Bioengineering Laboratory, Riken Institute, Saitama, Japan; Department of Biomedical Engineering, Dongguk University, Seoul, Republic of Korea.
Anal Chim Acta. 2013 Sep 24;796:108-14. doi: 10.1016/j.aca.2013.06.041. Epub 2013 Jul 3.
Enriching the surface density of immobilized capture antibodies enhances the detection signal of antibody sandwich microarrays. In this study, we improved the detection sensitivity of our previously developed P-Si (porous silicon) antibody microarray by optimizing concentrations of the capturing antibody. We investigated immunoassays using a P-Si microarray at three different capture antibody (PSA - prostate specific antigen) concentrations, analyzing the influence of the antibody density on the assay detection sensitivity. The LOD (limit of detection) for PSA was 2.5 ng mL(-1), 80 pg mL(-1), and 800 fg mL(-1) when arraying the PSA antibody, H117 at the concentration 15 μg mL(-1), 35 μg mL(-1), and 154 μg mL(-1), respectively. We further investigated PSA spiked into human female serum in the range of 800 fg mL(-1) to 500 ng mL(-1). The microarray showed a LOD of 800 fg mL(-1) and a dynamic range of 800 fg mL(-1) to 80 ng mL(-1) in serum spiked samples.
通过增加固定化捕获抗体的表面密度，可以增强抗体夹心微阵列的检测信号。在这项研究中，我们通过优化捕获抗体的浓度，改进了我们之前开发的 P-Si（多孔硅）抗体微阵列的检测灵敏度。我们使用 P-Si 微阵列研究了三种不同捕获抗体（PSA-前列腺特异性抗原）浓度下的免疫分析，分析了抗体密度对检测灵敏度的影响。当 PSA 抗体 PSA-H117 的浓度分别为 15μg/mL、35μg/mL 和 154μg/mL 时，阵列上的 PSA 的 LOD（检测限）分别为 2.5ng/mL、80pg/mL 和 800fg/mL。我们进一步研究了 PSA 在 800fg/mL 至 500ng/mL 范围内掺入人女性血清中的情况。微阵列在掺入血清的样品中显示出 800fg/mL 的 LOD 和 800fg/mL 至 80ng/mL 的动态范围。
