Prommapan Plengchart, Brljak Nermina, Lowry Troy W, Van Winkle David, Lenhert Steven
Department of Physics, Florida State University, Tallahassee, FL 32306, USA.
Department of Chemistry, Florida State University, Tallahassee, FL 32306, USA.
Nanomaterials (Basel). 2020 Dec 5;10(12):2433. doi: 10.3390/nano10122433.
Lipid multilayer gratings are promising optical biosensor elements that are capable of transducing analyte binding events into changes in an optical signal. Unlike solid state transducers, reagents related to molecular recognition and signal amplification can be incorporated into the lipid grating ink volume prior to fabrication. Here we describe a strategy for functionalizing lipid multilayer gratings with a DNA aptamer for the protein thrombin that allows label-free analyte detection. A double cholesterol-tagged, double-stranded DNA linker was used to attach the aptamer to the lipid gratings. This approach was found to be sufficient for binding fluorescently labeled thrombin to lipid multilayers with micrometer-scale thickness. In order to achieve label-free detection with the sub-100 nm-thick lipid multilayer grating lines, the binding affinity was improved by varying the lipid composition. A colorimetric image analysis of the light diffracted from the gratings using a color camera was then used to identify the grating nanostructures that lead to an optimal signal. Lipid composition and multilayer thickness were found to be critical parameters for the signal transduction from the aptamer functionalized lipid multilayer gratings.
脂质多层光栅是很有前景的光学生物传感器元件，能够将分析物结合事件转化为光信号的变化。与固态换能器不同，与分子识别和信号放大相关的试剂可以在制造前掺入脂质光栅墨水中。在此，我们描述了一种用针对凝血酶蛋白的DNA适配体对脂质多层光栅进行功能化的策略，该策略允许进行无标记分析物检测。一种双胆固醇标记的双链DNA连接体被用于将适配体连接到脂质光栅上。结果发现，这种方法足以使荧光标记的凝血酶与微米级厚度的脂质多层结合。为了利用厚度小于100 nm的脂质多层光栅线实现无标记检测，通过改变脂质组成提高了结合亲和力。然后使用彩色相机对从光栅衍射的光进行比色图像分析，以识别能产生最佳信号的光栅纳米结构。发现脂质组成和多层厚度是来自适配体功能化脂质多层光栅的信号转导的关键参数。
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