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항체칩을 이용하여 지유에 의한 HUVEC 세포의 세포주기 단백질 발현 프로파일 연구
김응윤,방지영,강인철 호서대학교 기초과학연구소 2009 기초과학연구 논문집 Vol.17 No.1
지유의 신생혈관 형성 억제 효능의 기전을 규명하기 위하여 단백질 칩 기반 항체 microarray를 이용하여 HUVEC 세포의 증식 관련 세포 주기 신호전달 단백질들의 발현 패턴을 연구하였다. 세포 주기 단백질에 대한 항체 45가지를 고정 시킨 단백질 칩 을 이용하여 지유를 처리한 HUVEC 세포 파쇄액에서의 단백질 발현 정도를 지유를 처리하지 않은 세포의 파쇄액과 비교하였을 때 차이가 나는 것을 발견하였다. 지유(S)를 처리한 샘플에서는 cyclin E와,BRCA1 이 감소하고, STAT3가 증가한 반면에 지유(M)을 처리한 샘플에서는 Rafl,MEK-1, MEK kinase-1 이 증가하고,cdc25A 감소하였다. 항체칩을 이용한 프로테오믹스 연구는 지유 추출물에 의한 HUVEC 세포에서 의 증식억제 및 기능성을 갖는 물질을 찾아내고 기전 연구를 하는데 중요한 실마리를 제공해 줄 수 있다.
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김응윤,이미영,황승용,강인철 한국바이오칩학회 2010 BioChip Journal Vol.4 No.3
We have developed an antibody microarray chip for biomarker analysis in rat livers exposed to five different toxic pollutants: volatile organic compounds (VOCs), dichloromethane, ethylbenzene, and trichloroethylene; and polycyclic aromatic hydrocarbons (PAHs), benzoanthracene and phenanthrene. The antibody chip consisted of antibodies against different biomarkers: alpha-2 μ globulin, glutathione S transferase (GST)-α, Hemoglobin β, and Phenylalanine hydroxylase. VOC- and PAH-treated rat livers showed differential expression patterns of the biomarker proteins. These results demonstrate that a protein chip based antibody array is useful for the analysis of biomarkers induced by environmental toxic pollutants and can be a powerful tool to study the mechanisms of pollutants in biological samples.
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Detection Technology for Antibody-Antigen Interaction on ProteoChip using Quantum Dot
김응윤,장수익,강인철 한국바이오칩학회 2007 BioChip Journal Vol.1 No.2
Quantum Dot (CdSe-ZnS nanocrystals, QD or Qdot) nanometer-size particles have a number of potential applications in a variety of fields. Qdots have already been put into use as an alternative to organic fluorescent dyes and fluorescent proteins. In this study, the applications of Qdots were explored in protein microarray techniques, using Angiogenin (ANG) and Anti-ANG antibody on a ProteoChip. We compared the signal-to-noise ratios between the Cy5 labeling and Qdot labeling methods. The Qdot method evidenced a higher signal-to-noise ratio than was observed in the Cy5 labeling method. This indicates that Qdots can be used as an enabling detection material for the specific quantification of proteins and profiling in protein chips.
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Protein Chip-based Dipeptidyl Peptidase IV Assay System
김응윤,강인철 한국바이오칩학회 2009 BioChip Journal Vol.3 No.3
Dipeptidyl peptidase IV (DPP-IV) is known to be a serine amino-peptidase that cleaves N-terminal X-Ala or X-Pro from target polypeptides. The enzyme plays a pivotal role in fatal diseases such as diabetes mellitus, obesity, tumor growths, and HIV infection. Inhibition of DPP-IV is currently being explored as a novel therapy for type II diabetes. Thus, we have developed a protein chip-based DPP-IV assay system for the screening of enzyme inhibitors. A biotin-labeled Gly-Pro sequence as a substrate was designed for immobilization on a ProteoChipTM. DPP-IV was incubated with the substrate-immobilized ProteoChip. Also, Cy5-labeled streptavidin was applied to the chip, and the fluorescence intensities on the spots were detected using a laser scanner. DPP-IV activity on the chip was determined using data obtained from the fluorescence intensities. Galvus (vildagliptin) and virtually screened leads were tested using the DPP-IV assay chip for identification as novel diabetes inhibitors.
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방지영,김응윤,심태경,유화승,이연월,김용수,조종관,최용진,정현자,강인철 한국바이오칩학회 2010 BioChip Journal Vol.4 No.4
The inhibitory effects of the water extract of HangAmDan-B(WEHAD-B), which is a crude extract of eight Korean medicinal animals and plants on bFGF-induced neovascularization were investigated. WEHAD-B significantly prevented bFGF-induced HUVE cell proliferation, adhesion, migration, and capillary-like tubular network formation. Half-maximal inhibition of proliferation on the endothelial cells by WEHAD-B was observed at a concentration of approximately 250 μg/mL. Our antibody microarray-based ProteoChip data showed that WEHAD-B increased the expression of STAT1 and Rb2, which are involved in cell growth, apoptosis, and controlling the cell cycle in bFGF-induced HUVECs. These results indicate that the inhibition of bFGF-induced angiogenesis by WEHAD-B may be due to upregulation of cell signaling proteins, STAT1 and Rb2. The blood vessel formation in a chick chorioallantoic membrane (CAM) treated with WEHAD-B was markedly reduced in length compared with a PBS-treated control group. Taken together, these data suggest that antibody-arrayed ProteoChip technology may be an useful tool for determining molecular mechanism of natural products in biological samples.
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