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RISS 인기검색어
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- 저자 : 한백상 (검색결과 4 건)
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Purification and characterization of monoclonal IgG antibodies recognizing Ebola virus glycoprotein
한백상,장호영,Trina Racine,Xiangguo Qiu,신정임 대한백신학회 2018 Clinical and Experimental Vaccine Research Vol.7 No.2
Purpose: The goal of this study was to purify and characterize Ebola virus glycoprotein (GP)-specific IgG antibodies from hybridoma clones. Materials and Methods: For hybridoma production, mice were injected by intramuscular-electroporation with GP DNA vaccines, and boosted with GP vaccines. The spleen cells were used for producing GP-specific hybridoma. Enzyme-linked immunosorbent assay, Western blot assay, flow cytometry, and virus-neutralizing assay were used to test the ability of monoclonal IgG antibodies to recognize GP and neutralize Ebola virus. Results: Twelve hybridomas, the cell supernatants of which displayed GP-binding activity by enzyme-linked immunosorbent assay and the presence of both IgG heavy and light chains by Western blot assay, were chosen as a possible IgG producer. Among these, five clones (C36-1, D11-3, D12-1, D34-2, and E140-2) were identified to secrete monoclonal IgG antibodies. When the monoclonal IgG antibodies from the 5 clones were tested for their antigen specificity, they recognized GP in an antigen-specific and IgG dose-dependent manner. They remained reactive to GP at the lowest tested concentrations (1.953-7.8 ng/mL). In particular, IgG antibodies from clones D11-3, D12-1, and E140-2 recognized the native forms of GP expressed on the cell surface. These antibodies were identified as IgG1, IgG2a, or IgG2b kappa types and appeared to recognize the native forms of GP, but not the denatured forms of GP, as determined by Western blot assay. Despite their GP-binding activity, none of the IgG antibodies neutralized Ebola virus infection in vitro, suggesting that these antibodies are unable to neutralize Ebola virus infection. Conclusion: This study shows that the purified IgG antibodies from 5 clones (C36-1, D11-3, D12-1, D34-2, and E140-2) possess GP-binding activity but not Ebola virus-neutralizing activity.
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Molecular Epidemiological Study of Measles Cases in Gyeonggi Province
한백상,문수경,박신희,유경신,김은비 대한미생물학회 2022 Journal of Bacteriology and Virology Vol.52 No.4
Measles is one of the highly contagious infectious diseases in which about 90% of the groups that have no immunity against measles are infected when exposed to the measles virus. Even after the Republic of Korea was certified as a ‘national measles eradication’ by the WHO in March 2014, two import-related outbreaks of measles occurred in 2014 and 2019. In this study, we analyzed epidemiological characteristics and genetic distribution of measles cases. Ninety-eight nasopharyngeal or oropharyngeal swab samples that confirmed cases of measles in Gyeonggi province in 2019 were conducted genotyping assay and sequence analysis. Fifty cases were classified as group outbreak-related cases and Forty-eight cases were classified as sporadic cases. As a result of genetic analysis, these cases were classified into D8, B3, and A genotypes, and except for the A type which was known as the vaccine type, D8 and B3 types were identified as import-related measles viruses. As with these results, for the complete eradication of measles, not only a high-quality surveillance system but also molecular epidemiological analysis data must be supported. The results of this study are expected to be used as important data for establishing a quarantine system through the accumulation of measles virus genotyping data and case reports data in the future when measles occurs in South Korea.
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이시형,한백상,최종선,신정임 대한백신학회 2017 Clinical and Experimental Vaccine Research Vol.6 No.2
Purpose: The goal of this study was to investigate the utility of DNA vaccines encoding Ebola virus glycoprotein (GP) as a vaccine type for the production of GP-specific hybridomas and antibodies. Materials and Methods: DNA vaccines were constructed to express Ebola virus GP. Mice were injected with GP DNA vaccines and their splenocytes were used for hybridoma production. Enzyme-linked immunosorbent assays (ELISAs), limiting dilution subcloning, antibody purification methods, and Western blot assays were used to select GP-specific hybridomas and purify monoclonal antibodies (MAbs) from the hybridoma cells. Results: Twelve hybridomas, the cell supernatants of which displayed GP-binding activity, were selected by ELISA. When purified MAbs from 12 hybridomas were tested for their reactivity to GP, 11 MAbs, except for 1 MAb (from the A6-9 hybridoma) displaying an IgG2a type, were identified as IgM isotypes. Those 11 MAbs failed to recognize GP. However, the MAb from A6-9 recognized the mucin-like region of GP and remained reactive to the antigen at the lowest tested concentration (1.95 ng/mL). This result suggests that IgM-secreting hybridomas are predominantly generated by DNA vaccination. However, boosting with GP resulted in greater production of IgG-secreting hybridomas than GP DNA vaccination alone. Conclusion: DNA vaccination may preferentially generate IgM-secreting hybridomas, but boosting with the protein antigen can reverse this propensity. Thus, this protein boosting approach may have implications for the production of IgG-specific hybridomas in the context of the DNA vaccination platform. In addition, the purified monoclonal IgG antibodies may be useful as therapeutic antibodies for controlling Ebola virus infection.
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경기도 내 환자에서 분리된 뎅기 바이러스의 분자생물학적 연구
이현경,문수경,이소현,조한길,한백상,이한나,문수진,김은비,김영숙,오조교 한국미생물학회 2021 미생물학회지 Vol.57 No.1
Dengue fever is an acute febrile illness infected by dengue virus, mediated by mosquitoes. To investigate the detection pattern, serotype and genotype distribution of dengue virus in Gyeonggi-do during 2016~2019, serums isolated from suspected patients who had oversea trip were collected and analyzed with RT-PCR. Dengue virus was detected in 111 cases (17.4%) from 638 samples. About 98.9% of dengue virus infections originated from endemic countries in Asia. Serotype was detected in order as DENV-2 (38.7%), DENV-1 (36.1%), DENV-3 (20.7%), and DENV-4 (4.5%). Genotype distribution of isolated dengue virus was analyzed by sequencing of the Envelope (E) region. Genotype analysis showed that dengue virus had a local specific genotype depending on visit-country, but no genetic variation occurred in the same serotype. This study suggests that genotype analysis of dengue virus be a useful tool for epidemiological investigation of dengue fever patients. 뎅기열은 대표적인 모기 매개 감염병이며, 뎅기 바이러스 에 의해 발생하는 급성 발열성 감염병이다. 본 연구에서는 2016년부터 2019년까지 해외 여행 후, 발열 등 뎅기열 의심 증 상이 나타나 경기도 내 병의원을 방문한 환자에서 분리한 뎅 기 바이러스의 검출 양상 및 혈청형과 유전형 분포 양상을 조 사하였다. 총 638건의 검체를 분석한 결과 111건(17.4%)에서 뎅기 바이러스를 검출하였다. 뎅기열 환자들의 약 98.9%가 뎅기열이 많이 발생하는 아시아를 방문하였다. 혈청형 분석 결과 DENV-2 (38.7%), DENV-1 (36.1%), DENV-3 (20.7%), DENV-4 (4.5%) 순으로 분리되었다. Envelope (E) 부위 유전 자의 염기 서열 분석을 통한 유전형 분석 결과 뎅기 바이러스 는 지역 특이적 유전형이 존재하였으나, 동일 혈청형 내 분리 주들 사이에 유전적 변이는 거의 없었다. 유전형 분석법은 뎅 기열 환자 발생 시 유입 경로 추적에 매우 유용하게 사용될 것 으로 생각된다.
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