Borrebaeck C A, Ekström S, Hager A C, Nilsson J, Laurell T, Marko-Varga G
Lund University, Lund, Sweden.
Biotechniques. 2001 May;30(5):1126-30, 1132. doi: 10.2144/01305dd05.
With the human genome in a first sequence draft and several other genomes being finished this year, the existing information gap between genomics and proteomics is becoming increasingly evident. The analysis of the proteome is, however, much more complicated because the synthesis and structural requirements of functional proteins are different from the easily handled oligonucleotides, for which a first analytical breakthrough already has come in the use of DNA chips. In comparison with the DNA microarrays, the protein arrays, or protein chips, offer the distinct possibility of developing a rapid global analysis of the entire proteome. Thus, the concept of comparing proteomic maps of healthy and diseased cells may allow us to understand cell signaling and metabolic pathways and will form a novel base for pharmaceutical companies to develop future therapeutics much more rapidly. This report demonstrates the possibilities of designing protein chips based on specially constructed, small recombinant antibody fragments using nano-structure surfaces with biocompatible characteristics, resulting in sensitive detection in the 600-amol range. The assay readout allows the determination of single or multiple antigen-antibody interactions. Mass identity of the antigens, currently with a resolution of 8000, enables the detection of structural modifications of single proteins.
随着人类基因组初步测序草图的完成以及今年其他几个基因组测序工作的结束，基因组学和蛋白质组学之间现有的信息差距日益明显。然而，蛋白质组分析要复杂得多，因为功能性蛋白质的合成和结构要求不同于易于处理的寡核苷酸，针对寡核苷酸，DNA芯片的使用已带来了首个分析突破。与DNA微阵列相比，蛋白质阵列或蛋白质芯片提供了对整个蛋白质组进行快速全局分析的独特可能性。因此，比较健康细胞和患病细胞的蛋白质组图谱这一概念可能使我们了解细胞信号传导和代谢途径，并将为制药公司更快地开发未来疗法奠定新基础。本报告展示了基于特殊构建的小重组抗体片段，利用具有生物相容性的纳米结构表面设计蛋白质芯片的可能性，可实现600阿摩尔范围内的灵敏检测。该检测读数可确定单一或多种抗原 - 抗体相互作用。目前抗原的质谱鉴定分辨率为8000，能够检测单个蛋白质的结构修饰。
