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분자 비컨을 이용한 살아 있는 세포에서 단일클론항체 경쇄와 중쇄 mRNA 검출에 의한 세포주 선별방법
정승아(Seunga Jeong),이원종(Won Jong Rhee) 한국생물공학회 2016 KSBB Journal Vol.31 No.1
Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.
선미정(Mi-Jeong Seon),정승애(Seungae Jung) 한국복지경영학회 2022 복지경영학연구 Vol.11 No.-
사회적경제교육의 효과는 사회적 가치와 경제적 가치가 내면화 및 태도화되어 직접적 실행되어야 효과성이 있으므로 초기 아동기(초등학교 1-2학년)부터 사회적경제교육이 누적될 필요가 있다. 그러나 현재 사회적경제교육은 성인 중심, 단기성, 선택적으로 진행되고 있고, 초기 아동 대상 커리큘럼은 매우 제한적이다. 이에 본연구는 초기 아동발달 특성을 고려한 사회적경제교육 프로그램을 제안하기 위하여 필수 구성요소를 파악하고, 이들을 위한 사회적 경제교육 프로그램을 제시하는 것에 목적이 있다. 구체적인 연구방법은 사회적경제교육 학계 및 현장 전문가 8인을 대상으로 FGI(Focus Group Interview)를 실시하고 반복적 비교분석방법(Constant Comparison Method)을 통해 사회적경제 교육의 필요성, 운영구성, 내용구성, 평가, 기타 5개 영역 11개 범주를 도출하였고, 본 결과를 바탕으로 프로그램을 제안하였다. 본 연구는 초기 아동의 발달 특성을 고려하여 사회적경제교육 커리큘럼 제안한 것에 의의가 있다. 향후 본 연구 내용을 기반으로 하여 활용가능한 콘텐츠 개발과 교육 실행을 통한 사전 사후 평가가 실행되기를 기대한다. 더불어 지속적인 사회적경제교육 프로그램 발전을 위하여 학년별 모듈화와 인력 전문화, 교과과정 내 편재를 통해 초기 아동기부터 사회적경제교육 경험이 반복되기를 희망한다. Since the effect of social economy education is effective only when social and economic values are internalized and directly implemented, the education needs to be accumulated from early childhood (1st and 2nd grades of elementary school). However, social economy education is currently being conducted adult-centered, short-term, and selectively, so the curriculum for early childhood is very limited. Therefore, the research aims to identify essential components to propose social economy education programs that consider the characteristics of early childhood development and suggest social economy education programs for them. As a result of a focus group interview(FGI) with eight experts, not only academic but also field professionals, and implementing the Constant Comparison Method, 11 components―a necessity, operation composition, content composition, evaluation, and five other areas―were derived, and programs proposed based on this result. This study is significant in that it proposes a social economic education curriculum for early children. And it is expected that content development and program pre-post evaluation based on this will be carried out in the future. In addition, for the continuous development of social economy education programs, it is hoped that social economy education experiences will be repeated from early childhood through modularization by grade, human resource development, and distribution by school curriculum.
Simultaneous and multiplexed detection of exosome microRNAs using molecular beacons
Lee, Ji Hye,Kim, Jeong Ah,Jeong, Seunga,Rhee, Won Jong Elsevier 2016 Biosensors & bioelectronics Vol.86 No.-
<P><B>Abstract</B></P> <P>Simultaneous and multiplexed detection of microRNAs (miRNAs) in a whole exosome is developed, which can be utilized as a PCR-free efficient diagnosis method for various diseases. Exosomes are small extracellular vesicles that contain biomarker miRNAs from parental cells. Because they circulate throughout bodily fluids, exosomal biomarkers offer great advantages for diagnosis in many aspects. In general, PCR-based methods can be used for exosomal miRNA detection but they are laborious, expensive, and time-consuming, which make them unsuitable for high-throughput diagnosis of diseases. Previously, we reported that single miRNA in the exosomes can be detected specifically using an oligonucleotide probe or molecular beacon. Herein, we demonstrate for the first time that multiple miRNAs can be detected simultaneously in exosomes using miRNA-targeting molecular beacons. Exosomes from a breast cancer cell line, MCF-7, were used for the production of exosomes because MCF-7 has a high level of miR-21, miR-375, and miR-27a as target miRNAs. Molecular beacons successfully hybridized with multiple miRNAs in the cancer cell-derived exosomes even in the presence of high human serum concentration. In addition, it is noteworthy that the choice of fluorophores for multiplexing biomarkers in an exosome is crucial because of its small size. The proposed method described in this article is beneficial to high-throughput analysis for disease diagnosis, prognosis, and response to treatment because it is a time-, labor-, and cost-saving technique.</P> <P><B>Highlights</B></P> <P> <UL> <LI> In situ multiplexed detection of microRNAs in exosome is developed. </LI> <LI> Exosome microRNAs can be detected by molecular beacons in the presence of human serum. </LI> <LI> The selection of fluorescent dyes for multiplexed detection in exosomes is crucial. </LI> </UL> </P>