Gering John P, Quaroni Luca, Chumanov George
Department of Chemistry, Clemson University, Clemson, South Carolina 20634, USA.
J Colloid Interface Sci. 2002 Aug 1;252(1):50-6. doi: 10.1006/jcis.2002.8415.
This work characterizes substrates for immunoassays obtained through the immobilization of vectorially oriented antibodies on glass. The method of preparation is based on the condensation reaction between an aldehyde group on the F(c) portion of antibodies and the hydrazide group on the modified glass surface. Light microscopy and AFM imaging in height and phase modes were used to assess the properties of the modified surface. Both techniques are consistent with a fairly uniform antibody coverage on the micrometer and submicrometer scales. ELISA tests were used to evaluate the activity and surface distribution of immobilized antibodies as well as nonspecific binding to surfaces after various modification steps. It was shown that exposure of the surfaces to a BSA solution minimized nonspecific binding to undetectable levels.
这项工作对通过将定向排列的抗体固定在玻璃上而获得的免疫分析底物进行了表征。制备方法基于抗体F(c)部分上的醛基与修饰玻璃表面上的酰肼基之间的缩合反应。使用光学显微镜以及原子力显微镜的高度和相位模式成像来评估修饰表面的性质。这两种技术均表明在微米和亚微米尺度上抗体覆盖相当均匀。酶联免疫吸附测定(ELISA)试验用于评估固定化抗体的活性和表面分布,以及在各种修饰步骤后与表面的非特异性结合。结果表明,将表面暴露于牛血清白蛋白(BSA)溶液可将非特异性结合降至不可检测的水平。