建立一種可應用於流式細胞儀上檢測之螢光微脂體複合體及其特性探討

Abstract

用於檢測與定量生物標記之技術已被快速發展及應用於研究與臨床診斷上。一般而言，生物標記是利用抗體-抗原為主之免疫檢測系統進行偵測，抗體會專一性地與特定抗原蛋白結合並可藉由標定物來定量。此篇研究建立之螢光物質標定之微脂體可作為探針應用於免疫檢測系統上。Lipo-PEI-PEG complex (LPPC)為本實驗室開發出來之新型微脂體，它具有穩定且快速吸附各種蛋白之能力且蛋白本身可保有原本之生物活性，此外LPPC亦具有容易被脂溶性螢光物質標定之特性。

這篇研究成功建立了螢光標定之LPPC，並測試其在流式細胞儀上偵測細胞激素(cytokine)之可行性。研究結果顯示其螢光強度與細胞激素之濃度成正比。同時此研究亦比較了與傳統用來偵測細胞激素之酵素連結免疫吸收分析（ELISA）間的差異性。因此，以螢光標定之LPPC作為檢測探針可提供作為另一種免疫檢測之方式。

The techniques to identify and quantify biomarkers have been developed rapidly on research and clinical diagnosis. Generally, detections for biomarkers are performed using the antibody-based immunoassay system, in which the targeting proteins are specifically bound by their antibodies and the complex can be detected via some signal-reporting labels. Here, an alternative is the fluorescent dyes incorporated liposomes as immunoassay probes. Lipo-PEI-PEG complex (LPPC) is a novel liposome, composed of lipids and two polymers. LPPC can strongly and quickly adsorb various proteins and maintain their own functions on surface. Furthermore, LPPC are easily incorporated lipophilic fluorescent dyes.

In this study, fluorescent dye-labeled LPPC was constructed and its ability for detection of soluble proteins by the flow cyotometer was examined. At the field of soluble cytokines, the total fluorescence intensity is proportional to the concentrations of target cytokines. The limitations were also evaluated compared with traditional ELISA assay. Thus, LPPC as a probe may provide an alternative method to immunoassay.