生物晶片學原與應用介紹怡杏 Ph.D. 2009, Aug, 6th威健股份有限公司威健股份有限公司 Welgene Biotech. 台市南港區園區街3號10之五 Tel: 02-66160001內 容 微體晶片 微陣技術 生物醫學之應用微陣技術與生物晶片微陣技術生物晶片微陣技術生物晶片相同點:將技術平台微小化微小化優點檢體減小 試劑減小 單一試驗之可檢測項目增多現有檢驗技術平台微小化 之可性 蛋白質 測酵素活性測酵素活性 電泳分偵測電泳分偵測 免疫之抗原抗體偵測免疫之抗原抗體偵測 核酸 (DNA, RNA) DNA雜配 電泳分偵測電泳分偵測 PCR毛細管電泳晶片毛細管電泳晶片毛細管電泳晶片毛細管電泳晶片PCR晶片晶片蛋白質微陣晶片蛋白質微陣晶片DNA微陣晶片微陣晶片生化反應晶片生化反應晶片AGCTGSouthern blottracer: Isotope Enzyme colorimetryHistory: Edward Southern developed technique of transferring DNA from a blot to a nitrocellulose sheet (JMB 98:503-517 1975)CACGCGtracerDNA微陣晶片-組成1. 載體載體2. 表面塗劑表面塗劑3. DNA 陣陣4. 偵測方法偵測方法: DNA-DNA 雜配雜配5. 訊號偵測訊號偵測Components of Proten microarrayVectorVectorSurface chemistrySurface chemistryDetection mode, eg. Ag-AbDetection mode, eg. Ag-AbDetection MethodDetection MethodScannerScannerLab-on-a-chip TechnologyCourtesy of Caliper Technologies Corp., Mountain View, CA, USA毛細管電泳晶片電泳毛細管檢體注入孔檢體至毛細管通道電泳毛細管檢體注入孔檢體至毛細管通道From : www.agilent.comHow Chips are MadeCover plate with holesGlass or QuartzMaskLight2. Develop3. Etch4. Bond1. ExposeEtched Channel PlateBonded Chip 5. Dice chips6. Mount onto caddyCourtesy of Caliper Technologies Corp., Mountain View, CA, USAPhotoresistScript Step I of IV On-chip gel electrophoresisLoading (ladder) 1.5 minLadder well current drivenWaste well voltage drivenSeparation ChannelPoint of detectionDetermination of PCR ProductDetermination of PCR ProductDNA ELISA Dai, Hongyue; van de Vijver, Marc J.*; He, Yudong D.; Hart, Augustinus A. M.*; Mao, Mao; Peterse, Hans L.*; van der Kooy, Karin*; Marton, Matthew J.; Witteveen, Anke T.*; Schreiber, George J.; Kerkhoven, Ron M.*; Roberts, Chris; Linsley, Peter S.; Bernards, Ren*; Friend, Stephen H. * Divisions of Diagnostic Oncology, Radiotherapy and Molecular Carcinogenesis and Center for Biomedical Genetics, The Netherlands Cancer Institute, 121 Plesmanlaan, 1066 CX Amsterdam, The Netherlands Rosetta Inpharmatics, 12040 115th Avenue NE, Kirkland, Washington 98034, USA癌症分型: 70個基因癌症 可治癒組癌症 可治癒組癌症 治癒失敗或發組癌症 治癒失敗或發組Agendias mammal printSignature isolated from Agilent 25K arrays (Merck), successfully transferred to 8-packFirst FDA approved IVDMIACharts path for other customers looking to develop diagnostic/ prognostic testsOther applications Array CGH Detection of mutations/single nucleotide polymorphismgeneral aspects of (molecular) cytogeneticsLimitation of traditional cytogenetic stain Need to culture cell lines Limited resolution (10M) Laborious Technicians experience is importantComparative Genomic HybridizationNormal Genomic DNACancer Genomic DNAConcept of Array-based CGHPeter Lichter, Berlin, GermanConcept of Array-based CGHMapping the 15q duplication基因分型 GenotypingP53之基因分型基因分型DNA polymeraseprimerdNTPDenature and hybridize to the chip基因分型 GenotypingC G T AProbe sequenceThe EndThe EndAny Question?Any Question?