Department of Physics and Nanotechnology, Aalborg University, Aalborg, Denmark.
Protein Sci. 2010 Sep;19(9):1751-9. doi: 10.1002/pro.461.
We here report for the first time the creation of prostate specific antigen (PSA) and Fab anti-PSA biosensor arrays using UV light-assisted molecular immobilization (LAMI), aiming at the detection and quantification of PSA, a cancer marker. The technology involves formation of free, reactive thiol groups upon UV excitation of protein aromatic residues located in spatial proximity of disulphide bridges, a conserved structural feature in both PSA and Fab molecules. The created thiol groups bind onto thiol reactive surfaces leading to oriented covalent protein immobilization. Protein activity was confirmed carrying out immunoassays: immobilized PSA was recognized by Fab anti-PSA in solution and immobilized Fab anti-PSA cross-reacted with PSA in solution. LAMI technology proved successful in immobilizing biomedically relevant molecules while preserving their activity, highlighting that insight into how light interacts with biomolecules may lead to new biophotonic technologies. Our work focused on the application of our new engineering principles to the design, analysis, construction, and manipulation of biological systems, and on the discovery and application of new engineering principles inspired by the properties of biological systems.
我们在这里首次报告了使用紫外光辅助分子固定化 (LAMI) 技术创建前列腺特异性抗原 (PSA) 和 Fab 抗 PSA 生物传感器阵列,旨在检测和定量 PSA,这是一种癌症标志物。该技术涉及在 PSA 和 Fab 分子中保守的二硫键结构附近的蛋白质芳香残基的 UV 激发下形成游离的、反应性的硫醇基团。生成的硫醇基团与硫醇反应性表面结合,导致蛋白质的定向共价固定化。通过进行免疫测定来证实蛋白质的活性:溶液中的 Fab 抗 PSA 识别固定化 PSA,溶液中的 PSA 与固定化 Fab 抗 PSA 发生交叉反应。LAMI 技术成功地固定了生物医学相关分子,同时保持了它们的活性,这表明对光与生物分子相互作用的深入了解可能会带来新的生物光子技术。我们的工作重点是将新的工程原理应用于生物系统的设计、分析、构建和操作,以及发现和应用受生物系统特性启发的新工程原理。