• Biotechnology and Bioprocess Engineering:BBE
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• 제1권1호
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• pp.13-17
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• 1996

The protein disulfide isomerase(PDI) reaction kinetics has been studied to evaluate its effect on the monoclonal antibody(MAb) refolding and assembly which accompanies disulfide bond formation The MAb in vitro assembly experiments showed that the assembly rate of heavy and light chains can be greatly enhanced in the presence of PDI as compared to the rate of assembly obtained by the air-oxidation. The reassembly patterns of MAb intermediates were identical for both with and without PDI, suggesting that the PDI does not determine the MAb assembly pathway, but rather facilitates the rate of MAb assembly by promoting PDI catalyzed disulfide bond formation. The effect of growth rate on PDI activities for MAb production has also been examined by using continuous culture system. The specific MAb productivity of hybridoma cells decreased as the growth rate increased. However, PDI activities were nearly constant for a wide range of growth rates except very high growth rate, indicating that no direct correlation between PDI activity and specific MAb productivity exists.

• 대한수의학회지
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• 제40권1호
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• pp.130-137
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• 2000

In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

• 한국축산식품학회지
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• 제34권6호
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• pp.822-828
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• 2014

In this study, the existence of skeletal muscle troponin I (smTnI), well-known as a muscle protein in fat tissues, and the utilization of smTnI as a biomarker for the identification of fat adulteration were investigated. A commercial antibody (ab97427) specific to all of animals smTnI was used in this study. Fat and meat samples (cooked and non-cooked) of pork and beef, and chicken considered as representative meats were well minced and extracted by heating and non-heating methods, and the extracts from fat and meat tissues were probed by the antibody used in both enzyme-linked immunosorbent assay (ELISA) and immunoblot. The antibody exhibited a strong reaction to all meat and fat extracts in ELISA test. On the other hand, the results of immunoblot analsis revealed a 23 kDa high intensity band corresponding to the molecular weight of smTnI (23786 Da). These results demonstrate that the existence of smTnI in all animal fat tissues. Since there are monoclonal antibodies specific to each species smTnI, smTnI in fat tissues could be used as a biomarker to identify or determine animal species adulterated in meat products. Therefore, an analytical method to identify fraudulent fat adulteration can be developed with an antibody specific to each species smTnI.

• Food Science and Biotechnology
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• 제17권1호
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• pp.15-19
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• 2008

This research was conducted to develop a quick and sensitive method of detecting carbamate residues using the immobilization of antibody-antigen interactions with surface plasmon resonance (SPR). We have used commercialized surface plasmon resonance equipment (Biacore 3000). The antibody used for the immunoassay was specific for glutathione-s-transferase (GST) and the antigens included several carbamate pesticides (carbofuran, carbaryl, and benfuracarb). When antigens were applied to the protein GST, the detection limit was 2 ng/mL of carbamate pesticide. The fabricated protein GST maintained its activity for over 200 measurements. Thus we determined that the SPR biosensors could detect the specific reversible binding of a reactant in solution to a binding partner immobilized on the surface of the sensor and allow real-time detection and monitoring.

• Journal of Plant Biology
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• 제39권2호
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• pp.99-105
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• 1996

Immunocytochemistry utilizes the specificity of the antigen-antibody reaction to localize specific antigens in cells or cellular organelles. Here we report the use of monoclonal antibodies, in conjunction with gold-labeled second antibodies to study the ultrastructural localization and tissue distribution of the Mr 98, 000 anionic peroxidase and other wall antigens. The antibody specific for this wall peroxidase, mWP3, labeled mainly the cell wall area. At the tissue level, the Mr 98, 000 peroxidase is located predominantly in the leaf mesophyll, internal coleoptile and sieve elements, but not in the root, as assayed with these procedures. The coleoptile walls were less heavily stained than the walls of leaf mesophyll cells. At the subcellular level, it is localized mainly in intercellular regions of the cell walls. A similar staining pattern was revealed by mWP19, one of anti-$\beta$ glucosidase antibody, though it looked less heavily stained than one with mWP3. In order to serve as a control wall staining using IgM monoclonal antibodies, mWP18 was used. Most of the label is localized over wall regions of cells of the young leaf mesophyll and coleoptile.

• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.279-282
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• 1996

간흡충 특이 단세포군항체인 CsHyb 0605-23과 반응하는 간흡충 항원의 특성을 밝히기 위하여 간흡충의 성충 조항원을 당지질 당질 단백으로 각각 분리한 후 각각 항원으로 사용하여 단세포 군항체와 효소떤역흡착검사법을 실시하였다. 그 결과. 오직 당질분획만이 단세포군항체와 반응하였다. 당질항원을 sodiumperiodate를 사용하여 약하게 산화시키자 단세포군항체와의 반응도가 떨어졌다. 따라서 이 단세포군항체에 반응하는 간흡충의 항원 및 항원결정기는 당질로 생각된다.

• 대한수의사회지
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• 제17권3호
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• pp.47-60
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• 1981

Lymphocytes that secrete antibodies can be made immortal by fusing them with myeloma tumor cells (cell fusion) and cloning the hybrids (hybridomas). Each clone is a long-term source of substantial quantities of a single highly specific antibody (monoclona

• 한국수산과학회지
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• 제50권5호
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• pp.547-552
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• 2017

The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

• 대한수의학회지
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• 제13권1호
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• pp.39-46
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• 1973

Cultural method, dark field microscopy & fluorescent antibody technique were compared for their sensitivity of the detection of leptospires from experimentally infected mice. Two groups of mice were infected with L. icterohemorrhagiae (M20) and L. australis (Ballico), and the infected blood, urine and a number of organs were subjected to the bacterial isolation. The results obtained were summarized as follows: 1. L. icterohemorrhagiae (M20) and L. australis (Ballico) in blood, urine and various tissues of experimentally infected mice were detected with a negrigible non specificity, by the fluorescent antibody technique. 2. The fluorescent antibody technique, as applied to detection of leptospires in blood, urine and various infected tissue, proved to be better than cultural method and dark-field microscopy. 3. Early detection of leptospires by fluorescent antibody technique were possible in blood at 2 days after inoculation, whereas detection of organisms in liver, spleen, lung and kidney were observed later. By means of fluorescent antibody technique, the detection of leptospires in kidney and urine was possible up to 34 days postinoculation, whereas those in other parts were impossible. 4. Fluorescent antibody reaction of leptospires were highly specific to homologous antigen rather than to heterologous one. 5. Fluorescent antibody technique may be of value in the application for the demonstration of leptospira from clinical specimens.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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• pp.432-437
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• 2002

Immunosuppressive potential of 65 kDa buffalo placental protein (bPP65) on B-cell proliferation in vitro and antibody response in vivo was evaluated. B-cell proliferation was estimated by measuring incorporation of tritiated thymidine in buffalo lymphocytes while primary antibody responses against phytohaemagglutinin (PHA) or keyhole limpet haemocyanin (KLH) were evaluated in mice. bPP65 suppressed proliferation of lipopolysaccharide (a B-cell specific mitogen)-stimulated buffalo lymphocytes in vitro indicating suppression of B-cells. This suppression was dose dependent over the protein concentration range $25-100 {\mu}g/ml$. Primary antibody responses in mice against PHA and KLH in presence of bPP65 were lower as compared to in its absence but these were not statistically significant. Amino acid composition data of bPP65 and BSA suggested that bPP65 is different from BSA.