• Korean Journal of Veterinary Service
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• v.37 no.3
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• pp.143-150
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• 2014

Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiological agent of PRRS characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRSV is a small enveloped virus containing a positive-sense, single-stranded RNA genome and divided into two genotype, type 1 (European) and type 2 (North American), respectively, by nucleotide identity. In this study, ORF7 gene of the type 1 and type 2 PRRSV was cloned and expressed in Baculovirus expression system. Also, monoclonal antibodies (MAbs) against ORF7 were produced and characterized. The expressed ORF7 proteins in the recombinant virus were confirmed by indirect fluorescence antibody (IFA) test using His6 and PRRSV-specific antiserum. A total of eight MAbs were produced and characterized. One (3G12) MAb was type 1 PRRSV ORF7-specific and two (6B10 and 16H8) were type 2 PRRSV ORF7-specific. Other five (1A1, 2A4, 4B4, 12C4 and 13F11) MAbs reacted with both type 1 and type 2 PRRSV. Some PRRSV ORF7-specific MAbs recognized the porcine tissues infected with PRRSV by IFA or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PRRSV ORF7-specific and could be used as reliable reagents for type 1/type 2 PRRSV detection.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1170-1178
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• 2023

Food allergy represents a severe problem for many societies, including sensitive populations, academies, health authorities, and the food industry. Peanut allergy occupies a special place in the food allergy spectrum. To prevent consumption by consumers suffering from a peanut allergy, a rapid and sensitive detection method is essential to identify unintended peanut adulteration in processed foods. In this study, we produced four monoclonal antibodies (MAbs; RO 3A1-12, PB 4C12-10, PB 5F9-23, and PB 6G4-30) specific to thermo-stable and soluble proteins (TSSPs) of peanut and developed an enzyme-linked immunosorbent assay (ELISA) based on the MAbs. Among them, PB 5F9-23 MAb was firmly bound to Ara h 1, and other MAbs strongly reacted to Ara h 3 in the Western blot analysis. An antibody cocktail solution of the MAbs was used to enhance the sensitivity of an indirect ELISA, and the limit of detection of the indirect ELISA based on the antibody cocktail solution was 1 ng/ml and improved compared to the indirect ELISA based on the single MAb (11 ng/ml). The cross-reaction analysis revealed the high specificity of developed MAbs to peanut TSSPs without cross-reaction to other food allergens, including nuts. Subsequently, analyzing processed foods by indirect ELISA, all foods labeled as containing peanuts in the product description were confirmed to be positive. The results indicate that the developed antibodies exhibit high specificity and sensitivity to peanuts and can be used as bio-receptors in immunoassays or biosensors to detect intentional or unintentional adulteration of peanuts in processed foods, particularly heat-processed foods.

• Proceedings of the KIEE Conference
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• 2006.07b
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• pp.849-850
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• 2006

최근 자동차 브레이크의 전기적 시스템에 대한 연구가 활발히 진행되고 있다. 이와 관련하여 본 논문에서는 자기 엑추에이터 브레이크 시스템(MABS)을 분석하였다. 주행 도중 차량의 이상 및 긴급 상황 발생시 MABS는 전기적 메커니즘을 통해 차량을 제동할 수 있다. MABS는 회전하고 있는 휠에 초당 수십회 작용하여 점차적이고 효율적으로 휠을 제동한다. 이는 제동시간의 단축과 안정성에 있어 향상된 성능을 보인다. 본 연구에서는 유한요소법을 적용한 시뮬레이션을 통해 MABS의 동작 특성을 분석하고, 실제 자동차의 상황을 가정하여 제동하는데 소요되는 시간 및 작동 회수 등에 대해 분석하였다.

• Journal of the Korean Wood Science and Technology
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• v.21 no.2
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• pp.31-37
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• 1993

Composites of styrene polymers with woody fibers were prepared, and the effect of compatibilizers on their mechanical properties was evaluated. To improve the compatibility of wood fibers and the matrix polymers, styrene-maleic anhydride copolymer(SMA) and maleic anhydride-modified polymers were used as compatibilizers. As results, maleic anhydride-modified polystyrene and SMA were proved to improve the tensile strength of the molded composites, and also were evaluated as good compatibilizers for the wood fiber polystyrene composite. Cellulosic fiber (dissolving pulp) provided better reinforcement than lignocellulosic fiber(thermomechanical pulp). On the contrary in the case of the composite of wood fiber and acrylonitrile-butadiene styrene copolymer(ABS), SMA and maleic anhydride-modified acrylonitrile-butadiene-styrene copolymer(MABS) did not act as compatibilizers. However, MABS was evaluated as a good polymer matrix to make wood fiber reinforced composite. The tensile properties of the composites of wood fiber and MABS were superior than those of wood fiber-ABS composites.

• The Korean Journal of Zoology
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• v.38 no.3
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• pp.382-387
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• 1995

To check the possible application of our anti-AFP monocional antibodies (MAbs) as immunodiagnostic reagents for hepatocellular carcinoma, ELISA and immunohistochemical assay were performed on the sera and liver biopsy specimens from the patients of hepatocellular carcinoma and other non-malignant hepatic disease. By non-competitive ELISA using anti-AFP MAbs, the highest incidence of AFP value was found only in the sera of hepatocellular carcinoma patients, i.e., more than 54% of patients had serum AFP levels of more than 500 ng/mi. By immunoperoxidase and indirect immunofluorescence techniques, anti-AFP MAbs were found to react with cytoplasm of hepatoceliular carcinoma cells. However immunohistochemical reactIvity to AFP in hepatocellular carcinoma cells was lower than that in non-neoplastic liver cells adjacent to the hepatocellular carcinoma. From these results with the similar findings from other studies, we suggest that AFP antigen is appropriate in the diagnosis assay (ELISA) but is not by immunohistochemical detection.

• Journal of Food Hygiene and Safety
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• v.32 no.1
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• pp.75-81
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• 2017

For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without cross-reaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

• The Korean Journal of Zoology
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• v.36 no.4
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• pp.522-528
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• 1993

Monoclonal antibodies (MAbs) against human alpha-fetoprotein (AFPI was produced by hybridizing SP 210-Ag 14 mouse myeloma cells with spleen cells of Balb/c mice immunized with purified AFP. Two subclones (D-6 and I-6) were expanded as ascite tumors in svngenic mice, and from ascitic fluid immunoglbulins were Pruified. Each anibodv was identified to be homogeneous by several criteria, and the affinity constant of D-6 and I-6 MAb to AEP was calculated to be 4.2${\times}$10-8 and 6.4${\times}$10-8 M-1, respectively. With these MAbs sensitive and accurate enzyme linked immunosorbent assay method was established.

• BMB Reports
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• v.37 no.3
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• pp.314-319
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• 2004

Flounder (Paralichthys olivaceus) Immunoglobulins (Igs) were purified from the serum of mouse IgG-immunized flounder by using affinity chromatography. Under denaturing conditions in SDS-PAGE, the flounder Igs appeared to be composed of 2 heavy (H) chains (72 and 77 kDa) and two light (L) chains (26 and 28 kDa). Monoclonal antibodies (MAbs) were produced by the fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells that were previously sensitized against affinity-purified flounder Igs. In a Western blot analysis, the produced MAbs, FIM511, FIM519, and FIM562 recognized both the 72 and 77 kDa H chains, 26 kDa, and 28 kDa L chain, respectively. Mouse antiserum against flounder Igs reacted more strongly with the L chain of 28 kDa than with 26 kDa, suggesting that the 28 kDa molecule is more immunogenic than the 26 kDa L chain molecule. In a FACS analysis, the ratios of the Ig+ cell population in the flounder head kidney and spleen cells were 49% and 24%, respectively. Unexpectedly, however, the ratios of the Ig+ B-like cell population in the flounder were not significantly augmented, even after the immunization of an immunogenic antigen. This suggests that the humoral immune response in fish could be considerably different from that in mammals. The produced MAbs in this study would be useful in characterizing flounder Ig+ B-like cells and in developing flounder Ig detecting an immunoassay system.

• The Plant Pathology Journal
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• v.19 no.5
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• pp.260-265
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• 2003

Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

• The Korean Journal of Zoology
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• v.32 no.4
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• pp.412-419
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• 1989

Monoclonal antibodies (MAbs) reacting with the plasmalemma of Amoeba proteus were produced. Specificity of the 3 MAbs was determined by transfer blotting of the SDS polvacryfamide gel. AMS antibody reacted with the mucopolysaccharide bands of the spacer gel, 220 KD and 50 KD proteins of the resolving gel. The maior glycoprotein bands (175 KD, 165 KD) and 50 KD protein of the plasmalemma were recognized by AUG antibody. A third, AMP antibody reacted with the 50 KD protein only. In immunofluorescence microscopy of the enzyme treated cells, the antigens of these MAbs were sensitive to proteases, but not sensitive to neuraminidase. In the assay of cell to substratum attachment after binding with the antibody, AMG and AMP antibodies exerted no effect, but AMS hindered the attachment and cell spreading. Thus the effective components of the plasmalemma in cell to substratum attachment appear to be the mucopolysaccharides and 220 KD protein. The membranes of latex particle infested phagosomes did not show any distinction from the plasmalemma in fluorescence microscopy. Phagosome membranes of amoebae appear to be derived from the plasma membrane without selection in terms of the antigen composition. Amoeba Proteus의 세포막과 반응하는 단세포군 항체를 생산하였다. SDS polyacrylamide gel을 transfer blotting하여 이들 항체의 반응 특이성을 조사해 본 결과 AMS 단항체는 PAS로 염색되는 spacer gel의 mucopolysaccharide 린드, resolving gel의 220 KD 및 50 KD 단백질과 반응하였으며, 세포막의 주요 당단백질인 175 KD 및 165 KD 빈드와 50 KD 단백질은 AMG 단항체에 의해서 인지되었다. 그리고 AMP단항체는 공통인 50 KD 단백질과 특이하게 반응하였다. 효소처리한 아메바의 면역형광칠미경적 조사에서 이들 항체에 대한 항원분자들은 모두 단백질분해효소에 민감하였으며 neuraminidase에 대해서는 변화가 없었다. 이들 항체를 결합시킨 아메바의 용기표면 부착 가능성을 분석한 결과 AMP 및 AMG 단항체는 아무런 영향을 미치지 못하였으며 AMS 단항체는 세포의 용기표면 부착 및 세포의 펴짐을 저해하였다. 따라서 아메바의 용기표면 부착은 mucopolysaccharide 및 220 KD 단백질에 의해서 매게되는 것으로 나타났다. 그리고 latex particle을 담고 있는 식포막은 면역형광형미경적 조사에서 세포막과 차이가 없었다. 따라서 겐포막은 항원 조성에 있어서 비 선택적으로 세포막에서 유도되는 것으로 나타났다.