• 한국어병학회지
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• 제6권2호
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• pp.87-92
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• 1993

1. 1990년 2월 강원도 삼척군에 소재한 송어양식장에서 전염성 조혈기 괴사증 증상으로 산천어 자치어가 대량폐사하였다. 2. 폐사한 산천어 치어로부터 세포배양법에 의해 전염성 조혈기 괴사증 바이러스가 분리되었다. 3. 분리된 바이러스는 구조단백의 크기와 항원성에 있어 미국에서 분리된 RB-76 바이러스주(electro-pherotype 1)와 유사하였다.

• The Plant Pathology Journal
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• 제26권1호
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• pp.25-31
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• 2010

Papaya ringspot virus (PRSV) causes the most widespread and devastating disease in papaya. Isolates of PRSV originating from different geographical regions in south India were collected and maintained on natural host papaya. The entire coat protein (CP) gene of Papaya ringspot virus-P biotype (PRSV-P) was amplified by RTPCR. The amplicon was inserted into pGEM-T vector, sequenced and sub cloned into a bacterial expression vector pRSET-A using a directional cloning strategy. The PRSV coat protein was over-expressed as a fusion protein in Escherichia coli. SDS-PAGE gel revealed that CP expressed as a ~40 kDa protein. The recombinant coat protein (rCP) fused with 6x His-tag was purified from E.coli using Ni-NTA resin. The antigenicity of the fusion protein was determined by western blot analysis using antibodies raised against purified PRSV. The purified rCP was used as an antigen to produce high titer PRSV specific polyclonal antiserum. The resulting antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) assay and compared its sensitivity levels with ELISA based assays for detection of PRSV isolates. IC-RT-PCR was shown to be the most sensitive test followed by dot-blot immunobinding assay (DBIA) and plate trapped ELISA.

• BMB Reports
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• 제38권6호
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• pp.717-724
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• 2005

The nontoxic B subunit of cholera toxin (CTB) can significantly increase the ability of proteins to induce immunological tolerance after oral administration, when it was conjugated to various proteins. Recombinant CTB offers great potential for treatment of autoimmune disease. Here we firstly investigated the feasibility of silkworm baculovirus expression vector system for the cost-effective production of CTB under the control of a strong polyhedrin promoter. Higher expression was achieved via introducing the partial non-coding and coding sequences (ATAAAT and ATGCCGAAT) of polyhedrin to the 5' end of the native CTB gene, with the maximal accumulation being approximately 54.4 mg/L of hemolymph. The silkworm bioreactor produced this protein vaccine as the glycoslated pentameric form, which retained the GM1-ganglioside binding affinity and the native antigenicity of CTB. Further studies revealed that mixing with silkworm-derived CTB increases the tolerogenic potential of insulin. In the nonconjugated form, an insulin : CTB ratio of 100 : 1 was optimal for the prominent reduction in pancreatic islet inflammation. The data presented here demonstrate that the silkworm bioreactor is an ideal production and delivery system for an oral protein vaccine designed to develop immunological tolerance against autoimmune diabetes and CTB functions as an effective mucosal adjuvant for oral tolerance induction.

• BMB Reports
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• 제30권6호
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• pp.403-409
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• 1997

Studies were undertaken to develop a competitive radioimmunoassay (RIA) and indirect antigen capture enzyme-linked immunosorbent assay (ELISA) for the determination of 5'-deoxy-5'-methylthioadenosine (MTA), which is formed from decarboxylated S-adenosylmethionine by spermidine and spermine synthase. Specific antiserum against MTA was raised in rabbits by immunization with MTA-BSA which was prepared by coupling BSA to oxidized MTA with periodate. Since MTA is oxidized easily to the sulfoxide, the sulfhydryl reagent, DTT. was added to the immunogen. For RIA, immunocomplexes were separated from free MTA by using ammonium sulfate precipitation. The antiserum showed almost no cross-reactivity with a variety of other nucleotides and riboses. But, the level of cross-reactivity of 5'-isobutylthioadenosine (SIBA) was high. These results showed the importance of hydrophobicity adjacent to the 5'-OH for determining antigenicity. The lower limit of detection by this assay was 100 fmol of MTA per tube. Using this assay. MTA levels were more easily and precisely determined in biological samples when compared with HPLC analysis. The RIA procedure is less time consuming. More than 24 analyses can be carried out in 2 h and required only a very small amount of sample ($20{\mu}l$ serum). In ELISA, biotin conjugated MTA-BSA was used as the labelled MTA. The sensitivity limit of this assay was lower than 100 pmol.

• 한국미생물·생명공학회지
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• 제30권4호
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• pp.320-324
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• 2002

우리나라 연안에 주로 발생되는 적조인 Cochlodinium polykrikoides를 빠르고 정밀한 생화학적인 방법으로 측정하기 위한 일종의 maker로서 C. polykrikoides의 세포막에 존재하는 특이 항원성을 가진 막 단백질을 분리하였다. 먼저, C. polykrikoides와 gymnodium sangineum의 세포를 삼투압으로 터뜨린 후 원심분리하여 세포막을 모았다. 그 후 양 적조생물의 세포막은 1 mM DTT가 함유된 50 mM Na-carbonate로 용해하고 SDS-PAGE행하여 용해된 막 단백질을 분리하였다. SDS-PAGE로 분리된 막 단백질은 C. polykrikoides의 세포 막 단백질로 제조한 항혈청을 사용하여 immune-blot한 결과 C. polykrikoides의 120 kDa의 단백질이 G. sangineum의 동일한 크기의 막 단백질과는 서로 다른 항원성을 나타내었다.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.129-136
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• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• 대한치과의사협회지
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• 제25권5호통권216호
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• pp.465-477
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• 1987

This is an experimental and clinical study on the use of an autogenous frozen mandibular bone. Two aspects must be considered in the treatment of mandibular tumors: the eradication of the disease, and the maintenamce of the mandibular continuity for a good functional and esthetic results. The authors used the cryosurgical method in the dogs and the tumor of the mandible in human, which consists of partial mandibular resection; immersion of the bone in the liquid nitrogen at -196℃, followed by reimplantation of the mandibular bone in its bed. In experimental study, the mandibular segment showed favorable healing process without rejection during the cohole experimental periods. In clinical study, the frozen bone showed no antigenicity and excellent cosmetic results.

• Parasites, Hosts and Diseases
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• 제37권3호
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• pp.157-162
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• 1999

Serum from mouse orally ingested with tissue cyst forming stain (Me49) of Toxoplasma gondii was assayed by Western blot and immunofluorescene assay (IFA) to establish early responses in antigenicity of the parasite in mouse model of foodborne toxoplasmosis. Sera were collected weekly to blot the RH antigen transferred onto nitrocellulose paper after being separated by 12% SDS-PAGE. With the second week serum, 34 kDa protein (p34) was detected uniquely, and all antigens of T.gondii were detected with the sera from 3 or 4 weeks. p34 was not a member of the major surface membrane proteins and confirmed to be localized in the rhoptry by IFA. It was secreted into parasitophorous vacuolar membrane (PVM) during the entry into host cells. 10.3% of sera detected p34, while all the ELISA positive sera detected the band. It has diagnostic usefulness of presumed T.gondii infection. We suggest the name of the p34 protein as ROP9.

• Parasites, Hosts and Diseases
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• 제53권6호
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• pp.665-673
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• 2015

After invasion of red blood cells, malaria matures within the cell by degrading hemoglobin avidly. For enormous protein breakdown in trophozoite stage, many efficient and ordered proteolysis networks have been postulated and exploited. In this study, a potential interaction of a 60-kDa Plasmodium falciparum (Pf)-heat shock protein (Hsp60) and Pf-calpain, a cysteine protease, was explored. Pf-infected RBC was isolated and the endogenous Pf-Hsp60 and Pf-calpain were determined by western blot analysis and similar antigenicity of GroEL and Pf-Hsp60 was determined with anti-Pf-Hsp60. Potential interaction of Pf-calpain and Pf-Hsp60 was determined by immunoprecipitation and immunofluorescence assay. Mizoribine, a well-known inhibitor of Hsp60, attenuated both Pf-calpain enzyme activity as well as P. falciparum growth. The presented data suggest that the Pf-Hsp60 may function on Pf-calpain in a part of networks during malaria growth.

• 한국수의병리학회지
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• 제7권1호
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• pp.11-15
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• 2003

Antigen retrieval (AR) techniques were widely used to recover the antigenicity from the fixed tissues, which were guided by the philosophy of rendering immunohistochemistry (IHC) applicable to routine formalin-fixed, paraffin-embedded tissues for wide application of IHC in research and clinical filed for morphological observation like as anatomy, histology and pathology. Protease antigen recovery (PAR) is an AR technique, which is obtained the antigen retrieve by using enzyme digestion, and commonly used in IHC field. However, during the IHC for the detection of ovine herpesvirus 2 (OvHV-2) antigen, we noted lymphocyte-like cells-specific staining in the infiltrated cells into various organs like as liver and kidney, which was also shown in the IHC tissues with isotype control. However, those signals were not observed in the tissues conducted with in situ hybridization. Therefore, we analyzed the specificity of the IHC detection results. We found that PAR may induce false-positive result during IHC in lymphocyte-like cells, which were infiltrated mainly around vessels and in interstitial tissues. Through the Phenotyping, we realized that those false-positive cells were B-cell-related cells. These results suggest that PAR, a AR using protease, may induce non-specific false-positive reactions during IHC.