Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Universitat Politècnica de València, Universitat de València, 46022 Valencia, Spain.
Department of Applied Physics and Electromagnetism-ICMUV, Universitat de València, 46100 Burjassot, Spain.
ACS Appl Mater Interfaces. 2022 Sep 14;14(36):41640-41648. doi: 10.1021/acsami.2c12808. Epub 2022 Sep 1.
The nanostructuration of biolayers has become a paradigm for exploiting nanoscopic light-matter phenomena for biosensing, among other biomedical purposes. In this work, we present a photopatterning method to create periodic structures of biomacromolecules based on a local and periodic mild denaturation of protein biolayers mediated by UV-laser irradiation. These nanostructures are constituted by a periodic modulation of the protein activity, so they are free of topographic and compositional changes along the pattern. Herein, we introduce the approach, explore the patterning parameters, characterize the resulting structures, and assess their overall homogeneity. This UV-based patterning principle has proven to be an easy, cost-effective, and fast way to fabricate large areas of homogeneous one-dimensional protein patterns (2 min, 15 × 1.2 mm, relative standard deviation ≃ 16%). This work also investigates the implementation of these protein patterns as transducers for diffractive biosensing. Using a model immunoassay, these patterns have demonstrated negligible signal contributions from non-specific bindings and comparable experimental limits of detection in buffer media and in human serum (53 and 36 ng·mL of unlabeled IgG, respectively).
生物层的纳米结构化已成为利用纳米级光物质现象进行生物传感的范例，这在其他生物医学用途中也是如此。在这项工作中，我们提出了一种光图案化方法，通过紫外激光照射介导的局部和周期性温和变性来创建基于生物大分子的周期性结构。这些纳米结构由蛋白质活性的周期性调制组成，因此沿图案没有形貌和组成变化。在此，我们介绍了该方法，探索了图案化参数，对所得结构进行了表征，并评估了它们的整体均一性。事实证明，这种基于 UV 的图案化原理是一种简单、具有成本效益且快速的方法，可以制造大面积的均匀一维蛋白质图案（2 分钟，15×1.2mm，相对标准偏差 ≃ 16%）。这项工作还研究了将这些蛋白质图案用作衍射生物传感器的换能器。使用模型免疫测定法，这些图案表明非特异性结合的信号贡献可以忽略不计，并且在缓冲液介质和人血清中的实验检测限相当（分别为 53 和 36ng·mL 未标记 IgG）。
