應用奈米流道電動力濃縮現象於腫瘤標誌免疫檢測晶片之研究

開發一適用於人類檢體的晶片已是未來生物晶片的發展趨勢，在檢測之前，將樣本進行預濃縮動作以提高濃度，以增加後續檢測之準確性是相當重要的一個步驟，尤其在低濃度的蛋白質檢測中，更顯得濃縮步驟的必要性。本研究旨在探討生物樣本液於不同pH環境濃縮效果分析，利用濃縮裝置增強免疫反應。流道是以低成本的聚二甲基矽氧烷(Polydimethylsiloxane, PDMS)轉印經由黃光微影製成的晶片圖形，並與印有黃金電極圖形之玻片接合，再施以高電壓形成電擊穿現象，製作奈米電動力濃縮裝置，此濃縮裝置不需稀釋或置換生物樣本液即可進行濃縮實驗，黃金電極圖形為免疫反應區，本研究測試生物樣本液Cy3螢光標記T24膀胱腫瘤蛋白於不同pH值緩衝溶液中進行濃縮的最佳化成果，並與市售螢光胎牛血清蛋白的實驗成果相互比較。研究結果顯示，當pH值為7.4及9時，T24膀胱腫瘤蛋白之濃縮效果有明顯提升，選擇人類生理pH值環境下進一步進行濃縮免疫電訊號實驗，修飾於電極上的Galectin-1 Ab可成功抓取T24細胞裂解液中的Galectin-1蛋白。

關鍵字

免疫分析； pH值；奈米通道；濃縮；蛋白質

並列摘要

Sample preconcentration is an important step that increases the accuracy of subsequent detection, especially for samples with extremely low concentrations.This study investigated the performance of sample preconcentration proteins in cell lysate at buffer solutions with different pH values and estimated nanochannels dimensions. The preconcentration nanofluidic chip for proteins was mainly fabricated by simple standard lithograph with replica of polydimethylsiloxane (PDMS) and fast nanofissures formation utilizing electric breakdown phenomenon. The performance of preconcentration for Cy3 labeled proteins in the cell lysate of T24 bladder tumor cells increased with the pH value when it was less than 9. When the pH value was increased to 10, the performance of protein preconcentration decreased significantly. The commercial available fluorescein isothiocynate labeled bovine serum albumin, FITC-BSA, was also demostrated in this study and the results with the same trend with T24 cell lysate were obtained. Choose human physiological pH environment for further preconcentrating immunoassays experiment. The experimental results indicate that Galectin-1 Ab successfully grab T24 cell lysates of Galectin-1 protein.

並列關鍵字

pH value ； nanochannels ； concentration ； protein ； immunoassay

參考文獻

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[5] S. W. Yeung and I.-M. Hsing, "Manipulation and extraction of genomic DNA from cell lysate by functionalized magnetic particles for lab on a chip applications," Biosensors and Bioelectronics, vol. 21, pp. 989–997, 2006.

國際替代計量

應用奈米流道電動力濃縮現象於腫瘤標誌免疫檢測晶片之研究

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