製造疼痛蛋白質微陣列晶片表面比較之研究

Comparison of Slide Surfaces for the Fabrication of Pain-Related Message Molecule Antibody Microarray

目前在治療病態性疼痛的原則仍以階段性治療為主，所得到的療效卻是不穩定且無 法令人滿意的。真正原因是在於沒有全盤瞭解疼痛。以前受限於研究工具，只能分析某些蛋白質的變化，對於複雜的蛋白質體的波動，則是束手無策。然而，想要有所突破非得從完整的調控網路著手。幸好，在科技進步之下，已有新的技術可以同時分析上百上千種蛋白質，藉由這樣的工具，我們就可以宏觀的蛋白質體的角度，觀察真正疼痛的機制。而抗體微陣列晶片正是這樣有力的工具。因為它的快速，已經開始運用在基礎及臨 床的研究上。同時它也能夠大量平行分析且進行定量的觀察，對於臨床研究上，尤其可 以搭配臨床數據，創造出卓越的研究價值。所以，它的成效不只在快速檢驗個別蛋白質 的變化，最重要的是可以建構調控路徑。因此，我們在實驗中檢驗了四種常見不同表面 的抗體微陣列晶片，搭配了四種疼痛傳遞因子（SP， 的前，NGF， 及TNF-a）。同時為了 改善效能，我們將蛋白質G被覆在晶片表面。最終結果顯示路基晶片如上蛋白質G最適合 從事疼痛蛋白分析所需。

Current therapeutic principles are step-by-step trials with multiple drugs for pathological pain. Such treatment plans often make physicians or patients uncertain and discouraged. The real problem is focused on the limited view of pain network. It was impossible for past researchers to observe so multiplexed proteins changes within different pain syndromes. Conventional methods for protein expression focus only on one or a few targets. However, the investigation of new therapeutic targets to pain needs a better realization of the global regulation network. Fortunately, with advancement of new technologies, it is possible to able to examine hundreds and thousands of protein simultaneously right now. Such large-scale studies have the promise to assemble individual pieces together to gain insights into the overall picture of proteome-wide modifications on pain mechanisms. Antibody microarrays are an emerging technology that promises to be a powerful tool. It has significant applications in basic and clinical researches, particularly because of the rapidity of the experiments. Such analysis is possible, if many experiments with highly parallel with quantitative information can be performed. Especially for clinical researches, the results from antibody microarray could be correlated with clinical information to assess the clinical value of multiple proteins or sets of proteins. The microarray format facilitates not only the rapid evaluation of many proteins individually but also the evaluation of coordinate patterns of expression. Therefore, we tested four commercial antibody microarray surfaces printing on substance P (SP), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), tumor necrosis factor-alpha (TNF- a). To improve the performance, protein G coating method was introduced into slide surface. The results showed the best performance can be achieved in aldehyde-derivatized slide by protein G coating method.