蛋白质微阵列检测抗原抗体相互作用

The Analysis of Antigen-antibody Interactions on Protein Microarrays

为了制备蛋白质微阵列和研究芯片表面抗原 抗体的相互作用 ,研究了如何在玻片表面固化蛋白质和用荧光染料 (Cy3,Cy5 )对蛋白质进行标记 .结果表明 ,在醛基修饰的玻璃表面 ,通过共价偶联的方法将抗原或抗体固定到芯片表面 ,能使二者保持其特异性结合能力 .同时 ,荧光标记后的抗原或抗体仍然具有特异性结合能力 .蛋白质微阵列是通过机械手在玻片表面排阵制作的 .芯片上的荧光信号获取采用了激光共焦荧光扫描系统 .用不同浓度的抗原探针阵列 ,对其相应的抗体靶分子的特异性结合进行了分析和研究 .此外 ,还通过在玻片表面固定兔IgG和固定鼠IgG ,对羊抗兔和羊抗鼠抗体与其相应抗原的特异性相互作用进行了检测 .

The probing proteins were immobilized onto the surface of glass slides modified by aldehyde. The target proteins were labeled by either Cy5 or Cy3 for specific cases. A high-precision robot was designed to print protein samples onto glass slides to form the microarrays. The activity and the specific affinity interaction of the proteins were not compromised after fluorescent dye labeling process and covalent immobilization. To detect the antigen, arrays of antibodies attached to the glass substrate was used. Likewise, for the assessment of the property of antibody, antigen-based array was utilized. The concentration of the proteins immobilized onto slides is an important factor fur determining the sensitivity in detecting the interaction of antigen-antibody. The interactions of goat-anti-mouse and goat-anti-rabbit antibodies with their corresponding antigens were proven specific as revealed by the results generated from the present protein arrays.