• Title/Summary/Keyword: ELISA antibody

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Enzyme Immunoassay for Rapid Detection of the Fungicide Iprovalicarb Residues (살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 효소면역분석법)

  • Cho, H.K.;Kyung, K.S.;Lee, E.Y.
    • Journal of Biosystems Engineering
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    • v.31 no.6 s.119
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    • pp.535-540
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    • 2006
  • For a biosensor development, an enzyme-linked immunosorbent assay (ELISA) of the fungicide iprovalicarb was developed by minimizing the processing time. The time for whole incubation process was reduced from 135 minutes to 15 minutes. The concentration of antibody was varied to improve sensitivity. The total processing time was reduced from 2.5 hours to 20 minutes, the final sensitivity ($IC_{50}$ value) of 7.93 ng/mL and the lowest detection limit of 0.045 ng/mL were obtained. This ELISA was applied to potatoes and onions, and the recoveries were in the range of 98.85 $\sim$ 101.20% and 87.97 $\sim$ 102.70%, respectively. Accordingly, this method can be used as basis for a biosensor for rapid monitoring of iprovalicarb residues in crops.

Adhesive Properties of Lactobacillus brevis FSB-1 In Vivo

  • Kim, Seong-Yeong;Shin, Kwang-Soon;Lee, Ho
    • Food Science of Animal Resources
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    • v.30 no.4
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    • pp.560-567
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    • 2010
  • This study was conducted to evaluate the in vivo gastrointestinal survival and adhesive properties of orally administered Lactobacillus brevis FSB-1. ELISA conducted using polyclonal antibodies specific for L. brevis FSB-1 was able to detect the organism in feces; therefore, we used ELISA to determine the concentration of lactic acid bacteria in feces collected from Wister rats that had been administered $10^{10}$ cells/rat/d orally for 20 d. The mean recovery of L. brevis FSB-1 was approximately $10^{7.22}$ cells/g of wet feces during the oral administration period, and $10^{7.50}$ and $10^{7.46}$ at 8 and 10 d after the end of oral administration, respectively. These results indicate that L. brevis FSB-1 was able to survive in the gastrointestinal tract of rats, and that it had a high adhesive property in rat colons.

Development of an Enzyme-Linked Immunosorbent Assay for the Organophorus Insecticide Bromophos

  • Park, Won-Cheol;Cho, Young-Ae;Kim, Yoo-Jung;Hammock, Bruce D.;Lee, Yong-Tae;Lee, Hye-Sung
    • Bulletin of the Korean Chemical Society
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    • v.23 no.10
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    • pp.1399-1426
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    • 2002
  • A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos. Three bromophos analogues (haptens) were synthesized and were coupled to carrier proteins to use as immunogens or coating antigens. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin (BSA) for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin (OVA). Using the serum with highest specificity and an enzyme tracer, an antibody-coated ELISA was developed, which showed an $IC_{50}$ of 40 ng/mL with a detection limit of 7 ng/mL. The antibodies in this assay showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides chlorpyrifos and fenitrothion.

Production of Monoclonal Antibody against Sulfamethazine and Development of ELISA

  • C.S. Chae;Lee, M.H.;Lee, H.;S.Y. Yoon;P.D. Ryu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.176-176
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    • 1996
  • 축산식품중에 잔류하고 있는 sulfamethazine을 검출하기 위하여 sulfamethazine에 대한 단클론항체를 생산하고 이를 이용하여 효소면역측정법을 개발하였다. 면역원은 sulfamethazine에 KLH를 그리고 흡착항원은 BSA를 glutaraldehyde법으로 결합시켰다. 면역원으로 Balb/c mouse를 면역시킨 다음 비장 형질세포률 얻어 myeloma cell과 융합하여 융합잡종세포를 만들었다. Sulfamethazine에 대한 항체를 분비하는 융합잡종세포를 단계회석법과 ELISA를 이용하여 cloning하여 D2, A9, B8, Bl 클론을 얻었다. 이들 클론에서 얻어진 단클론항체를 사용하여 indirect competitive ELISA를 실시하여 표준곡선을 작성하여 본 결과 농도의존성 곡선을 얻을 수 있었다. 4클론중에서 A9 클론을 사용하여 다른 유사한 sulfonamide듣과 p-aminobenzoic acid와 교차반응을 조사한 결과 sulfamerazine에 12.5%의 교차반응을 보였으나 다른 설파제에 대해서는 교차 반응을 보이지 않았다.

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Diagnosis and gI antibody dynamics of pseudorabies virus in an intensive pig farm in Hei Longjiang Province

  • Wang, Jintao;Han, Huansheng;Liu, Wanning;Li, Shinian;Guo, Donghua
    • Journal of Veterinary Science
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    • v.22 no.2
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    • pp.23.1-23.10
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    • 2021
  • Background: Pseudorabies (PR), caused by the pseudorabies virus (PRV), is an endemic disease in some regions of China. Although there are many reports on epidemiological investigations into pseudorabies, information on PRV gI antibody dynamics in one pig farm is sparse. Objectives: To diagnose PR and analyze the course of PR eradication in one pig farm. Methods: Ten brains and 1,513 serum samples from different groups of pigs in a pig farm were collected to detect PRV gE gene and PRV gI antibody presence using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Results: The July 2015 results indicated that almost all brain samples were PRV gE gene positive, but PRV gI antibody results in the serum samples of the same piglets were all negative. In the boar herd, from October 2015 to July 2018 three positive individuals were culled in October 2015, and the negative status of the remaining boars was maintained in the following tests. In the sow herd, the PRV gI antibody positive rate was always more than 70% from October 2015 to October 2017; however, it decreased to 27% in January 2018 but increased to 40% and 52% in April and July 2018, respectively. The PRV gI antibody positive rate in 100-day pigs markedly decreased in October 2016 and was maintained at less than 30% in the following tests. For 150-day pigs, the PRV gI antibody positive rate decreased notably to 10% in April 2017 and maintained a negative status from July 2017. The positive trend of PRV gI antibody with an increase in pig age remarkably decreased in three tests in 2018. Conclusions: The results indicate that serological testing is not sensitive in the early stage of a PRV infection and that gilt introduction is a risk factor for a PRV-negative pig farm. The data on PRV gI antibody dynamics can provide reference information for pig farms wanting to eradicate PR.

Characterization of Mouse Hybridoma Producing Monoclonal Antibody Paragonimus westermani (폐디스토마에 대한 Mmonoclonal Antibody생산 세포주개발에 대한 연구)

  • 고광삼;이숙영;이근배
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.64-68
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    • 1985
  • In this study, hybridoma techniques were applied to produce monoclonal antobodies to Paragonimus westermani, commonly known as lung distoma. Balb/c mice were immunized intraperitoneally every week with increasing doses of purfied protein of Paragonimus westermani adult worms begining with 0.3/mg/mouse/7 days and ending with 0.5mg at 28th day. Spleen cells from these immunized mice were hybridized with myeloma cells (NS-1) and the hybridized cells were selected in HAT media. The antibody secreting cells among the hybrid cells were initially selected by ELISA. Those initially selected cells were further screened by the criteria of antibody producing activity, and seneral cell lines among them were further tested with immunodiffusion method. One hybridoma clone produced IgM and another clone produced IgG1. The supernatant of the hybridoma clone producing IgM had titer 1:64 and the hybridoma clone producing IgG1 had titer 1:256 measured by immunofluorescence technique.

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The prevalence of rubella antibodies in the women of childbearing age (서울시내 일개 대학병원에 내원한 가임 여성의 풍진항체 보유율)

  • Uhm Jin Woen
    • Journal of Korean Public Health Nursing
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    • v.13 no.1
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    • pp.88-96
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    • 1999
  • This study was conducted in May of 1996 to December of 1996 in order to investigate the status of rubella antibodies in the women of childbearing age. The subjects were 543 fertile women (Ages 21-42 years). ELISA method was used for the detection of rubella antibodies and then questionaire survey was performed to know about the variables of past history of rubella. past rubella immunization. parity and cognition. The results were as follows: 1. The Positive rate of rubella Ig G antibody in total subjects was $65.0\%$. The positive rate of rubella Ig G antibody was $72.2\%$ in 21-25 age group. $71.4\%$ in 26-30 age group. $54.5\%$ in 31-35 age group, $52.6\%$ in 36-42 age group. As age increased, the positive rate of rubella Ig G antibody was decreased. There was statistically significant difference by age group(P=0.00l). In the subjects with a history of rubella immunization, the positive rate of rubella IgG antibody was $81.8\%$, and in those with past history of rubella was $83.3\%$ of positive rate. 2, Cognition rate about rubella immunization showed $50.8\%$ in total subjects. and there was no significant difference between parity and cognition rate of rubella immunization(P=0.85l). observed a low positive rate of rubella IgG antibody as compaired with other studies. Therefore, to prevent congenital rubella infection, rubella immunization was needed for unmarried women.

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B3(Fab)-streptavidin Tetramer Has Higher Binding Avidity than B3(scFv)-streptavidin Tetramer

  • Won, Jae-Seon;Kang, Hye-Won;Nam, Pil-Won;Choe, Mu-Hyeon
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1101-1106
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    • 2009
  • Multivalent and multi-specific antibodies can provide valuable tools for bio-medical research, diagnosis and therapy. In antigen-antibody interactions, the avidity of antibodies depends on the affinity and the number of binding sites.$^1$ As artificial multivalent antibody agents, single chain Fv-streptavidin fusion tetramer proteins $(scFv-SA)_4$ have been previously tested.$^{1,\;2}$ Although, the Fab domain is known to be more stable than scFv in animal models,$^{3,\;4}$ it has never been used to make a multivalent agent with a streptavidin fusion. In this study, we prepared tetra-valent $(Fab-cSA)_4$ by fusing Fab with core streptavidin (cSA). This molecule was made using inclusion body production, refolding and chromatography purification. Affinities of the Fab-cSA tetramer and a scFv-cSA tetramer to a cell surface antigen were compared by ELISA using biotin-HRP. The Fab-cSA tetramer showed higher binding avidity than the scFv-cSA tetramer. The higher binding avidity of the Fab-cSA tetramer demonstrates its potential as a therapeutic agent for target-specific antibody therapy.

Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.