親和分析技術之開發與應用

Abstract

一般而言，細菌感染及細胞癌化是可以治療的疾病，但如果無法及時診斷細菌感 染及細胞癌化情形而施以適當治療可能會對病人造成無法逆轉的傷害甚至會導致死 亡。因此本計晝將開發以親和力為基礎而可用於快速檢測致病細菌和癌細胞的新穎親 和方法為研究目標，而胜肽適配體將被選擇可以標靶特定細胞的探針分子。胜肽適配 體具有類似抗體的分子辨識功能，儘管胜肽適配體的尺寸遠小於抗體，但具有可比擬 的親和常數。並且相對於製備抗體的時間及花費，胜肽適配體具有可大量合成製造的 優點。由於目前具有對致病細菌和癌細胞親和力的胜肽適配體仍然相當有限，在本計 晝中我們將建立以奈米探針為基礎的開發平台以探尋可標靶致病細菌和癌細胞的胜肽 適配體。並且，我們將使用新發現的胜肽適配體為探針分子用於建立快速診斷致病細 菌和癌細胞的分析技術。在第一年和第二年的計晝，我們將致力於探尋可和致病細菌 親合的胜肽適配體；此外，這兩年之中，我們將利用所發現的新胜肽適配體於發展新 的親和技術：即發展組合侷限式表面電漿共振和大氣壓下表面輔助雷射脫附/游離質 譜法的親和感測技術，並致力於大氣壓下表面輔助雷射脫附/游離質譜影像技術的開 發。在第三年和第四年，將致力於發現可標靶癌症細胞的胜肽適配體，將選用乳癌細 胞為樣本；此外，組合侷限式表面電漿共振和無接觸大氣壓化學游離法親和感測技術 及親合無接觸大氣壓化學游離法的發展也將是這兩年致力的研究方向。我們期望藉著 胜肽適配體的開發及親和技術的發展下，可以對於快速診斷細菌感染和癌症細胞的研 究有所助益。

In general, bacterial infections and cancer diseases are treatable. However, patients suffering from bacterial infections or cancer diseases without conducting prompt diagnosis and medical treatment may be irreversibly damaged and even die. Thus, the core of this proposal is to develop new affinity-based methods for rapid detection of pathogenic bacteria and cancer cells. Peptide aptamers will be used as the probe molecules to recognize these target cells. Peptide aptamers have similar capabilities to antibodies in terms of affinity and specificity toward their target species although their sizes are much smaller than antibodies. Additionally, it is less difficult to generate peptide aptamers in a large quantity than to generate antibodies in terms of production time and cost. Since the peptide aptamers specific toward pathogenic bacteria and cancer cells are still limited, nanoprobe-based platforms for explore new peptide aptamers for target cells will be established in this project. We will use the newly discovered peptide aptamers as the probe molecules to establish new affinity methods for rapid diagnosis for pathogenic bacteria and cancer cells. The combination of affinity techniques and ambient MS will be studied in this project. In the first and second years, efforts will be devoted in the exploration of new peptide aptamers for pathogenic bacteria. Furthermore, the discovered peptide aptamers will be used in the development of affinity techniques at the same time. The combination of localized surface plasmon resonance (LSPR) and ambient surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) and the development of ambient SALDI imaging are the main affinity techniques, which will be established in these two years. In the third and fourth years, efforts will be made to discover new peptide aptamers for cancer cells. Breast cancer cells will be used as the model samples. Furthermore, the combination of LSPR with contactless-atmospheric pressure ionization (C-API) and affinity-based C-API will be developed in the last two years. We expect the peptide aptamers explored in this proposal and affinity methods developed accordingly can be potentially used to assist the rapid diagnosis of bacterial infections and cancer cells.