近來螢光分子探針，大部分是用在酶蛋白偵測上，如醣苷酶(glycosidases)、蛋白酶(proteases)等。其方法是透過酶的催化使探針上的非螢光基團發生結構變化，使其轉變成具螢光性質的基團。本論文開發以Cy5螢光探針為主的非酶蛋白偵測方法，在聚次甲基的γ位有苯基取代的Cy5螢光分子對蛋白質之親和性配體結合後，可不經由酶的催化，選擇性的偵測蛋白質。進一步的研究發現，此螢光探針具自組裝性質，並會形成J-型聚集使螢光淬熄。當目標分析物存在時，因配體和目標物的結合使得螢光探針的聚集拆卸，並發出強烈的近紅外螢光。基於探針的自組裝/拆卸使得螢光顯著增益的性質，本論文設計了一系列螢光探針，並可成功標記癌細胞表面之標記物：跨膜型碳酸酐酶IX。

Abstract
Currently most of the fluorogenic probes are designed for the detection of enzymes which work by converting the nonfluorescence substrate into the fluorescence product via an enzymatic reaction. On the other hand, the design of fluorogenic probes for non-enzymatic proteins remains a great challenge. Herein, we report a general strategy to create near-IR fluorogenic probes, where a small molecule ligand is conjugated to a novel γ-phenyl-substituted Cy5 fluorophore, for the selective detection of proteins through a non-enzymatic process. Detail mechanistic studies reveal that the probes self-assemble to form fluorescence-quenched J-type aggregate. In the presence of target analyte, bright fluorescence in the near-IR region is emitted through the recognition-induced disassembly of the probe aggregate. This Cy5 fluorophore is a unique self-assembly/disassembly dye as it gives remarkable fluorescence enhancement. Based on the same design, three different fluorogenic probes were constructed and one of them was applied for the no-wash imaging of tumor cells for the detection of hypoxia-induced cancer-specific biomarker, transmembrane-type carbonic anhydrase IX.

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