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인플루엔자 바이러스 검출을 위한 종이 기반 neuraminidase 효소 활성 평가 센서 개발
황철환 ( Cheol-hwan Hwang ),정성근 ( Seong-geun Jeong ),박한규 ( Han-kyu Park ),이창수 ( Chang-soo Lee ),김윤곤 ( Yun-gon Kim ) 한국화학공학회 2016 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.54 No.3
본 연구에서는 인플루엔자 바이러스 표면에 존재하는 neuraminidase 효소의 활성을 평가 할 수있는 종이칩 기반의 분석 시스템을 구축하였다. 종이칩의 장점을 살려 분석 전문가와 장비 없이 현장 진단(Point-of-care)이 가능하도록X-Neu5Ac 기질을 이용한 비색분석법을 통해 시료 내 neuraminidase 효소의 존재를 정량적으로 확인 할 수 있도록 설계 및 제작하였다. Neuraminidase 효소의 활성을 확인할 수 있는 종이칩 센서(Paper-based neuraminidase assay sensor;PNAS) 성능 실험 결과 neuraminidase를 0.004 U/mL 농도부터 검출 가능하였으며, 인간 혈청에 각기 다른 농도로 존재하는 neuraminidase 효소의 양을 활성 평가를 통해 정량적으로 검출할 수 있음을 입증하였다(R2 > 0.99). 또한, 보관기간에 따른 종이칩의 안정성 평가 결과 빛이 차단 된 4 oC 환경에서 보관 시 70일까지 초기 성능이 안정하게 유지됨을 확인하였다. 마지막으로, PNAS 상에서 효소 반응의 신뢰성 평가를 위해 미카엘리스-멘텐 동역학 (Michaelis-Menten kinetics)을 적용하여 X-Neu5Ac 기질에 대한 neuraminidase의 동역학 분석 결과 Km 값은 8.327×10.3 M으로 확인되었으며, 이 값은 용액상에서의 효소 반응 속도 계산으로부터 산출된 값과(Km=0.89×10.3 M) 근사한 수치임을 확인하였다. 본 연구로부터 개발된 종이칩 기반의 neuraminidase 효소 활성 평가 시스템은 인플루엔자 바이러스의 신속하고 안전한 검출에 다양하게 응용 될 수 있을 것으로 생각된다. In this study, we described a paper-based neuraminidase assay sensor (PNAS) which can be applied to detect the infection by influenza viruses. The PNAS was designed and manufactured to quantitatively identify the levels of neuraminidase in the sample, which is based on colorimetric analysis using the X-Neu5Ac substrate. The limit of detection of the PNAS was determined as 0.004 U/mL of neuraminidase. According to the amount of neuraminidase in human serum, the PNAS could monitor the enzyme activity with a good linearity (R2 > 0.99). In addition, the initial performance of the PNAS has been maintained up to 70 days in the 4 ℃. Finally, we demonstrated whether the Michaelis- Menten kinetics is applied to the PNAS, which can show the reliability of the enzyme reactions. The kinetic studies indicated that the PNAS provides the good condition for enzyme reactions (Km=8.327×10.3 M), but they were performed on paper chip nonetheless. The paper-based neuraminidase assay sensor may be useful in a wide range of rapid and safe detection of influenza virus.
황철환 ( Cheol Hwan Hwang ) 한국사진학회 2010 AURA Vol.0 No.23
Text and image have been in competitive relation to each other as a way to express human emotion, showing their actual existences in history. After Renaissance, Text having taken more steps from simply explaining Image has been developed as a variety of means to describe over the collage of the text cubism and futurism and the language experiment like dadaism and surrealism as well as the concept art. Text which is a sentence of "The images flooded the era of the digital" comes into contact as much. Now the text which was the old way of transferring communication into images of the development of a variety and emerging visual media with communicating a clear and strong message. However, the text is not disappearing. Text becomes image, and the combination of text and image is found easier in everyday life. Moreover, the emotion in the era of the digital is the text, which is the image. The text plays an important role in the relationship to languages which are exchanging between the human characters. Initially, the symbol has been created from text in the changes of history. The birth of today`s many kind of texts has an important position for the preservation of our culture. In particular, the role of modern text is more important in that it refers to the communication for us as well as fun and beauty of aesthetic art in our live. Therefore, the basic starting point for this paper research is on the character of the text and the meaning of the text use in the painting representations of modern arts. In other words, this study researches the expression of visual art with text, as well as meaning of the text in the modern arts. In addition to that, this study tries to add from geometric of text the basic geometric of the object of Paul Cezanne who analyzed about the object with "sphere, cone, cylinder". Text of the character in the image gives a sign of the communication, and the autonomous and communication of sing. Image representation in the humans five senses has been in relation to the visual, but it can be concluded that another viewpoint of sing and arts will be needed. Thanks.
질량분석 시스템을 이용한 극지 토양 유래 신규 미생물의 지질 A 화학적 구조 분석
황철환(Cheol-hwan Hwang),박한규(Han-Gyu Park),김윤곤(Yun-Gon Kim) 한국생물공학회 2016 KSBB Journal Vol.31 No.4
For decades, the microorganisms in arctic soils have been newly discovered according to the climate change and global warming. In this study, the chemical structure of a lipid A molecule from Pseudomonas sp. strain PAMC 28615 which was newly discovered from arctic soils was characterized by mass spectrometric approaches such as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and MALDI multi-stage tandem mass spectrometry (MS). First, lipopolysaccharide (LPS) from Pseudomonas sp. strain PAMC 28615 was extracted and subsequently hydrolyzed to obtain the lipid A. The parent ion peak at m/z 1632 was determined by MALDI-TOF MS, which also can validate our lipid A purification method. For detailed structural determination, we performed the multiple-stage tandem mass analysis (MS<SUP>4</SUP>) of the parent ion, and subsequently the abundant fragment ions in each MS stage are tested. The fragment ions in each MS stage were produced from the loss of phosphate groups and fatty acyl groups, which could be used to confirm the composition or the position of the lipid A components. Consequently, the mass spectrometry-based lipid A profiling method could provide the detail chemical structure of lipid A from the Pseudomonas sp. strain PAMC 28615 as an arctic bacterium from the frozen arctic soil.