【正文】
f the array spot was proportional to the antigen concentration ( Fig. 3 ). This result demonstrated the potential application of singlemolecule,sandwichtype, antibody nanoarrays for the detection of individual protein molecule markers at the singlemolecule level. A representative calibration curve based on the plot of the spot fluorescence intensity against the human leptin standard in the range of concentrations of 100 zM– pM using the nanoarray protein chip and TIRFM is shown in Fig. 4 A. The linear range was 100 zM–400 aM (correlation coefficient, R = ) in the assay of the nanoarray protein chip ( Fig. 4B). The spot intensity values were calculated after background subtractions. The LOD of the leptin protein was 100 zM in the nanoarray leptin protein chip assay. Comparison of ELISA and nanoarray protein chip 說明納米陣列蛋白質(zhì)芯片與酶聯(lián)免疫吸附測定相比具有更低的檢測限,需要更少的樣品量和檢測時間。 說明在低濃度范圍內(nèi)納米陣列芯片上的響應(yīng)信號與待測物的濃度更成比例關(guān)系 存在的問題 成本過高 , 需一系列昂貴的尖端儀器 芯片的標(biāo)準(zhǔn)化問題 提高芯片的特異性、簡化樣品制備和標(biāo)記操作程序、增加信號檢測的靈敏度和消除芯片背景對于結(jié)果分析的影響等等 展望 — 蛋白質(zhì)芯片未來的發(fā)展重點(diǎn) 建立快速、廉價(jià)、高通量的蛋白質(zhì)表達(dá)和純化方法,高通量制備抗體并定義每種抗體的親和特異性。 改進(jìn)基質(zhì)材料的表面處理技術(shù)以減少蛋白質(zhì)的非特異性結(jié)合。 提高芯片制作的點(diǎn)陣速度;提供合適的溫度和濕度以保持芯片表面蛋白質(zhì)的穩(wěn)定性及生物活性。 研究通用的高靈敏度、高分辨率檢測方法,實(shí)現(xiàn)成像與數(shù)據(jù)分析一體化。 謝謝!