An improved design to capture magnetic microparticles for capillary electrophoresis based immobilized microenzyme reactors

Abstract

In this paper, we demonstrate the effectiveness of a new 3D printed magnet holder that enables capture of magnetic microparticles in commercially available capillary electrophoresis equipment with a liquid or air based coolant system. The design as well as the method to capture magnetic microparticles inside the capillary are discussed. This setup was tested at temperature and pH values suitable for performing enzymatic reactions. To demonstrate its applicability in CE‐ immobilized microenzyme reactors (IMER) development, human flavin‐containing monooxygenase 3 and bovine serum albumin were immobilized on amino functionalized magnetic microparticles using glutaraldehyde. These microparticles were subsequently used to perform in‐line capillary electrophoresis with clozapine as a model substrate. This setup could be used further to establish CE‐IMERs of other drug metabolic enzymes in a commercially available liquid based capillary coolant system. The CE‐IMER setup was successful, although a subsequent decrease in enzyme activity was observed on repeated runs.

Capillary electrophoresis integrated immobilized enzyme reactor for kinetic and inhibition assays of β‐secretase as the Alzheimer's disease drug target*

“…So in order to economize on chemicals consumption, the combination with lower concentrations was chosen as the optimum for further development. The final optimization step in terms of the composition of the immobilization mixture investigated the possible beneficial effect of BSA co‐immobilization, as described in our previous collaboration as well as other previously published immobilization protocols . Interestingly, up to a threefold increase in activity was observed when 20 μg/mL BSA was co‐immobilized with BACE1, see Figure D.…”

“…As has been shown in our previous work [30,32], the composition of the BGE and the overall settings of the separation method may greatly affect the performance of an IMER. Thus, mutual compatibility of all respective steps of the analysis is of utmost importance.…”

“…With the goal to further automate the IMER formation, a novel protocol was developed based on experience from our previous collaboration . This protocol does not require any additional equipment and can be carried out entirely in a commercial instrument, as specified in section .…”

Immobilizing sulfotransferase 1A1 on magnetic microparticles and their evaluation using capillary electrophoresis

“…The first objective was to find a separation medium compatible with the conditions of enzymatic incubation. Indeed, it was seen in earlier work that a medium applicable for both separation and enzymatic reaction, would be beneficial during the final construction of a CE‐IMER . Henceforth, 25 mM phosphate buffer at pH 7.4 was used for the separation of paracetamol and its metabolite paracetamol sulfate.…”

Recent advances in nanomaterial‐based capillary electrophoresis

“…These microparticles (MPs) were subsequently used to perform in‐line CE with clozapine as a model substrate. The CE‐IMER setup was successful . Teicoplanin modified Fe 3 O 4 MPs employing PDA adsorbent was developed and used as chiral adsorbents.…”