Zubtsov D A, Savvateeva E N, Rubina A Yu, Pan'kov S V, Konovalova E V, Moiseeva O V, Chechetkin V R, Zasedatelev A S
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov str., 32, 119991 Moscow, Russia.
Anal Biochem. 2007 Sep 15;368(2):205-13. doi: 10.1016/j.ab.2007.04.040. Epub 2007 Apr 29.
Protein microchips are designed for high-throughput evaluation of the concentrations and activities of various proteins. The rapid advance in microchip technology and a wide variety of existing techniques pose the problem of unified approach to the assessment and comparison of different platforms. Here we compare the characteristics of protein microchips developed for quantitative immunoassay with those of antibodies immobilized on glass surfaces and in hemispherical gel pads. Spotting concentrations of antibodies used for manufacturing of microchips of both types and concentrations of antigen in analyte solution were identical. We compared the efficiency of antibody immobilization, the intensity of fluorescence signals for both direct and sandwich-type immunoassays, and the reaction-diffusion kinetics of the formation of antibody-antigen complexes for surface and gel-based microchips. Our results demonstrate higher capacity and sensitivity for the hydrogel-based protein microchips, while fluorescence saturation kinetics for the two types of microarrays was comparable.
蛋白质微芯片旨在对各种蛋白质的浓度和活性进行高通量评估。微芯片技术的迅速发展以及众多现有技术带来了对不同平台进行评估和比较的统一方法问题。在此，我们将用于定量免疫测定的蛋白质微芯片的特性与固定在玻璃表面和半球形凝胶垫上的抗体的特性进行比较。用于制造这两种类型微芯片的抗体点样浓度以及分析物溶液中抗原的浓度是相同的。我们比较了抗体固定效率、直接免疫测定和夹心型免疫测定的荧光信号强度，以及基于表面和凝胶的微芯片形成抗体 - 抗原复合物的反应扩散动力学。我们的结果表明，基于水凝胶的蛋白质微芯片具有更高的容量和灵敏度，而两种类型微阵列的荧光饱和动力学相当。
