Microfluidic technology is of interest in the study of enzymes and their inhibitors, which greatly improves the analytical performance in terms of reducing the amount of reagents, shortening the analysis time and automation. Creative Biolabs is a world-leading expert who is skilled in employing microfluidic technologies to analyze various biomolecules including proteins, nucleic acid, enzymes and their inhibitors as well as small molecules. We are dedicated to providing one-stop high-quality microfluidic chip development services to accelerate research on enzymes and their inhibitors.
Enzymes are powerful biocatalysts regulating all the biological processes in all living entities. They are valuable in both commercial and industrial processes and have been used in many different industries such as food, medicine, chemical, and biotechnology. Especially, enzymes can be used as a drug or drug target for the treatment of various diseases. For example, IgA1 protease treatment may reverse mesangial deposits and hematuria in a model of IgA nephropathy. Besides, enzymes can be used as disease markers, and detecting and quantifying levels of biomarker enzyme activity help to disease diagnosis.
Microfluidic chip technologies including lab-on-a-chip technology, three-dimensional (3D) cell culture, organs-on-chip and droplet techniques are applied for the high-throughput screening and detection study of enzymes, coupled with various kinds of detection techniques. Compared to other conventional screening methods, microfluidic chips are inexpensive to prepare, allowing for a library size with a throughput limit of 107 mutants/novel enzymes. In particular, water-in-oil droplets combining with fluorescent-activated droplet sorting (FADS) extend the application range, enabling detection enzyme activity in a wide range of expression systems such as lysed cells or excreted enzymes contained in the oil compartment.
Table.1 Reported use of microfluidic chips for directed evolution or discovery of novel enzymes. (Scheele, 2018)
|Entry||Aim||Compartmentalization||Expression system||Incubation type||Incubation time||Screening method||Screening rate (cells/second)||Original library size|
|1||Directed Evolution||Polydisperse droplets||Variable||Variable||Variable||FACS||20000||Variable|
|2||Directed Evolution||Monodisperse droplets||Cell display||Channel||5 min.||FADS||2000||107|
|3||Directed Evolution||Monodisperse droplets||Lysed cells||Syringe||1 hr.||FADS||1000||107|
|5||Directed Evolution||Monodisperse droplets||Secreted enzymes||Capillary||2 hr.||FADS||400||3×106|
|6||Discovery Enzymes||Monodisperse droplets||Lysed cells||Syringe||48 hr.||FADS||2000-2500||1.25×106|
|7||Directed Evolution||Monodisperse GSB´s||Lysed cells||Capillary||5 min||FACS||20000||5×105|
Bruyneel et al. (2005) designed and developed a microfluidic-based enzyme assay that integrates microfluidic chips and capillary reversed-phase HPLC with ESI-MS-based detection systems for bioactivity screening. In this detection system, the inhibition of cathepsin B was detected by monitoring three product traces, obtained by cleavage of the substrate, with detection limits of 0.17-2.6 micromol L-1. Another study showed three-layer polydimethylsiloxane (PDMS)-based microfluidic chip fabricated with sensitive fluorescence detection for detecting urease inhibition with advantages of low reagent consumption, reduced analysis time and ease of manipulation. In addition, scientists integrated microfluidic droplet systems with sensitive fluorescence detection systems for high-throughput measurement of enzyme activity. The droplet-based bioreactors allow the synthesis of the enzyme from cognate DNA and reactions with its substrate. The fluorescence detection systems are used to monitor the fluorescence released by enzymatic reactions. These microfluidic chip technologies provide a potential platform for the screening of inhibitory compounds in drug discovery.
Fig.2 A sample photograph of droplet formation (A) and droplet separation before detection (B). (Wu, 2011)
Creative Biolabs offers a wide range of microfluidic chip development services from scheme design and optimization to high-quality data delivery. We have more than a decade of microfluidic chip development experience and ensure to deliver optimal solutions to promote your success. For research on enzymes and their inhibitors, we support a series of microfluidic chip-based methods for the detection and screening of enzymes and their inhibitors via the combination of fluorescence and surface-enhanced Raman scattering (SERS) spectroscopies as well as MS. We work with customers to provide optimized solutions for the development, manufacturing, and functionality of microfluidic chips. If you are interested in our services, please contact us to discuss your project.
For Research Use Only. Not For Clinical Use.