利用表面電漿共振感測微奈米濃縮晶片的蛋白質免疫分析

人體內微量的蛋白質變化能用於早期癌症的診斷，如前列腺特異性抗原。奈米等級的流道具有離子篩選的功能，配合微-奈-微流道的結構設計，就能利用電壓源的施加讓微米流道內產生蛋白質的濃縮。蛋白質的預濃縮能增加偵測系統的靈敏度，並降低人體血清中微量的蛋白質分析的時間和生物樣本的消耗。利用濃縮晶片能將抗原濃縮後藉由濃度的提高增加免疫分析中抗體-抗原鍵結反應的靈敏度。MIT的Dr. Jongyoon Han團隊在2005年利用這樣的流道設計配合半導體技術將濃縮晶片製作出來，達到高濃度的蛋白質濃縮(Applied Optics 40, 3810-3821)。我的實驗主要目標是進一步將濃縮晶片搭配架設好的表面電漿共振系統進行免螢光標靶的蛋白質免疫分析。為了優化奈米流道濃縮晶片，我利用電流計測量微奈米流道的導電率常數，進而建立電阻式電路模型。運用模型計算出微奈米濃縮晶片流道的參數，利用半導體製程將晶片完成。架設表面電漿共振系統，量測不同樣式、尺寸金膜的表面電漿共振訊號。架設正立式螢光顯微鏡，配合微奈米濃縮晶片與表面電漿共振系統測量前列腺特異性抗原PSA(prostate – specific antigen)的免疫分析。電阻式電路模型首創應用在濃縮晶片的設計上，成功的建立流道尺寸對應濃縮產生和濃縮倍率的關係，並藉由模擬結果優化最後濃縮晶片的表現。首次利用表面電漿共振系統測量經濃縮過後的蛋白質免疫分析，證實能利用濃縮增加蛋白質抗體-抗原的鍵結。

關鍵字

微流道電路模型；微奈米濃縮晶片；預濃縮；免疫分析；表面電漿共振

並列摘要

Some concentration variation of antigens can be considered as cancer marker. Nanochannels between two microchannels can achieve protein preconcentrator due to ion selectivity of nanochannels. Using nanochannels and microchannels, Professor Han’s group developed a million-fold preconcentrator in 2004(Applied Optics 40, 3810-3821). I set up surface plasmon resonance (SPR) system as a label-free sensor to detect variation of antigen-antibody conjugate, and combined with the preconcentrator to enhance sensitivity of immunoassays by upper antigen concentration which is preconcentrated by the preconcentrator. To optimize the performance of preconcentrator, Picoammeter was used to identify the parameters of micro/nanochannels. The resistive network electrical model was built by the identified parameters of micro/nanochannels to calculate and design the channel size of preconcentration chip, then fabricate chips using semiconductor technologies and equipment. The surface plasmon resonance (SPR) platform was set up to measure SPR signal and design the size and shape of the thin film gold in the preconcentration chip. The fluorescence microscopy was set up to integrated with SPR platform and preconcentration chip to do immunoassays of preconcentrated prostate – specific antigen (PSA). Originally, the resistive network circuit model was developed. And the preconcentration chip design rule was built. The SPR system combined with preconcentration chip to be applied to immunoassays, and the antigen-antibody conjugate enhanced by preconcentration.

並列關鍵字

preconcentration ； microfluidic ； nanofluidic ； surface plasmon resonance (SPR) ； immunoassay ； resistive network electrical model

參考文獻

24. 留連晉, 表面電漿共振感測系統應用於微奈米流體晶片之蛋白質檢測, in 臺灣大學電機工程學研究所學位論文2008, 臺灣大學. p. 1-62.

28. 于念中, 利用表面電漿共振感測微米流體晶片中的蛋白質免疫分析反應, in 臺灣大學生醫電子與資訊學研究所學位論文2010, 臺灣大學. p. 1-63.

1. Lee, K.H., et al., Microfluidic systems integrated with two-dimensional surface plasmon resonance phase imaging systems for microarray immunoassay. Biosens Bioelectron, 2007. 23(4): p. 466-72.

2. Jeong Hoon Lee, Y.-A.S., Steven R. Tannenbaum, Jongyoon Han, Increase of Reaction Rate and Sensitivity of Low-Abundance Enzyme Assay Using Micro/Nanofluidic Preconcentration Chip. Anal. Chem, 2008(80): p. 3198-3204.

4. Pu, Q.S., et al., Ion-enrichment and ion-depletion effect of nanochannel structures. Nano Letters, 2004. 4(6): p. 1099-1103.

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利用表面電漿共振感測微奈米濃縮晶片的蛋白質免疫分析

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