生物芯片技术、产业现况与趋势

1、Sequencing costs have dropped 100-fold over the last 10 years, corresponding to a roughly twofold decrease every 18 months. This rate is similar to Moores law concerning improvements in semiconductor manufacture. In both sequencing and semiconductors, such improvement does not happen automatically,

2、but requires aggressive technological innovation fuelled by major investment.Nature 409, 860 - 921 (15 February 2001)1 Proteome sizes and number of genes shared with each of the human protein sets(with a Blast cutoff of 1010 ) Organisms Number of proteins Number matching Ensembl proteome Number matc

3、hing Celera proteome Human 31,780 26,544 Bacteria/Archaea 85,824 4,388 3,915 Yeast 9,030 7,508 7,103 nematode worm 19,400 13,770 12,660 fruit fly 14,080 15,324 14,302 mustard weed 25,470 9,151 9,081 Parasites 11,606 5,146 4,756 SCIENCE VOL 292 8 JUNE 2001, Microbial Genes in the Human Genome: Latera

4、l Transfer or Gene Loss?, p19032基因序列的特性 是一种信息。 所有生物具有共通的语法。 具有跨平台执行的能力。 具有自我演化、自我繁殖、自我扩充的能力。3 生物芯片是微小化、整合化 之风潮，表现在生技领域之结果。 运用分子生物学、分析化学、生化反应等原理； 以硅芯片、玻璃或高分子为基材； 配合微机电、自动化或其他精密加工技术，所制作之高科技元件与系统； 用于进行快速之生化分析检验。4生物芯片范畴mTASmicro Total Analysis SystemLab-on-a-ChipcDNA/DNA microarrayGeneChipMicrofluidic C

5、hipBio MEMS(Bio NEMS)ProteinChipHigh Throughput ScreeningChip5生物芯片(Biochips)6生物芯片应用范围基础研究、新药开发医疗诊断、环境检测食品安全、军事预警法医鉴定、制程控制7改变相关产业的竞争力改变相关产业的竞争力改变生活方式改变生活方式生物芯片的重要性89分子检测与治疗(Molecular Diagnostics and Therapy)个人化医疗(Personalized Medicine)预防医学(Preventive Healthcare)检测工具将由配角地位，成为产业主角之一。生物芯片将是新世代医疗之基础！10A

6、AA AA AC C C CT TC CG GT TT TT TC CG GG GG GA AA AA AA AC C C CT TC CG GT TT TT TC CG GG GG GA A11应用领域 基础研究、新药开发与筛选、生技研发 基因表现程度监测 基因型鉴定 毒理、药理、治疗标的与机制 医疗诊断 个人医药、预防医学 食品安全、环境检测 法医鉴定 农渔畜牧 居家生活12基因表现程度检测基因表现程度检测Gene Expression ProfilingGene Expression ProfilingDNADNARNARNAProteinProteinBiochemicalBioche

7、micalReactionsReactionsDNADNAReplicationReplication复制复制TranscriptionTranscription转录转录TranslationTranslation转译转译ReverseReverseTranscriptionTranscription逆转录逆转录cDNAcDNA13个人化医疗未来未来血压血压胆固醇胆固醇运动习惯运动习惯+ +基因型（基因型（SNP profileSNP profile）风险评估风险评估药物反应药物反应+ +基因表现（基因表现（Expression profileExpression profile）生理状态生理

8、状态目前目前 血压血压胆固醇胆固醇运动习惯运动习惯+ +家族病史家族病史基因型基因型疾病疾病14AffymetrixGlaxoWellcomeAffymaxGlaxoSmithKlineAffymetrix1988成立，新药开发1995-2001，并购隶属于Glaxo Wellcome2001，再度成为独立公司核心技术Combinatorial solid phase chemistry1993成立，研发生物芯片Glaxo Wellcome占有30%以上股权，于1996 IPO核心技术：Photolithography15ATCAATTCCACTTTCATCGADNA chipA AA AA

9、AA AA AA AA AA AA AA AA AA AC CA AA AA AC C1 13 32 24 45 56 6A AA AA AC C7 7A AA AA AC C8 8A AA AA AA AC C9 9A AAffymetrix16Affymax TechnologyAffymax TechnologyUSP 5,571,639, November 5, 1996 (May 24, 1994)USP 5,571,639, November 5, 1996 (May 24, 1994)Computer-aided engineering system for design of

10、sequence arrays Computer-aided engineering system for design of sequence arrays and lithographic masks and lithographic masks AffymetrixAffymetrix United States Patent 5,744,305, April 28, 1998 (June 6, United States Patent 5,744,305, April 28, 1998 (June 6, 1995)1995)Arrays of materials attached to

11、 a substrateArrays of materials attached to a substrateAffymetrix重要专利分析17United States Patent 5,744,305United States Patent 5,744,305An array of oligonucleotides, the array comprising: An array of oligonucleotides, the array comprising: a planar, non-porous solid support having at least a a planar,

12、non-porous solid support having at least a first surface; and a plurality of different first surface; and a plurality of different oligonucleotides attached to the first surface of the oligonucleotides attached to the first surface of the solid support at a density exceeding 400 different solid supp

13、ort at a density exceeding 400 different oligonucleotides/cm2, wherein each of the different oligonucleotides/cm2, wherein each of the different oligonucleotides is attached to the surface of the oligonucleotides is attached to the surface of the solid support in a different predefined region, has a

14、 solid support in a different predefined region, has a different determinable sequence, and is at least 4 different determinable sequence, and is at least 4 nucleotides in length. nucleotides in length. Affymetrix重要专利分析18Affymetrix R&D ExpansesYear 2000 1999 1998 1997 1996 Total revenues $200,83

15、0 $109,074 $52,413 $19,765 $11,972 R&D expenses $57,384 $43,524 $38,433 $28,590 $18,762 Net loss $(53,990) $(25,504) $(26,800) $(22,780) $(12,227) Net loss per share $(0.98) $(0.54) $(0.62) $(0.50) $(0.30) (in thousands, except per share amounts) 19Affymetrix, 1998-2000核心技术：光罩法核心技术：光罩法/Oligonucl

16、eotide 4-20 bases/Oligonucleotide 4-20 bases重点产品：感染性疾病、新药开发重点产品：感染性疾病、新药开发产业策略：封闭系统产业策略：封闭系统重要客户：重要客户：EasyAccessEasyAccess专案专案Warner-Lambert CompanyWarner-Lambert CompanyAmerican Home Products/Genetics InstituteAmerican Home Products/Genetics InstituteEos Biotechnology, Inc.Eos Biotechnology, Inc.Ho

17、ffmann-La Roche Ltd.Hoffmann-La Roche Ltd.Hoechst Marion Roussel, Inc.Hoechst Marion Roussel, Inc.Howard Hughes Medical InstituteHoward Hughes Medical InstituteRhone-Poulenc RorerRhone-Poulenc Rorer20基因芯片专利诉讼High DensityArray, 1998/15,445,9345,744,3055,800,992Sequence byHybridization5,202,2315,525,4

18、645,695,940Chemical Synthesis2000/7Southern DNA ArrayAffymetrixAffymetrixOxford Gene Tech.PEHyseqIncyte/SynteniSequence byHybridization5,795,716High DensityArray5,744,305侵权抵触无效侵权无效侵权21基因芯片量产制程原位合成原位合成in-situin-situ synthesis synthesis微影光罩法微影光罩法PhotolithographyPhotolithography(Light-directed in-situ

19、synthesis)(Light-directed in-situ synthesis)四色印刷法四色印刷法4- 4-base ink-jet in-situ synthesisbase ink-jet in-situ synthesis預先合成預先合成ex-situex-situ synthesis synthesis噴墨法噴墨法Ink-jet printing Ink-jet printing 點陣法點陣法Blot-printing/SpottingBlot-printing/Spotting探针密度探针密度高高(50,000/cm2)(50,000/cm2)低低(6,000/cm2)(6

20、,000/cm2)探针长度探针长度短短(10-50 mers)(10-50 mers)长长(10-100 mers)(10-100 mers)or higheror higher22Affymetrix, 2001- 专利诉讼互有胜负。 Affymetrix-Oxford Gene technology，负 Affymetrix-Hyseq，和 Affymetrix-Incyte，未定 市场成长缓慢，且占有率蚕食鲸吞，节节下滑。 制程技术瓶颈无法突破，成本居高不下。 产品设计错误，造成客户损失。 封闭系统，客户群有限。 无法与客户分享信息，抱怨连连。23Affymetrix, 2000- 购入

21、Genetic MicroSystems (GMS)，提供客户自制芯片服务 努力改善客户关系。 转移公司运行方向 成立Perlegen Sciences建立SNP资料库 与Hyseq和解，并成立Callida Genomics, Inc专责芯片业务 Ayymetrix可能转型成为新药开发公司244-Base Ink-Jet in-situ Synthesis4-Base Ink-Jet in-situ Synthesis 富有弹性 适合小量试产 可以发展成实验室使用之产品 Rosetta Inphamatics + AgilentFlexJetTM DNA MicroarraysChange

22、array design at will. Synthesize different length oligonucleotides on the same array. Move from design to array in hours. Work on newly characterized organisms and have the array design change and grow with your knowledge of the genome. 254-base Inkjet in-situ SynthesisACGATCGATCGATCGATCGATCGAACATCG

23、ATCGATCGATCGATCGA26微流体生物芯片mTAS/Lab-on-a-ChipsBiochipBiochipSystemSystemInformationSpecimensReagents/SubstratesInformation平行检测：提供完整信息。迅速正确：节省时间，降低人为错误。降低成本：容易普及化。“sample-in-answer-our27mTAS1990Andreas Manz, NovartisMiniaturized Total Analysis Systems: A Novel Concept for Chemical Sensors, Sensors and

24、 Actuators B1, 244-248, 19901995International Symposium on Miniaturised Total Analysis Systems TAS 95 2001Sep. 2001, Lab on a ChipMiniaturisation for Chemistry and Biology, Published quarterly by the Royal Society of Chemistry.28Mixer/Separator/valve29Micro-PCRElectrophoresis30Separating via mobilit

25、y, t=0 1000 mm1000 mm50 mm50 mm31Separating via mobility, t=t1 100 cm100 cm5 cm5 cm32聚合脢連鎖反應 PCR( DNA Amplification: 105 X )60 oC70 oC90 oC Thermal cycle33Mixer/Separator/valve+-Electrophoresisthe movement of a charged particle relative to the liquid it it suspended in under the influence of an appl

26、ied electric field34Mixer/Separator/valve+-ElectroosmosisThe walls of the capillary cell carry a surface charge so the application of the electric field causes the liquid to flow along the cell walls. This is called electroosmosis -+-+-35Mixer/Separator/valveDielectrophoresisThe translational motion

27、 induced when particles are exposed to stationary electrical fields. +-+-36产业发展走向孕育孕育萌芽萌芽研发研发整合整合成长成长稳定稳定衰退衰退同业同业合并、并购合并、并购产能集中产能集中研发公司研发公司数量增加数量增加智权纷争智权纷争增加增加产业策略产业策略联盟谈判联盟谈判密集密集合并、并购合并、并购集中智权集中智权市场规模市场规模快速成长快速成长产品世代产品世代发展迅速发展迅速主要研发主要研发能量集中能量集中于学术、国防、于学术、国防、能源、太空等能源、太空等研究机构研究机构革命性技术革命性技术进入市场进入市场生物芯

28、片生物芯片合作研究合作研究共同开发共同开发策略联盟策略联盟分工整合分工整合技术转让技术转让代工代销代工代销时间时间产业活动产业活动371994Michael Ramsey, U.S. DOEs Oak Ridge National Lab.6,001,229 (19940801-19991214)MolecularToollicensedOrchidBioSciencesacquired Caliper Technologiessub-licensed 38Caliper Technologies专利(60)6267858 High throughput screening assay sys

29、tems in microscale fluidic devices 6274089 Microfluidic devices, systems and methods for performing integrated reactions and separations 6274337 High throughput screening assay systems in microscale fluidic devices 6287520 Electropipettor and compensation means for electrophoretic bias 6287774 Assay

30、 methods and system 6251343 Microfluidic devices and systems incorporating cover layers 6235175 Microfluidic devices incorporating improved channel geometries 6235471 Closed-loop biochemical analyzers 6238538 Controlled fluid transport in microfabricated polymeric substrates 6233048 Fourier detectio

31、n of species migrating in a microchannel 6221226 Methods and systems for monitoring and controlling fluid flow rates in microfluidic systems 6171067 6012902 Micropump 6171850 Integrated devices and systems for performing temperature controlled reactions and analyses 6172353 System and method for mea

32、suring low power signals 6174675 Electrical current for controlling fluid parameters in microchannels 6182733 Methods of manufacturing microfabricated substrates 6186660 Microfluidic systems incorporating varied channel dimensions 6011252 Method and apparatus for detecting low light levels 6042709 M

33、icrofluidic sampling system and methods 6042710 Methods and compositions for performing molecular separations 39 1994Zoya Soane,5,750,015 (19960313-19980512)Soane TechnologiesAclara BioSciencesSpun-off40Aclara专利6284113 Apparatus and method for transferring liquids 6176962 Methods for fabricating enc

34、losed microchannel structures 6054034 Acrylic microchannels and their use in electrophoretic applications 6056860 Surface modified electrophoretic chambers 6074827 Microfluidic method for nucleic acid purification and processing 6093296 Method and device for moving molecules by the application of a

35、plurality of electrical fields 6103199 Capillary electroflow apparatus and method 6103537 Capillary assays involving separation of free and bound species 5858188 Acrylic microchannels and their use in electrophoretic applications 5883211 Thermoreversible hydrogels comprising linear copolymers and th

36、eir use in electrophoresis 6007690 Integrated microfluidic devices 5858188 Acrylic microchannels and their use in electrophoretic applications 41微流体芯片公司 各家公司，各有独门绝活 Caliper (Agilent)：电泳分离、micro PCR Aclara (ABS)：电泳分离 Nanogen：电场分离，电场杂交控制、电场探针固定化 Cepheid：micro PCR Affymetrix：System integration 各家皆推出产品，

37、但仍以技术研发为重心。42October 21 25, 2001October 21 25, 2001Monterey Conference Center Monterey Conference Center Monterey, CaliforniaMonterey, CaliforniaMicrofluidics & Nanotechnology Microfluidics & Nanotechnology World-to-chip Interfacing World-to-chip Interfacing Fluid Dispensing Fluid Dispensing

38、 Fluid Mechanics and Modeling ToolsFluid Mechanics and Modeling ToolsNanofluidic Components Nanofluidic Components Nanoengineered Materials Nanoengineered Materials Materials Materials Surface ModificationSurface ModificationChemical and Biochemical Arrays Chemical and Biochemical Arrays Interfacial

39、 Characterization Interfacial Characterization Application Application Clinical Diagnostics Clinical Diagnostics Genomics and Proteomics Genomics and Proteomics Environmental Assays Environmental Assays Separation Science Separation Science Cellular Analysis Cellular Analysis Drug Discovery Drug Discovery Micromachining Micromachining Micropumps and Valves Micropumps and Valves Microfluidic Components Microfluidic Components Micromachining MethodsMicromachining MethodsP