Development for Advanced Bio-nanodevice for Functional Genomics and Proteomics

随着人类基因组测序的完成，人类基因组计划将迅速进入后基因组测序时代，包括单核苷酸多态性(SNP)分析、功能基因组学、突变分析、转录组分析、蛋白质组分析、代谢组分析和系统生物学。由96-384根毛细管组成的毛细管阵列电泳在基因组测序时代起着至关重要的作用，但在后人类基因组测序时代，进一步发展DNA、mRNA、蛋白质和代谢物的分析技术是未来医学和基于功能基因组学和蛋白质组学的新药发现技术的高度要求。我们研制了用于DNA和基因组分析的NaN-BioDevice。用微芯片技术在10-120个S范围内分离出1000bp的DNA片段，90年代内进行Y染色体多态性分析。利用微芯片技术在2分钟内完成了人血标本的单倍型分析。宽2 0 0 nm，高4 0 0 nm的纳米柱阵列是成功的…更适用于10-25s内DNA的快速分离。这种纳米生物装置对于在15秒内快速分离从几kDa到200 kDa的蛋白质样品非常有用。利用该技术，从人T细胞系--淋巴母细胞Jurkat细胞中提取的复杂蛋白质混合物在15s内得到分离，重复性高。该方法能在较短的时间内检测和鉴定Jurkat细胞热休克后增加的靶蛋白，而常规的双向凝胶电泳法需要数小时的时间。这种微芯片上的新技术将取代蛋白质组分析的高通量筛选系统的巨大优势，特别是在快速检测少量目标蛋白质方面，这些蛋白质是细胞或组织特定生理或病理状态的独特型。该纳米生物装置可用于糖蛋白糖链的快速分析。我们开发了用于纳米生物器件的高灵敏度LED诱导荧光检测系统，并应用于荧光标记糖链的分析。从糖蛋白(AGP和Ig G)中消化的糖链的复杂混合物在60 S范围内被高分辨地分离和检测。该纳米生物装置将适用于单个基因组DNA分子的SNP分析。较少

Since human genome sequencing has been completed, human genome project will quickly move on to the post genome sequencing era, including single nucleotide polymorphism (SNP) analysis, functional genomics, mutation analysis, transcriptome analysis, proteome analysis, metabolome analysis, and systems biology. Capillary array electrophoresis with 96-384 capillaries plays a vital role in the genome sequencing era, but in the post Human Genome sequencing era, further development of analytical technology for DNA, mRNA, protein, and metabolites is highly required for future medicine and new drug discovery technology based on functional genomics and proteomics. We developed nan-biodevice for DNA and genomic analysis. DNA fragments up to 1000 bp is separated within 10-120 s by microchip technology, Y-chromosomal polymorphism is analyzed within 90s. Haplotyping from human blood sample was achieved within 2 min by microchip. Array of nano-pillar, which is 200 nm wide and 4000 nm tall, is successf … More ully applicable to fast separation of DNA within 10-25s. The nano-biodevice is extremely useful for fast separation of protein samples from several kDa to 200 kDa within 15 seconds. Utilizing this technique, complex protein mixture extracted from human T cell line, lymphoblastic Jurkat cells, were separated within 15s with high reproducibility. The target proteins of the Jurkat cells, which increase after heat-shock apoptosis, wore detected and identified within by this method, while several hours are required for conventional 2-DE analysis. This novel technique on a microchip will ofter the enormous advantages for high through-put screening systems of proteome analysis, especially for rapid detection of small amount of target proteins, those are idiotypic for a specific physiologic or pathologic state of cells or tissues. The nano-biodevice will be applicable to fast-analysis of sugar chain on the glycoproteins. We developed high-sensitive LED induced fluorescence detection system for nano-biodevice and applied to analysis of fluorescent labeled sugar chain. Complex mixture of sugar chains digested from glycoproteins (AGP and IgG) is separated and detected within 60 s with high-resolution. The nano-biodevice will be applicable to the SNP analysis of a single genomic DNA molecule. Less