Automating functional enzyme screening & characterization
Files
2 page manuscript accepted in the 10th IWBDA workshop.
Date
2018-08-03
DOI
Authors
Ortiz, Luis
Lashkaripour, Ali
Densmore, Douglas
Version
OA Version
Citation
L. Ortiz, A. Lashkaripour, D. Densmore. (2018). Automating Functional Enzyme Screening & Characterization. In The Proceedings of the 10th International Workshop on Bio-Design Automation.
Abstract
Microfluidics continue to gain traction as an inexpensive alternative to standard multi-well plate-based, and flow cytometry- based, assay platforms. These devices are especially useful for the types of ultra-high throughput screens needed for enzyme discovery applications where large numbers (>106) of unique samples must be screened rapidly1. Coupled with cell-free protein synthesis2, microfluidics are being used to identify novel enzymes useful for a variety of applications with unprecedented speed. However, these devices are typically produced using PDMS, and require considerable infrastructure and artisanal skill to fabricate, limiting their accessibility. Likewise, enzyme hits obtained from a screen are often validated manually and would benefit from automation of downstream validation processes. To address these limitations, we propose a workflow which leverages software tools to automate the rapid design and fabrication of low-cost polycarbonate microfluidic devices for use as high-throughput screening platforms for enzyme discovery, as well as an automated DNA assembly tool to streamline validation of screening candidates. Using this workflow, we aim to identify novel oxidoreductase enzymes from environmental metagenomic DNA libraries, for use in electrochemical biosensors.
Description
This work has been presented in the 10th IWBDA workshop.
License
Attribution-NonCommercial-NoDerivs 3.0 United States