基于滤纸微孔板的酶联免疫吸附测定方法研究  

Study on Enzyme Linked Immunosorbent Assay Using Paper-based Micro-zone Plates

作　　者：白鹏[1] 罗雁[1] 李英[1] 余晓冬[1] 陈洪渊[1] 

机构地区：[1]南京大学化学化工学院,生命分析化学国家重点实验室,南京210093

出　　处：《分析化学》2013年第1期20-24,共5页Chinese Journal of Analytical Chemistry

基　　金：国家重大科学仪器设备开发专项子课题(No.2011YQ17006711);973计划课题(No.2011CB911003);国家自然科学基金创新研究群体项目(No.21121091)资助

摘　　要：用光刻法在普通定量滤纸上制作出了疏水图案,得到了滤纸微孔板。将表面修饰后的SiO2微球负载在滤纸表面,以提高滤纸纤维吸附蛋白质的能力,基于此建立了以间接酶联免疫吸附法测定羊抗兔免疫球蛋白的方法。以酶标兔抗羊IgG为例,负载SiO2微球后,滤纸微孔板吸附该蛋白质的量提高了20%～700%;将SiO2负载法应用于羊抗兔IgG的检测时,可使信号增强20%～150%。在本实验条件下,测定羊抗兔IgG的线性范围为3×10-12～3×10-8mmol/孔。本方法使生物免疫试剂消耗量减少至3～5μL/孔、检测时间降至20 min/板、滤纸微孔板制作成本可低至约每板1.1美元,为研制准确、快速和低成本的疾病检测设备提供了新的思路和可能性。Photolithography was used to pattern hydrophobic design on common quantitative filter paper, based on which we successfully fabricated the paper-based micro-zone plates. In order to improve its protein- absorbing capability for further signal magnification, modified silicon dioxide （SiO2 ） micro-beads were deposited onto the surface of paper. Then indirect-ELISA for goat anti-rabbit IgG was applied on the 36-well plates. Taking enzyme linked goat anti-rabbit IgG as an example, loading SiO2 beads could remarkably increase the quantity of the absorbed protein by 20% -700%. When this method was used for goat anti-rabbit IgG detection, signals were magnified by 20% -150% , and the linear range of detection was from 3 x 10-~2 mmol/zone to 3 x 104 mmol/zone under given conditions. In this manner, bio-reagent consumption was reduced to 3-5 pL per zone, test time dropped to 25min per plate and the cost of fabrication reduced to S1.1 per plate, which indicated a promising application foreground for new disease diagnosis devices.

关 键 词：滤纸 微孔板 光刻 酶联免疫分析 二氧化硅微球 

分 类 号：O651[理学—分析化学] R446.6[理学—化学]