以氧化鋅奈米線晶片檢測肺癌細胞分泌之角質蛋白基因片段21 - 1

細胞角質蛋白（cytokeratin）是上皮細胞內的結構性蛋白，是細胞骨架的成分之一。在全身各種上皮細胞內廣泛分佈，共有20種。某些特定的角質蛋白僅存在於特定組織中，如角質蛋白19（cytokeratin 19）主要表達於支氣管上皮細胞內，而在腫瘤中也會出現特異性表達，因此具有成為肺部腫瘤標記的潛力。角質蛋白19本身不溶於水，但其分解後的片段具有可溶性，可以在人體中進行檢測。而角質蛋白19的基因片段21-1（CYFRA21-1）即為檢測此片段的腫瘤標記物，其致病量為 3.8 ng/ml。本研究使用氧化鋅奈米線基於指叉電極方式檢測肺癌細胞分泌的CYFRA21-1在唾液中的濃度。首先使用掃描式電子顯微鏡、能量散射X射線光譜儀和X光繞射光譜儀對氧化鋅奈米線進行物性分析；接著使用APTES和GA對氧化鋅奈米線進行改質，並在其上鍵結抗體，使其具有結合CYFRA21-1的能力。並使用田口直交表對其抓取能力進行三水準實驗優化，最後以電性方式對唾液中的CYFRA21-1進行線性濃度量測。在量測中，2.5至 12.5ng/ml範圍內各濃度皆呈現良好的線性趨勢。人體唾液中含有各種物質，因此檢測平台的抗干擾性為其十分重要的指標；而干擾性實驗也說明本論文中外泌體感測器具優秀的選擇性和抗干擾性。最後再驗證含生物材料之檢測平台的有效期限，結果顯示在抗體保存期限內，檢測靈敏度皆保持在良好的範圍內。

關鍵字

田口法；電性量測；氧化鋅奈米線；角質蛋白19基因片段21-1

並列摘要

Cytokeratins are structural proteins found in epithelial cells, constituting the cytoskeleton. There are 20 different types of cytokeratins, distributed throughout various epithelial cells in the body. Specific cytokeratins are found in certain tissues, such as cytokeratin 19 (CK19) expressed in bronchial epithelial cells, and exhibit specific expression in tumors, making them potential markers for lung tumors. CK19 itself is insoluble in water, but its soluble fragments can be detected in the human body. The gene fragment 21-1 of CK19 (CYFRA21-1) serves as a tumor marker to detect these fragments, with a pathological level of 3.8 ng/ml. In this study, zinc oxide nanowires were used to detect the concentration of CYFRA21-1 secreted by lung cancer cells in saliva, employing a forked electrode setup. The physical properties of zinc oxide nanowires were analyzed using scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction spectroscopy. The nanowires were then modified with APTES and GA, and antibody binding was facilitated to enable the specific binding of CYFRA21- 1. The capturing capability was optimized using a three-level experiment based on the Taguchi orthogonal array, and the linear concentration of CYFRA21-1 in saliva was measured electrically. All concentrations within the range of 12.5 to 2.5 ng/ml exhibited a good linear trend.As human saliva contains various substances, the anti-interference ability of the detection platform is crucial. The interference experiments demonstrate the selectivity and anti-interference capability of the exosome sensor in this study. Finally, the validity period of the detection platform containing biological materials was verified, and the results showed that the detection sensitivity remained within a good range throughout the antibody storage period.