• 한국동물학회지
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• 제34권1호
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• pp.44-53
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• 1991

Distribution of the antigens during the cell cycle of amoebae was followed by immunof-luorescence microscopy using a monoclonal antibody against the nucleus as a probe. While the cells were in the interphase, the antigen was localized on the nucleus membrane. But it was dispersed all over the cytoplasm during mitosis and cytokinesis. The molecular weights of the immunoreacted antigens were 210 KD and 190 KD as determined by SDS PAGE and western blotting of the purified nuclei. The antigens were not soluble in non-ionic detergent, but were released from the nucleus by incubation with 0.05 M sodium carbonate, pH 10.6 or with 8 M urea at serial chemical extraction. Thus the antigens appeared to be peripheral proteins of the nurBeus envelope. The isoelectic point of both antigens was 7.64 as determined by 2 D PAGE and transfer blotting. Considering the peiipherd association with the nucleus membrane and the dispersed distribution during mitosis, the antigens could be lamin like proteins. Hourever, it appears also possible that they are the component molecules of the unusually structured aurous lamina of amoeba nucleus since they have the large molecular weight and the basic pl.

• 미생물학회지
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• 제28권1호
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• pp.6-12
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• 1990

adr 아형 B형 간염바이러스의 preS2염기서열을 베타갈락토시다세 유전자에 연결시켜 클론닝한 후, 융합단백질의 형태로 대장균에서 발현시켰다. 한국인 간염환자로부터 분리된 표면항원으로 유도한 단일 클론항체 H8는 preS2에 특이성을 지니고 있으며, 상기의 융합단백질과 특이적으로결합하였다. 이 단일 클론항체는 adw2아형 preS2 융합단백질과는 단지 낮은 수준의 결합성을 보여주었는데, 이와 같은 단일클론항체 H8의 adr 아형과 adw2아형에 대한 결합성의 차이는 preS2부위의 아미노산 차이에 기인한다고볼 수 있다.

• 대한수의학회지
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• 제36권2호
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• pp.395-403
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• 1996

Monoclonal antibodies(MAbs) against field isolates of the bovine rotavirus A strain(G6), V strain(G10) and reference I-801 strain(G8) were produced and characterized. Six MAbs(4C2, 4D9, 5E1, 5E7, 5D5, 3E4) against A strain had neutralizing activity and reacted only with the G6 bovine rotaviruses determined by fluorescence focus neutralization (FFN) test. Otherwise, five neutralizing MAbs(1G2, 2G6, 5E2, 5E12, 5H7) against I-801 strain neutralized the G6 and G8 bovine rotaviruses. Five non-neutralizing MAbs(5F12, 7F12, 5E11, 2A11, 2B12) were VP6-specific and cross-reacted with all bovine and porcine rotaviruses examined by fluorescence antibody(FA) test. None of the MAbs reacted with bovie viral diarrhea virus(BVDV) and bovine coronavirus(BCV) determined by FA and FFN test.

• Clinical and Experimental Reproductive Medicine
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• 제17권1호
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• pp.87-91
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• 1990

Development of a solid-phase chemiluminescence immunoassay for the detection of progesterone in serum extract was described. The chemiluminescence immunoassay was establised utilizing anti-progesterone monoclonal antibody that coated on polystyrene tubes and progesterone-ABEI conjugate as tracer. The light yield generated from antibody bound conjugate was counted on clinilumat luminometer by oxidation with microperoxidase and peroxide. The chemiluminescence immunoassay was high specific and accurate and detects as little as 3.9ng/ml of progesterone. The intra-assay CV ranged from 6% to 11.5% and inter-assay CV ranged from 13.6% to 18.7%. This assay system was good correlated with conventional kit radioimmunoassay system (r=0.98).

• 한국동물위생학회지
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• 제37권3호
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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.

• 한국어병학회지
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• 제32권1호
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• pp.45-48
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• 2019

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against recombinant VP28 structural protein (rVP28) of white spot syndrome virus (WSSV). We established six hybridoma clones secreting MAbs against rVP28: 15A11, 20G6, 31H2, 34H6, 38D1 and 43A1. All six MAbs recognized the 25 kDa of protein in gill homogenates of WSSV-infected shrimp by western blot analysis, while no reactivity was observed in gill homogenates of normal shrimp. Moreover, high enzyme-linked immunosorbent assay (ELISA) optical density (OD) values (0.8-2.68) were observed in the hemolymphs from WSSV-infected shrimp, while low OD values (less than 0.24) were recorded in the hemolymphs from normal shrimp, by using these six MAbs produced in this study. These results suggest that these six MAbs are useful for the detection of WSSV.

• 한국식품위생안전성학회지
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• 제18권1호
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• pp.25-29
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• 2003

리스테리아의 p60단백질은 Listeria속 고유의 단백질로써, 주요 세포외 단백질이기에, 식품에서 이 세균의 존재를 밝혀주는 중요한 지시단백질로 사용된다. 본 연구는 재조합 DNA 방법으로 재조합-p60 단백질을 대장균에서 생산하였으며, 아밀로오스 컬럼 크로마토그라피의 방법으로 정제하여, Listeria spp.의 p60에 특이적 항체를 생산하는 단일클론을 선별하였다. 생산된 항p60 항체 1H4는 병원성 Listeria 균주인 L. monocytogenes, L. ivanovii, L. weishi meri II 특이적으로 강한 항체반응을 보였으며, Listeria속의 비병원성균인 L. innocua등과는 상대적으로 매우 약한 항체반응을 보였으며, 타 세균의 단백질과는 거의 반응하지 않았다. 1H4항체는 복수생산법에 의해 대량생산되었으며, 이 항체의 특이성은 면역학적 방법에 의한 리스테리아 검출키트의 개발에 유용하게 사용될 수 있을 것이다.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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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.

• 한국동물학회지
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• 제35권1호
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• pp.1-7
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• 1992

Seven monoclonal antibodies were produced by fusing splenocytes from Balb/C mouse immunized with partially purified human interferon-a (HUIFN-a) with NSO plasmacytoma cells. aery were identified as five IgG class (432.22: IgG2b/n, 460.52: IgG2b/a , 548.46: IgG2a/n , 573.10: IgG2b/h , 625.12: IgG2b/n ), one IgA class (460.50: IgA/n ) and one IsM class (465.27: IgA/n ), and all of them revealed highly sensitive to HUIFN- a IgG class monoclonal antibodies have pts ranged from 8.2 to 8.6. Ascites fluids produced from primed Balb/c mice and were purified through column chromatography. The cytopathic effect (CPE) inhibition assay to examine neutralization of HuIFU-a by IgG class monoclonal antibodies, gave that MAbs 460.52, 548.46, 573.10 can neutralize HUIFU- a arith varying degrees except 432.22. Therefore, it is deduced that these various monoclonal antibodies may recognize the distinct epitopes on HUIFN-a.

• IMMUNE NETWORK
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• 제3권1호
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• pp.23-28
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• 2003

Background: Intracellular antibody specific to hepatitis B virus X protein (HBx) might be useful for studying the role of HBx in hepatocellular carcinogenesis and HBV replication. Methods: With variable region genes for H7 monoclonal anti-HBx Ab, we constructed a vector for bacterial expression of single chain Ab (scFv) and a vector for eukaryotic cell expression of it. The expression of H7 scFv and its binding activity against HBx was examined by immunoblotting and immunofluorescence microscopy. Results: H7 scFv expressed in bacterial cells retained reactivity to HBx. We demonstrated its intracytoplasmic expression in CosM6 eukaryotic cells. Conclusion: This is the first study showing the expression of intracellular anti-HBx Ab in eukaryotic cells. H7 scFv may be a good tool to study the function of HBx in HBV infection.