-80^{\circ}C$, respectively. Antibodies (Abs) were produced from rabbits immunized with myosin whole molecule (MWM) isolated from beef round, heavy meromyosin S-1 (S-1) and light meromyosin (LMM) prepared by digestion of MWM. Each immunoglobulin G (IgG) was separated from antiserum. At 6 month storage, IA of anti-MWM IgG for myosin was decreased to 32.67, 32. 23, 51.52 and 34.27% in Exp-1 and to 14.82, 15.61, 25.3 and 23.7% in Exp-2 at -10, -20, -50 and $-80^{\circ}C$, respectively (P<0.05). In Exp-1, the reactivities of anti-LMM IgG were decreased to 25.12, 21.42, 49.05 and 28.96%, and those of Exp-2 were to 11.88, 9.56, 20.63 and 12.64% at -10, -20, -50 and $-80^{\circ}C$, respectively, at 6 times thawing (P<0.05). Conclusively, myosin was denaturated by freezing treatment and LMM or myosin rod part might have suffered from more extreme demage than HMM S-1, and samples at $-50^{\circ}C$ were slightly injured less than others by freezing treatment.

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ELISA detection of IgG antibody against a recombinant major surface antigen (Nc-p43) fragment of Neospora caninum in bovine sera

  • Ahn, Hye-Jin;Kim, Sera;Kim, Dae-Yong;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.41 no.3
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    • pp.175-177
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    • 2003
  • An ELISA was established to measure bovine IgG directed against the recombinant antigenic determinant of Nc-p43, a major surface antigen of Neospora caninum. In a previous study, two thirds of the C-terminal of the molecule was expressed as a $6{\;}{\times}{\;}His$ tagged protein (Ncp43p) for ELISA using 2/3 of the N-terminal of SAG1 from Toxoplasma gondii as a control (TgSAG1A). Among 852 cattle sera collected from stock farms scattered nation-wide, 103 sera (12.1%) were found to react with Ncp43p positively, but no positive reaction was observed with TgSAG1A. This study shows that Ncp43p could be available as an efficient antigen for the diagnosis of neosporosis in cattle. Furthermore, it together with TgSAG1A, could be useful for the differential diagnosis of N. caninum and T.gondii infections in other mammals.

Effect of CpG Oligodeoxynucleotides on Airways of Mice with Established Airways Inflammation (기도 염증이 유발된 생쥐에서 CpG Oligodeoxynucleotides가 미치는 효과)

  • Hwang, Hei-Won;Kim, Su-Jin;Kim, Won-Duk;Cho, Sung-Min;Lee, Dong-Suk;Choi, Sung-Min
    • Clinical and Experimental Pediatrics
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    • v.45 no.7
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    • pp.875-883
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    • 2002
  • Purpose : Airways eosinophilia and increased IgE, characteristic features of asthma, result from a predominant Th2 response. In this study, we investigated the effect of CpG oligodeoxynucleotides (ODNs) on the inhibition of airways eosinophilia in mice with established airway inflammation. We also investigated the immunological mechanisms involved. Methods : Groups of BALB/c mice were sensitized intradermally with ovalbumin(OVA). At week 10, airway inflammation was induced by intranasal challenge of the mice with OVA. At week 14, the mice were challenged intranasally again with OVA in the presence and without the presence of CpG ODNs. Mice with saline administration served as negative controls. Bronchoalveolar lavage fluids(BALF) were obtained and eosinophils were counted. Th1 and Th2 cytokines in the spleen cell cultures were measured by ELISA. Serum OVA-specific IgE and IgG2a antibodies were also measured by ELISA. Results : BALF eosinophils were significantly inhibited in the CpG ODNs-treated mice(P<0.01). IgE and IgG2a levels increased significantly in both CpG ODNs-treated and untreated groups as compared to the negative control group; there was, however, no significant difference between the two groups four days after intranasal administration of CpG ODNs. Cytokine analysis revealed decreased production of IL-4, IL-5, and IL-13 and increased production of IL-12 in the CpG ODNs-treated group as compared to the untreated group. Interestingly, $IFN-{\gamma}$ levels were not upregulated in the CpG ODNs-treated group. Conclusion : CpG ODNs vaccination is a potentially useful approach for reversing airways eosinophilia in mice with established airways inflammation.

Serological Cross-Reactivity between Sarcocystis and Toxoplasma in Pigs (돼지에 있어서 Sareocustis와 Toxoplusma 감량의 혈청학적 교차반응 시험)

  • 문무홍
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.188-194
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    • 1987
  • The development of antibody titers and crossreaction between Sarcocystis and Toxoplasma were investigated by means of IF A test and ELISA in pigs experimentally infected with $1.5{\times}10^6$ S. suicanis sporocysts and 10,000 T. gondii oocysts, respectively. The intact and soluble Sarcocystis antigens were prepared from the bradyzoites harvested by peptic digestion of infected pork. The intact and soluble Toxoplasma antigens were prepared from the tachyzoites in mouse peritoneal cavity. IgG antibodies in pigs infected with Sarcocystis and Toxoplasma, respectively were detected first at 2 weeks post infection on both IF A test and ELISA. The antibody titer to Toxoplasma reached its maximum at 6 weeks post infection and decreased thereafter. The antibody titer to Sarcocystis reached its maximum terminally. The cross-reaction titer in pigs infected with Toxoplasma against Sarcocystis antigen was up to 1 : 16 in IFA test and up to 1 : 32 in ELISA. The titer in control group was below 1 : 4 in both reactions.

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Fabrication of a paper-based ELISA to detect polygalacturonase (Polygalacturonase를 검출하기 위한 종이 기반의 효소결합 면역반응 센서 제작)

  • Hwang, Young-Kug;Kim, Ji-Kwan;Lee, Young Hwan;Choi, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.30 no.5
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    • pp.337-341
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    • 2021
  • In this paper, we describe the fabrication of a paper-based enzyme-linked immunosorbent assay (ELISA) to detect polygalacturonase (PG), which is used as a biomarker to determine whether a plant is infected with a disease. The proposed paper-based ELISA can analyze the concentration of PG in a short time using a small sample compared to the traditional ELISA, which is generally performed using a well plate. To increase the resolution of the sensor, we optimized the dilution ratio of the HRP-conjugated goat anti-rabbit IgG antibody and the dilution ratio of the anti-PG and HRP-conjugated goat anti-rabbit IgG antibodies. Furthermore, for quantitative analysis of PG concentration, Delta RGB analysis was conducted to detect color changes in the sensing window displayed by the PG samples at various concentrations. Based on the experiment, the fabricated paper-based ELISA could measure at least 0.25 ㎍ of PG and the measurement range was 0.25-2 ㎍. Therefore, the paper-based ELISA for detecting PG is expected to be able to determine the presence or absence of disease in crops at the infection stage in the future.

Changes of Bovine Colostral Immunoglobulin G on Processing Conditions (가공처리조건이 초유 Immunoglobulin G의 변화에 미치는 영향)

  • 이수원;양동훈;황보식;이승환
    • Food Science of Animal Resources
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    • v.21 no.3
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    • pp.265-271
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    • 2001
  • We investigated changes of immunoglobulin G (IgG) concentrations by heating and drying condition. Also it is performed to group for commercial product by promoting of IgG preservation and reducing of protein denaturation. The result was that content of IgG in colostrum was higher than normal milk. Especially, IgG content of colostrum within 12 hrs after parturition was over 44.67mg/ml and it is 60 times of normal milk. IgG contents was reduced rapidly according as passage of the time. IgG content of the sample heating at 30min at 65$^{\circ}C$ was still a little higher that heating for 10sec at 72$^{\circ}C$. IgG denaturation of heat treatment at 100$^{\circ}C$ for 10sec was lower than at 85$^{\circ}C$ for 30min. We investigated the changes of IgG concentrations of kinds of market milk different with heating processing. This result showed that IgG denaturation ratio by ultra high temperature pasteurization (UHT) was higher than long time low temperature pasteurization (LTLT). On the other hands, IgG content by spray drying was 14.5mg/g and freezing drying was 10.8mg/g. It showed that denaturation of protein content by freezing drying was more than spray drying.

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