Abstract
In the course of their development, industrial biocatalysis processes have to be optimized in small-scale, e. g., within microfluidic bioreactors. Recently, we introduced a novel microfluidic reactor device, which can handle defined reaction compartments of up to 250 μL in combination with magnetic micro carriers. By transferring the magnetic carriers between subsequent compartments of differing compositions, small scale synthesis, and bioanalytical assays can be conducted. In the current work, this device is modified and extended to broaden its application range to the screening and optimization of bioprocesses applying immobilized enzymes. Besides scaling the maximum compartment volume up to 3 mL, a temperature control module, as well as a focused infrared spot were integrated. By adjusting the pump rate, compartment volumes can be accurately dosed with an error <5% and adjusted to the requested temperature within less than a minute. For demonstration of bioprocess parameter optimization within such compartments, the influence of pH, temperature, substrate concentration, and enzyme carrier loading was automatically screened for the case of transferring UDP-Gal onto N-acetylglucosamine linked to a tert-butyloxycarbonyl protected amino group using immobilized β1,4-galactosyltransferase-1. In addition, multiple recycling of the enzyme carriers and the use of increased compartment volumes also allows the simple production of preparative amounts of reaction products.
Highlights
Microfluidic reaction devices are widely used for analysis, synthesis, diagnostics, biosensing, and a broad range of other applications in the pharmaceutical and healthcare industries
All modules are controlled by the QmixElements automation software and are interconnected by fluorinated ethylene propylene (FEP) tubing with an outer diameter of 1/8" purchased from CS-Chromatographie Service (Langerwehe, Germany)
The original reactor device was designed for analytical purposes using small sample and reaction compartment volumes of 10– 250 μL
Summary
Microfluidic reaction devices are widely used for analysis, synthesis, diagnostics, biosensing, and a broad range of other applications in the pharmaceutical and healthcare industries. They enable the miniaturization, integration, and automation of biochemical assays with real time or end-of-line analytical measurements (Haeberle and Zengerle, 2007; Zhang and Haswell, 2007; Malic et al, 2010). Magnetic particles are well-suited because of their easy separation by magnetic fields, resulting in an enzyme free product without time-consuming and expensive purification steps. Magnetic particles with immobilized enzyme are already used in analytical biotechnology as biochemical detectors for sugar (Nomura et al, 2004) and protein (Choi et al, 2002). The magnetic particles placed in these compartments are controlled by permanent- and electromagnetic fields
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