冷光蛋白質微陣列平台技術之研究

Development of a Chemiluminescent Protein Microarray Platform

劉茂隆(Mao-Lung Liu)

指導教授 ： 游吉陽

大同大學/工程學院/生物工程學系所/碩士(2005年)

摘要

蛋白質微陣列已成為蛋白質體學研究中的重要工具。蛋白質微陣列的優勢在於可同時進行大量分析，但所需成本較高，故降低成本有其必要性。
本研究目的在於發展一低成本之冷光蛋白質微陣列之技術平台。以MicroCaster arrayer手動式打點機，將anti-human interleukin-2 polyclonal antibody及anti-human interleukin-4 polyclonal antibody點製於nitrocellulose membrane，以標定horseradish peroxidase的streptavidin結合帶有biotinylated anti-human interleukin-2及biotinylated anti-human interleukin-4 monoclonal antibody應用類似sandwich type之enzyme-linked immunosorbent assay反應，以偵測recombinant human interleukin-2及recombinant human interleukin-4。使用horseradish peroxidase產生之chemiluminescence以charge-coupled device拍攝，藉
由ScanAlyze軟體分析。
本技術平台以冷光基質反應產生chemiluminescence，由於產生發光
之反應中間物會向外擴散，故本系統最高點製密度為192點。本系統反
應條件確立後，recombinant human interleukin-4與recombinant human interleukin-2偵測極限分別為0.125 ng/ml和0.5 ng/ml，檢量線跨越2個

級數。
由於蛋白質微陣列缺乏類似polymerase chain reaction的反應提高靈敏度，故應用streptavidin-peroxidase polymer與streptavidin-biotin complex兩種方式，藉由提高反應訊號，以求更低的偵測極限。結果顯示雖然有
使蛋白質微陣列之偵測信號增加，但未降低偵測極限。

關鍵字

蛋白質微陣列 ； 偵測極限

並列摘要

A low-cost, low-density protein microarray on nitrocellulose membrane for the detection of human cytokines was developed. The detection was based on chemiluminescent sandwich-type immunoassay. The printed spots were uniform and consistent in size. Under optimized condition, the limits of detection for human interleukin 2 and interleukin 4 were 0.5 and 0.125 ng/ml, respectively. The linear range of the calibration curves spanned
over two orders of magnitude.
The effects of several potential signal enhancers were also tested.

並列關鍵字

LOD. ； Protein microarray

參考文獻

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