Article
Continuous microfluidic platform combining cell lysis and protein extraction for screening overall process conditions
Journal of Chemical Technology and Biotechnology
— 2023 — Wiley
Key information
Authors:
Published in
November 20, 2023
Abstract
BACKGROUND Microfluidic devices have been increasingly used as tools to accelerate the development of new biomanufacturing methods, given their ability to test a large set of variables in a short amount of time with minimal reagent consumption. However, while being able to expedite a bioprocess design, the modular aspect of these devices has not been systematically explored, with most applications focusing on single unitary operations. This paper explores the modular design of these devices through the development of an integrated microfluidic chip. RESULTS Two separate unitary operations were included into a single device for continuous operation: product release by chemical cell lysis and product concentration by aqueous two-phase system. In this way, it is possible to screen multiple conditions for each operation and evaluate their combined effect on the final product, making this a great tool in the development and optimization of a process. A recombinant Escherichia coli (E. coli) strain producing green fluorescent protein (GFP) was used as model system, which allows for lysis efficiency and partition coefficient to be evaluated by fluorescence microscopy. A chemical lysis solutions was used in the lysis step, presenting lysis efficiencies of about 100%, while a polyethylene glycol (PEG)/phosphate system was screened for the separation stage, presenting partition coefficients of about 4. CONCLUSIONS The integrated device allowed for the continuous screening of multiple combinations of operation conditions, highlighting, for example, conditions where improvements on the lysis efficiency subsequently impaired the separation, decreasing the partition of the target product, thus demonstrating the need for the evaluation of a process in its entirety. In this way, the microfluidic device delivered a rapid process screening and optimization, with very low reagent consumption, and using a simple fluorescence microscope for data collection. © 2023 Society of Chemical Industry (SCI).
Publication details
Authors in the community:
Ricardo Gil Fradique
ist426748
Publication version
AM - Accepted manuscript
Publisher
Wiley
Link to the publisher's version
https://doi.org/10.1002/jctb.7564
Title of the publication container
Journal of Chemical Technology and Biotechnology
First page or article number
618
Last page
625
Volume
99
Issue
3
Fields of Science and Technology (FOS)
chemical-engineering - Chemical engineering
Keywords
- cell lysis
- microfluidics
- ATPS
- E.coli
- bioprocess integration
Publication language (ISO code)
eng - English
Rights type:
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