Cranfield University, Cranfield Health, Cranfield, Bedfordshire MK43 0AL, UK.
Talanta. 2011 Oct 30;86:377-83. doi: 10.1016/j.talanta.2011.09.031. Epub 2011 Sep 22.
An immunoassay in optimised conditions with a highly sensitive surface plasmon resonance (SPR) based biosensor was developed for the detection of the cancer biomarker carcinoembryonic antigen (CEA). Different formats of the immunoassay were initially investigated on the surface of the gold sensor chip. A self-assembled monolayer (SAM) was formed on the gold chip using 11-mercaptoundecanoic acid (MUDA), before the immobilisation of the antibodies was conducted. The assay was then formed in a direct capture and a sandwich assay. In order to increase the sensor signal the CEA antigen was incubated with the detection/capture antibody before it was injected to the sensor chip surface and the results were recorded in real-time using the Biacore 3000 instrument. A detection limit of 3 ng ml(-1) CEA was obtained with a dynamic detection range from 3 ng ml(-1) to 400 ng ml(-1) with correlation coefficients of 1.00 and 0.99 for the sandwich and rabbit anti-mouse (RAM) capture assay. Kinetic data analysis was performed for the standard capture test and subsequently for the developed assays and R(max) showed an increase from 215 RU for the standard capture test to 428 RU for the RAM-capture assay and 734 RU for the sandwich assay, respectively. The developed SPR immunosensor using the sandwich assay format showed high sensitivity and reproducibility for CEA detection which makes it a promising procedure for cancer biomarker analysis.
本研究开发了一种基于表面等离子体共振(SPR)的高灵敏度生物传感器的免疫分析方法,用于检测癌胚抗原(CEA)这一癌症生物标志物。最初在金传感器芯片的表面上研究了不同格式的免疫分析。在金芯片上使用 11-巯基十一烷酸(MUDA)形成自组装单层(SAM),然后进行抗体固定。随后形成直接捕获和三明治夹心免疫分析。为了提高传感器信号,将 CEA 抗原与检测/捕获抗体孵育,然后将其注入传感器芯片表面,并使用 Biacore 3000 仪器实时记录结果。使用夹心免疫分析和兔抗鼠(RAM)捕获免疫分析,获得了 3ngml(-1)CEA 的检测限,动态检测范围分别为 3ngml(-1)至 400ngml(-1),相关系数分别为 1.00 和 0.99。对标准捕获测试和随后开发的测试进行了动力学数据分析,R(max)分别从标准捕获测试的 215RU 增加到 RAM 捕获测试的 428RU 和夹心免疫分析的 734RU。使用夹心免疫分析格式开发的 SPR 免疫传感器对 CEA 检测具有高灵敏度和重现性,这使其成为癌症生物标志物分析的一种很有前途的方法。