11:10-12:00, Rm 103

Launching new technologies from the industrial sectorZ-01Unravelling biomolecular mechanisms in the lipid environmentU. Devi, R.C. Hutton¹, M. J. Swann¹, G Ronan¹& I Barsukov²¹Farfield Group, Voyager, West Wing Level 7, Chicago Avenue, Manchester, M90 3DQ, UnitedKingdom, ²School of Biological Sciences, The University of Liverpool, Liverpool L69 3BX, UKDual Polarisation Interferometry (DPI) serves as a platform with unique capabilities formeasuring conformational changes in proteins and to unravel biomolecular mechanismsin the lipid environment. DPI is able to measure the bilayer birefringence in real timewhich is a direct measurement of the degree of alignment and order within lipid bilayermembrane.Z-04A novel diagnostic method for human immunodeficiency virus (HIV) 1using nanoporous sol-gel based protein microarray and applicationsMinjoung Jo, Seram Lee, Byoung Don HanPCL, Inc., 83421 Research Complex 2, Sungkyunkwan University, Suwon, KoreaUsing sol-gel technology, various molecules can be encapsulated not covalently bondedinside of dual-nanoporous structure. Features of sol-gel chip like encapsulation and 3Dstructurecan make it more sensitive (femto-molar level) than other protein chip. So solgelchip has high specificity and low background signal. Sol-gel can be applied asuniversal chip material for oligonucleotides, proteins and chemicals, etc. Sol-gel chip canbe used to generate an array for interaction assay. A novel antigen microarray in 96-wellplate format was designed for HIV1 antibody detection using sol-gel. Serum samples of145 patients were tested with this novel system which had been previously tested forHIV1 by ELISA and western blot (122 negative and 23 positive). Our method yieldedsignificantly fewer false-positive results than ELISA, and similar false-positive and falsenegativeresults in comparison to western blot. This new, inexpensive method willimprove the specificity and sensitivity for massive sample diagnosis. Additionally, this newtechnology can also be applied for multiple disease diagnostics.This work was supportedby the International Collaborative Research and Development Program (# GT-2009-ME-DI-0076).Z-02Drug target identification of a natural anticancer agent plumbagin usingGPScreen ; an innovative technology for drug target discovery usingdrug-induced haploinsufficiency in S. pombe genome-wideheterozygous deletion mutant libraryDong-Myung Kim, Ju-Hee Lee, Ji-Hyun Yeon, Pyoung-Oh Yoon, Whijae Roh andHan-Oh ParkS. pombe Research Team, Gene-to-Drug Division, Bioneer Corporation, Daejeon 306-220, KoreaDIH(Drug-induced haploinsufficiency) in yeast is a valuable tool for identification of drugtarget and its mode-of-action. In this study, we have tried to identify its detailed moleculartarget of a natural anticancer agent plumbagin using an innovative DIH-based drug targetscreening system GPScreen . Plumbagin induced a potent anti-proliferative activity andthe cell elongation and septum formation. Interestingly, DIH was found in its3-deletedmutant, suggesting that its3 might be a molecular target of the agent. In mode of actions,plumbagin induced a significant increase of ROS and pretreatment of an ROS scavengerNAC protected the growth inhibition by plumbagin, suggesting that ROS might be acritical mediator for the cytotoxicity. Plumbagin also induced DIH in TOR2-deleted mutant.Plumbagin in decreased PI-5 kinase-1B mRNA in human cancer cells, which is a humanortholog of its3. These results show that PI-5 kinase-1B is another molecular target ofplumbagin and GPScreen ‚ drug target screening system is a valuable tool foridentification of both the drug target and mode-of-action of drug candidates.Z-05Miniaturizing RNAi assays: acoustic droplet ejection enables efficientand reproducible transfer of siRNA molecules at low concentrationsCarl Jarman¹, Anne Marie Byrne², Anthony Mitchell Davies²and Siobhan Pickett¹¹Labcyte Inc., Sunnyvale, CA, U.S.A., ²Institute of Molecular Medicine, Trinity College, Dublin,IrelandTo facilitate large-scale, high-throughput functional genomics studies using RNAi, wehave developed acoustic droplet ejection (ADE) technology to meet the current demandsof a successful siRNA library based screen. High throughput applications require reliableand reproducible transfections to be performed in high density well plate formats. Thesuccess of an siRNA experiment relies on the effective delivery of the siRNA molecule.Using ADE technology, a unique tipless and touchless liquid transfer technology used inthe Echo liquid handlers (Labcyte Inc.), we have successfully transfected esophagealcancer cells using a liposome-mediated transfection method. We have also demonstratedthe use of ADE technology for reverse transfection. Here, ADE was used to depositsiRNAs in a transfection matrix onto glass slides, then overlaid with a monolayer ofadherent cells and the slides incubated to allow reverse transfection. The effects of genesilencing were then assessed by digital image analysis at a single cell level.Z-03Quantitative long-term live-cell imaging of cell migration andangiogenesis using IncuCyteJe-Ik KimEmaGene Science, Daewoon Plaza B/D Rm 404-1, 818-5, Yangcheong-ri, Ochang-eup,Cheongwon-gun, Chungbuk 363-883, KoreaCell migration is a multistep process that is a fundamental component of many importantbiological and pathological processes such as embryonic development, angiogenesis,immune cell trafficking, and tumor metastasis. The Essen BioScience Team designedand built a highly reproducible and quantitative 96-well scratch wound cell migrationassay. The assay includes a unique 96-pin WoundMaker equipped with PTFE tips. Cellmigration is quantified in IncuCyte, which is a compact long-term, live-cell imaginginstrument that resides in a standard CO2 incubator. The 96-well cell migration assay runsautomatically and acquires images at regular intervals without removing the cells from theincubator environment. In all cases, kinetic data are analyzed using custom algorithmswhich provide a sensitive and temporal description of the effect being studied. TheEssen BioScience also developed a new in vitro angiogenesis assay that consists of aco-culture of human fibroblasts and human endothelial cells. GFP-labeling of HUVECsprovides the means to visualize and quantify all stages of endothelial cell morphogenesisin a time-dependent manner. IncuCyte automatically collects data for the duration of theassay (14-21 days) and custom algorithms quantify tube length, area, and branching.Z-06Sensing pH in vivo using fluorescence lifetimeGuobin Ma, Dao Chao Huang, Niculae Mincu, Muriel Jean-Jacques, and MarioKhayatART Advanced Research Technologies, Inc., Montreal, CanadaDetection of alterations of physiological parameters(e.g. pH and temperature) resultingfrom malignant transformation of tissue can be a powerful diagnostic tool for earliercancer detection and prognosis. Fluorescent lifetime imaging(FLIM) of targetedfluorescent labeled antibodies can be sensitive to such variations and provide functionalimages of the regions of interest. We present a method of in vivo animal imaging to sensethe local pH microenvironment by FLIM using a time-resolved fluorescence imagingplatform. The imaging agent is a pH sensitive fluorescent dye,SNARF-1. SNARF-1 mixedwith matrigel at different biological relevant pH levels were implanted. Then, the micewere imaged using a time-resolved imaging system(OptixMX). The intensities from pHsamples are slightly different while the signal from matrigel-only is comparable to thetissue autofluorescence. However, the fluorescence lifetimes from the 3 pH variedsamples are clearly different. These results demonstrate that FLIM can be used in vivo todifferentiate the local pH microenvironment that could find many applications in cancerresearch and other fields. Further studies using pathological models with varied pHmicroenvironment caused by physiological effect are under the way.350 Korean Society for Biochemistry and Molecular Biology