Silina Y E, Morgan B
Institute for Biochemistry, Zentrum für Human und Molekularbiologie (ZHMB), Campus B 2.2, University of Saarland, 66123, Saarbrücken, Germany.
Institute for Biochemistry, Zentrum für Human und Molekularbiologie (ZHMB), Campus B 2.2, University of Saarland, 66123, Saarbrücken, Germany.
Talanta. 2021 Feb 1;223(Pt 1):121688. doi: 10.1016/j.talanta.2020.121688. Epub 2020 Sep 24.
Amperometric biosensors have been widely utilized for the cost-effective and rapid analysis of various bioanalytes, for example glucose. However, a lack of standardization and validation procedures remains a major limitation in biosensor development. Therefore, despite rapid advances in material science driving the development of amperometric biosensors, to date only a few biosensors, detecting a limited range of analytes, are available on the market. It is believed, once this issue is addressed, it can significantly facilitate the next step in the overall concept "go to the market" production and implementation of amprerometric biosensors for a large industrial scale. Herein, we report on the use of laser desorption ionization mass spectrometry (LDI-MS) for the standardization of amperometric biosensors, based upon a complete and non-destructive characterization and validation of layer-by-layer (LbL) biosensors at each fabrication step. We reveal that specific ionization pathways of mediators, polymers and enzymes from the biosensor surface allows for robust quality control during LbL biosensor manufacture. Furthermore, this LDI-MS approach can also be used to monitor, and therefore ensure, the encapsulation of enzymes in one-step nanobiosensors. Specifically, we show that LDI-MS can be used for the rapid chemical profiling of LbL biosensors and one-step synthesized nanobiosensors, as well as to assess their synthesis quality and to monitor for batch-to-batch and intra- and inter-day changes in their function and behavior. Our novel approach will thus contribute to the future development, improved design and fine tuning of both conventional LbL-fabricated amperometric biosensors and one-step designed nanobiosensors.
安培生物传感器已被广泛用于各种生物分析物（例如葡萄糖）的经济高效且快速的分析。然而，缺乏标准化和验证程序仍然是生物传感器开发中的一个主要限制。因此，尽管材料科学的快速发展推动了安培生物传感器的开发，但迄今为止，市场上仅有少数能检测有限范围分析物的生物传感器。人们认为，一旦这个问题得到解决，它可以显著促进“推向市场”这一整体概念的下一步发展，即大规模生产和应用安培生物传感器。在此，我们报告基于在逐层（LbL）生物传感器制造的每个步骤进行完整且无损的表征和验证，使用激光解吸电离质谱（LDI-MS）对安培生物传感器进行标准化。我们发现，生物传感器表面的介质、聚合物和酶的特定电离途径允许在LbL生物传感器制造过程中进行稳健的质量控制。此外，这种LDI-MS方法还可用于监测并因此确保酶在一步法纳米生物传感器中的封装。具体而言，我们表明LDI-MS可用于对LbL生物传感器和一步合成的纳米生物传感器进行快速化学分析，以及评估它们的合成质量，并监测其功能和行为在批次间以及日内和日间的变化。因此，我们的新方法将有助于传统LbL制造的安培生物传感器和一步设计的纳米生物传感器的未来发展、改进设计和精细调整。
