Low-cost and user-friendly biosensor to test the integrity of mRNA molecules suitable for field applications
File(s)RNA sensor manuscript final.docx (1.25 MB) Supplementary Information .docx (622.66 KB)
Accepted version
Supporting information
Author(s)
Moya-Ramirez, Ignacio
Kontoravdi, Kleio
Polizzi, Karen
Type
Journal Article
Abstract
The use of mRNA in biotechnology has expanded with novel applications such as vaccines and therapeutic mRNA delivery recently demonstrated. For mRNA to be used in patients, quality control assays will need to be routinely established. Currently, there is a gap between the highly sophisticated RNA integrity tests available and broader application of mRNA-based products by non-specialist users, e.g. in mass vaccination campaigns. Therefore, the aim of this work was to develop a low-cost biosensor able to test the integrity of a mRNA molecule with low technological requirements and easy end-user application. The biosensor is based on a bi-functional fusion protein, composed by the λN peptide that recognizes its cognate aptamer encoded on the 5’ end of the RNA under study and β-lactamase, which is able to produce a colorimetric response through a simple test. We propose two different mechanisms for signal processing adapted to two levels of technological sophistication, one based on spectrophotometric measurements and other on visual inspection. We show that the proposed λN-βLac chimeric protein specifically targets its cognate RNA aptamer, boxB, using both gel shift and biolayer interferometry assays. More importantly, the results presented confirm the biosensor performs reliably, with a wide dynamic range and a proportional response at different percentages of full-length RNA, even when gene-sized mRNAs were used. Thus, the features of the proposed biosensor would allow to end-users of products such as mRNA vaccines to test the integrity of the product before its application in a low-cost fashion, enabling a more reliable application of these products.
Date Issued
2019-07-01
Date Acceptance
2019-05-03
Citation
Biosensors and Bioelectronics, 2019, 137, pp.199-206
ISSN
0956-5663
Publisher
Elsevier
Start Page
199
End Page
206
Journal / Book Title
Biosensors and Bioelectronics
Volume
137
Copyright Statement
© 2019 Elsevier B.V. All rights reserved. This manuscript is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence http://creativecommons.org/licenses/by-nc-nd/4.0/
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Grant Number
EP/K038648/1
Subjects
Biosensor
Low-cost
Low-tech
Point-of-care diagnostics
RNA degradation
RNA integrity
0301 Analytical Chemistry
0903 Biomedical Engineering
1007 Nanotechnology
Bioinformatics
Publication Status
Published
Date Publish Online
2019-05-07