本研究為設計一高分子材料以偵測目標中的鼠疫抗原(Antigen of Yersinia pestis)。基材為線/間距比率為1：1.5的圖案化光阻矽晶圓，其線寬為500 nm、600 nm、700 nm、800 nm的三維光柵。以此結構利用原子轉移自由基聚合(Atom Transfer Radical Polymerization, ATRP)製備甲基丙烯酸(Poly(sodium methacrylate)，PMAS)的線型高分子刷，進行12、18、24、30小時的聚合成長。接著利用PMAAS末端有-COOH官能基，透過EDC/NHS的耦合Protein G與鼠疫F1抗原之單株抗體(Mouse-Anti-F1)，進行鼠疫抗原抓取。由於檢驗原理是用雷射會因表面規律的微結構產生光柵繞射，對比微結構受到破壞所產生的能量散失進行偵測，但抗原的三維尺寸較小，對微結構的破壞有限，使的偵測受限。需額外建立起三明治結構以增強訊號。三明治結構由聚苯乙烯微球為基材，改質上Protein G與鼠疫F1抗原之多株抗體(Rabbit-Anti-F1)後與被晶片抓取之抗原做結合，達到破壞微結構，增強光學訊號之目的。
接著對於100 μg/mL-10 μg/mL做鼠疫抗原濃度靈敏度測試，並透過螢光顯微鏡觀察螢光佐證，結果顯示即使在鼠疫抗原濃度較高時，晶片所抓取之抗原對雷射繞射強度並無太大影響，而當聚苯乙烯微球與晶片表面抗原結合時，雷射繞射強度呈線性下降的趨勢，其線性範圍可達到濃度10 ng/ mL，而此濃度下螢光顯微鏡已無法發現鼠疫抗原的螢光存在，說明雷射檢測鼠疫桿菌的靈敏度較螢光顯微鏡來的高。說明利用雷射能量檢測鼠疫桿菌的靈敏度較螢光顯微鏡高。而線型500nm的圖案化高分子晶片對於不同濃度的鼠疫桿菌經過光學訊號放大後所作出的線性回歸之相關係數值為0.9589代表鼠疫桿菌濃度有95.89%的概率能以雷射解釋，期望利用此線性規律，能以雷射能量值快速得到抗原濃度，達到快速檢測抗原的作用，以此模型作為未來具前瞻發展性的快篩微縮基材，取代過去對抗原的檢測方式，發展為拋棄式(disposable)的快篩微縮晶片(Lab-on-a-chip)於抗原之檢測領域。

This study designed a polymer material to detect the antigen of Yersinia pestis. The substrate is a patterned photoresist silicon wafer with a line/pitch ratio of 1:1.5, a three-dimensional grating with 500 nm, 600 nm, 700 nm, and 800 nm. With this structure, a linear polymer brush of sodium methacrylate (PMAS) was prepared by atom transfer radical polymerization (ATRP), and polymerization growth was carried out for 12, 18, 24, and 30 hours. In order to capture the antigen,we use EDC/NHS coupling Protein G and the monoclonal antibody (Mouse-Anti-F1 Yp) to the the carboxylic acid group at PMAA,. Because the principle of detection is that the laser has grating diffraction due to the regular microstructures on the surface,and campare it to the energy loss caused by the destruction of the microstructure.But the three-dimensional size of the antigen is small, and the destruction to the microstructure is limited.Make detection unaccuracy. An additional sandwich structure is needed to construct to enhance the optical signal. The sandwich structure is made of polystyrene microspheres, modified with protein G and Polyclonal Antibody(Rabbit-anti-F1 Yp). Then combined with the antigen captured by the chip to destroy the microstructure and enhance the optical signal.
Then, 100 μg/mL-10 μg/mL was tested for the sensitivity of the antigen concentration, combine with fluorescent microscope. The results showed that even when the antigen concentration was high, the laser energy does not have much effect, and when the polystyrene microspheres are combined with the antigen on the surface of the chip, the laser diffraction intensity decreases linearly, and the linear range can reach 10 ng/mL. At this concentration, the fluorescence of antigen can no longer be observed. Indicating that the sensitivity of laser detection of antigen is higher than that of a fluorescent microscope. The 500nm patterned polymer chip for different concentrations of Yersinia pestis after optical signal enhancing has correlation coefficient value of 0.9589, representing 95.89% probability of antigem concentration can be explained by laser energy, which has great potential as an rapid screening techniques for detecting antigens.

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