蛋白质微阵列芯片技术及其在抗体筛选中的应用

Protein Microarray Technology and Its Application in Antibody Screening

以兔IgG为模式蛋白质,对其在醛基修饰玻片表面的固定浓度、固定时间和温度等条件进行了优化,结果表明:在室温下,当固定蛋白质的浓度为 1g/L、固定时间为 4h时,可获得理想的蛋白质固定效果;蛋白质的定量检测范围为 1μg/L^10mg/L。按优化的蛋白质微阵列芯片制作条件将规模化制备的抗体制作成抗体微阵列芯片,通过与荧光标记的人球蛋白和人白蛋白的相互作用,实现了对不同抗体株抗球蛋白和抗白蛋白活性的快速筛选与比较。

The rabbit IgG was immobilized onto the surface of glass slide modified with aldehyde, and then fluorescein isothiocyanate labeled goat anti-rabbit IgG was applied to the array at room temperature. The protein microarray was manufactured by a manual arrayer, fluorescence images were collected by biochip scanner and analyzed with GenePix 3. 0 software. The optimal immobilization concentrations of protein molecules, immobilization time, the detection limit and the reproducibility were examined. On the base of these results, an antibody microarray with twenty kinds of antibodies was manufactured and used to screen the specific affinity interaction between antibodies and albumin or globulin, the results were in accordance with those obtained by the traditional enzyme linked immunosorbent assay.