Shakurov Ruslan, Sizova Svetlana, Dudik Stepan, Serkina Anna, Bazhutov Mark, Stanaityte Viktorija, Tulyagin Petr, Konopsky Valery, Alieva Elena, Sekatskii Sergey, Bespyatykh Julia, Basmanov Dmitry
Lopukhin Federal Research and Clinical Center of Physical Chemical Medicine of Federal Medical Biological Agency, 1A Malaya Pirogovskaya Street, 119435 Moscow, Russia.
Research Institute for Systems Biology and Medicine (RISBM), Nauchniy Proezd 18, 117246 Moscow, Russia.
Polymers (Basel). 2023 Jun 8;15(12):2607. doi: 10.3390/polym15122607.
We propose and demonstrate dendrimer-based coatings for a sensitive biochip surface that enhance the high-performance sorption of small molecules (i.e., biomolecules with low molecular weights) and the sensitivity of a label-free, real-time photonic crystal surface mode (PC SM) biosensor. Biomolecule sorption is detected by measuring changes in the parameters of optical modes on the surface of a photonic crystal (PC). We describe the step-by-step biochip fabrication process. Using oligonucleotides as small molecules and PC SM visualization in a microfluidic mode, we show that the PAMAM (poly-amidoamine)-modified chip's sorption efficiency is almost 14 times higher than that of the planar aminosilane layer and 5 times higher than the 3D epoxy-dextran matrix. The results obtained demonstrate a promising direction for further development of the dendrimer-based PC SM sensor method as an advanced label-free microfluidic tool for detecting biomolecule interactions. Current label-free methods for small biomolecule detection, such as surface plasmon resonance (SPR), have a detection limit down to pM. In this work, we achieved for a PC SM biosensor a Limit of Quantitation of up to 70 fM, which is comparable with the best label-using methods without their inherent disadvantages, such as changes in molecular activity caused by labeling.
我们提出并展示了用于敏感生物芯片表面的基于树枝状聚合物的涂层,该涂层可增强小分子(即低分子量生物分子)的高性能吸附以及无标记实时光子晶体表面模式(PC SM)生物传感器的灵敏度。通过测量光子晶体(PC)表面光学模式参数的变化来检测生物分子的吸附。我们描述了生物芯片的逐步制造过程。以寡核苷酸作为小分子并在微流控模式下进行PC SM可视化,我们表明聚酰胺胺(PAMAM)修饰芯片的吸附效率比平面氨基硅烷层高近14倍,比3D环氧 - 葡聚糖基质高5倍。所获得的结果表明基于树枝状聚合物的PC SM传感器方法作为一种用于检测生物分子相互作用的先进无标记微流控工具具有进一步发展的前景。当前用于检测小生物分子的无标记方法,如表面等离子体共振(SPR),检测限低至皮摩尔。在这项工作中,我们实现了PC SM生物传感器高达70飞摩尔的定量限,这与最佳的使用标记的方法相当,且没有它们固有的缺点,如标记引起的分子活性变化。