Development for Advanced Bio-nanodevice for Functional Genomics and Proteomics

随着人类基因组测序的完成，人类基因组计划将很快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。96-384根毛细管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，基于功能基因组学和蛋白质组学的未来医学和新药发现技术需要进一步发展DNA、mRNA、蛋白质和代谢物的分析技术。我们开发了用于DNA和基因组分析的纳米生物器件。微芯片技术可在10 ~ 120s内分离出长度达1000bp的DNA片段，90秒内分析y染色体多态性。用微芯片在2分钟内完成人血单倍型分型。该纳米柱阵列宽200nm，高4000nm，可在10-25s内实现DNA的快速分离。该纳米生物装置对于在15秒内从几kDa到200 kDa的蛋白质样品的快速分离非常有用。利用该技术，从人T细胞系Jurkat淋巴母细胞中提取的复杂蛋白混合物可在15s内分离，具有较高的重复性。热休克凋亡后Jurkat细胞的靶蛋白增加，该方法可在细胞内检测和鉴定，而传统的2-DE分析需要数小时。这种基于微芯片的新技术将为蛋白质组分析的高通量筛选系统提供巨大的优势，特别是对于快速检测少量靶蛋白，这些靶蛋白是细胞或组织的特定生理或病理状态的独特型。该纳米生物装置将适用于糖蛋白上糖链的快速分析。研制了高灵敏度LED诱导荧光检测系统，并将其应用于荧光标记糖链的分析。从糖蛋白（AGP和IgG）中消化的复杂糖链混合物在60秒内分离和高分辨率检测。该纳米生物装置将适用于单个基因组DNA分子的SNP分析。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