Towards microbioprocess control: an inexpensive 3D printed microbioreactor with integrated online real-time glucose monitoring
Bioprocessing is of crucial importance in pharmaceutical, biofuel, food and other industries. Miniaturization of bioprocesses into microbioreactors allows multiplexing of experiments as well as reduction of reagent consumption and labour-intensity. A crucial part of the research within microbioreactors is biochemical analysis of product, byproduct and substrate concentrations that currently heavily relies on large analytical equipment. Biosensors are a promising analytical tool, however, integration into a microbioreactor is associated with challenges in ensuring sterility, appropriate sensing range, control of matrix effects and stability. In this work we present a novel biosensor integrated analytical chip that features an internal, actuated buffer flow in contact with a biosensor downstream and a diffusion limiting membrane exposed to the sample upstream. The technology was developed and tested using an electrochemical glucose oxidase biosensor and was found to successfully surmount the aforementioned challenges including the extension of the linear range of sensitivity to more than 20 g L−1 for online, real time monitoring of glucose. The biosensor integration chip with the glucose biosensor was then mounted onto a 3D printed microbioreactor with 1 mL of internal volume. The system successfully monitored the consumption of glucose of Saccharomyces cerevisiae in real time for more than 8 h. The developed technology and measurement methodologies are transferrable to other biosensors and microbioreactors as well as large scale applications.
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