Abstract

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.

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